Method for determining sensitivity to 2,2'-dithio-bis-ethanesulfonate

Abstract

To provide methods for treating cancer. [Solution] A method comprising: (a) obtaining a cancer sample from a patient; (b) treating the sample with a 2,2'-dithio-bis-ethanesulfonate analog; (c) obtaining the expression level of one or more of a plurality of biomarkers in the sample, wherein the plurality of biomarkers include NRF2 (SEQ ID NO: 1); (d) determining from the expression level of NRF2 (SEQ ID NO: 1) that the sample is sensitive to treatment with disodium 2,2'-dithio-bis-ethanesulfonate; and (e) if the expression level of NRF2 (SEQ ID NO: 1) is greater than 1x compared to non-cancer cells, administering a cancer treatment to the subject including a disodium 2,2'-dithio-bis-ethanesulfonate analog.

Description

Technical Field

[0001] This application relates to determining the presence and / or level of biomarkers for detecting a patient's sensitivity to 2,2'-dithio-bis-ethanesulfonate as part of cancer treatment. This application relates to a method for detecting the expression level of a gene encoding a biomarker in a cancer patient and predicting the responsiveness of the cancer patient to sodium 2,2'-dithio-bis-ethanesulfonate as part of cancer therapy.

Background Art

[0002] Cancer chemotherapy is an area where scientific efforts are expanding and is an important element of cancer treatment along with surgery and radiotherapy. Chemotherapy was once only accepted as a means to extend the survival period of patients diagnosed as incurable by surgery and / or radiotherapy, but now it is a treatment method observed in almost all mutated cancers.

[0003] Modern cancer chemotherapy usually involves a combination of two or more different drugs, and with the progress of technology and medical knowledge, the possibility of patient recovery in many forms of cancer has been significantly improved. The role of chemotherapy agents in cancer therapy varies greatly depending on the form of cancer. For example, chemotherapy is often the main treatment method for cancers such as ovarian, testicular, breast, bladder cancers, leukemia, and lymphoma, and is generally used in combination with radiotherapy in the treatment of many sarcomas, melanomas, myelomas, and other treatments.

[0004] Chemotherapy agents are classified into several diverse groups. Most of these drugs act as cytotoxic agents, and each member of a particular group is generally assumed to exert its cytotoxic effect through a similar biological mechanism. The complete understanding of the biological and biochemical mechanisms of the action of anti-tumor drugs is not fully known.

[0005] Therefore, there is always a need for a method to determine or predict whether cancer is responsive to chemotherapeutic agents containing 2,2'-dithio-bis-ethanesulfonate or its analogues. [Overview of the project]

[0006] This application discloses a method for treating cancer in a patient, wherein a tumor or cancer sample is obtained from the patient and screened for responsiveness to 2,2'-dithio-bis-ethanesulfonate or an analogue. The sample is treated with 2,2'-dithio-bis-ethanesulfonate or an analogue to determine whether one or more of several biomarkers are expressed in the sample. The biomarker or several biomarkers may be nuclear factor erythrocyte-related factor 2 (NRF2) or other biomarkers. If the expression level of NRF2 (and / or other biomarkers) in the sample is greater than 1x the non-cancer cell or reference level, it indicates that the patient may be responsive to 2,2'-dithio-bis-ethanesulfonate or an analogue.

[0007] One embodiment further includes a method in which multiple biomarkers are selected from the group consisting of NQO1 (SEQ ID NO: 2), PHGDH (SEQ ID NO: 3), HMOX1 (SEQ ID NO: 4), SLC7A11 (SEQ ID NO: 5), SRXN1 (SEQ ID NO: 6), SOX2 (SEQ ID NO: 7), GPX2 (SEQ ID NO: 8), GPX3 (SEQ ID NO: 9), and GPX7 (SEQ ID NO: 10). Non-cancer cells define the reference level of the biomarkers.

[0008] Cancer treatment includes the administration of one or more chemotherapeutic agents selected from platinum complexes and taxanes. Cancer treatment also includes radiation therapy.

[0009] A method for testing tumor samples from patients with a known cancer type that is resistant to one or more cancer therapies and has an unknown response to 2,2'-dithio-bis-ethanesulfonate or its analogues. The method comprises the steps of contacting the sample with 2,2'-dithio-bis-ethanesulfonate or its analogues, and (a) a first single-stranded nucleic acid molecule that can specifically hybridize with the nucleotides of several biomarkers, wherein the first biomarker is NFR2, and (b) NQO1 (SEQ ID NO: 2), PHGDH (SEQ ID NO: 3), HMOX1 (SEQ ID NO: 4), SLC7A11 (SEQ ID NO: 5), SRXN1 (SEQ ID NO: 6), SOX2 (SEQ ID NO: 7), GPX2 (SEQ ID NO: 8), GPX3 (SEQ ID NO: 9) The method comprises a device comprising a second single-stranded nucleic acid molecule that can specifically hybridize with nucleotides of multiple biomarkers selected from GPX7 (SEQ ID NO: 10), and the steps of contacting a sample, detecting the expression levels of the multiple biomarkers, and administering 2,2'-dithio-bis-ethanesulfonate or its analogue to patients determined to be responsive to 2,2'-dithio-bis-ethanesulfonate or its analogue from the expression of the biomarkers from a reference level.

[0010] As will be apparent, features and characteristics of one aspect of this application are applicable to many other aspects of this application. The present invention is described below with reference to the accompanying drawings by the following non-limiting embodiments. [Brief explanation of the drawing]

[0011] [Figure 1] This shows the induction of Nrf2 nuclear expression in cells treated with 30 μM t-BHQ and cells treated with 2,2'-dithio-bis-ethanesulfonate. [Modes for carrying out the invention]

[0012] [Sequence List] The amino acid sequence of sequence number 1--NRF2(NFE2L2),

[0013] Amino acid sequence of SEQ ID NO: 2 - NQ01,

[0014] Amino acid sequence of SEQ ID NO: 3 - PHGDH,

[0015] Amino acid sequence of SEQ ID NO: 4 - HMOX1

[0016] Amino acid sequence of SEQ ID NO: 5 - SLC7A11,

[0017] Amino acid sequence of SEQ ID NO: 6 - SRXN1

[0018] Amino acid sequence of SEQ ID NO: 7 - SOX2,

[0019] Amino acid sequence of SEQ ID NO: 8 - GPX2,

[0020] Amino acid sequence of SEQ ID NO: 9 - GPX3, and Amino acid sequence of SEQ ID NO: 10 - GPX7.

