Methods of treating solid tumor cancer using cryptocephalosporin and biomarkers

Abstract

Disclosed herein are methods for determining the likelihood that an individual suffering from solid tumor cancer will benefit from treatment with cryptosteglitin. Additionally, there are treatment methods based on such determinations. In several embodiments, the markers prostaglandin reductase 1 (PTGR1), protein tyrosine phosphatase non-receptor type 14 (PTPN14), aspartate beta-hydroxylase (ASPH) together with one or more genes or alone can be used to enhance or guide treatment with cryptosteglitin. In certain embodiments, the protein or gene may be expressed or methylated.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 745,382, filed on October 14, 2018, which is incorporated herein by reference in its entirety. Technical Field

[0003] The present invention relates to one or more markers for determining the sensitivity of a cancer patient to an anticancer agent (e.g., illudin) to be administered to the patient, which markers can determine whether the patient's cancer is responsive to treatment with the anticancer agent and the application of the markers. Background Art

[0004] Cryptostegans have been chemically modified and used to treat cancer. Irofulven, a cryptostegans, is a chemically modified form of the fungal toxin cryptostegans S. It is a DNA alkylating agent with an unusual mechanism. This cryptostegans is a DNA and protein destroyer that targets rapidly dividing malignant tumor cells. It enters tumor cells where it interferes with DNA replication and cell division by binding to DNA and protein targets. This causes the tumor cells to shut down and therefore die (apoptosis). A study determined that at certain doses, tumor cells are very susceptible to the dual destructive activity of Irofulven, while normal cells only show marginal responses to the cytocidal drug. Not all solid cancers are sensitive to cryptostegans, and methods to determine individuals who are sensitive to cryptostegans are elusive.

[0005] Despite recent advances in diagnostic and treatment strategies for individuals with solid tumors, particularly prostate, ovarian, liver, and kidney cancers, many critical knowledge gaps remain regarding their screening and treatment. There is a poor understanding of patient characteristics, including genetic characteristics, that best stratify individuals into response groups at diagnosis. There is also a poor understanding of risk factors for the development of or death from these cancers, and a lack of effective application of real-world evidence into clinical practice. This lack of understanding means that predictions of which individuals will have the best outcome with a specific treatment are suboptimal. Additionally, current predictions of which individuals may be harmed by unnecessary or inappropriate treatment or be safely managed without treatment remain poor.

[0006] Therefore, there is a need for a method for treating and determining whether an individual will or is more or less likely to respond to cryptocephalosporin treatment. The present application is particularly directed to this need. Summary of the invention

[0007] The present application relates to methods for determining the likelihood that an individual suffering from solid tumor cancer will benefit from treatment with cryptocephalosporin. In addition, there are methods of treatment based on such determinations. In several embodiments, the markers prostaglandin reductase 1 (PTGR1), protein tyrosine phosphatase non-receptor type 14 (PTPN14), aspartate beta-hydroxylase (ASPH) are used in the method together with one or more genes or alone. In certain embodiments, the protein or gene may be expressed or methylated.

[0008] A method of treating solid tumor cancer with targeted drug therapy, comprising:

[0009] Identifying patients with cancer that is susceptible to treatment with a compound of the formula:

[0010]

[0011] wherein R1, R2 and R3 are independently (C1-C4) alkyl, methyl or hydroxy.

[0012] definition

[0013] The term "subject" refers to an animal, including but not limited to primates (e.g., humans), cows, pigs, sheep, goats, horses, dogs, cats, rabbits, rats, or mice. The terms "subject" and "patient" are used interchangeably herein, e.g., referring to a mammalian subject, such as a human subject, in one embodiment a human.

[0014] The terms "treat," "treating," and "treatment" are intended to include alleviating or eliminating a condition, or one or more symptoms associated with a condition; or alleviating or eradicating the cause of the condition itself.

[0015] The terms "manage", "managing", and "management" encompass preventing the recurrence of a given disease, disorder, or condition in a patient already suffering from the disease, disorder, or condition, and / or prolonging the time a patient suffering from the disease, disorder, or condition remains in remission. The terms encompass modulating the threshold, development, and / or duration of a disease, disorder, or condition, or changing the way a patient responds to a disease, disorder, or condition.

