Biomarkers for cancer therapy

Abstract

To provide biomarkers for cancer therapy.SOLUTION: The disclosure provides a method for using MYC gene as a biomarker for predicting therapeutic efficacy of survivin inhibitors such as YM155 monobromide in cancer therapy, and a related kit, a composition, and a method for conducting the method in a subject in need of cancer diagnosis and treatment. In some embodiments, the method comprises administering a chemotherapeutic agent except for (or other than) YM155 monobromide when the copy number of the MYC gene in the cancer tissue is not substantially increased relative to the reference MYC gene copy number, or when the MYC gene chromosomal locus in the cancer tissue is not translocated relative to the reference MYC gene chromosomal locus.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to PCT / CN2018 / 100206, filed August 13, 2018, and PCT / CN2018 / 115826, filed November 16, 2018, each of which is incorporated by reference in its entirety. [Background technology]

[0002] Embodiments of the present disclosure relate to the use of the MYC gene as a biomarker for predicting the therapeutic efficacy of survivin inhibitors, such as YM155 monobromide, in cancer treatment, as well as related kits, compositions, and methods for diagnosing and treating cancer in subjects in need thereof.

[0003] 2. Description of Related Art YM155 monobromide is a small molecule survivin inhibitor that induces downregulation of survivin and exhibits potent antitumor activity (e.g., Minematsu et al., Drug Metabolism and Disposition, 37:619-628, 2008). YM-155 exerts antitumor effects in various in vivo cancer models, including prostate, pancreatic, and lung cancer (see, e.g., Nakahara et al., Cancer Research 67:8014-8021, 2007, and Na et al., PLoS One 7(6), 2012). However, there is a need in the art to better predict the anti-cancer therapeutic efficacy of YM155 monobromide, and thereby identify patients who will most likely benefit from treatment with this and other therapies. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Minematsu et al.,Drug Metabolism and Disposition,37:619-628,2008 [Non-Patent Document 2] Nakahara et al., Cancer Research 67:8014-8021,2007 [Non-Patent Document 3] Na et al.,PLoS One 7(6),2012 Summary of the Invention [Means for solving the problem]

[0005] Embodiments of the present disclosure include methods for treating cancer in a subject in need thereof, comprising: (a) determining the MYC gene copy number or MYC gene chromosomal location site in a sample of cancer tissue from a subject; (b) if the MYC gene copy number in the cancer tissue is increased compared to the MYC gene copy number reference, or if the MYC gene chromosomal location site in the cancer tissue is translocated compared to the MYC gene chromosomal location site reference, administering to the subject YM155 monobromide [1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(pyrazin-2-ylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide], or an analog or derivative thereof, thereby treating cancer in a subject in need thereof.

[0006] Some embodiments include administering to the subject a chemotherapeutic agent that excludes (or is not limited to) YM155 monobromide if the MYC gene copy number in the cancer tissue is not substantially increased compared to a reference MYC gene copy number or if the MYC gene chromosomal location site in the cancer tissue is not translocated compared to a reference MYC gene chromosomal location site.

[0007] Also included is a method for predicting therapeutic response to YM155 monobromide [1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(pyrazin-2-ylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide], or an analog or derivative thereof, in a cancer subject, (a) determining the MYC gene copy number or MYC gene chromosomal location site in a sample of cancer tissue from a subject; (b)(i) characterizing the subject as responsive to YM155 monobromide therapy if the MYC gene copy number in the cancer tissue is increased compared to a reference MYC gene copy number, or if the MYC gene chromosomal location site in the cancer tissue is translocated compared to a reference MYC gene chromosomal location site; or (ii) characterizing the subject as non-responsive to YM155 monobromide therapy if the MYC gene copy number in the cancer tissue is not substantially increased compared to the reference MYC gene copy number or if the MYC gene chromosomal location site in the cancer tissue is not translocated compared to the reference MYC gene chromosomal location site, thereby predicting a therapeutic response to YM155 monobromide in the cancer subject.

[0008] Some embodiments include administering YM155 monobromide to the subject if the control is characterized as responsive to YM155 monobromide therapy.Some embodiments include administering a chemotherapeutic agent to the subject that excludes YM155 monobromide if the subject is characterized as non-responsive to YM155 monobromide therapy.

[0009] In some embodiments, the MYC gene copy number in the cancer tissue is increased by about or at least about 1.5, 2, 3, 4, 5, 6, 7, 8, 9, or 10 fold compared to a reference MYC gene copy number.

[0010] Some embodiments include determining the MYC gene copy number in cancer tissue by array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) array, copy number variation (CNV) sequencing, or multiplex ligation-dependent probe amplification (MLPA). Some embodiments include determining the MYC gene chromosomal location site in cancer tissue by in situ hybridization (ISH), fluorescent in situ hybridization (FISH), next generation sequencing (NGS), or comparative genomic hybridization (CGH). Some embodiments include obtaining the MYC gene copy number reference from a database, or determining the MYC gene copy number reference from non-cancerous tissue from a control, optionally by aCGH, SNP array, CNV sequence, or MLPA. Some embodiments include obtaining the MYC gene chromosomal location site from a database, or determining the MYC gene chromosomal location site reference from non-cancerous tissue from a control, optionally by ISH, FISH, NGS, or CGH.

[0011] Some embodiments include obtaining a sample of cancer tissue from a subject. In certain embodiments, the sample of cancer tissue is a surgical sample, a biopsy sample, a pleural effusion sample, or an ascites sample obtained from the subject, optionally selected from one or more of lung, blood, breast, gastrointestinal tract (stomach, colon, rectum), ovary, pancreas, liver, bladder, cervix, nerve, uterus, salivary gland, kidney, prostate, thyroid, or muscle tissue.

[0012] In certain embodiments, the subject is a human subject.

[0013] In certain embodiments, the cancer is a carcinoma, a rhabdomyosarcoma, e.g., a sarcoma such as alveolar rhabdomyosarcoma (including sarcomas derived from bone, tendon, cartilage, muscle, fat, fibrous, vascular, adipose, and / or connective tissue), a neuroblastoma, a medulloblastoma, an astrocytoma, a glioblastoma multiforme, a retinoblastoma, a myeloma, a leukemia, a lymphoma (including Hodgkin's lymphoma and non-Hodgkin's lymphoma), an adenosquamous carcinoma, The cancer is selected from one or more of carcinosarcoma, mixed mesodermal tumor, teratocarcinoma, lung cancer (including non-small cell lung cancer, small cell lung cancer, adenocarcinoma, and squamous cell lung cancer), breast cancer (including metastatic breast cancer), gastrointestinal cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, cervical cancer, glioblastoma, uterine cancer, salivary gland cancer, kidney or renal cancer (e.g., Wilm's tumor), prostate cancer, thyroid cancer, and head and neck cancer.

[0014] In certain embodiments, MYC gene is selected from MYCC and MYCN.In certain embodiments, MYC gene is MYCC, and cancer is selected from lung cancer and blood cancer, optionally leukemia and lymphoma.In certain embodiments, MYC gene is MYCN, and cancer is selected from neuroblastoma, small cell lung cancer, prostate cancer, alveolar rhabdomyosarcoma, medulloblastoma, glioblastoma multiforme, retinoblastoma, and breast cancer.

[0015] Also included is the use of a diagnostic kit for determining therapeutic response to YM155 monobromide [1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(pyrazin-2-ylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide], or an analogue or derivative thereof, therapy in a cancer subject, comprising means for measuring MYC gene copy number or MYC gene chromosomal location site in a sample of tissue from the subject, including cancerous tissue and non-cancerous tissue.

[0016] In certain embodiments, the means for measuring MYC gene copy number comprises reagents for performing a diagnostic assay selected from one or more of array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) array, copy number variation (CNV) sequencing, and multiplex ligation-dependent probe amplification (MLPA) on the human MYC gene.

[0017] In certain embodiments, the means for measuring the MYC gene chromosomal location site comprises a reagent for performing a diagnostic assay selected from one or more of in situ hybridization (ISH), fluorescent in situ hybridization (FISH), next generation sequencing (NGS), and comparative genomic hybridization (CGH) on the human MYC gene. Some embodiments comprise a MYC gene copy number reference value obtained from a database or determined from non-cancerous tissue from a control. Some embodiments comprise a MYC gene chromosomal location site reference obtained from a database or determined from non-cancerous tissue from a control.

[0018] It also includes a patient care kit. (a) a means for measuring MYC gene copy number or MYC gene chromosomal location site in a sample of tissue from a subject, including cancerous tissue and non-cancerous tissue; (b) YM155 monobromide [1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(pyrazin-2-ylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide], or an analogue or derivative thereof.

[0019] In certain embodiments, the means for measuring MYC gene copy number comprises reagents for performing diagnostic assays on the human MYC gene selected from one or more of array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) array, copy number variation (CNV) sequencing, and multiplex ligation-dependent probe amplification (MLPA). In certain embodiments, the means for measuring MYC gene chromosomal location comprises reagents for performing diagnostic assays on the human MYC gene selected from one or more of in situ hybridization (ISH), fluorescent in situ hybridization (FISH), next generation sequencing (NGS), and comparative genomic hybridization (CGH).

[0020] Some embodiments include a MYC gene copy number reference value obtained from a database or determined from non-cancerous tissue from a control.Some embodiments include a MYC gene chromosomal location site reference obtained from a database or determined from non-cancerous tissue from a control.