[0021] [Definitions] Unless otherwise specified, all technical and scientific terms used in this specification shall be construed to have the same meaning as commonly understood by one of ordinary skill in the art (e.g., in cell culture, molecular genetics, immunology, immunohistochemistry, protein chemistry, and biochemistry).

[0022] An amino acid sequence that aligns with the amino acid sequence set forth in SEQ ID NO: X (when referring to a variant polypeptide) means that the variant amino acid sequence and the amino acid sequence set forth in SEQ ID NO: X are aligned by an appropriate method that enables identification of positions in the variant amino acid sequence where the same amino acid is present (at the same position), or a different amino acid is present (substitution), or one or more extra amino acids are present (insertion or extension), or no amino acid is present (deletion or truncation).

[0023] As used herein, the term “antibody” includes intact molecules, as well as molecules containing or composed of fragments thereof, such as Fab, F(ab')2, Fv, and scFv, and engineered variants, including diabodies, triabodies, minibodies, and single-domain antibodies that can bind to epitope determinants. Thus, antibodies may exist as intact immunoglobulins or as various forms of modifications.

[0024] The term "biomarker" refers to any molecule such as a gene, gene transcript (e.g., mRNA), peptide or protein or a fragment thereof, produced by a subject, that is useful for identifying a subject in order to predict the responsiveness of the subject to a treatment, including sodium 2,2'-dithio-bis-ethanesulfonate or an analog thereof. A biomarker that is differentially present (i.e., increased or decreased) in a biological sample from a subject or group of subjects has a first phenotype (e.g., having a disease) as compared to a biological sample from a subject or group of subjects having a second phenotype (e.g., not having the disease). A biomarker may be differentially present at any level, generally at a level of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, or more increased, or generally at a level of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% (i.e., absent) decreased. A biomarker is preferably differentially present at a statistically significant level (e.g., a p-value of less than 0.05 and / or a q-value of less than 0.10 determined using either Welch's T-test or Wilcoxon's rank sum test).

[0025] The terms "cancer" and "cancerous" typically refer to or describe a physiological condition in mammals (e.g., humans) characterized by uncontrolled cell proliferation.Examples of cancers include prostate cancer, ovarian cancer (e.g., ovarian adenocarcinoma or fetal cancer), liver cancer (e.g., hepatocellular carcinoma (HCC) or liver cancer), myeloma (e.g., multiple myeloma), colorectal cancer (e.g., colon and rectal cancer), leukemia (e.g., acute myeloid leukemia, acute lymphomatous leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, acute myeloblastic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroleukemia, and chronic leukemia), myelodysplastic syndromes, and lymphoma (e.g., diffuse large B-cell lymphoma). Lymphoma, cutaneous T-cell lymphoma, peripheral T-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, Waldenstrom macroglobulinemia, lymphocytic lymphoma), cervical cancer, esophageal cancer, melanoma, glioma (e.g., oligodendroglioma), pancreatic cancer (e.g., adenosquamous cell carcinoma, signet ring cell carcinoma, hepatoid carcinoma, colloidal carcinoma, islet cell carcinoma, pancreatic neuroendocrine carcinoma), gastrointestinal stromal tumors, sarcomas (e.g., fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, angiosarcoma, endosarcoma, lymphangiosarcoma, lymphangiosarcoma, endolymphatic sarcoma, leiomyoma, Ewing's sarcoma) , rhabdomyosarcoma), breast cancer (medullary carcinoma), ER-positive cancer, bladder cancer, head and neck cancer (e.g., head and neck squamous cell carcinoma), lung cancer (e.g., non-small cell lung cancer, large cell carcinoma, bronchogenic carcinoma, papillary carcinoma), metastatic cancer, oral cancer, uterine cancer, testicular cancer (e.g., seminoma and fetal cancer), skin cancer (e.g., squamous cell carcinoma and basal cell carcinoma), thyroid cancer (e.g., papillary carcinoma and medullary carcinoma), brain cancer (e.g., astrocytoma and craniopharyngioma), stomach cancer, carcinoma in situ, bone cancer, biliary tract cancer, eye cancer, laryngeal cancer, kidney cancer (e.g., kidney Examples of neoplasms include, but are not limited to, cell carcinomas and Wilms' tumors, gastric cancer, blastomas (e.g., nephroblastoma, medulloblastoma, hemangioblastoma, neuroblastoma, and retinoblastoma), polycythemia vera, chordoma, synovial tumors, mesothelioma, adenocarcinoma, sweat gland cancer, sebaceous gland cancer, cystadenocarcinoma, cholangiocarcinoma, choriocarcinoma, epithelial carcinoma, ependymoma, pineal glandoma, acoustic neuroma, Schwann cell tumors, meningiomas, pituitary adenomas, nerve sheath tumors, small intestine cancer, endocrine cancer, penile cancer, urethral cancer, melanoma of the skin or eye, gynecological tumors, solid tumors in childhood, and neoplasms of the central nervous system.The term cancer includes solid tumors (e.g., prostate cancer, ovarian cancer, or hepatocellular carcinoma (HCC)) and blood cancers.

[0026] The term “diagnosis,” and its variations such as “to diagnose,” “diagnosed,” or “diagnosis,” should be interpreted as including the diagnosis of recurrent illness, but not limited to the primary diagnosis of a clinical condition.

[0027] The term "subject" refers to any animal that may develop cancer, and includes animals such as mammals (e.g., humans) or non-human mammals (e.g., cats and dogs), laboratory animals (e.g., mice, rats, rabbits or guinea pigs, etc.), and livestock. In preferred embodiments, the subject is human.

[0028] The “sample” can be any suitable type and may refer to, for example, a material in which the presence or level of a biomarker can be detected. Preferably, the sample is obtained from the subject so that the detection of the presence and / or level of the biomarker can be performed in vitro. Alternatively, the presence and / or level of the biomarker can be detected in vivo. The sample can be used directly from the source or after at least one (partial) purification step. The sample can be prepared in any readily available medium that does not interfere with the method of the present invention. Typically, the sample is an aqueous solution, biological fluid, cells, or tissue. Preferably, the sample is blood, plasma, serum, urine, platelets, or other substances. Pretreatment may include, for example, preparing plasma from blood, diluting viscous fluids, etc. Treatment methods may include filtration, distillation, separation, concentration, inactivation of interfering components, and addition of reagents. The selection and pretreatment of biological samples before testing are well known in the art and do not need further explanation.

[0029] As used herein, the terms “expression level” and “expression level” refer to the amount of a gene product, such as DNA, RNA (e.g., messenger RNA (mRNA)), or protein corresponding to a particular gene in a cell, tissue, biological specimen, organism, or patient.