[0016] The terms "prevent," "preventing," and "prevention" are intended to include methods of delaying and / or arresting the onset of a disorder, disease or condition and / or its attendant symptoms; inhibiting a subject from developing a disorder, disease or condition; or reducing a subject's risk of developing a disorder, disease or condition.

[0017] The term "sensitivity" or "sensitive" when referring to treatment with a compound is a relative term that refers to the degree of effectiveness of the compound in alleviating or reducing the progression of the disease, disorder or condition being treated. For example, when used in reference to the treatment of a disease, disorder or condition associated with a compound, the term "increased sensitivity" or "sensitive to treatment" refers to an increase in the effectiveness of the treatment by at least 3%, specifically at least 5%, or more.

[0018] As used herein, and unless otherwise specified, the term "therapeutically effective amount" of a compound is an amount sufficient to provide a therapeutic benefit in the treatment or management of a disease, disorder, or condition, or to delay or minimize one or more symptoms associated with the presence of a disease, disorder, or condition. A therapeutically effective amount of a compound means an amount of a therapeutic agent, alone or in combination with other therapies, that provides a therapeutic benefit in the treatment or management of a disease, disorder, or condition. The term "therapeutically effective amount" may encompass an amount that improves overall therapy, reduces or avoids symptoms or causes of a disease, disorder, or condition, or enhances the therapeutic efficacy of another therapeutic agent.

[0019] As used herein, an "effective patient response" refers to any increase in therapeutic benefit to a patient. An "effective patient response" can be, for example, a 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 75%, 80%, 90%, or 100% decrease in the rate of progression of a disease, disorder, or condition. An "effective patient response" can be, for example, a 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 75%, 80%, 90%, or 100% decrease in the physical symptoms of a disease, disorder, or condition. An "effective patient response" can also be, for example, an increase in the patient's response by 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 75%, 80%, 90%, 100%, 120%, 140%, 150%, 170%, 180%, 190%, 200% or more, as measured by any suitable method, such as gene expression, cell counts, analytical results, etc.

[0020] Improvement of a disease, disorder or condition can be characterized as a complete or partial response. A "complete response" refers to the absence of clinically detectable disease with normalization of any previously abnormal radiographic studies, bone marrow and cerebrospinal fluid (CSF) or abnormal monoclonal protein measurements. A "partial response" refers to a reduction of at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% in all measurable disease, disorder or condition load (i.e., the number of malignant cells present in an individual, or the volume of a measured tumor mass or the amount of an abnormal monoclonal protein) in the absence of new lesions. The term "treatment" encompasses complete and partial responses.

[0021] The term "refractory or resistant" refers to a situation where a patient does not respond to treatment even after intensive treatment. For example, a patient may have residual cancer cells (e.g., leukemia or lymphoma cells) in their lymphatic system, blood, and / or blood-forming tissues (e.g., bone marrow).

[0022] As used herein, the terms "determine," "measure," "evaluate," "assess," and "analyze" generally refer to any form of measurement, and include determining whether an element is present or not. These terms include quantitative and / or qualitative determinations. Assessments may be relative or absolute. "Assessing the presence of..." may include determining the amount of something that is present, as well as determining whether it is present or not.

[0023] The terms "isolated" and "purified" refer to the separation of a substance (such as DNA / mRNA or protein) such that the substance accounts for a substantial portion of the sample in which it resides, i.e., greater than the substance normally found in its native or unisolated state. Typically, a substantial portion of a sample accounts for, for example, greater than 1%, greater than 2%, greater than 5%, greater than 10%, greater than 15%, greater than 20%, greater than 25%, greater than 30%, greater than 40%, greater than 50% or more of the sample, typically up to about 90%-100%. For example, a sample of isolated DNA / mRNA may typically account for at least about 1% total mRNA. Techniques for purifying polynucleotides are well known in the art and include, for example, gel electrophoresis, ion exchange chromatography, affinity chromatography, flow sorting, and sedimentation according to density.