[0021] In some uses or kits, the cancer is a carcinoma, a rhabdomyosarcoma, a sarcoma, such as, for example, alveolar rhabdomyosarcoma (including sarcomas derived from bone, tendon, cartilage, muscle, fat, fibrous, vascular, adipose, and / or connective tissue), a neuroblastoma, a medulloblastoma, an astrocytoma, a glioblastoma multiforme, a retinoblastoma, a myeloma, a leukemia, a lymphoma (including Hodgkin's lymphoma and non-Hodgkin's lymphoma), adenoma, squamous cell carcinoma, or a combination thereof. The cancer is selected from one or more of skin cancer, carcinosarcoma, mixed mesodermal tumor, teratocarcinoma, lung cancer (including non-small cell lung cancer, small cell lung cancer, adenocarcinoma, and squamous cell lung cancer), breast cancer (including metastatic breast cancer), gastrointestinal cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, cervical cancer, glioblastoma, uterine cancer, salivary gland cancer, kidney cancer or renal cancer (e.g., Wilm's tumor), prostate cancer, thyroid cancer, and head and neck cancer.

[0022] In some uses or kits, MYC gene is selected from MYCC and MYCN.In some examples, MYC gene is MYCC, and cancer is selected from lung cancer and blood cancer, optionally leukemia and lymphoma.In some examples, MYC gene is MYCN, and cancer is selected from neuroblastoma, small cell lung cancer, prostate cancer, alveolar rhabdomyosarcoma, medulloblastoma, glioblastoma multiforme, retinoblastoma, and breast cancer. [Brief description of the drawings]

[0023] [Figure 1] The chemical structure of YM155 monobromide (CAS 781661-94-7) is shown. [Diagram 2] Figure 1 shows that YM155 inhibits cell proliferation in human lung cancer cell lines. NCI-H1975, HCC827, NCI-H226, and HCC4006 cells were cultured in 96-well plates and treated with the indicated doses (nM) of YM155. Cell proliferation was detected by EdU proliferation assay. Data presented are mean ± SEM. [Diagram 3] Figure 1 shows that YM155 inhibits cell proliferation of human acute myeloid leukemia (AML) cell lines. U937 and HL-60 cells were cultured in 96-well plates and treated with the indicated doses (nM) of YM155. Cell proliferation was detected by XTT cell viability assay. Data presented are mean ± SEM. [Figure 4] Figure 1 shows that YM155 inhibits cell proliferation in U937 and RAMOS cell lines. U937 and RAMOS cells were cultured in 96-well plates and treated with YM155 at the indicated doses (nM). Cell proliferation was detected by XTT cell viability assay. Data presented are mean ± SEM. [Diagram 5] Provides genomic information for the human MYC gene (see http: / / uswest.ensembl.org / Homo_sapiens / Location / View?db=core;g=ENSG00000136997;r=8:127735434-127742951). [Figure 6]Provides genomic information for the human MYC gene (see http: / / uswest.ensembl.org / Homo_sapiens / Location / View?db=core;g=ENSG00000136997;r=8:127735434-127742951). [Figure 7] We show that YM155 inhibits cell proliferation in human neuroblastoma cell lines, especially those characterized by increased copy number of N-MYC (see also Table E3). Cell proliferation was detected by EdU assay. Data presented are mean ± SD. [Figure 8] The location of the MYCN gene on the short (p) arm of chromosome 2 at position 24.3 is shown. [Figure 9] Figure 9 shows the antitumor effect of YM155 in HCC827 xenografts in mice. Figure 9A shows the tumor growth curve, and Figure 9B shows the data as the mean tumor volume ± SD (n=5). **p<0.01 compared with the vehicle group. [Figure 10] 10 shows the antitumor effect of YM155 in HCC4006 xenografts in mice. Figure 10A shows the tumor growth curves, and Figure 10B shows the data as mean tumor volume ± SD (n=8). [Figure 11] Figure 11 shows the antitumor effect of YM155 in RAMOS xenografts in mice. Figure 11A shows the tumor growth curve, and Figure 11B shows the data as the mean tumor volume ± SD (n=3). **p<0.01 compared to the vehicle group. [Figure 12] Figure 12 shows the antitumor effect of YM155 in U937 xenografts in mice. Figure 12A shows the tumor growth curves, and Figure 12B shows the data as mean tumor volume ± SD (n=3). [Figure 13] Figure 13 shows the antitumor effect of YM155 in IMR-32 xenografts in mice. Figure 13A shows the tumor growth curves, and Figure 13B shows the data as mean tumor volume ± SD (n=3). [Figure 14]Figure 14 shows the antitumor effect of YM155 in SH-SY5Y xenografts in mice. Figure 14A shows the tumor growth curves, and Figure 14B shows the data as mean tumor volume ± SD (n=2 for vehicle, n=3 for YM155 treatment). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0024] The present disclosure relates to the surprising discovery that MYC gene amplification and / or translocation in human malignancies is associated with increased anti-cancer efficacy of survivin inhibitors, such as YM155 monobromide, and thus may be used as a biomarker(s) for optimizing cancer treatment with these and other agents. Without wishing to be bound by any theory, it is believed that expression of the MYC oncogene in MYC-amplified or MYC-translocated cancer cells is otherwise pro-apoptotic, except that its pro-apoptotic signaling is negatively controlled by survivin, a member of the inhibitor of apoptosis (IAP) protein family that inhibits caspases and blocks cell death. It is therefore expected that survivin inhibitors, such as YM155 monobromide, can be used to block the apoptosis inhibitory activity of survivin in the context of MYC amplification and / or translocation, favoring the pro-apoptotic signaling of MYC to increase cancer cell death, thereby providing optimal therapeutic efficacy in this context.

[0025] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. Although any methods, materials, compositions, reagents, cells similar or equivalent to those described herein can be used to carry out or test the subject matter of this disclosure, preferred methods and materials are described. All publications and references cited herein, including but not limited to patents and patent applications, are incorporated herein by reference in their entirety as if each individual publication or reference was specifically and individually indicated to be incorporated herein by reference as if fully set forth. Any patent application to which this application claims priority is incorporated herein by reference in its entirety in the manner described above for publications and references.

[0026] For purposes of this disclosure, the following terms are defined below.

[0027] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0028] "Antagonist" or "inhibitor" refers to a biological structure or chemical agent that prevents or otherwise reduces the physiological action of another molecule, such as a protein (e.g., survivin). In some cases, the antagonist or inhibitor specifically binds to the other molecule and / or a functional ligand of the other molecule. In some cases, the antagonist or inhibitor downregulates the expression of the other molecule (e.g., survivin). Full antagonists and partial antagonists are included.

[0029] "Agonist" or "activator" refers to a biological structure or chemical agent that increases or enhances the physiological action of another agent or molecule. In some cases, agonists specifically bind to other agents or molecules. Full agonists and partial agonists are included.

[0030] "About" means a quantity, level, value, number, frequency, percentage, dimension, size, amount, weight, or length that differs by about 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1% from a reference quantity, level, value, number, frequency, percentage, dimension, size, amount, weight, or length.

[0031] Throughout this disclosure, unless the context requires otherwise, the terms "comprise", "comprises" and "comprising" are understood to imply the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step or element or group of steps or elements.

[0032] "Consisting of" means including and limited to whatever follows the phrase "consisting of". Thus, the phrase "consisting of" indicates that the recited elements are required or essential, and that no other elements may be present. "Consisting essentially of" means including any elements recited after the phrase, and is limited to other elements that do not interfere with or contribute to the activity or action set forth in this disclosure for the recited elements. Thus, the phrase "consisting essentially of" indicates that the recited elements are required or essential, but that other elements are optional and may or may not be present depending on whether they materially affect the activity or action of the recited elements.

[0033] "50% inhibitory concentration" (or "IC 50" is a measure of the potency of a drug to inhibit a particular biological or biochemical function. This quantitative measure indicates how much of a particular drug (inhibitor) is required to half-inhibit a given biological process (or a component of the process, i.e., an enzyme, cell, cell receptor, or microorganism). Values ​​are typically expressed as molar concentrations. Concentrations are commonly used as a measure of antagonist drug potency in pharmacological studies. In some cases, the IC 50 represents the concentration of drug required for 50% inhibition in vitro. IC 50 can be determined by constructing a dose-response curve and examining the effect of different concentrations of the agent on the desired activity, e.g., inhibition of tumor cell proliferation, tumor cell killing.

[0034] An "increased" or "enhanced" amount is typically a "statistically significant" amount, and is about or at least about 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10, 15, 20, 30, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, or 1000 fold, or about or less than about and may include increases that are about 5%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000% (including all integers and ranges in between). A "decreased" or "reduced" amount is typically a "statistically significant" amount, and is about or at least about 1.2, 1.4, 1.6, 1.8, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 1000 times, or about or at least about 5%, 10%, 11%, 12%, 14%, 16%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 50%, 50%, 50%, 50%, 60%, 60%, 70%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, 170%, 180%, 190%, 200%, 210%, 220%, 230%, 240%, 250%, 260%, 270%, 280%, 290%, 300%, 315%, 320%, 335%, 340%, 350%, 365%, 370%, 380%, 390%, 400%, 410%, 420%, %, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000% (including all integers and ranges in between).

[0035] The terms "polynucleotide" and "nucleic acid" include mRNA, RNA, cRNA, cDNA, and DNA, including genomic DNA. The terms typically refer to a polymeric form of nucleotides (either ribonucleotides or deoxynucleotides, or modified forms of either type of nucleotide) of at least 10 bases in length. The terms include single- and double-stranded forms of DNA.

[0036] "Gene" refers to a hereditary unit consisting of a sequence of DNA that occupies a specific location on a chromosome and codes for a functional molecule or protein. The structure of a gene consists of numerous elements of which the actual protein coding sequence is often only a small part. These elements include non-transcribed DNA regions, as well as non-translated regions of RNA. In addition, genes can have expression modifying control regions that extend for many kilobases upstream or downstream of the coding sequence. The information in a gene can also be represented by (or found in) the sequence of the RNA or the encoded protein.