[0030] The term “healthy individual” is interpreted to mean an individual known not to have cancer, and such knowledge is derived from the individual’s clinical data, including but not limited to diagnostic assays different from those described herein.

[0031] "Reference level" means a level of the compound or additional biomarker of the present invention that indicates a specific disease state, phenotype, or lack thereof, as well as a combination of disease states, phenotypes, or lack thereof.

[0032] A "reference sample" refers to a sample containing the reference level of a biomarker. For example, a reference sample can be obtained from a subject that does not have a specific disease, condition, or phenotype such as cancer or acute injury.

[0033] Detailed explanation This application includes a method for detecting patients with cancer, for example, patients with cancer resistant to one or more cancer therapies other than 2,2'-dithio-bis-ethanesulfonate or its analogues (e.g., patients with lung cancer, prostate cancer, ovarian cancer, or hepatocellular carcinoma (HCC) resistant to 2,2'-dithio-bis-ethanesulfonate or its analogues alone, or in combination with one or more cancer therapies), and for determining the response of cancer patients (e.g., patients with lung cancer, prostate cancer, ovarian cancer, or HCC) to treatment with 2,2'-dithio-bis-ethanesulfonate or its analogues alone or in combination with other therapies. This application also features a method for treating cancer in patients in need of treatment (e.g., patients with lung cancer, prostate cancer, ovarian cancer, or HCC or treatment-resistant forms thereof), comprising administering 2,2'-dithio-bis-ethanesulfonate or its analogues to patients who are responsive to or have been determined to be responsive to 2,2'-dithio-bis-ethanesulfonate or its analogues according to the diagnostic methods described herein. Responsiveness may be a protective effect (protection against the effects of anticancer agents) indicated by 2,2'-dithio-bis-ethanesulfonate or its analogues and / or anticancer activity. Responsiveness may be indicated by one or more biomarkers.

[0034] According to one embodiment, a method for testing tumor samples from patients with known cancer types, whose responsiveness to 2,2'-dithio-bis-ethanesulfonate or its analogues is known or unknown, comprising: (a) contacting the sample with 2,2'-dithio-bis-ethanesulfonate or its analogues; and (b) after contact with 2,2'-dithio-bis-ethanesulfonate or its analogues, i) a nucleotide that can specifically hybridize with a plurality of sensitivity biomarkers selected from the NFR2 / NFE2L2 (SEQ ID NO: 1) biomarkers. The method includes (i) contacting a sample with a device containing a single-stranded nucleic acid molecule and / or (ii) a single-stranded nucleic acid molecule that can specifically hybridize with nucleotides of multiple resistance biomarkers selected from the biomarkers NQO1 (SEQ ID NO: 2), PHGDH (SEQ ID NO: 3), HMOX1 (SEQ ID NO: 4), SLC7A11 (SEQ ID NO: 5), SRXN1 (SEQ ID NO: 6), SOX2 (SEQ ID NO: 7), GPX2 (SEQ ID NO: 8), GPX3 (SEQ ID NO: 9), and GPX7 (SEQ ID NO: 10), and (c) detecting the expression levels of the multiple biomarkers. If the biomarkers are present in amounts exceeding normal levels in non-tumor cells or reference cells, the patient may be responsive to treatment containing 2,2'-dithio-bis-ethanesulfonate or an analogue thereof.

[0035] In some cases, cancer is selected from a group consisting of prostate cancer, ovarian cancer, hepatocellular carcinoma (HCC), cervical cancer, renal cell carcinoma (RCC), esophageal cancer, melanoma, glioma, pancreatic cancer, gastrointestinal stromal tumor (GIST), sarcoma, estrogen receptor-positive (ERpos) breast cancer, non-small cell lung cancer (NSCLC), colon cancer, bladder cancer, head and neck squamous cell carcinoma (SCCHN), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), myelodysplastic syndrome (MDS), chronic myeloid leukemia - chronic phase (CMLCP), diffuse large B-cell lymphoma (DLBCL), cutaneous T-cell lymphoma (CTCL), peripheral T-cell lymphoma (PTCL), and Hodgkin lymphoma.

[0036] Patients treated or tested for responsiveness to treatment (e.g., 2,2'-dithio-bis-ethanesulfonate) by the methods described herein may be patients diagnosed with cancers such as those described herein, e.g., prostate cancer, ovarian cancer, or hepatocellular carcinoma (HCC). Diagnosis may be made by any method or technique known in the art, such as X-ray, MRI, or biopsy, and may be confirmed by a physician. To minimize the patient's exposure to potentially ineffective drug treatments, patients may be determined, either responsive or inresponsive, to cancer treatments such as 2,2'-dithio-bis-ethanesulfonate according to the methods described herein. In specific examples, samples may be selected from the group consisting of tissue, blood, plasma, serum, urine, urine supernatant, urine cell pellets, semen, prostatic secretions, and prostate cells.

[0037] Another embodiment includes a method for treating or determining the sensitivity of non-small cell lung cancer to 2,2'-dithio-bis-ethanesulfonate therapy by evaluating NRF2 (SEQ ID NO: 1) expression and at least one gene selected from the group consisting of NQO1 (SEQ ID NO: 2), PHGDH (SEQ ID NO: 3), HMOX1 (SEQ ID NO: 4), SLC7A11 (SEQ ID NO: 5), SRXN1 (SEQ ID NO: 6), SOX2 (SEQ ID NO: 7), GPX2 (SEQ ID NO: 8), GPX3 (SEQ ID NO: 9), and GPX7 (SEQ ID NO: 10). For illustrative purposes, non-small cell lung cancer cell line HCC827 was treated with 2,2'-dithio-bis-ethanesulfonate alone at concentrations of 1 mM and 15 mM for 2 hours, and changes in gene expression levels were measured by whole transcriptome profiling using RNA-seq. Untreated control samples for the same period were used as baseline gene expression levels. A change in expression level of more than 1.5 to 2 times may indicate sensitivity to 2,2'-dithio-bis-ethanesulfonate.

[0038] In another embodiment, NRF2 (Sequence ID 1) may be an alias for NFE2L2.

[0039] In another embodiment, detection of NRF2 associated with one or more of the following genes: JAG1, IGF1, NANOG, GPX6, GPX5, PRDX4, TXN, TERT, TXNRD2, FGF2, PRDX2, PRDX3, PRDX1, BMPR1A, PDGFC, GPX1, PSPH, SHMT1, TXN2, SHMT2, PRDX6, NPNT, PSAT1, NOTCH1, GLRX2, VEGFC, GSR, ADAM10, PRDX5, TXNRD1, GLRX, ATF4, SIRT1, ITGB2, G6PD, GPX4, GPX3, GPX2, NQO1, PHGDH, GPX7, SLC7A11, SOX2, SRXN1, HMOX1, BCL2, BMI1, HEBP1, NFE2L2, PGK1, POU5F1, and TALDO1.