[0024] The term "sample" as used herein relates to a material or mixture of materials, typically but not necessarily in fluid form, which contains one or more components of interest.

[0025] As used herein, "biological sample" refers to a sample obtained from an individual organism, including a sample of biological tissue or fluid origin obtained, reached or collected in vivo or in situ. Biological samples also include samples from areas of an individual organism containing precancerous or cancerous cells or tissues. Such samples may be, but are not limited to, organs, tissues, fractions, and cells isolated from mammals. Exemplary biological samples include, but are not limited to, cell lysates, cell cultures, cell lines, tissues, oral tissues, gastrointestinal tissues, organs, organelles, biological fluids, blood samples, urine samples, skin samples, and the like. Biological samples include, but are not limited to, whole blood, partially purified blood, PBMCs, tissue biopsies, and the like.

[0026] As used herein, and unless otherwise indicated, the term "optically pure" means a composition comprising one optical isomer of a compound and being substantially free of other isomers of the compound. For example, an optically pure composition of a compound having one chiral center will be substantially free of the opposite enantiomer of the compound. An optically pure composition of a compound having two chiral centers will be substantially free of other diastereomers of the compound. Typical optically pure compounds contain greater than about 80% by weight of one enantiomer of the compound and less than about 20% by weight of the other enantiomers of the compound, or greater than about 90% by weight of one enantiomer of the compound and less than about 10% by weight of the other enantiomers of the compound, or greater than about 95% by weight of one enantiomer of the compound and less than about 5% by weight of the other enantiomers of the compound, or greater than about 97% by weight of one enantiomer of the compound and less than about 3% by weight of the other enantiomers of the compound, or greater than about 99% by weight of one enantiomer of the compound and less than about 1% by weight of the other enantiomers of the compound.

[0027] As used herein, the term "cryptocephalides" includes compounds having Formula I:

[0028] (I)

[0029]

[0030] R1, R2 and R3 are independently (C1-C4) alkyl, methyl or hydroxyl. The term cryptocephalin may include hydroxymethylacylfulvene (HMAF, ilofofen), which has the following formula II:

[0031] (II) The term cryptocephalin may also include (-)-hydroxyureamethylacylfulvene, which has the following formula III

[0032] (III) BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 The relative importance of an exemplary gene signature for hydroxycarbamidomethylfulvene is shown.

[0034] Figure 2 PTGR1 and PTPN14 were shown to be associated with a positive response to hydroxyureamethylacylfulvene. DETAILED DESCRIPTION

[0035] One member of the anticancer agent sensitivity determination markers includes the gene markers prostaglandin reductase 1 (PTGR1), protein tyrosine phosphatase non-receptor type 14 (PTPN14), aspartate beta-hydroxylase (ASPH), used together with one or more genes or alone. In certain embodiments, the protein gene may be expressed or methylated. Prostaglandin reductase 1 (PTGR1) is a highly inducible enzyme with enone reductase activity. The marker PTGRI is associated with sensitivity to cryptosteglitin-based therapy or with true responders to cryptosteglitin-based therapy. The use of markers alone or in combination with other markers can reduce the gap in cancer (e.g., prostate cancer) treatment by developing screening tests based on predictive biomarkers to enable accurate drug-based treatment of patients. The present inventors have discovered that the detection of circulating solid tumor-associated mutant nucleic acid or protein sequences in individuals with solid tumor cancers can accurately determine whether the individual is sensitive to treatment with cryptosteglitin.

[0036] In a specific embodiment, the DNA of PTGR1, PTPN14, and ASPH genes is methylated. DNA methylation is the process of adding methyl groups to DNA molecules. Methylation can change the activity of DNA fragments without changing the sequence. When located in a gene promoter, DNA methylation is generally used to inhibit gene transcription.

[0037] A method of treating or managing solid tumor cancers (e.g., prostate cancer, ovarian cancer, liver cancer, kidney cancer, and thyroid cancer) comprising

[0038] Identifying patients with cancer that is sensitive to treatment with: Cryptocephalosporin of the formula:

[0039]

[0040] Cryptostegansin or a pharmaceutically acceptable salt, solvate or stereoisomer thereof; and administering a therapeutically effective amount of cryptostegansin to the patient.