[0037] A "subject" or "subject in need thereof" includes a mammalian subject, such as a human subject.

[0038] "Statistically significant" means that the result is unlikely to have occurred by chance. Statistical significance can be determined by any method known in the art. Commonly used measures of significance include p-value, which is the frequency or probability that the observed event occurs if the null hypothesis is true. If the p-value obtained is less than the significance level, the null hypothesis is rejected. In simple cases, the significance level is defined as a p-value of 0.05 or less.

[0039] "Substantially" or "essentially" means nearly entirely or completely, e.g., 95% or more of a given quantity.

[0040] "Therapeutic response" refers to an improvement in symptoms (whether sustained or not) upon administration of a therapeutic response.

[0041] As used herein, the term "therapeutically effective amount," "therapeutic dose," "prophylactically effective amount," or "diagnostically effective amount" is the amount of agent required to elicit a desired biological response following administration.

[0042] As used herein, "treatment" of a subject (e.g., a mammal, such as a human) or cell is any type of intervention used to change the natural course of a subject or cell. Treatment includes, but is not limited to, administration of a pharmaceutical composition, and can be performed either prophylactically or after the onset of a pathological event or contact with a pathogen. Also included are "prophylactic" treatments, which can be directed to reducing the rate of progression of the disease or condition being treated, delaying the onset of the disease or condition, or reducing the severity of its onset. "Treatment" or "prevention" does not necessarily indicate complete eradication, cure, or prevention of the disease or condition or its associated symptoms.

[0043] The term "wild type" refers to a gene or gene product (eg, a polypeptide) that is most frequently observed in a population and is thus arbitrarily designated the "normal" or "wild" form of the gene.

[0044] Each embodiment herein is intended to apply to all other embodiments unless expressly stated otherwise.

[0045] This disclosure relates, in part, to the surprising discovery that amplification and / or translocation of the MYC gene in human malignancies is associated with increased anti-cancer efficacy of the chemotherapeutic agent YM155 monobromide and therefore may be used as a biomarker for optimizing cancer treatment with that and other agents.

[0046] Accordingly, embodiments of the present disclosure include a method for treating cancer in a subject in need thereof, comprising: (a) determining a MYC gene copy number or a MYC gene chromosomal location site in a sample of cancer tissue from the subject; and (b) if the MYC gene copy number in the cancer tissue is increased compared to a reference MYC gene copy number, or if the MYC gene chromosomal location site in the cancer tissue is translocated compared to a reference MYC gene chromosomal location site, administering YM155 monobromide [1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(pyrazin-2-ylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide] to the subject, thereby treating cancer in the subject in need thereof. Certain embodiments include administering to the subject a chemotherapeutic agent that excludes (or is not limited to) YM155 monobromide if the MYC gene copy number in the cancer tissue is not substantially increased compared to a reference MYC gene copy number (e.g., is the same as or is increased by less than about 1.1 fold) or if the MYC gene chromosomal location site in the cancer tissue is not translocated compared to a reference MYC gene chromosomal location site.

[0047] Also included is a method for predicting a therapeutic response to YM155 monobromide [1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(pyrazin-2-ylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide] in a subject with cancer, comprising: (a) determining a MYC gene copy number or a MYC gene chromosomal location site in a sample of cancer tissue from the subject; and (b) (i) determining whether the MYC gene copy number in the cancer tissue is increased compared to a reference MYC gene copy number or whether the MYC gene chromosomal location site in the cancer tissue is increased compared to a reference MYC gene copy number. characterizing the subject as responsive to YM155 monobromide therapy if the MYC gene copy number in the cancer tissue is translocated compared to the reference; or (ii) characterizing the subject as non-responsive to YM155 monobromide therapy if the MYC gene copy number in the cancer tissue is not substantially increased compared to the MYC gene copy number reference (e.g., the same or increased by less than about 1.1 fold), or if the MYC gene chromosomal location site in the cancer tissue is not translocated compared to the MYC gene chromosomal location site reference, thereby predicting the therapeutic response to YM155 monobromide in the cancer subject. Certain embodiments include administering YM155 monobromide to the subject if the control is characterized as responsive to YM155 monobromide therapy. Some embodiments include administering a chemotherapeutic agent other than (or other than) YM155 monobromide to the subject if the subject is characterized as non-responsive to YM155 monobromide therapy.

[0048] "MYC gene" or "MYC oncogene" refers to a family of proto-oncogenes that encode transcription factors, examples of which include c-Myc (also MYCC) and N-myc (also MYCN).

[0049] The MYCC gene encodes a nuclear phosphoprotein involved in cell cycle progression, apoptosis, and cell transformation. The encoded protein forms a heterodimer with the associated transcription factor MAX. This complex binds to E-box DNA consensus sequences and controls the transcription of specific target genes. Evidence indicates that translation initiates from both the upstream in-frame non-AUG (CUG) and downstream AUG start sites, resulting in the production of two isoforms with distinct N-termini. In the human genome, the MYCC gene is located on chromosome 8:127, 735, 434-127, 741, 434, forward strand (see, e.g., Figures 5 and 6, and Gene:MYC ENSG00000136997).

[0050] The MYCN gene encodes a protein with a basic helix-loop-helix (bHLH) domain. It is located in the cell nucleus and dimerizes with another bHLH protein to bind to DNA. MYCN is overexpressed in many different types of cancer, including, for example, neuroblastoma, rhabdomyosarcoma, medulloblastoma, astrocytoma, glioblastoma, retinoblastoma, prostate cancer, breast cancer, Wilm's tumor, and small cell lung cancer (see, for example, Beltran, Mol Cancer Res. 12: 815-822, 2014). In fact, MYCN amplification is an adverse prognostic factor in neuroblastoma. Amplicons (materials co-amplified with MYCN) vary between subjects and, in certain instances, include, for example, the DDX1 gene. In some cases, MYCN amplification correlates with 1p36 deletions and chromosome 17q gains. In the human genome, the MYCN gene is located on the short (p) arm of chromosome 2 at position 24.3 (cytogenetic location of 2p24.3; molecular location of base pairs 15,940,438 to 15,947,007 on chromosome 2; see also Figure 8).

[0051] Thus, in certain embodiments, the MYC gene is selected from MYCC and MYCN.

[0052] "YM155 monobromide" has the molecular formula C 20 H 19Refers to the small molecule 1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(pyrazin-2-ylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide, having N4O3·Br and CAS number 781661-94-7, including its pharma- ceutically acceptable salts and acids. Also includes biologically active or equivalent analogs and / or derivatives of YM155 monobromide.

[0053] As mentioned above, in some cases, the MYC gene copy number in cancer tissue is increased compared to the MYC gene copy number reference.In certain embodiments, the MYC gene copy number in cancer tissue is increased by a statistically significant amount compared to the MYC gene copy number reference.In some embodiments, the MYC gene copy number in cancer tissue is increased by about or at least about 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9 or 10 times (or more) compared to the MYC gene copy number reference.

[0054] The MYC gene copy number in cancer tissue can be determined by any of a variety of methods.For example, in some embodiments, the MYC gene copy number is determined by array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) array, copy number variation (CNV) sequencing, or multiplex ligation-dependent probe amplification (MLPA).Therefore, certain embodiments include determining or detecting the copy number of the MYC gene in a sample of cancer tissue from a subject in need thereof.Also include comparing the copy number of the MYC gene in the sample of cancer tissue with a MYC gene copy number reference.

[0055] The MYC gene chromosomal location site in cancer tissue can be determined by any of a variety of methods.For example, in some embodiments, the MYC gene chromosomal location site in cancer tissue is determined by in situ hybridization (ISH), fluorescent in situ hybridization (FISH), next generation sequencing (NGS), or comparative genomic hybridization (CGH).Therefore, certain embodiments include determining or detecting the MYC gene chromosomal location site in a sample of cancer tissue from a subject in need thereof.Also include comparing the MYC gene chromosomal location site in cancer tissue with a MYC gene chromosomal location reference.

[0056] CGH refers to a molecular cytogenetic method for analyzing copy number changes (CNVs) in DNA of a test sample compared to a reference sample, without the need to culture cells. This technique allows for a fast and efficient comparison between two genomic DNA samples originating from two sources that are often most closely related, since they are suspected to contain differences in terms of either gain or loss of either whole chromosomes or subchromosomal regions (parts of whole chromosomes). This technique was originally developed to evaluate the differences between the chromosomal complements of solid tumors and normal tissues (see, for example, Kallioniemi et al., Science. 258(5083):818-821, 1992). The use of DNA microarrays in conjunction with CGH techniques has led to the development of a more specific form of array CGH (aCGH), allowing locus-by-loci CNV measurements with a resolution as low as 100 kilobases (see, for example, Pinkel, Annu Rev Genom Hum Genet. 6:331-354, 2005). CNVs are a common form of significant genetic variation that leads to abnormal copy numbers of large genomic regions in cells, and high-resolution sequence data can be analyzed by next-generation sequencing (NGS) to identify genes that are involved (see, e.g., Zhao et al., BMC Bioinformatics. 14 Suppl 11:S1, 2013). MLPA refers to a variation of multiplex polymerase chain reaction that allows the amplification of multiple targets with only a single primer pair (see, e.g., Schouten et al., Nucleic Acids Res. 30(12):e57, 2002). In situ hybridization (ISH) and fluorescent in situ hybridization (FISH) refer to types of hybridization that use labeled complementary DNA, RNA, or modified nucleic acid strands (i.e., probes) to localize specific DNA or RNA sequences in a tissue section or section (in situ) (see, e.g., Parra & Windle, Nature Genetics. 5:17-21, 1993, and Gall & Pardue, PNAS USA. 63:378-383, 1969).Thus, in some cases, the methods and kits described herein use and / or include reagents for carrying out any one or more of the foregoing techniques.