[0040] One embodiment includes a method for treating cancer in a patient requiring administration, comprising the step of administering 2,2'-dithio-bis-ethanesulfonate to a patient who has been determined to be responsive to 2,2'-dithio-bis-ethanesulfonate by a method disclosed herein. In particular, the patient may have cancer that is resistant to one or more cancer therapies other than 2,2'-dithio-bis-ethanesulfonate, or in combination with it (e.g., a patient with prostate cancer, ovarian cancer, or HCC).

[0041] Another embodiment is a method for treating a solid tumor cancer in a subject, comprising the steps of (a) obtaining expression levels of a plurality of biomarkers in a sample from the subject, or preparing expression levels of the obtained plurality of biomarkers in a sample from the subject, wherein the plurality of biomarkers include (1) NRF2 (SEQ ID NO: 1) and at least one biomarker selected from the group consisting of NQO1 (SEQ ID NO: 2), PHGDH (SEQ ID NO: 3), HMOX1 (SEQ ID NO: 4), SLC7A11 (SEQ ID NO: 5), SRXN1 (SEQ ID NO: 6), SOX2 (SEQ ID NO: 7), GPX2 (SEQ ID NO: 8), GPX3 (SEQ ID NO: 9), and GPX7 (SEQ ID NO: 10), (b) determining that the subject is sensitive to treatment with 2,2'-dithio-bis-ethanesulfonate, and (c) administering cancer treatment comprising 2,2'-dithio-bis-ethanesulfonate.

[0042] In one embodiment, the patient or subject is treated with 2,2'-dithio-bis-ethanesulfonate in conjunction with one or more other cancer treatments. For example, such treatments may include cisplatin, paclitaxel, and other available treatments.

[0043] The method involves administering an effective amount of 2,2'-dithio-bis-ethanesulfonate, a pharmaceutically acceptable salt thereof, and / or an analog thereof to a subject who is receiving or is scheduled to receive a chemotherapeutic agent, at a rate of approximately 0.1 g / min to approximately 2.0 g / min.

[0044] In another embodiment, an effective amount of 2,2'-dithio-bis-ethanesulfonate, a pharmaceutically acceptable salt thereof, and / or an analogue thereof is administered to the subject at a rate of about 0.2 g / min to about 1.0 g / min.

[0045] In another embodiment, an effective amount of 2,2'-dithio-bis-ethanesulfonate, a pharmaceutically acceptable salt thereof, and / or an analogue thereof is administered to the subject at a rate of about 0.7 g / min.

[0046] In one embodiment, a dose of 2,2'-dithio-bis-ethanesulfonate, a pharmaceutically acceptable salt thereof, and / or an analogue thereof is administered to a subject over a period of about 45 minutes.

[0047] In another embodiment, the total dose of 2,2'-dithio-bis-ethanesulfonate, its pharmaceutically acceptable salt, and / or analogues administered to the subject is approximately 4.0 g / m². 2 Approximately 35g / m 2 The single dose is approximately 18.4 g / m². 2 The administration of one or more of the aforementioned doses of the compound to the subject may take place over a period of approximately 45 minutes.

[0048] The present invention also relates to a method for reducing, preventing, mitigating, delaying the onset, reducing the severity, and / or facilitating the resolution of chemotherapy-related toxicity in a subject receiving a chemotherapy agent, comprising an effective amount of 2,2'-dithio-bis-ethanesulfonate, a pharmaceutically acceptable salt thereof, and / or an analogue thereof, at a rate of about 0.1 g / min to about 4.6 g / min, with a total dose of about 4 g / m². 2 ~about 35gm 2 The method includes a step of administering to the target. Approximately 18.4 g / m² 2 It is preferable to administer the total dose to the subject at a rate of approximately 0.1 g / min to approximately 4.6 g / min. Approximately 0.4 g / min 2 At a rate of one minute, approximately 18.4 g / m² is administered over approximately 45 minutes. 2 It is particularly preferable to administer the drug targeting the total dose.

[0049] In certain examples, the methods disclosed herein may further include the step of isolating a biomarker or probe from a sample using a machine. Alternatively, the methods disclosed herein may further include the step of contacting a sample with a label that specifically binds to a biomarker, probe, or a combination thereof. In some embodiments, the methods disclosed herein may further include the step of contacting a sample with a label that specifically binds to a biomarker selected from the genes described herein. In some embodiments, the methods disclosed herein may further include amplifying a biomarker, probe, or any combination thereof. The methods disclosed herein may further include the step of sequencing a target, probe, or any combination thereof. In some cases, the method may further include the step of quantifying the expression levels of multiple biomarkers. In some embodiments, the method may further include the step of labeling multiple biomarkers.

[0050] In some embodiments, diagnosing, predicting, and / or monitoring the state or outcome of cancer may include determining a 2,2'-dithio-bis-ethanesulfonate regimen for treatment. Determining a treatment regimen may include administering anticancer drugs. Alternatively, determining a treatment for cancer may include modifying a treatment regimen to include or reduce 2,2'-dithio-bis-ethanesulfonate. Modifying a treatment regimen may include increasing, decreasing, or discontinuing the treatment regimen.

[0051] In another embodiment, the 2,2'-dithio-bis-ethanesulfonate is a disodium salt.

[0052] In other embodiments, the 2,2'-dithio-bis-ethanesulfonate may be analogs such as monosodium 2,2'-dithio-bis-ethanesulfonate, sodium potassium 2,2'-dithio-bis-ethanesulfonate, dipotassium 2,2'-dithio-bis-ethanesulfonate, calcium 2,2'-dithio-bis-ethanesulfonate, magnesium 2,2'-dithio-bis-ethanesulfonate, monopotassium 2,2'-dithio-bis-ethanesulfonate, or manganese 2,2'-dithio-bis-ethanesulfonate; ammonium 2,2'-dithio-bis-ethanesulfonate.

