Methods of treating cancer

Abstract

Disclosed herein is a method for predicting whether a subject with cancer will exhibit a beneficial response to arginine deficiency therapy. The method includes the step of determining the plasma concentration of arginine in the subject, followed by administering arginine deficiency therapy to the subject alone or in combination with an anticancer agent, based on the determined plasma concentration of arginine. According to some embodiments of this disclosure, the anticancer agent is selected from the group consisting of FOLFOX, docetaxel, cisplatin, pemetrexed, pembrolizumab, and combinations thereof.

Description

[Technical Field]

[0001] This disclosure generally relates to the field of cancer treatment. More specifically, this disclosure relates to a method of treating cancer by arginine deficiency therapy. [Background technology]

[0002] Cancer is a group of diseases characterized by the development of abnormal cells that divide uncontrollably and exhibit the ability to invade and destroy normal tissue. Cancer is the second leading cause of death worldwide, accounting for nearly 10 million deaths annually. The most common causes of cancer death are lung cancer, colorectal cancer, liver cancer, stomach cancer, and breast cancer. Cancer treatment varies depending on the type and stage of cancer. Major cancer treatments include surgery, chemotherapy, radiation therapy, immunotherapy, hormone therapy, and molecular targeted therapy. However, none of these treatments are always satisfactory, and a variety of side effects are observed in cancer patients. Therefore, there is ongoing interest in identifying and developing alternative methods for treating cancer.

[0003] Some cancers have nutritional requirements for a specific amino acid (e.g., arginine), and amino acid deficiency (e.g., arginine deficiency) may offer a potential treatment for these cancers. Arginine can be broken down by several enzymes, including arginine deiminase (ADI), a mycoplasma-derived microbial enzyme that exhibits high affinity for arginine and catalyzes its conversion to citrulline and ammonia. Citrulline can be recycled back to arginine in normal cells expressing argininosuccinate synthase 1 (ASS1). A PEGylated (polyethylene glycol-modified) form of ADI (ADI-PEG20) has been prepared and is reported to be in clinical trials to target arginine-dependent tumors with arginine deficiency therapy, but resistance to ADI often occurs due to reactivation or upward regulation of ASS1. Furthermore, it has been reported that prolonged treatment with an ADI can activate various cellular pathways related to resistance to apoptosis, which may limit the overall duration of effectiveness of ADI treatment.

[0004] Therefore, in related technologies, there is a demand for therapeutic strategies to improve treatment outcomes in cancer patients by optimizing arginine deficiency therapy. [Overview of the Initiative] [Means for solving the problem]

[0005] The following is a simplified summary of this disclosure to provide readers with a basic understanding of this specification. This summary is not a comprehensive overview of this disclosure, nor does it identify key elements of the invention or limit its scope. Its purpose is merely to provide a simplified description of some of the ideas disclosed herein, serving as a prelude to the more detailed descriptions that follow.

[0006] The present disclosure is based at least in part on the finding that the plasma concentration of arginine is associated with the response of cancer patients to therapy based on the PEGylated form of ADI (ADI-PEG20). Accordingly, the plasma concentration of arginine can be used to predict the therapeutic response of cancer patients to arginine deprivation therapy and also as a guide for individually formulating a suitable therapy regimen.

[0007] Thus, the present disclosure provides a method for predicting whether a subject suffering from cancer (e.g., hepatocellular carcinoma) will exhibit a beneficial response to arginine deprivation therapy. The method comprises (a) determining the plasma concentration of arginine in the subject, and (b) based on the plasma concentration of arginine determined in step (a), performing arginine deprivation therapy on the subject alone or in combination with an anti-cancer agent .

[0008] According to embodiments of the present disclosure, arginine deprivation therapy comprises administering to the subject an arginine-depleting agent selected from the group consisting of difluoromethylornithine (DFMO), recombinant arginine deiminase (rADI), recombinant arginase (rArg), recombinant arginine decarboxylase (rADC), the PEGylated form of rADI (hereinafter "PEGylated rADI"), the PEGylated form of rArg (hereinafter "PEGylated rArg"), the PEGylated form of rADC (hereinafter "PEGylated rADC"), and combinations thereof. According to some embodiments of the present disclosure, the anti-cancer agent is selected from the group consisting of FOLFOX, docetaxel, cisplatin, pemetrexed, pembrolizumab, and combinations thereof.

[0009] According to some embodiments, PEGylated rADI and FOLFOX are administered and performed individually on the subject when the plasma concentration of arginine in step (a) is 34 micromoles per liter or more (≧34 μmol / L). In some exemplary embodiments, PEGylated rADI is administered in an amount of about 36 mg per square meter of body surface area per week, and FOLFOX is performed every other week. 2 per and FOLFOX is performed every other week.