[0057] Examples of "references" (e.g., MYC gene copy number "reference", MYC gene chromosomal site "reference") include values ​​or amounts or locations obtained from a database, e.g., "wild type" MYC gene copy number values ​​or amounts, or "wild type" MYC gene chromosomal site sites (see, e.g., Figures 5 and 6 for human MYCC gene chromosomal site references, and Figure 8 for human MYCN gene chromosomal site references). "References" also include values ​​or amounts or locations obtained from one or more controls, e.g., non-cancer tissues from one or more healthy or non-cancer control subjects (e.g., a population of healthy or non-cancer control subjects), or one or more corresponding non-cancer control tissues from the subject being tested. Typically, the "corresponding" non-cancer control tissues are obtained from the same type of tissue as the cancer tissue being tested. As with cancer tissues, the MYC gene copy number reference from a non-cancer control can be determined by any of a variety of methods, including, e.g., aCGH, SNP arrays, CNV sequences, and / or MLPA (see above). Similarly, MYC gene chromosomal location site references from non-cancer controls can be determined by any of a variety of methods, including, for example, ISH, FISH, NGS, and / or CGH (see above).

[0058] In some embodiments, the sample of cancer tissue (or non-cancer control tissue) is a surgical sample, a biopsy sample, a pleural effusion sample, or an ascites sample from the subject.Specific examples of the sample of cancer tissue (or non-cancer control tissue) include lung, blood, breast, gastrointestinal tract (stomach, colon, rectum), ovary, pancreas, liver, bladder, cervix, nerve, uterus, salivary gland, kidney, prostate, thyroid, or muscle tissue.Certain embodiments include, for example, obtaining a sample of cancer tissue (or non-cancer control tissue) from the subject before determining MYC gene copy level or MYC gene chromosomal location site.

[0059] In some embodiments, the subject is a human subject.

[0060] As mentioned above, certain embodiments include administering to the subject an anti-cancer agent that excludes (or is not limited to) YM155 monobromide when the subject is characterized as being non-responsive to YM155 monobromide therapy, for example, when the MYC gene copy number in the cancer tissue is not substantially increased compared to the MYC gene copy number reference, or when the MYC gene chromosomal location site in the cancer tissue is not translocated compared to the MYC gene chromosomal location site reference. Exemplary anti-cancer agents (other than YM155 monobromide) for administration to a subject characterized as being non-responsive to YM155 monobromide therapy include, for example, small molecules such as cytotoxic agents, chemotherapeutic agents, and antiangiogenic agents that are considered to be useful in the treatment of various cancers. General classes of anti-cancer agents include, but are not limited to, alkylating agents, antimetabolites, anthracyclines, antitumor antibiotics, platinums, type I topoisomerase inhibitors, type II topoisomerase inhibitors, vinca alkaloids, and taxanes.

[0061] Specific examples of anti-cancer drugs for administration to subjects characterized as non-responsive to YM155 monobromide therapy include chlorambucil, cyclophosphamide, cilengitide, lomustine (CCNU), melphalan, procarbazine, thiotepa, carmustine (BCNU), enzastaurin, busulfan, daunorubicin, doxorubicin, gefitinib, erlotinib. These include idarubicin, temozolomide, epirubicin, mitoxantrone, bleomycin, cisplatin, carboplatin, oxaliplatin, camptothecin, irinotecan, topotecan, amsacrine, etoposide, etoposide phosphate, teniposide, temsirolimus, everolimus, vincristine, vinblastine, vinorelbine, vindesine, CT52923, and paclitaxel, as well as pharmaceutically acceptable salts, acids, or derivatives of any of the above. Further examples of anticancer drugs include imatinib, crizotinib, dasatinib, sorafenib, pazopanib, sunitinib, vatalanib, geftinib, erlotinib, AEE-788, dichloroacetate, tamoxifen, fasudil, SB-681323, and semaxanib (SU5416) (Chico et al. al., Nat Rev Drug Discov. 8:829-909, 2009).

[0062] Further examples of anti-cancer agents for administration to subjects characterized as non-responsive to YM155 monobromide therapy include alkylating agents, such as thiotepa, cyclophosphamide (CYTOXAN™); alkyl sulfonates, such as busulfan, improsulfan, and piposulfan; aziridines, such as benzodopa, carboquone, meturedopa, and uredopa; altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylthio. ethylenimines and methylamelamines, including trimethylolomelamine; nitrogen mustards, such as chlorambucil, chlornaphazine, clophosfamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nobuenbiquine, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas, such as carmustine, chlorozo tosine, fotemustine, lomustine, nimustine, ranimustine; antibiotics, such as aclacinomycin, actinomycin, ausramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid , nogalamycin, olivomycin, peplomycin, potfilomycin, puromycin, queramycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine;Pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, 5-FU; androgens such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; antiadrenals such as aminoglutethimide, mitotane, trilostane; folic acid supplements such as frolinic acid acid);acegrastone;aldophosphamide glycoside;aminolevulinic acid;amsacrine;bestrabucil;bisantrene;edatraxate;defofamine;demecolcine;diazicon;elformitin;elliptinium acetate;ethoglucide;gallium nitrate;hydroxyurea;lentinan;lonidamine;mitoguazone;mitoxantrone;mopidamol;nitracrine;pentostatin;phenamet;pirarubicin;podophylline Acid; 2-ethylhydrazide; Procarbazine; PSK; Razoxane; Sizofiran; Spirogermanium; Tenuazonic acid; Triazicon; 2,2',2"-Trichlorotriethylamine; Urethane; Vindesine; Dacarbazine; Mannomustine; Mitobronitol; Mitolactol; Pipobroman; Gacytosine; Arabinoside ("Ara-C"), cyclophosphamide; Thiotepa; Taxoids, such as paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, NJ), and docetaxel (TAXOTERE®, Rhone-Poulenc Rorer, Antony, France); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitors RFS 2000; difluoromethylomithine (DMFO);Retinoic acid derivatives, such as Targretin™ (bexarotene), Panretin™ (alitretinoin); ONTAK™ (denileukin diftitox); esperamycin; capecitabine; and pharma- ceutically acceptable salts, acids, or derivatives of any of the above.

[0063] Further examples of anti-cancer agents for administration to subjects characterized as non-responsive to YM155 monobromide therapy include antiestrogens, including, for example, tamoxifen, raloxifene, aromatase-inhibiting 4(5)-imidazole, 4-hydroxytamoxifen, trioxyphene, ketoxifene, LY117018, onapristone, and toremifene (Fareston); and antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and antihormonal agents that act to regulate or inhibit hormone action on tumors, such as pharmacologic acceptable salts, acids, or derivatives of any of the above.

[0064] Further examples of anti-cancer agents for administration to subjects characterized as non-responsive to YM155 monobromide therapy include 3F8, 8H9, abagovomab, adecatumumab, afutuzumab, alemtuzumab, alacizumab (pegol), amatuximab, apolizumab, bavituximab, bectumomab, belimumab, bevacizumab, bivatuzumab (mertansine), brentuximab vedotin, cantuzumab (mertansine), cantuzumab (ravtansine), capromab (pendetide), catumaxomab, cetuximab, sitatuzumab (bogatox), situatuzumab (situatuzumab ... Custumumab, clivatuzumab (tetraxetan), conatumumab, dacetuzumab, dalotuzumab, detumomab, drozitumab, ecromeximab, edrecolomab, elotuzumab, enabatuzumab, ensituximab, epratuzumab, ertumaxomab, etaracizumab, farletuzumab, FBTA05, figitumumab, framvotumab, galiximab, gemtuzumab, ganitumab, gemtuzumab (ozogamicin), girentuximab, glenbatumumab (vedotin), ibritumomab tiuxetan, icrucumab, igovomab, indatuxima Bravtansine, Intetumumab, Inotuzumab ozogamicin, Iprimumab (MDX-101), Iratumumab, Labetuzumab, Lexatumumab, Lintuzumab, Lorvotuzumab (Mertansine), Lukatumumab, Rumiriximab, Mapatumumab, Matuzumab, Milatuzumab, Mitumomab, Mogamulizumab, Moxetumomab (Pasudotox), Nacolomab (Tafenatox), Naptumomab (Estafenatox), Narunatumumab, Necitumumab, Nimotuzumab, Nivolumab, Neuradiab® (with or without radioactive iodine), NR-LU-10, ofatumumab, olaratumumab, onartuzumab, oportuzumab (monatox), oregovomab, panitumumab, patritumab, pemtumomab, pertuzumab, pritumumab, racotumomab, radletumab, ramucirumab, rilotumumab, rituximab, lobatumumab, samalizumab, sibrotuzumab, siltuximab, tabalumab, taplitumomab (paptox), tenatumomab, teprotumumab, TGN1412, ticilimumab, tremelimumab, tigatuzumab, TNX-650, tositumomab, TRBS07, trastuzumab,Anti-cancer antibodies such as tucotuzumab (celmoleukin), ublituximab, urelumab, veltuzumab, volociximab, votumumab, and zalutumumab are included. Also included are fragments, variants, and derivatives of these antibodies.