[0053] In certain embodiments, 2,2'-dithio-bis-ethanesulfonates are administered with chemotherapeutic agents, such as fluoropyrimidines, pyrimidine nucleosides, purine nucleosides, antifolic acid agents, platinum analogs, anthracyclines / anthracendions, epipodophyllotoxins, camptothecin, hormones, hormone analogs, antihormone drugs, enzymes, proteins, peptides, or polyclonal and monoclonal antibodies, vinca alkaloids, taxanes, epothyrons, antimicrotubule agents, alkylating agents, antimetabolites, topoisomerase inhibitors, antivirals, or other cytotoxic and / or cell proliferation inhibitors. Examples of fluoropyrimidines include 5-fluorouracil (5-FU), S-1 capecitabine, Futraful, 5'-deoxyfluorouridine, UFT, and enyluracil. Pyrimidine nucleosides include, for example, cytarabine, deoxycytidine, 5-azacytosine, gemcitabine, 5-azacytosine, and 5-azadeoxycytidine. Purine nucleosides include, for example, fludarabine, 6-mercaptopurine, thioguanine, allopurinol, cladribine, and 2-chloroadenosine. Antifolic acid agents include, for example, methotrexate (MTX), trimethrexate, aminopterin, and methylene-10-deazaminopterin (MDAM). Platinum analogs include, for example, cisplatin, carboplatin, oxaliplatin, satoraplatin, picoplatin, tetraplatin, platinum-DACH, and their analogs. Anthracyclines / anthracendions include, for example, doxorubicin, daunorubicin, epirubicin, and idarubicin. Epipodophyllotoxin derivatives include, for example, etoposide, phosphate etoposide, and teniposide. Camptothecins include, for example, irinotecan, topotecan, 9-aminocamptothecin, 10,11-methylenedioxycamptothecin, calenitecin, 9-nitrocamptothecin, and TAS103.Hormones and hormone analogs may include, for example, estrogens and estrogen analogs including anastrozole, diethylstilbestrol, estradiol, Premarin, and raloxifene; progesterone, progesterone analogs, and progestins including progesterone, norethinodrel, estisterone, dimestisterone, megestrol acetate, medroxyprogesterone acetate, hydroxyprogesterone caproate, and norethisterone; androgens including fluoxymesterone, methyltestosterone, and testosterone; as well as adrenaline corticosteroids including dexamthazone, prednisone, cortisol, and solu-medrol. Antihormones include, for example, (i) antiestrogens including tamoxifen, fulvestrant, and toremifene; aminoglutethimide, testotractone, droloxifen, and anastrozole; (ii) antiandrogens including bicalutamide, flutamide, nilutamide, and goserelin; (iii) antitestosterones including flutamide, leuprolide, and triptorelin; (iv) adrenal steroid inhibitors including aminoglutethimide and mitotane; and anti-leuteinizing hormones including goserelin. Enzymes, proteins, peptides, and polyclonal and / or monoclonal antibodies may include, for example, asparaginase, cetuximab, erlotinib, bevacizumab, rituximab, gefitinib, trastuzumab, interleukin, interferon, leuprolide, and pegasparanase. Vinca alkaloids include, for example, vincristine, vinblastine, vinorelbine, and vindesine. Taxanes include, for example, paclitaxel, docetaxel, and their preparations and analogues. Alkylating agents may include, for example, dacarbazine, procarbazine, temozolamide, thiotepa, nitrogen mustards (e.g., mechloretamine, chlorambucil, L-phenylalanine mustard, melphalan, etc.), oxazaphosphorines (e.g., ifosfamide, cyclophosphamide, mephosfamide, perphosfamide, trophosfamide, etc.), alkyl sulfonates (e.g., busulfan), and nitrosoureas (e.g., carmustine, lomustine, semustine, etc.).Epothiron includes, for example, epothiron AE. Antimetabolites include, for example, tomdex and methotrexate, 6-mercaptopurine, and 6-thioguanine. Topoisomerase inhibitors include, for example, irinotecan, and topotecan, calenitecin, amsacrin, etoposide, etoposide phosphate, teniposide, and doxorubicin, daunorubicin, and other analogs. Antiviral agents include, for example, acyclovir, valacyclovir, ganciclovir, amantadine, rimantadine, lamivudine, and zidovudine. Monoclonal antibody agents include, for example, bevacizumab, trastuzumab, and rituximab, as well as cell proliferation inhibitors such as erlotinib. In general, cell proliferation inhibitors are mechanism-based drugs that slow the progression of neoplastic diseases.

[0054] <Detection of biomarkers for sensitivity to 2,2'-dithio-bis-ethanesulfonate treatment> From the above description, it will be clear that the diagnostic method of the present invention may include some degree of quantification for determining the level of biomarkers in a patient sample. Such quantification can be easily provided by including a suitable control sample.

[0055] In one embodiment, the internal control is included in the method of the present invention. A preferred internal control is one or more samples taken from one or more healthy individuals.

[0056] As is known to those skilled in the art, if each assay performed does not include an internal control, the control may be derived from an established dataset.

[0057] Data for control subjects may be selected from the following groups: 1. datasets containing measurements of the presence or expression levels of biomarkers for a typical population of subjects known to have cancer or a specific cancer; 2. datasets containing measurements of the presence or levels of biomarkers for subjects under test, including previously taken measurements, such as when the subject is known to be healthy, or when the subject has cancer, or when the subject has been diagnosed or is in the early stages of disease progression; 3. datasets containing measurements of the presence or levels of biomarkers for healthy individuals or populations of healthy individuals; and 4. datasets containing measurements of the presence or levels of biomarkers for normal individuals or populations of normal individuals.

[0058] Those skilled in the art can, based on the teachings provided herein, readily determine a baseline for comparison in any diagnostic assay of the present invention without excessive experimentation.

[0059] Compounds that bind to biomarkers when used diagnostically can be linked to diagnostic reagents such as detectable labels to enable easy detection of binding events in vitro or in vivo. Suitable labels include radioisotopes, dye markers, or other imaging reagents for the detection and / or localization of target molecules. Compounds linked to detectable labels can be used with appropriate in vivo imaging techniques such as radiology, fluoroscopy, magnetic resonance imaging (MRI), CAT scans, positron emission tomography (PET), and computed tomography.

[0060] Certain methods can detect susceptibility to cancer treatments containing 2,2'-dithio-bis-ethanesulfonate. As will be understood by those skilled in the art, susceptibility refers to the actual percentage of positives in a test that are correctly identified as susceptible to treatments containing 2,2'-dithio-bis-ethanesulfonate or its analogues. In one embodiment, the methods of the present invention can detect 2,2'-dithio-bis-ethanesulfonate with susceptibility of at least 50%, 60%, or 66%, or at least 75%, 80%, 85%, 88%, 89%, 90%, or at least 95%. In another embodiment, the methods of the present invention can diagnose or detect cancer with susceptibility of at least 80%, or at least 85%, or at least 90%, or at least 95%.