[0010] According to some embodiments, PEGylated rADI and pembrolizumab are administered individually to a subject when the plasma concentration of arginine in step (a) is 60.2 micromoles per liter or more (≥60.2 μmol / L). In some exemplary embodiments, PEGylated rADI is administered in an amount of about 36 mg per square meter of body surface area per week, and pembrolizumab is administered in an amount of about 200 mg every three weeks. 2 According to some embodiments, PEGylated rADI, pemetrexed and cisplatin are administered individually to a subject when the plasma concentration of arginine in step (a) is 68.2 micromoles per liter or more (≥68.2 μmol / L). In some exemplary embodiments, PEGylated rADI is administered in an amount of about 36 mg per square meter of body surface area per week, pemetrexed is administered in an amount of about 500 mg per square meter of body surface area every three weeks, and cisplatin is administered in an amount of about 75 mg per square meter of body surface area every three weeks.

[0011] According to some embodiments, PEGylated rADI is administered alone to a subject when the plasma concentration of arginine in step (a) is 84.2 micromoles per liter or more (≥84.2 μmol / L). In some preferred embodiments, PEGylated rADI is administered in an amount of about 18 mg per square meter of body surface area per week. 2 According to some embodiments, PEGylated rADI and docetaxel are administered individually to a subject when the plasma concentration of arginine in step (a) is 97.5 micromoles per liter or more (≥97.5 μmol / L). In some exemplary embodiments, PEGylated rADI is administered in an amount of about 36 mg per square meter of body surface area per week, and docetaxel is administered in an amount of per square meter of body surface area every three weeks. 2 2 2 According to some embodiments, PEGylated rADI and docetaxel are administered individually to a subject when the plasma concentration of arginine in step (a) is 97.5 micromoles per liter or more (≥97.5 μmol / L). In some exemplary embodiments, PEGylated rADI is administered in an amount of about 36 mg per square meter of body surface area per week, and docetaxel is administered in an amount of per square meter of body surface area every three weeks.

[0012] According to some embodiments, PEGylated rADI is administered alone to a subject when the plasma concentration of arginine in step (a) is 84.2 micromoles per liter or more (≥84.2 μmol / L). In some preferred embodiments, PEGylated rADI is administered in an amount of about 18 mg per square meter of body surface area per week. 2 According to some embodiments, PEGylated rADI and docetaxel are administered individually to a subject when the plasma concentration of arginine in step (a) is 97.5 micromoles per liter or more (≥97.5 μmol / L). In some exemplary embodiments, PEGylated rADI is administered in an amount of about 36 mg per square meter of body surface area per week, and docetaxel is administered in an amount of per square meter of body surface area every three weeks.

[0013] According to some embodiments, PEGylated rADI and docetaxel are administered individually to a subject when the plasma concentration of arginine in step (a) is 97.5 micromoles per liter or more (≥97.5 μmol / L). In some exemplary embodiments, PEGylated rADI is administered in an amount of about 36 mg per square meter of body surface area per week, and docetaxel is administered in an amount of per square meter of body surface area every three weeks. 2 According to some embodiments, PEGylated rADI and docetaxel are administered individually to a subject when the plasma concentration of arginine in step (a) is 97.5 micromoles per liter or more (≥97.5 μmol / L). In some exemplary embodiments, PEGylated rADI is administered in an amount of about 36 mg per square meter of body surface area per week, and docetaxel is administered in an amount of per square meter of body surface area every three weeks. 2It is administered in an amount of about 75 mg per area.

[0014] According to some embodiments, PEGylated rADI and cisplatin are administered individually to a subject when the plasma concentration of arginine in step (a) is 122 micromoles per liter or more (≧122 μmol / L). In some exemplary embodiments, PEGylated rADI is administered in an amount of about 36 mg per body surface area of 1 m 2 per week, and cisplatin is administered in an amount of about 30 mg per body surface area of 1 m 2 per week for 3 weeks, followed by a 1-week drug-free period.

[0015] Examples of cancers treatable by this method (i.e., arginine deprivation therapy alone or in combination with an anti-cancer agent) include, but are not limited to, breast cancer, brain tumor, colorectal cancer, head and neck squamous cell carcinoma, hepatocellular carcinoma (HCC), leukemia (e.g., acute myeloid leukemia (AML), lymphoma, lung cancer, melanoma, mesothelioma (e.g., malignant pleural mesothelioma (MPM)), neuroblastoma, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, and sarcoma.

[0016] Subjects treatable by this method are mammals, preferably humans.

[0017] Many of the relevant features and advantages of the present disclosure will be better understood by reference to the following detailed description taken in conjunction with the accompanying drawings.

[0018] This description will be better understood from the following detailed description read in light of the accompanying drawings.