[0065] The methods described herein may be used to treat and / or diagnose any of a variety of cancers or tumors. In some embodiments, the cancer is a primary cancer, i.e., a cancer that grows at the anatomical site where tumor progression began and gave rise to a cancer mass. In some embodiments, the cancer is a secondary or metastatic cancer, i.e., a cancer that has spread from the primary site or tissue to one or more different sites or tissues. In some cases, the cancer is a carcinoma, rhabdomyosarcoma, sarcoma, such as alveolar rhabdomyosarcoma (including sarcomas derived from bone, tendon, cartilage, muscle, fat, fibrous, blood vessels, adipose, and / or connective tissue), neuroblastoma, medulloblastoma, astrocytoma, glioblastoma multiforme, retinoblastoma, myeloma, leukemia, lymphoma (including Hodgkin's lymphoma and non-Hodgkin's lymphoma), adenosquamous carcinoma, carcinoma. In some embodiments, the cancer is selected from one or more of: myeloma, mixed mesodermal tumor, teratocarcinoma, lung cancer (including non-small cell lung cancer, small cell lung cancer, adenocarcinoma, and squamous cell lung cancer), breast cancer (including metastatic breast cancer), gastrointestinal cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, cervical cancer, glioblastoma, uterine cancer, salivary gland cancer, kidney or renal cancer (e.g., Wilm's tumor), prostate cancer, thyroid cancer, and head and neck cancer.

[0066] In certain embodiments, the MYC gene is MYCC and the cancer is selected from lung cancer and blood cancer, optionally leukemia and lymphoma. In certain embodiments, the MYC gene is MYCN and the cancer is selected from neuroblastoma, small cell lung cancer, prostate cancer, alveolar rhabdomyosarcoma, medulloblastoma, glioblastoma multiforme, retinoblastoma, and breast cancer.

[0067] In some embodiments, as described above, the cancer or tumor is metastatic cancer.In addition to the above-mentioned cancer, exemplary metastatic cancers include, but are not limited to, bladder cancer that metastasizes to bone, liver, and / or lung; breast cancer that metastasizes to bone, brain, liver, and / or lung; colorectal cancer that metastasizes to liver, lung, and / or peritoneum; renal cancer that metastasizes to adrenal gland, bone, brain, liver, and / or lung; lung cancer that metastasizes to adrenal gland, bone, brain, liver, and / or other lung sites; melanoma that metastasizes to bone, brain, liver, lung, and / or skin / muscle; ovarian cancer that metastasizes to liver, lung, and / or peritoneum; pancreatic cancer that metastasizes to liver, lung, and / or peritoneum; prostate cancer that metastasizes to adrenal gland, bone, liver, and / or lung; gastric cancer that metastasizes to liver, lung, and / or peritoneum; thyroid cancer that metastasizes to bone, liver, and / or lung; and uterine cancer that metastasizes to bone, liver, lung, peritoneum, and / or vagina.

[0068] In certain embodiments, the methods described herein are sufficient to cause a statistically significant reduction in viable tumor mass, such as at least 10%, 20%, 30%, 40%, 50% or more tumor mass reduction, or tumor regression as indicated by altered (e.g., statistically significantly reduced) scan dimensions.In certain embodiments, the methods described herein are sufficient to cause stable disease.In certain embodiments, the methods described herein are sufficient to cause a clinically meaningful reduction in symptoms of a particular disease manifestation known to those skilled in the art.

[0069] The method for treating cancer can be combined with other therapeutic modalities.For example, the combination therapy described herein can be administered to a subject before, during, or after other therapeutic interventions, including symptom care, radiation therapy, surgery, transplantation, hormone therapy, photodynamic therapy, antibiotic therapy, or any combination thereof.Symptom care includes administration of corticosteroids to reduce cerebral edema, headache, cognitive impairment, and vomiting, and administration of anticonvulsants to reduce seizures.Radiation therapy includes whole brain irradiation, fractionated radiation therapy, and radiosurgery, such as stereotactic radiosurgery, which can be further combined with conventional surgery.

[0070] Methods for identifying a subject having one or more of the diseases or conditions described herein are known in the art.

[0071] For in vivo use, eg, for the treatment or testing of disease in humans, the agents described herein will generally be incorporated into one or more therapeutic or pharmaceutical compositions prior to administration.

[0072] To prepare a therapeutic or pharmaceutical composition, an effective or desired amount of one or more agents is typically mixed with any pharmaceutical carrier(s) or excipients known to those skilled in the art to be suitable for the particular agent and / or method of administration. Pharmaceutical carriers may be liquid, semi-liquid, or solid. Solutions or suspensions used for parenteral, intradermal, subcutaneous, or topical application may include, for example, sterile diluents (such as water), saline (e.g., phosphate buffered saline, PBS), fixed oils, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; antimicrobial agents (such as benzyl alcohol and methylparaben); antioxidants (such as ascorbic acid and sodium bisulfite) and chelating agents (such as ethylenediaminetetraacetic acid (EDTA)); buffers (acetates, citrates, and phosphates). When administered intravenously (e.g., by intravenous infusion), suitable carriers include solutions containing saline or phosphate buffered saline (PBS), as well as thickeners and solubilizers such as glucose, polyethylene glycol, polypropylene glycol, and mixtures thereof.

[0073] The administration of the agents described herein is carried out in pure form or in a suitable therapeutic or pharmaceutical composition, via any of the acceptable administration methods for the agent to serve similarly.Therapeutic or pharmaceutical compositions can be prepared by combining the agent-containing composition with suitable physiologically acceptable carriers, diluents, or excipients, and can be formulated into preparations in solid, semi-solid, liquid, or gaseous form, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols.In addition, other pharmacologic active ingredients (including other small molecules as described elsewhere herein) and / or suitable excipients such as salts, buffers, and stabilizers can be present in the composition, but do not have to be.

[0074] Administration can be accomplished by a variety of different routes, including oral, parenteral, nasal, intravenous, intradermal, intramuscular, subcutaneous, or topical. The preferred method of administration will depend on the nature of the condition being treated or prevented. Certain embodiments include administration by intravenous infusion.

[0075] Carriers can include, for example, pharma- ceutically or physiologically acceptable carriers, excipients, or stabilizers that are non-toxic to cells or mammals exposed at the dosages and concentrations used. Often, the physiologically acceptable carrier is an aqueous pH buffered solution. Examples of physiologically acceptable carriers include buffers, such as phosphate, citrate, and other organic acids; antioxidants, including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrins; chelating agents, such as EDTA; sugar alcohols, such as mannitol or sorbitol; salt-forming counterions, such as sodium; and / or non-ionic surfactants, such as polysorbate 20 (TWEEN®), polyethylene glycol (PEG), and poloxamer (PLURONICS®).

[0076] In some embodiments, one or more agents may be encapsulated in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), for example, in microcapsules prepared by coacervation techniques or interfacial polymerization (e.g., hydroxymethylcellulose or gelatin-microcapsules and poly-(methyl methacrylate) microcapsules, respectively). Such techniques are disclosed in Remington's Pharmaceutical Sciences, 16th edition, Oslo, A., Ed., (1980). The particle(s) or liposomes may further comprise other therapeutic or diagnostic agents.

[0077] The exact dosage and duration of treatment are a function of the disease being treated and can be determined empirically using known test protocols or by testing the composition in model systems known in the art and extrapolating therefrom. Controlled clinical trials can also be carried out. Dosage can also vary according to the severity of the condition to be alleviated. Pharmaceutical compositions are generally formulated and administered to exert a therapeutically useful effect while minimizing undesirable side effects. The composition can be administered once or divided into multiple smaller doses and administered at intervals. For any particular subject, the particular administration regimen can be adjusted over time according to individual needs.

[0078] Thus, typical routes of administration of these and related therapeutic or pharmaceutical compositions include, but are not limited to, oral, topical, transdermal, inhalation, parenteral, sublingual, buccal, rectal, vaginal, and intranasal. As used herein, the term parenteral includes subcutaneous injection, intravenous, intramuscular, intrasternal injection or infusion techniques. The therapeutic or pharmaceutical compositions according to certain embodiments of the present disclosure are formulated to allow the active ingredients contained therein to be bioavailable upon administration of the composition to a subject or patient. The composition administered to a subject or patient may take the form of one or more dosage units, e.g., a tablet may be a single dosage unit, and a container of a drug described herein in aerosol form may hold multiple dosage units. The actual methods of preparing such dosage forms will be known or will be apparent to those skilled in the art. See, for example, Remington: The Science and Practice of Pharmacy, 20th Edition (Philadelphia College of Pharmacy and Science, 2000). The composition administered typically contains a therapeutically effective amount of a drug described herein for the treatment of a disease or condition of interest.

[0079] The therapeutic or pharmaceutical composition may be in solid or liquid form. In one embodiment, the carrier(s) are particulate, so that the composition is, for example, in tablet or powder form. The carrier(s) may be liquid, so that the composition is, for example, oral oil, injectable liquid, or aerosol, which is, for example, useful for inhalation administration. When intended for oral administration, the pharmaceutical composition is preferably in either solid or liquid form, with semi-solid, semi-liquid, suspension, and gel forms being included within the forms considered herein as either solid or liquid. Certain embodiments include sterile injectable solutions.

[0080] As a solid composition for oral administration, the pharmaceutical composition can be formulated into powder, granules, compressed tablets, pills, capsules, chewing gum, wafers, etc. Such solid compositions typically contain one or more inert diluents or edible carriers. In addition, one or more of the following may be present: binders, such as carboxymethylcellulose, ethylcellulose, microcrystalline cellulose, tragacanth gum, or gelatin; excipients, such as starch, lactose, or dextrin; disintegrants, such as alginic acid, sodium alginate, Primogel, corn starch, etc.; lubricants, such as magnesium stearate or Sterotex; glidants, such as colloidal silicon dioxide; sweeteners, such as sucrose or saccharin; flavorings, such as peppermint, methyl salicylate, or orange flavorings; and colorings. When the pharmaceutical composition is in the form of a capsule, such as a gelatin capsule, it may contain, in addition to the above-mentioned types of materials, liquid carriers, such as polyethylene glycol or oil.