[0041] A method of treating or administering a solid tumor comprises

[0042] Identifying patients with solid tumor cancers that are sensitive to treatment with cryptocephalin having the formula:

[0043]

[0044] or a pharmaceutically acceptable salt, solvate or stereoisomer thereof; and

[0045] A therapeutically effective amount of the compound is administered to the patient.

[0046] A method for predicting response to treatment in a patient with a solid tumor, comprising:

[0047] obtaining a biological sample from the patient;

[0048] measuring the expression level of PTGR1, PTPN14, ASPH or a combination thereof in the biological sample; and

[0049] comparing the expression level in the biological sample to the expression level in a biological sample from an individual not suffering from a solid cancer;

[0050] wherein an increased expression level in a biological sample from a patient relative to an individual not suffering from a solid tumor indicates the likelihood of an effective response to treatment with a compound of the formula:

[0051]

[0052] or a pharmaceutically acceptable salt, solvate or stereoisomer thereof.

[0053] A method for predicting response to treatment in a patient with a solid tumor, comprising:

[0054] obtaining a biological sample from the patient;

[0055] measuring the expression level of PTGR1, PTPN14, ASPH or a combination thereof in the biological sample; and

[0056] comparing the expression level of PTGR1, PTPN14, ASPH, or a combination thereof in the biological sample with the expression level in a biological sample from an individual not suffering from a solid tumor;

[0057] wherein a reduced expression level of PTGR1, PTPN14, ASPH, or a combination thereof in a biological sample from a patient relative to an individual not suffering from a solid tumor indicates a likelihood of an effective response to treatment with a compound of the formula:

[0058]

[0059] or a pharmaceutically acceptable salt, solvate or stereoisomer thereof.

[0060] In one embodiment, provided herein is a method of treating or managing solid tumor cancer, comprising: identifying a patient with a solid tumor cancer that is sensitive to treatment with cryptosteglitin or a pharmaceutically acceptable salt thereof; and administering to the patient a therapeutically effective amount of the compound. In certain embodiments, the methods provided herein further comprise administering a therapeutically effective amount of an additional active agent that is cryptosteglitin.

[0061] In one embodiment, cryptocephalosporin is administered once on day 1 of a 21 day cycle or another cycle.

[0062] In another embodiment, cryptocephalosporin may be administered with one or more second active ingredients. Such active ingredients may be other anti-cancer agents or agents used to treat cancer patients.

[0063] As stated above, the solid tumor cancer may be selected from the group consisting of colorectal cancer, pancreatic cancer, primary liver cancer, renal cancer, ovarian cancer, uterine cancer, lung cancer, breast cancer, prostate cancer, sarcoma, and adipose tissue cancer.

[0064] In one embodiment, one or more of the following genes can be used together with PTGR1 as a marker to determine sensitivity to cryptobactin-based anticancer agents: TNFRSF1B, LCK, TAL1, GNG12, S100A10, CD247, ATP1B1, LAMC1, PTPRC, PTPN7, PTPN14, CAPN2, ENAH, LCT, CXCR4, ITGA4, CASP 10. HDAC4, NUP210, WWTR1, PFN2, MLF1, CD38, RHOH, ITK, LCP2, HIST1H3B, MYB, IKZF1, PIK3CG, MET, NRF1, TRBC1, PTPRN2, PTK2B, ASPH, NFIB, CNTRL, PRKCQ, MYOF, CTBP2, LMO2 , CD3D, TWF1, NCKAP1L, CKAP4, LCP1, CFL2, TJP1, ALDH1A2, TLE3, TNFRSF12A, PRKCB, NFATC3, WWOX, CBFA2T3, ATP2A3, GIPC1, HAUS5, MAP4K1, SDC4, APP, SLC38A5, WAS and IL2RG.