[0061] <Protein detection technology> In one embodiment, a biomarker polypeptide is detected in a patient sample, and the presence and / or level of the polypeptide in the sample may indicate cancer. For example, this method may include the steps of contacting a biological sample derived from a subject with a compound that can bind to a biomarker polypeptide, and detecting the formation of a complex between the compound and the biomarker polypeptide. As used herein, the term “biomarker polypeptide” includes, for example, immunogenic fragments and fragments of biomarker polypeptides, including epitopes of biomarker polypeptides.

[0062] In one embodiment, the compound that binds to the biomarker is an antibody.

[0063] In another embodiment, an antibody against a biomarker polypeptide is detected in a patient sample, and the presence and / or level of the antibody in the sample indicates cancer.

[0064] Preferred detection systems contemplated herein include any known assays for detecting proteins or antibodies in biological samples isolated from human subjects, such as detection-based systems using antibodies or non-antibody compounds, such as small molecules (e.g., chemical compounds, agonists, antagonists, allosteric regulators, competitive inhibitors, or non-competitive inhibitors of proteins). According to these embodiments, antibodies or small molecules can be used in any standard solid-phase or liquid-phase assay format suitable for protein detection. Optical or fluorescent detection, such as using mass spectrometry, MALDI-TOF, biosensor technology, evanescent fiber optics, or fluorescence resonance energy transfer, is explicitly included in the present invention. Assay systems suitable for high-throughput screening of mass samples, such as high-throughput spectroscopy and resonance (e.g., MALDI-TOF, electrospray MS, or nanoelectrospray MS), are also considered. Another suitable protein detection technique involves the use of multiple reaction monitoring (MRM) in LC-MS (LC / MRM-MS) (Anderson and Hunter, 2006).

[0065] The immunoassay format is particularly suitable for selection from the group consisting of immunoblotting, Western blotting, dot blotting, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), and enzyme immunoassay. Modified immunoassays or protein chip technologies utilizing fluorescence resonance energy transfer (FRET), isotope-encoded affinity tagging (ICAT), matrix-assisted laser desorption / ionization time of flight (MALDI-TOF), electrospray ionization (ESI), biosensor technologies, and evanescent fiber optic technologies are also useful.

[0066] <Nucleic acid detection technology> Any suitable technique can be used to enable qualitative and / or quantitative assessment of the levels of biomarker polynucleotides in a sample. As used herein, the terms “nucleic acid molecule” or “polynucleotide” refer to oligonucleotides, polynucleotides, or any fragment thereof.

[0067] Comparisons can be made by referencing standard controls or control levels found in healthy tissue. For example, the level of a transcribed gene can be determined by Northern blotting and / or RT-PCR. With the advent of quantitative (real-time) PCR, quantitative analysis of gene expression can be performed by using appropriate primers for the gene of interest. Nucleic acids can be labeled and hybridized on a gene array, in which case the gene concentration will be directly proportional to the intensity of the radioactive or fluorescent signal produced on the array.

[0068] Methods for direct sequencing of nucleotide sequences are well known to those skilled in the art and can be found, for example, in Ausubel et al., eds., Short Protocols in Molecular Biology, 3rd ed., Wiley, (1995) and Sambrook et al., Molecular Cloning, 3rd ed., Cold Spring Harbor Laboratory Press, (2001). Sequencing can be carried out by any suitable method, such as dideoxy sequencing, chemical sequencing, or variations thereof. Direct sequencing has the advantage of determining any base pair change in a particular sequence.

[0069] For testing purposes, nucleic acids can be isolated from a sample. Appropriate methods will be known to those skilled in the art. For example, RNA can be isolated from the sample to be analyzed using conventional procedures, such as those provided by QIAGEN technology. This RNA is then reverse transcribed into DNA using reverse transcriptase, and the DNA molecule of interest is amplified by PCR technique using specific primers.

[0070] Diagnostic procedures can also be performed directly on patient samples. For example, hybridization or amplification assays such as Southern or Northern blot analysis, immunohistochemistry, single-stranded higher-order polymorphism analysis (SSCP), and PCR analysis are among the useful techniques in this regard. If necessary, target or probe nucleic acids can be immobilized on solid supports such as microtiter plates, membranes, polystyrene beads, glass slides, or other solid phases.

[0071] <Kit> The present invention provides a kit for the diagnosis or detection of cancer. Such a kit may be suitable for the detection of nucleic acid species, or, as described above, for the detection of polypeptide gene products.

[0072] In polypeptide detection, antibodies are most commonly used as components of the kit. However, any agent capable of specifically binding to a biomarker gene product would be useful in this embodiment of the present application. Other components of the kit typically include labels, secondary antibodies, substrates (if the gene is an enzyme), inhibitors, cofactors, and preparations of regulatory gene products, enabling the user to quantify expression levels and assess whether the diagnostic experiment worked correctly. Enzyme-linked immunosorbent assay-based (ELISA) tests and competitive ELISA tests are particularly suitable assays that can be readily performed by those skilled in the art using the kit components.

[0073] If necessary, the kit further includes means for detecting the binding of an antibody to a biomarker polypeptide. Such means include, for example, reporter molecules such as enzymes (e.g., horseradish peroxidase or alkaline phosphatase), dyes, radioactive nucleotides, luminescent groups, fluorescent groups, biotin, or colloidal particles such as colloidal gold or selenium. Preferably, such reporter molecules are directly bound to the antibody.

[0074] In yet another embodiment, the kit may further include a reference sample. In one embodiment, the reference sample includes a polypeptide to be detected by an antibody. Preferably, the polypeptide is at a known concentration. Such polypeptides are particularly useful as standards. Thus, various known concentrations of such polypeptides can be detected using the diagnostic assays described herein.

[0075] To detect nucleic acids, such kits may include a first container, such as a vial or plastic tube, or a microtiter plate containing oligonucleotide probes. The kit may optionally include a second container for holding primers. The probes can hybridize to DNA whose expression changes are associated with cancer, and the primers are useful for amplifying this DNA. Kits containing oligonucleotide probes immobilized on solid supports can also be developed, for example, using arrays (see Supplement to Nature Genetics, No. 21(1), 1999).

[0076] For PCR amplification of nucleic acids, the kit may include nucleic acid primers complementary to at least some of the biomarker genes described herein. The primer set typically includes at least two, preferably four, oligonucleotides capable of specific amplification of DNA. Fluorescently labeled oligonucleotides enabling quantitative PCR measurement may be included (e.g., TaqMan Chemistry, Molecular Beacons). Enzymes suitable for DNA amplification may also be included.

[0077] <Regression Algorithms and Statistics> To develop a panel of biomarkers suitable for the diagnosis or detection of cancer, the inventors analyzed numerous biomarkers in a statistical model. Such improvements in the performance of a test are sometimes referred to as "in-sample" performance. Fair evaluation of a test requires evaluation using extra-sample subjects, i.e., subjects not included in the construction of the initial predictive model. This is achieved by evaluating test performance using cross-validation.