[0081] The therapeutic or pharmaceutical composition may be in the form of a liquid, such as an elixir, syrup, solution, emulsion, or suspension. The liquid may be for oral administration or for delivery by injection, as two examples. If for oral administration, the preferred composition contains, in addition to the compound, one or more of a sweetener, a preservative, a dye / colorant, and a flavor enhancer. In the composition intended to be administered by injection, one or more of a surfactant, a preservative, a wetting agent, a dispersing agent, a suspending agent, a buffer, a stabilizer, and an isotonic agent may be included.

[0082] Liquid therapeutic or pharmaceutical compositions, whether in solution, suspension, or other similar form, may contain one or more of the following adjuvants: sterile diluents, such as water for injection, saline, preferably saline, Ringer's solution, isotonic sodium chloride, fixed oils, such as synthetic mono- or diglycerides, polyethylene glycols, glycerin, propylene glycol, or other solvents that may function as solvents or suspending media; antibacterial agents, such as benzyl alcohol or methylparabens; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers, such as acetates, citrates, or phosphates, and agents for adjusting tonicity, such as sodium chloride or dextrose. Parenteral preparations may be enclosed in glass or plastic ampoules, disposable syringes, or multiple dose vials. Saline is the preferred adjuvant. Injectable pharmaceutical compositions are preferably sterile.

[0083] Liquid therapeutic or pharmaceutical compositions intended for either parenteral or oral administration should contain an amount of drug such that a suitable dosage is obtained. Typically, this amount is at least 0.01% of the drug of interest in the composition. If intended for oral administration, this amount may vary from 0.1 to about 70% by weight of the composition. Certain oral therapeutic or pharmaceutical compositions contain from about 4% to about 75% of the drug of interest. In certain embodiments, therapeutic or pharmaceutical compositions and preparations according to the invention are prepared such that a parenteral dosage unit contains from 0.01 to 10% by weight of the drug of interest before dilution.

[0084] Therapeutic or pharmaceutical compositions may contain various materials that change the physical form of solid or liquid dosage units. For example, the composition may contain materials that form a coating shell around the active ingredient. The materials that form the coating shell are typically inert and may be selected from, for example, sugar, shellac, and other enteric coating agents. Alternatively, the active ingredient may be encapsulated in a gelatin capsule. Therapeutic or pharmaceutical compositions in solid or liquid form may contain components that bind to the drug and thereby aid in the delivery of the compound. Suitable components that may act in this capacity include monoclonal or polyclonal antibodies, one or more proteins, or liposomes.

[0085] The compositions described herein can be prepared with carriers that protect the drug from rapid excretion from the body, such as time-release formulations or coatings.Such carriers include, but are not limited to, release-controlled formulations, such as implants and microencapsulated delivery systems, and biodegradable, biocompatible polymers, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, polyorthoesters, polylactic acid, and others known to those skilled in the art.

[0086] The therapeutic or pharmaceutical composition may be prepared by methodology well known in the pharmaceutical art. For example, a therapeutic or pharmaceutical composition intended to be administered by injection may contain one or more of salts, buffers, and / or stabilizers together with sterile distilled water to form a solution. Surfactants may be added to facilitate the formation of a homogeneous solution or suspension. Surfactants are compounds that non-covalently interact with drugs to facilitate the dissolution or homogeneous suspension of the drug in an aqueous delivery system.

[0087] Certain embodiments include the use of a diagnostic kit for determining or predicting therapeutic response (or responsiveness) to YM155 monobromide [1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(pyrazin-2-ylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide] therapy in a cancer subject, comprising means for measuring MYC gene copy number or MYC gene chromosomal location site in a sample of tissue from the subject, including cancerous tissue and non-cancerous tissue. Also included is a patient care kit comprising (a) a means for measuring MYC gene copy number or MYC gene chromosomal location site in a sample of tissue from a subject, including cancerous and non-cancerous tissue; and (b) YM155 monobromide [1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(pyrazin-2-ylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide].

[0088] In some embodiments, the means for measuring MYC gene copy number comprises reagents for performing a diagnostic assay on the human MYC gene selected from one or more of array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) array, copy number variation (CNV) sequencing, and multiplex ligation-dependent probe amplification (MLPA). In some embodiments, the means for measuring MYC gene chromosomal location comprises reagents for performing a diagnostic assay on the human MYC gene selected from one or more of in situ hybridization (ISH), fluorescent in situ hybridization (FISH), next generation sequencing (NGS), and comparative genomic hybridization (CGH).

[0089] Certain diagnostic kits or patient care kits include a MYC gene copy number reference value obtained from a database or determined from non-cancerous tissue from a control.Some diagnostic kits or patient care kits include a MYC gene chromosomal location site reference obtained from a database or determined from non-cancerous tissue from a control.The kit may also include instructions on how to determine, for example, the MYC gene copy number and / or the MYC gene chromosomal location site of the cancerous tissue from the subject and / or the sample from the non-cancerous control.

[0090] In some embodiments, the diagnostic kit or patient care kit contains separate containers, dividers, or compartments for the composition(s) and informational material(s). For example, the composition(s) or reagents may be contained in a bottle, vial, or syringe, and the informational material(s) may be contained in association with the container. In some embodiments, the separate elements of the kit are contained within a single, non-separated container. For example, the composition(s) or reagents are contained in a bottle, vial, or syringe to which the informational material in the form of a label is attached. In some embodiments, the kit contains multiple (e.g., a pack) individual containers, each containing one or more compositions, reagents, and / or unit dosage forms of YM155 monobromide. For example, the kit contains multiple syringes, ampoules, foil packets, or blister packs, each containing a reagent or a single unit dose of YM155 monobromide. The containers of the kit may be airtight, waterproof (e.g., impermeable to changes in moisture or evaporation), and / or light-tight.

[0091] The patient care kit optionally includes a device suitable for administration of the agent(s), such as a syringe, inhaler, dropper (e.g., eye dropper), swab (e.g., cotton or wooden swab), or any such delivery device. In some embodiments, the device is an implantable device that dispenses a metered amount of the agent(s). Also included are methods of providing the kit, for example, by combining the components described herein.

[0092] In certain aspects, the diagnostic or therapeutic response test or method described herein is carried out in a diagnostic laboratory, and the results are then provided to the subject, or to the doctor or other medical provider involved in the subject's health care and cancer treatment.Therefore, certain embodiments include a method for providing the results of response test to the subject in need thereof, or to the doctor or other medical provider.These results or data may be in the form of hard copy or paper copy, or in electronic form, such as computer readable media.

[0093] All publications, patent applications, and issued patents cited in this specification are herein incorporated by reference to the same extent as if each individual publication, patent application, or issued patent was specifically and individually indicated to be incorporated by reference.

[0094] Although the foregoing invention has been described in some detail by way of illustration and examples for purposes of clarity of understanding, it will be readily apparent to those skilled in the art that certain changes and modifications may be made without departing from the spirit or scope of this description or the appended claims. The following examples are provided for illustrative purposes only, and not for limiting purposes. Those skilled in the art will readily recognize a variety of non-critical parameters that can be changed or modified to produce essentially similar results. EXAMPLES

[0095] Example 1 Efficacy of YM155 monobromide in cancer cell lines Studies were performed to evaluate the efficacy of YM155 monobromide in various cancer cell lines and to correlate the efficacy of YM155 monobromide with MYC gene copy number.

[0096] Lung cell culture. Human lung cancer cell lines NCI-H1975, HCC827, NCI-H226, and HCC4006 cells were cultured in RPMI 1640 (Hyclone™, SH30809.01B) supplemented with 10% fetal bovine serum (GEMINI, 900-108). Cultures were incubated at 37°C in 5% CO2. NCI-H1975, HCC827, and NCI-H226 were purchased from Cell Bank, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. HCC4006 was purchased from CoBioer Biosciences Corporation (Nanjing, China).

[0097] Cell treatment and proliferation assay. NCI-H1975, HCC827, NCI-H226, and HCC4006 cells were seeded in 96-well plates (Corning-Costar, 3603) at 2000 cells / well in 200 μL culture medium, respectively, for 24 h. The cells were then treated with YM155 (50, 30, 20, 10, 2, 0.4, 0.08, 0.016, and 0.0032 nM; APExBIO, A4221) or DMSO (0.1%; Amresco, 67-68-5). After 72 h of incubation, the cells were labeled with 5-ethynyl-2'-deoxyuridine (EdU, final concentration 1 uM; Sigma, 900584) for an additional 4 h under the same culture conditions.

[0098] After incubation with EdU, cells were fixed with formaldehyde (final concentration 4%, Thermo, 28908) for 30 min at room temperature. Cells were washed twice with PBS (Hyclone™, SH30256.01) and then fixed with 0.5% Triton in PBS. X-100 (T8787-250ML) overnight at 4° C. After discarding the supernatant, the cells were incubated with Hoechst 33342 (Invitrogen, H1399) for 1 hour at room temperature and then washed two more times with PBS.

[0099] The cells were incubated with staining mixture (Beijing Percans Oncology Medical Research Co., Ltd., RUO-00401#150T) at room temperature for 30 minutes, then washed three times with PBS. The cells were kept in PBS and protected from light throughout the experiment.

[0100] High-content imaging analysis. Treated cells were scanned for image acquisition on a CellInsight™ CX5 High-Content Screening (HSC) Platform (Thermo Fisher) equipped with filters for Hoechst 33342 (Ex: 386 nm) and EdU (Ex: 560 nm). The total cell number and EdU positive cell number were analyzed by measuring the signal intensity of the nuclear region.