[0065] In another embodiment, one or more of the following gene markers can be used: WWTR1, HIST1H3B, ASPH, MYOF, CTBP2, CBFA2T3, and SDC4. In another embodiment, one or more markers are combined with PTGR1, and one, more, or all of the following genes are combined with PTGR1: PTPN14, ITGA4, HDAC4, ITK, IKZF1, PIK3CG, NRF1, ASPH, and PRKCQ. In some cases, the sensitivity can be close to 100%.

[0066] In another embodiment, two markers PTGR1 and SDC4 can be used as markers.

[0067] In the case of using the PTGR1 gene combination, screening for anticancer drug sensitivity enhancers can be performed by using the expression change of the PTGR1 gene combination after exposure to a cryptostegliostigmine-based anticancer agent as an indicator. That is, a substance that reduces the level of the PTGR1 gene combination before exposure to a cryptostegliostigmine-based anticancer agent, or promotes expression changes or increased levels after exposure to a cryptostegliostigmine-based anticancer agent in vitro or in vivo enhances sensitivity to the anticancer agent. In addition, in the case of in vitro, a substance that promotes expression changes or increased levels after exposure to an anticancer agent corresponding to a target cancer cell can be used as a substance that enhances sensitivity to a cryptostegliostigmine-based anticancer agent (i.e., an anticancer drug sensitivity enhancer).

[0068] In order to carry out the method of determining the sensitivity of a sample to an anticancer agent, a kit containing a protocol for measuring any substance present in the sample is used. The kit contains reagents for measuring any of these substances, instructions for using the reagents, standards for determining whether there is sensitivity to a cryptosteganum-based anticancer agent, etc. The standards include (relative) standard levels, (relative) high threshold levels, (relative) low threshold levels, factors affecting measurement, degree of influence, etc. of these metabolism-related substances. These substance levels can be set so as to conform to the selected cryptosteganum-based anticancer agent. Sensitivity determination can be performed in the same manner based on the standards.

[0069] Screening of cryptostemonin-based anticancer agents can be performed with the aid of cryptostemonin-based anticancer agent sensitivity determination markers as indicators. That is, substances that can change the level of anticancer agent sensitivity determination markers in vitro or in vivo are evaluated as anticancer agents. For example, in vitro, substances that change the level of anticancer agent sensitivity determination markers in various cancer cells after exposure to the substance can act as anticancer agents. In addition, when the level of anticancer agent sensitivity determination markers is changed after the substance is administered to an animal carrying cancer, the substance can act as an anticancer agent. If the anticancer agent is expected to exhibit a pharmacological effect, an increase in the level of the anticancer agent sensitivity determination marker is observed before tumor shrinkage occurs or a cytocidal effect is achieved. Therefore, screening based on the level of anticancer agent sensitivity determination markers as an indicator can achieve the determination of whether the test substance acts as an applicable anticancer agent in a shorter period of time, thereby greatly expecting that the efforts and costs involved in the development of anticancer agents will be reduced.

[0070] When cancer is not sensitive to an anticancer agent, no pharmacological effect can be expected from the anticancer agent. If such an ineffective anticancer agent is continuously administered to a patient, cancer may progress and side effects may worsen. Therefore, anticancer agent sensitivity determination markers are useful not only for determining the therapeutic response to an anticancer agent, but also greatly contribute to preventing the worsening of side effects that would otherwise be caused by the continuous administration of a medically inactive anticancer agent.

[0071] By using the anticancer agent sensitivity enhancer thus obtained and the cryptostegliostigmine-based anticancer agent as a sensitivity enhancement target of the enhancer in a combined manner, the therapeutic effect of the cryptostegliostigmine-based anticancer agent is greatly enhanced. The combination of the cryptostegliostigmine-based anticancer agent sensitivity enhancer and the anticancer agent as a sensitivity enhancement target of the enhancer may be a composition containing both components, or a combination drug containing preparations of the individual components.

[0072] To aid in monitoring an individual, particularly if the reference value is obtained from the individual as part of an ongoing screening program. Alternatively, the reference value may be obtained from a population of individuals who do not have solid tumor cancer.