[0078] Tests for statistical significance include linear and nonlinear regression methods such as ANOVA, Kruskal-Wallis, Wilcoxon, Mann-Whitney, odds ratios, and Bayesian stochastic algorithms. However, as the number of biomarkers being measured increases, it may generally be more convenient to use more advanced methods such as random forests, simple logistic regression, and Bayesian networks, to name a few.

[0079] For example, Bayesian probabilities can be employed. In this situation, 10-fold cross-validation can be used to estimate the "out-of-sample" performance of the model in question. For each combination of biomarkers under consideration, the data can be randomly divided into 10 subsamples, each subsample having an equal ratio of healthy subjects to subjects at each stage of the disease. Next, each subsample can be excluded, and a logistic model can be built using the remaining 90% of subjects. This model can then be used to estimate the probability of cancer in the excluded subsamples, providing an estimate of the "out-of-sample" performance. By repeating this for the remaining 9 subsamples, the "out-of-sample" performance can be estimated from the survey data itself. These predicted out-of-sample probabilities can then be compared to the actual disease status of the subjects to create a receiver operated characteristic (ROC) curve, from which the cross-validated susceptibility with 95% specificity can be estimated.

[0080] Each “out-of-sample” performance estimate using cross-validation (or other methods) is unbiased but contains an element of variability. Therefore, ranking models (based on biomarker combinations) can only reflect the relative performance of such models. However, as demonstrated by “out-of-sample” performance assessments, a set of biomarkers that can be used in numerous combinations to generate diagnostic tests almost certainly includes biomarker combinations that can withstand repeated evaluations.

[0081] Therefore, in light of the teachings herein, those skilled in the art will understand that the sensitivity and specificity of a test for diagnosing cancer can be modified by selecting different combinations of the biomarkers described herein. [Examples]

[0082] [Example 1] Non-small cell lung cancer cell line HCC827 was treated with 1 mM and 15 mM disodium 2,2'-dithio-bis-ethanesulfonate alone for 2 hours, and changes in gene expression levels were measured by whole transcriptome profiling using RNA-seq. Untreated control samples for the same period were used as baseline gene expression levels. Using the threshold for multiplicative change of biomarker genes and NRF2 itself, nine genes were identified as increasing in response to exposure to disodium 2,2'-dithio-bis-ethanesulfonate. These nine genes include NQO1 (SEQ ID NO: 2), PHGDH (SEQ ID NO: 3), HMOX1 (SEQ ID NO: 4), SLC7A11 (SEQ ID NO: 5), SRXN1 (SEQ ID NO: 6), SOX2 (SEQ ID NO: 7), GPX2 (SEQ ID NO: 8), GPX3 (SEQ ID NO: 9), and GPX7 (SEQ ID NO: 10). A change in expression levels of more than 1.5 to 2 times from the reference level indicated sensitivity to disodium 2,2'-dithio-bis-ethanesulfonate.

[0083] [Example 2] Figure A demonstrates the induction of nuclear expression of nuclear factor erythroid 2-related factor 2 (Nrf2) by 2,2'-dithio-bis-ethanesulfonate in PC12 cells (rat adrenal medullary pheochromocytoma cell line) differentiated into neuron-like cells by nerve growth factor (NGF). PC12 cells were suspended in DMEM containing 7.5% inactivated fetal bovine serum and 7.5% inactivated horse serum. After seeding in 100 mm culture dishes coated with type I collagen, these cells were primed by incubation for 7 days in the presence of 100 ng / mL NGF. The medium was then replaced with Neurobasal® medium supplemented with 2 mM glutamine containing N-2 supplement and 30 ng / mL NGF, and the cells were incubated again for 3 days to differentiate into neuron-like cells. Next, these cells were treated with 6 mM 2,2'-dithio-bis-ethanesulfonate for 1, 2, 4, and 8 hours, or with 30 μM tert-butylhydroquinone (t-BHQ), a known Nrf2 activator, for 3 hours. Then, nuclear proteins were extracted from the cells for detection of Nrf2 nuclear expression by Western blotting.

[0084] Figure 1 shows the induction of Nrf2 nuclear expression in cells treated with 30 μM t-BHQ and cells treated with 2,2'-dithio-bis-ethanesulfonate. Nuclear expression of 6 mM Nrf2 began 1 hour after the start of treatment, and evident Nrf2 expression persisted up to 8 hours after the start of treatment. In control cells treated with 2,2'-dithio-bis-ethanesulfonate or t-BHQ alone, nuclear expression of Nrf2 was rarely observed at any treatment duration.

[0085] These results revealed that in PC12 cells differentiated into neuron-like cells, 2,2'-dithio-bis-ethanesulfonate led to nuclear expression of Nrf2, indicating sensitivity to 2,2'-dithio-bis-ethanesulfonate.

[0086] Protein sequences from the Uniprot database for NRF2 (SEQ ID NO: 1), NQO1 (SEQ ID NO: 2), PHGDH (SEQ ID NO: 3), HMOX1 (SEQ ID NO: 4), SLC7A11 (SEQ ID NO: 5), SRXN1 (SEQ ID NO: 5), SOX2 (SEQ ID NO: 7), GPX2 (SEQ ID NO: 8), GPX3 (SEQ ID NO: 9), and GPX7 (SEQ ID NO: 10):

[0087] [Sequence ID 1 (Sequence ID 1)] >sp|Q16236|NF2L2 / NFE2L2 or NRF2_Human Nuclear Factor Erythroid 2 Related Factor 2 OS=Homo sapiens OX=9606 GN=NFE2L2 PE=1 SV=3 [ka]

[0088] [Sequence 2] >sp|P15559|NQO1_Human NAD(P)H dehydrogenase [quinone] 1OS=Homo sapiens OX=9606 GN=NQO1 PE=1 SV=1 [ka]

[0089] [Sequence ID 3 (Sequence ID 3)] >sp|O43175|PHGDH or SERA_Human D-3-phosphoglycerate dehydrogenase OS=Homo sapiens OX=9606 GN=PHGDH PE=1 SV=4 [ka]

[0090] [Sequence 4 (Sequence 4)] >sp|P09601|HMOX1_Human Heme Oxygenase 1 OS=Homo sapiens OX=9606 GN=HMOX1 PE=1 SV=1 [ka]

[0091] [Sequence 5] >sp|Q9UPY5|SLC7A11 or XCT_Human Cystine / Glutamate Transporter OS=Homo sapiens OX=9606 GN=SLC7A11 PE=1 SV=1 [ka]