[0101] As shown in Figure 2, YM155 inhibits cell proliferation in human lung cancer cell lines. NCI-H1975, HCC827, NCI-H226, and HCC4006 were exposed to different concentrations of YM155 for 72 hours. However, the proliferation inhibitory effect of YM155 was different in these cell lines, and the calculated IC 50 are shown in Table E1 below. NCI-H1975 and HCC827 with MYC gene amplification (increased MYC gene copy number compared to reference) were significantly more sensitive to YM155 than NCI-H226 and HCC4006 without MYC gene amplification. [Table 1]

[0102] Blood cell culture. Human acute myeloid leukemia cells (U937 and HL-60) and human Burkitt's lymphoma cells (Ramos) were cultured in RPMI 1640 supplemented with 10% fetal bovine serum. Cultures were incubated at 37°C in 5% CO2. U937 and HL-60 cells were purchased from Cell Bank, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. Ramos cells were purchased from CoBioer Biosciences Corporation (Nanjing, China). Ramos cells carry an IgH-c-MYC translocation mutation.

[0103] Cell treatment and proliferation assay. U937, HL-60, and Ramos cells were seeded in 96-well plates (Corning-Costar, 3599) at 40000 cells / well in 200 μL culture medium treated with YM155 (200, 100, 50, 25, 12.5, 2.5, 1, 0.5 nM; APExBIO, A4221) or DMSO (0.1%; Amresco, 67-68-5). After 72 h of incubation, cell proliferation was measured by XTT assay (Cell Proliferation Kit II XTT, Sigma, 11465015001) according to the manufacturer's protocol. Briefly, after YM155 treatment, 50 μL of XTT labeling mixture was added to each well, and then the microplate was incubated for 4 h in a humidified atmosphere. The optical density (OD) at 490 nm (OD) was measured. 490 ) and 650 nm (OD 650 The absorbance at wavelengths of 1000 and 1500 nm was determined by SpectraMax 190. 50 OD 490- O.D. 650 was calculated using the value of

[0104] As shown in Figures 3 and 4, YM155 inhibits cell proliferation in human acute myeloid leukemia cells and human Burkitt's lymphoma cells. U937, HL-60, and Ramos were exposed to different concentrations of YM155 for 72 hours. However, the proliferation inhibitory effect of YM155 was different in these cell lines, and the calculated IC 50 are shown in Table E2 below. HL-60 cells with MYC gene amplification (increased MYC gene copy number compared to reference) and Ramos cells with MYC gene translocation were more sensitive to YM155 than U937 cells without MYC gene amplification or translocation. [Table 2]

[0105] Neuroblastoma cell culture. Neuroblastoma cell lines IMR-32, NB-1, KP-N-YN, SK-N-BE, SK-N-SH, and SH-SY5Y cells were cultured in 96-well plates and treated with the indicated doses (nM) of YM155. Human neuroblastoma cell line IMR-32 was cultured in MEM (Hyclone™ SH30024.01) supplemented with 10% fetal bovine serum (GEMINI, 900-108), and neuroblastoma cell lines NB-1 and KP-N-YN were cultured in RPMI supplemented with 10% fetal bovine serum. Neuroblastoma cell line SK-N-BE was cultured in F12 (Gibco, 11765-054) / MEM (1:1) (Hyclone™, SH30024.01) supplemented with 10% fetal bovine serum, neuroblastoma cell line SH-SY5Y was cultured in MEM / F12 (1:1) supplemented with 10% fetal bovine serum, and neuroblastoma cell line SK-N-SH was cultured in MEM supplemented with 10% fetal bovine serum. Cultures were incubated at 37° C. in 5% CO2.

[0106] IMR-32, SH-SY5Y, and SK-N-SH were purchased from Cell Bank, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. NB-1, KP-N-YN, and SK-N-BE cells were purchased from CoBioer Biosciences Corporation (Nanjing, China).

[0107] Cell treatment and proliferation assay. IMR-32, SH-SY5Y, SK-N-SH, NB-1, KP-N-YN, and SK-N-BE cells were seeded in 96-well plates (Corning-Costar, 3603) at 2000 cells / well in 200 μL culture medium for 24 h. Cells were then treated with YM155 (200, 100, 50, 25, 12.5, 6.25, 3.125, 1.5625, and 0.78125 nM; APExBIO, A4221) or DMSO (0.1%; Amresco, 67-68-5). After 72 h of incubation, cells were labeled with 5-ethynyl-2'-deoxyuridine (EdU, final concentration 1 uM; Sigma, 900584) for an additional 4 h under the same culture conditions.

[0108] After incubation with EdU, cells were fixed with formaldehyde (final concentration 4%, Thermo, 28908) for 30 min at room temperature. Cells were washed twice with PBS (Hyclone™, SH30256.01) and then fixed with 0.5% Triton in PBS. The cells were permeabilized with X-100 (T8787-250ML) at 4°C overnight. After discarding the supernatant, the cells were incubated with Hoechst 33342 (Invitrogen, H1399) at room temperature for 1 hour, then washed twice more with PBS. The cells were incubated with staining mixture (Beijing Percans Oncology Medical Research Co.,Ltd., RUO-00401#150T) at room temperature for 30 minutes, then washed three times with PBS. The cells were kept in PBS and protected from light throughout the experiment.

[0109] High-content imaging analysis. Treated cells were scanned for image acquisition on a CellInsight™ CX5 High-Content Screening (HSC) Platform (Thermo Fisher) equipped with filters for Hoechst 33342 (Ex: 386 nm) and EdU (Ex: 560 nm). The total cell number and EdU positive cell number were analyzed by measuring the signal intensity of the nuclear region.

[0110] As shown in Figure 7 and Table E3 below, YM155 inhibits cell proliferation of neuroblastoma cell lines, which is particularly useful for inhibiting cell proliferation of IMR-32, NB-1, KP-N-YN, and SK-N-BE neuroblastoma cell lines, which are characterized by increased copy number of NMYC. [Table 3]

[0111] These data indicate that MYC gene amplification (i.e., increased MYC gene copy number compared to a reference) and MYC gene translocations can be used to predict responsiveness to YM155 monobromide therapy in cancers, including lung cancer, hematopoietic cell carcinoma, neuroblastoma, and other cancers.

[0112] Example 2 HCC827 / HCC 4006 xenograft experiments Four to six week old female mice (Balb / C-nu) were purchased from Beijing HFK BIOSCIENCE Co., Ltd. Cell suspensions in cell culture medium (HCC827 (pulmonary epithelial adenocarcinoma): 5 × 10 per mouse) were cultured at 100 x 100 cells / mL. 6 / 0.1mL; HCC4006: 5 × 10 per mouse 6 The mice were subcutaneously inoculated with 0.1 mL of 1000 mM NaCl in the right flank of the mouse. 3 ~150mm 3 When tumor volume reached 100 μg / kg, mice were randomized into treatment and control groups. 5 mg / kg YM155 (Biochem partner, BCP01864) or vehicle (saline, Shijiazhuang No.4 Pharmaceutical) was administered subcutaneously once daily, 5 days per week for 3 weeks. Mice were assessed for body weight twice weekly, and tumor diameters (long and short diameters) were measured using standard calipers. Tumor volume = 1 / 2 * long diameter * (short diameter) 2 After 2-3 weeks of observation, the mice were sacrificed for analysis. The data are shown in Figures 9A-9B and 10A-10B.

[0113] Example 3 RAMOS / U937 xenograft experiments Four to six week old female mice (RAMOS: SCID; U937: NOD / SCID) were purchased from BEIJING HFK BIOSCIENCE Co., Ltd. Cell suspensions in cell culture medium (RAMOS: 1 × 10 per mouse) were cultured at 100 x 100 cells / mL. 7 / 0.1mL; U937: 2x10 per mouse 6 The mice were subcutaneously inoculated with 0.1 mL of 100x1000 / 0.1 mL of Matrigel into the right flank. 3 ~150mm 3 When tumor volume reached 100 mg / kg, mice were randomized into treatment and control groups. 5 mg / kg YM155 (Biochem partner, BCP01864) or vehicle (saline, Shijiazhuang No.4 Pharmaceutical) was administered subcutaneously once daily for 9 days. Mice were assessed for body weight twice weekly, and tumor diameters (long and short diameters) were measured using standard calipers. Tumor volume = 1 / 2 * long diameter * (short diameter) 2 After 2-3 weeks of observation, the mice were sacrificed for analysis. The data are shown in Figures 11A-11B and 12A-12B.