[0073] Cells for testing can be obtained by any method known in the art, including but not limited to surgical resection; biopsy, such as but not limited to needle biopsy, core biopsy or aspiration; or collected from fluid samples, such as blood, urine, cerebrospinal fluid, cystic fluid, etc. Methods for measuring DNA / mRNA include but are not limited to polymerase chain reaction, in situ hybridization, gel electrophoresis, sequence analysis and microarray analysis or a combination thereof. Methods for measuring proteins include but are not limited to mass spectrometry, 1-D or 2-D gel-based analysis systems, chromatography, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), enzyme immunoassay (EIA), Western blotting, immunoprecipitation and immunohistochemistry. Antibody arrays or protein chips can also be used. The agent is "likely" to produce an anti-cancer effect in an individual means that in one or more parameters tested (e.g., the level of PTGR1, PTPN14 and ASPH mRNA, the level of methylated DNA and / or protein, the expression level of the genes or exons listed in Example 1), the individual is more similar to other individuals in which the agent produces a significant anti-cancer effect than to other individuals in which the agent does not produce a significant anti-cancer effect. The agent is "unlikely" to produce an anti-cancer effect in an individual means that in one or more parameters tested, the individual is more similar to other individuals in which the agent does not produce a significant anti-cancer effect than to other individuals in which the agent produces a significant anti-cancer effect.

[0074] The pharmaceutical composition can be used to prepare individual single unit dosage forms. The pharmaceutical compositions and dosage forms provided herein include the immunomodulatory compounds provided herein, or pharmaceutically acceptable salts, solvates, hydrates, stereoisomers or prodrugs thereof. The pharmaceutical compositions and dosage forms provided herein may further include one or more excipients.

[0075] The pharmaceutical compositions and dosage forms provided herein may also include one or more second active ingredients. Thus, the pharmaceutical compositions and dosage forms provided herein include active ingredients disclosed herein (e.g., immunomodulatory compounds). Examples of optional second or additional active ingredients are disclosed herein.

[0076] Also contemplated are kits and compositions for carrying out the methods provided herein. In certain embodiments, kits are provided herein for determining the efficacy of anticancer compounds. In certain embodiments, kits are provided herein for evaluating the efficacy of compounds in treating patients. In certain embodiments, kits are provided herein for determining the effects of immunomodulatory compounds. The kits comprise a solid carrier and a component for detecting the gene, protein or glycoprotein expression of at least one biomarker in a biological sample. Such kits may be, for example, a measuring rod, a membrane, a chip, a disc, a test strip, a filter, a microsphere, a slide, a porous plate or an optical fiber. The solid carrier of the kit may be, for example, plastic, silicon, metal, resin, glass, film, particle, precipitate, gel, polymer, thin sheet, sphere, polysaccharide, capillary, film, plate or slide. The biological sample may be, for example, a cell culture, a cell line, a tissue, an oral tissue, a gastrointestinal tissue, an organ, an organelle, a biological fluid, a blood sample, a urine sample or a skin sample.

[0077] In one embodiment, the kit provided herein comprises a cryptocephalin compound provided herein, or a pharmaceutically acceptable salt, solvate or hydrate thereof. The kit may further comprise additional active agents, including but not limited to those disclosed herein.

[0078] Examples

[0079] Certain embodiments provided herein are illustrated by the following non-limiting examples. As shown in Example 1, analysis of in vitro cryptostegansin (LP-184) sensitivity data from a panel of cancer cell lines with known multi-omics molecular profiles generated a signature of 10 genes (ASPH, HDAC4, IKZF1, ITGA4, ITK, NRF1, PIK3CG, PRKCQ, PTGR1, and PTPN14) that were able to predict cryptostegansin sensitivity with up to or including 100% accuracy in blind testing. Gene expression, DNA mutations, DNA methylation, and protein expression status of these 10 genes were further calculated as correlations with cryptostegansin (LP-184) sensitivity. As shown in Table 1, of these 10 genes, 3 genes appeared to have highly significant correlations with cryptostegansin bioactivity at the multi-omics level. These genes are identified as PTGR1, PTPN14, and ASPH. In accordance with Figure 1 These three genes also had the highest gene weights in terms of relative importance when determining sensitivity to cryptocephalosporin (LP-184). Figure 2 As shown, it is presented as the most prominent genes in classifying samples as cryptosteglitin responders or non-responders. Overall, a novel gene set has been established that determines sensitivity to cryptosteglitin (LP-184) in cancer and predicts response to said cryptosteglitin.