[0092] [Sequence 6] >sp|Q9BYN0|SRXN1_Human Sulfiredoxin-1 OS=Homo sapiens OX=9606 GN=SRXN1 PE=1 SV=2 [ka]

[0093] [Sequence 7 (Sequence 7)] >sp|P48431|SOX2 Human Transcription Factor SOX-2OS=Homo sapiens OX=9606 GN=SOX2 PE=1 SV=1 [ka]

[0094] [Sequence ID 8 (Sequence ID 8)] >sp|P18283|GPX2_Human Glutathione Peroxidase 2OS=Homo sapiens OX=9606 GN=GPX2 PE=1 SV=3 [ka]

[0095] [Sequence ID 9 (Sequence ID 9)] >sp|P22352|GPX3_Human Glutathione Peroxidase 3OS=Homo sapiens OX=9606 GN=GPX3 PE=1 SV=2 [ka]

[0096] [Sequence ID 10] >sp|Q96SL4|GPX7_Human Glutathione Peroxidase 7OS=Homo sapiens OX=9606 GN=GPX7 PE=1 SV=1 [ka]

Claims

1. A method for treating a patient's cancer, (a) The step of obtaining a sample of the cancer from the patient, (b) The step of treating the sample with a 2,2'-dithio-bis-ethanesulfonate analog, (c) A step of obtaining the expression level of one or more of a plurality of biomarkers in the sample, wherein the plurality of biomarkers include NRF2 (SEQ ID NO: 1), (d) A step of determining whether the sample is sensitive to treatment with disodium 2,2'-dithio-bis-ethanesulfonate based on the expression level of NRF2 (SEQ ID NO: 1), (e) A method comprising the step of administering a cancer treatment to the subject containing a 2,2'-dithio-bis-ethanesulfonate disodium analog when the expression level of NRF2 (SEQ ID NO: 1) is greater than 1-fold compared to non-cancer cells.

2. The method according to claim 1, wherein the plurality of biomarkers further comprises one biomarker selected from the group consisting of NQO1 (SEQ ID NO: 2), PHGDH (SEQ ID NO: 3), HMOX1 (SEQ ID NO: 4), SLC7A11 (SEQ ID NO: 5), SRXN1 (SEQ ID NO: 6), SOX2 (SEQ ID NO: 7), GPX2 (SEQ ID NO: 8), GPX3 (SEQ ID NO: 9), and GPX7 (SEQ ID NO: 10).

3. The method according to claim 1, wherein the non-cancer cells are the level of expression in a biological sample obtained from a healthy patient, and are a reference level.

4. The method according to claim 1, wherein the 2,2'-dithio-bis-ethanesulfonate analog is disodium 2,2'-dithio-bis-ethanesulfonate.

5. The method according to claim 1, wherein the cancer treatment comprises the administration of one or more chemotherapeutic agents selected from the group consisting of platinum complexes and taxanes.

6. The method according to claim 1, wherein the cancer treatment includes radiation therapy.

7. The method according to claim 1, wherein one or more nucleic acid molecules from the sample are brought into contact with a device comprising (a) a first single-stranded nucleic acid molecule that can specifically hybridize with the nucleotide of the biomarker, and (b) a second single-stranded nucleic acid molecule that can specifically hybridize with the nucleotides of a plurality of biomarkers, wherein the biomarker is a nucleic acid.

8. A method for treating solid tumor cancer in the subject, (a) The step of treating the tumor with a 2,2'-dithio-bis-ethanesulfonate analog, (b) A step of obtaining the expression level of one of a plurality of biomarkers in a sample from a subject, wherein the plurality of biomarkers include NRF2 (SEQ ID NO: 1), (c) A step of determining whether the tumor is sensitive to treatment with a 2,2'-dithio-bis-ethanesulfonate disodium analog based on the expression level of NRF2 (SEQ ID NO: 1), (d) A method comprising the step of administering a cancer treatment comprising 2,2'-dithio-bis-ethanesulfonate disodium when the expression level of NRF2 (SEQ ID NO: 1) is greater than 1 times that of non-cancer cells.

9. The method according to claim 8, wherein the plurality of biomarkers further comprises one biomarker selected from the group consisting of NQO1 (SEQ ID NO: 2), PHGDH (SEQ ID NO: 3), HMOX1 (SEQ ID NO: 4), SLC7A11 (SEQ ID NO: 5), SRXN1 (SEQ ID NO: 6), SOX2 (SEQ ID NO: 7), GPX2 (SEQ ID NO: 8), GPX3 (SEQ ID NO: 9), and GPX7 (SEQ ID NO: 10).

10. The method according to claim 8, wherein the compound is a disodium 2,2'-dithio-bis-ethanesulfonate analog.

11. The aforementioned compound is present in approximately 14 g / m³ ２ ~Approx. 22g / m ２ The method according to claim 9, which is within the range of claim 9.

12. The method according to claim 10, wherein the chemotherapy treatment comprises the administration of one or more chemotherapeutic agents selected from the group consisting of platinum complexes and taxanes.

13. The method according to claim 11, wherein the platinum complex agent is selected from the group consisting of cisplatin, oxaliplatin, carboplatin, satraplatin, and derivatives and analogs thereof.

14. A method for testing tumor samples from a patient having a known cancer type, wherein the patient is resistant to one or more cancer therapies and has an unknown response to disodium 2,2'-dithio-bis-ethanesulfonate. The step of contacting the sample with disodium 2,2'-dithio-bis-ethanesulfonate, (a) a first single-stranded nucleic acid molecule that can specifically hybridize with the nucleotides of multiple biomarkers, wherein the first biomarker is NFR2; and (b) a second single-stranded nucleic acid molecule that can specifically hybridize with the nucleotides of multiple resistance biomarkers selected from the biomarkers NQO1 (SEQ ID NO: 2), PHGDH (SEQ ID NO: 3), HMOX1 (SEQ ID NO: 4), SLC7A11 (SEQ ID NO: 5), SRXN1 (SEQ ID NO: 6), SOX2 (SEQ ID NO: 7), GPX2 (SEQ ID NO: 8), GPX3 (SEQ ID NO: 9), and GPX7 (SEQ ID NO: 10); the step of bringing the sample into contact with a device comprising: The step of detecting the expression levels of the plurality of biomarkers, A method comprising the step of administering 2,2'-dithio-bis-ethanesulfonate disodium to a patient who has been determined to be responsive to 2,2'-dithio-bis-ethanesulfonate disodium.

15. A change in expression level of more than 1.5 to 2 times may indicate sensitivity to 2,2'-dithio-bis-ethanesulfonate.

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