[0114] Example 6 IMR-3 / SH-SY5Y xenograft experiments Four to six week old female mice (IMR-32: NOD / SCID; SH-SY5Y: Balb / C-nu) were purchased from BEIJING HFK BIOSCIENCE Co., Ltd. Cell suspensions in cell culture medium (IMR-32: 5x10 per mouse) were cultured at 100 x 100 cells / mL. 6 / 0.1mL + 0.1mL Matrigel; SH-SY5Y: 2x10 per mouse 6 The mice were subcutaneously inoculated with 0.1 mL of 100x1000 / 0.1 mL of Matrigel into the right flank. 3 ~150mm 3When tumor volume reached 100 μg / kg, mice were randomized into treatment and control groups. 5 mg / kg YM155 (Biochem partner, BCP01864) or vehicle (saline, Shijiazhuang No.4 Pharmaceutical) was administered subcutaneously once daily, 5 days per week for 3 weeks. Mice were assessed for body weight twice weekly, and tumor diameters (long and short diameters) were measured using standard calipers. Tumor volume = 1 / 2 * long diameter * (short diameter) 2 After 2-3 weeks of observation, the mice were sacrificed for analysis. The data are shown in Figures 13A-13B and 14A-14B. In certain embodiments, for example, the following items are provided: (Item 1) 1. A method for treating cancer in a subject in need thereof, comprising: (a) determining the MYC gene copy number or MYC gene chromosomal location site in a sample of cancer tissue from the subject; (b) if the MYC gene copy number in the cancer tissue is increased compared to a reference MYC gene copy number or if the MYC gene chromosomal location site in the cancer tissue is translocated compared to a reference MYC gene chromosomal location site, administering to the subject YM155 monobromide [1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(pyrazin-2-ylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide], or an analogue or derivative thereof, thereby treating cancer in the subject in need thereof. (Item 2) 2. The method of claim 1, further comprising administering to the subject a chemotherapeutic agent that excludes (or is not limited to) YM155 monobromide if the MYC gene copy number in the cancer tissue is not substantially increased compared to the reference MYC gene copy number or if the MYC gene chromosomal location site in the cancer tissue is not translocated compared to the reference MYC gene chromosomal location site. (Item 3) 1. A method for predicting therapeutic response to YM155 monobromide [1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(pyrazin-2-ylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide], or an analog or derivative thereof, in a cancer subject, comprising: (a) determining the MYC gene copy number or MYC gene chromosomal location site in a sample of cancer tissue from the subject; (b)(i) characterizing the subject as responsive to YM155 monobromide therapy if the MYC gene copy number in the cancer tissue is increased compared to a reference MYC gene copy number, or if the MYC gene chromosomal location site in the cancer tissue is translocated compared to a reference MYC gene chromosomal location site; or (ii) characterizing the subject as non-responsive to YM155 monobromide therapy if the MYC gene copy number in the cancer tissue is not substantially increased compared to the MYC gene copy number reference or if the MYC gene chromosomal location site in the cancer tissue is not translocated compared to the MYC gene chromosomal location site reference, thereby predicting a therapeutic response to YM155 monobromide in the cancer subject. (Item 4) 4. The method of claim 3, comprising administering YM155 monobromide to the subject if the subject is characterized as responsive to YM155 monobromide therapy. (Item 5) 4. The method of claim 3, comprising administering to the subject a chemotherapeutic agent that excludes YM155 monobromide if the subject is characterized as non-responsive to YM155 monobromide therapy. (Item 6) 6. The method of any one of items 1 to 5, wherein the MYC gene copy number in the cancer tissue is increased by about or at least about 1.5, 2, 3, 4, 5, 6, 7, 8, 9, or 10 fold compared to the reference MYC gene copy number. (Item 7) 7. The method of any one of items 1 to 6, comprising determining the MYC gene copy number in the cancer tissue by array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) array, copy number variation (CNV) sequencing, or multiplex ligation-dependent probe amplification (MLPA). (Item 8) The method according to any one of items 1 to 7, comprising determining the MYC gene chromosomal location site in the cancer tissue by in situ hybridization (ISH), fluorescent in situ hybridization (FISH), next generation sequencing (NGS), or comparative genomic hybridization (CGH). (Item 9) 9. The method of any one of items 1 to 8, comprising obtaining the MYC gene copy number reference from a database or, optionally, determining the MYC gene copy number reference from a non-cancerous tissue from a control by aCGH, SNP array, CNV sequence, or MLPA. (Item 10) 10. The method of any one of items 1 to 9, comprising obtaining the MYC gene chromosomal location site from a database or, optionally, determining the MYC gene chromosomal location site reference from a non-cancerous tissue from a control by ISH, FISH, NGS, or CGH. (Item 11) 11. The method according to any one of items 1 to 10, comprising obtaining a sample of the cancer tissue from the subject. (Item 12) 12. The method of any one of items 1 to 11, wherein the sample of cancer tissue is a surgical sample, a biopsy sample, a pleural effusion sample, or an ascites sample obtained from the subject, and is optionally selected from one or more of lung, blood, breast, gastrointestinal tract (stomach, colon, rectum), ovary, pancreas, liver, bladder, cervix, nerve, uterus, salivary gland, kidney, prostate, thyroid, or muscle tissue. (Item 13) 13. The method according to any one of items 1 to 12, wherein the subject is a human subject. (Item 14) The cancer may be a carcinoma, a sarcoma, such as rhabdomyosarcoma, e.g., alveolar rhabdomyosarcoma (including sarcomas derived from bone, tendon, cartilage, muscle, fat, fibrous, vascular, adipose, and / or connective tissue), a neuroblastoma, a medulloblastoma, an astrocytoma, a glioblastoma multiforme, a retinoblastoma, a myeloma, a leukemia, a lymphoma (including Hodgkin's lymphoma and non-Hodgkin's lymphoma), an adenosquamous carcinoma, a carcinosarcoma, a mixed mesodermal tumor, a teratocarcinoma, a sarcoma ... 14. The method of any one of items 1 to 13, wherein the cancer is selected from one or more of the following: lung cancer (including non-small cell lung cancer, small cell lung cancer, adenocarcinoma, and squamous cell lung cancer), breast cancer (including metastatic breast cancer), digestive cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, cervical cancer, glioblastoma, uterine cancer, salivary gland cancer, kidney cancer or renal cancer (e.g., Wilm's tumor), prostate cancer, thyroid cancer, and head and neck cancer. (Item 15) 15. The method according to any one of items 1 to 14, wherein the MYC gene is selected from MYCC and MYCN. (Item 16) 16. The method of claim 15, wherein the MYC gene is MYCC and the cancer is selected from lung cancer and hematological cancer, optionally leukemia and lymphoma. (Item 17) 16. The method of claim 15, wherein the MYC gene is MYCN and the cancer is selected from neuroblastoma, small cell lung cancer, prostate cancer, alveolar rhabdomyosarcoma, medulloblastoma, glioblastoma multiforme, retinoblastoma, and breast cancer. (Item 18) Use of a diagnostic kit for determining therapeutic response to YM155 monobromide [1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(pyrazin-2-ylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide], or an analogue or derivative thereof, in a cancer subject, comprising a means for measuring MYC gene copy number or MYC gene chromosomal location site in a sample of tissue from the subject, including cancerous tissue and non-cancerous tissue. (Item 19) 19. The use of item 18, wherein the means for measuring MYC gene copy number comprises reagents for performing a diagnostic assay selected from one or more of array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) array, copy number variation (CNV) sequencing, and multiplex ligation-dependent probe amplification (MLPA) on the human MYC gene. (Item 20) 20. The use according to item 18 or 19, wherein the means for determining the MYC gene chromosomal location site comprises reagents for performing a diagnostic assay selected from one or more of in situ hybridization (ISH), fluorescent in situ hybridization (FISH), next generation sequencing (NGS), and comparative genomic hybridization (CGH) on the human MYC gene. (Item 21) 21. The use according to any one of items 18 to 20, comprising a MYC gene copy number reference value obtained from a database or determined from non-cancerous tissue from a control. (Item 22) 21. The use according to any one of items 18 to 20, comprising a MYC gene chromosomal location site reference obtained from a database or determined from non-cancerous tissue from a control. (Item 23) The cancer may be a carcinoma, a sarcoma, such as rhabdomyosarcoma, e.g., alveolar rhabdomyosarcoma (including sarcomas derived from bone, tendon, cartilage, muscle, fat, fibrous, vascular, adipose, and / or connective tissue), a neuroblastoma, a medulloblastoma, an astrocytoma, a glioblastoma multiforme, a retinoblastoma, a myeloma, a leukemia, a lymphoma (including Hodgkin's lymphoma and non-Hodgkin's lymphoma), an adenosquamous carcinoma, a carcinosarcoma, a mixed mesodermal tumor, a teratocarcinoma, a sarcoma ... The use according to any one of items 18 to 22, wherein the cancer is selected from one or more of the following: cancer, lung cancer (including non-small cell lung cancer, small cell lung cancer, adenocarcinoma, and squamous cell lung cancer), breast cancer (including metastatic breast cancer), digestive cancer, gastric cancer, colorectal cancer, colon cancer, rectal cancer, ovarian cancer, pancreatic cancer, liver cancer, bladder cancer, cervical cancer, glioblastoma, uterine cancer, salivary gland cancer, kidney cancer or renal cancer (e.g., Wilm's tumor), prostate cancer, thyroid cancer, and head and neck cancer. (Item 24) 24. The use according to any one of items 18 to 23, wherein the MYC gene is selected from MYCC and MYCN. (Item 25) 25. The use according to item 24, wherein the MYC gene is MYCC and the cancer is selected from lung cancer and blood cancer, optionally leukemia and lymphoma. (Item 26) 25. The use according to item 24, wherein the MYC gene is MYCN and the cancer is selected from neuroblastoma, small cell lung cancer, prostate cancer, alveolar rhabdomyosarcoma, medulloblastoma, glioblastoma multiforme, retinoblastoma, and breast cancer. (Item 27) (a) a means for measuring MYC gene copy number or MYC gene chromosomal location site in a sample of tissue from a subject, including cancerous tissue and non-cancerous tissue; (b) YM155 monobromide [1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(pyrazin-2-ylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide], or an analogue or derivative thereof. (Item 28) 28. The patient care kit of item 27, wherein the means for measuring MYC gene copy number comprises reagents for performing a diagnostic assay on the human MYC gene selected from one or more of array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) array, copy number variation (CNV) sequencing, and multiplex ligation-dependent probe amplification (MLPA). (Item 29) 29. The patient care kit of item 27 or 28, wherein the means for determining the MYC gene chromosomal location site comprises reagents for performing a diagnostic assay selected from one or more of in situ hybridization (ISH), fluorescent in situ hybridization (FISH), next generation sequencing (NGS), and comparative genomic hybridization (CGH) on the human MYC gene. (Item 30) 30. The patient care kit of any one of items 27 to 29, comprising a MYC gene copy number reference value obtained from a database or determined from non-cancerous tissue from a control. (Item 31) 31. The patient care kit of any one of items 27 to 30, comprising a MYC gene chromosomal location site reference obtained from a database or determined from non-cancerous tissue from a control. (Item 32) 32. The patient care kit of any one of items 27 to 31, wherein the MYC gene is selected from MYCC and MYCN.

Claims

[Claim 1] The invention as depicted in the drawings.

Images