[0080] Example 1

[0081] Genes associated with LP-184 (cryptocephalin family) sensitivity in cancer

[0082]

[0083] Figure 1 The relative importance of an exemplary gene signature to hydroxycarbamidofulvene is shown. In this relative variable importance plot, a gene weight analysis was performed to analyze the relative ranking of the top 10 genes in the hydroxycarbamidofulvene signature in cancer. The data show that PTGR1 (prostaglandin reductase 1) followed by PTPN14 and ASPH is a gene signature for hydroxycarbamidofulvene sensitivity.

[0084] Figure 2 PTGR1 and PTPN14 are shown to be associated with a positive response to hydroxyureamethylfulvene. In this sensitivity analysis plot, the effect of gene expression on the response variable was investigated across the LP-184 gene signature using the lekprofile function. High expression of PTGR1 (prostaglandin reductase 1) was associated with a positive response to LP-184), with PTPN14 being the next most highly associated gene. This experiment was performed on a panel of cancer cells.

[0085] The above embodiments are intended to be illustrative only, and those skilled in the art will recognize or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific compounds, materials, and procedures. All such equivalents are considered to be within the scope of the embodiments and are covered by the appended claims.

Claims

1. Use of a means for detecting the expression level of at least one of PTGR1, PTPN14, ASPH or a combination thereof in a biological sample from an individual in the preparation of a reagent for predicting the response of a patient suffering from cancer to treatment with hydroxyureamethylacylfulvene or a pharmaceutically acceptable salt thereof, wherein: The hydroxyureamethylacylfulvene has the formula 2. The use according to claim 1, wherein The cancer is newly diagnosed, relapsed, or refractory.

3. The use according to claim 1, wherein PTGR1, PTPN14 or ASPH is methylated.

4. The use according to claim 1, wherein It contains: obtaining a biological sample from the patient; measuring the expression level of at least one of PTGR1, PTPN14, ASPH or a combination thereof in the biological sample; and comparing the expression level in the biological sample to the expression level in a biological sample from an individual not suffering from cancer; wherein an increase in the expression level in the biological sample from the patient relative to the individual not suffering from the cancer indicates a response to treatment with hydroxyureamethylacylfulvene or a pharmaceutically acceptable salt, solvate or stereoisomer thereof.

5. The use according to claim 1, wherein The biological sample is blood, plasma, serum or tissue biopsy.

6. The use according to claim 1, wherein The cancer is colorectal cancer, pancreatic cancer, primary liver cancer, kidney cancer, ovarian cancer, uterine cancer, lung cancer, breast cancer, prostate cancer, sarcoma or adipose tissue cancer.

7. The use according to claim 1, wherein The expression level is the expression level of mRNA, DNA and / or protein of at least one of PTGR1, PTPN14, ASPH or a combination thereof in one or more cells of the cancer.

8. A kit for determining whether an anticancer effect in cancer may be produced by hydroxyureamethylacylfulvene, comprising means for determining the expression level of PTGR1, PTPN14, ASPH or a combination thereof in cancer relative to a normal value in a biological sample from a patient, relative to the expression level of PTGR1, PTPN14, ASPH or a combination thereof from an individual suffering from cancer, and wherein the hydroxyureamethylacylfulvene has the formula 9. The kit according to claim 8, wherein The cancer is colorectal cancer, pancreatic cancer, primary liver cancer, kidney cancer, ovarian cancer, uterine cancer, lung cancer, breast cancer, prostate cancer, sarcoma or adipose tissue cancer.

10. The kit according to claim 8, wherein Means are included for determining the level of mRNA, DNA and / or protein corresponding to PTGR1, PTPN14, ASPH or a combination thereof in one or more cells of the cancer.

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