Brief Description of the Drawings

[0019] [Figure 1] FIG. 1 is a graph illustrating the treatment of cancer patients with ADI-PEG20 analyzed by finding the arginine cut-off value using the maximum log-rank statistic. [Figure 2]Figure 2 is a line graph illustrating the correlation between arginine plasma concentration and overall survival in cancer patients who ingested ADI-PEG20 alone according to Example 1 of this disclosure. [Figure 3] Figure 3 is a graph illustrating the treatment of cancer patients with ADI-PEG20 + docetaxel, analyzed by finding the arginine cleavage value using the largest log-rank statistic. [Figure 4] Figure 4 is a line graph illustrating the correlation between arginine plasma concentration and overall survival in cancer patients who ingested ADI-PEG20 in combination with docetaxel according to Example 1 of this disclosure. [Figure 5] Figure 5 is a graph illustrating the treatment of cancer patients with ADI-PEG20 + cisplatin, analyzed by finding the arginine cleavage value using the largest log-rank statistic. [Figure 6] Figure 6 is a line graph illustrating the correlation between arginine plasma concentration and overall survival in cancer patients who ingested ADI-PEG20 in combination with cisplatin according to Example 1 of this disclosure. [Figure 7] Figure 7 is a graph illustrating the treatment of cancer patients with ADI-PEG20 + FOLFOX, analyzed by finding the arginine cleavage value using the largest log-rank statistic. [Figure 8] Figure 8 is a line graph illustrating the correlation between arginine plasma concentration and overall survival in cancer patients who ingested ADI-PEG20 in combination with mFOLFOX6 according to Example 1 of this disclosure. [Figure 9] Figure 9 is a graph illustrating the treatment of cancer patients with ADI-PEG20 + pemetrexed + cisplatin, analyzed by finding the arginine cleavage value using the largest log-rank statistic. [Figure 10] Figure 10 is a line graph illustrating the correlation between arginine plasma concentration and overall survival in cancer patients who ingested ADI-PEG20 in combination with pemetrexed and cisplatin according to Example 1 of this disclosure. [Figure 11]Figure 11 is a graph illustrating the treatment of cancer patients with ADI-PEG20 + pembrolizumab, analyzed by finding the arginine cleavage value using the largest log-rank statistic. [Figure 12] Figure 12 is a line graph illustrating the correlation between arginine plasma concentration and overall survival in cancer patients who ingested ADI-PEG20 in combination with pembrolizumab according to Example 1 of this disclosure. [Modes for carrying out the invention]

[0020] The detailed description below, accompanied by the attached drawings, is intended to illustrate this embodiment, but is not intended to represent only the forms in which this embodiment may be configured or utilized. This description outlines the function of the embodiment and the steps for configuring and operating it. However, the same or equivalent functions and steps may be performed by different embodiments.

[0021] I. Definition In this specification, the term “hepatocellular carcinoma” (or abbreviated as “HCC”) refers to a malignant tumor of hepatocyte origin. HCC is a type of liver cancer. Because HCC lacks fibrous stroma, bleeding and necrosis may occur. According to the Barcelona Clinical Liver Cancer (BCLC) staging system updated in 2022, HCC can be classified into five stages, including: (1) Stage 0 (very early), defined as the presence of a single tumor mass less than 2 cm in size with no vascular invasion or extrahepatic dissemination; (2) Stage A (early), defined as the presence of one tumor mass of any size or up to three tumor masses of multiple HCC with no vascular invasion or extrahepatic dissemination; (3) Stage B (intermediate), defined as the presence of multiple HCC with no vascular invasion or extrahepatic dissemination; (4) Stage C (advanced), defined as patients with portal vein invasion and / or extrahepatic dissemination; and (5) Stage D (terminal), defined as patients with significant cancer-related symptoms and / or liver dysfunction. According to the BCLC system classification, the term "advanced hepatocellular carcinoma" or "advanced HCC" refers to locally advanced HCC or metastatic HCC (i.e., HCC that has spread from the liver to other parts of the body). Generally, advanced HCC is unresectable (i.e., it has spread to surrounding tissues and cannot be surgically removed) and is not treatable with localized treatments such as radiation therapy.

[0022] In arginine deficiency therapy, the term "arginine deficiency agent" refers to a compound or drug that disrupts the uric acid cycle, cutting off the supply of arginine to cancer cells, thereby stopping cancer growth and inducing cell death.

[0023] The term "FOLFOX" refers to a chemotherapy regimen comprising 5-fluorouracil (5-FU), folinic acid (leucovorin), and oxaliplatin. As used herein, the term "FOLFOX" is not intended to be limited to any specific amount or administration regimen of these compounds. Rather, as used herein, "FOLFOX" includes all combinations of these compounds in any amount and administration regimen. Several different FOLFOX regimens exist, known in the art, based on the dosage and method of administration in which the three drugs are administered, including FOLFOX-4, FOLFOX-6, modified FOLFOX-6 (mFOLFOX-6), and FOLFOX-7. According to some embodiments of this disclosure, FOLFOX is mFOLFOX-6.

[0024] In this specification, the term “survival” refers to the act of living or the fact of being alive. The phrase “overall survival” (OS) refers to the extended portion of life expectancy compared to an unmedicated or untreated individual or patient.

[0025] In this specification, the term "confidence interval" (CI) has the ordinary meaning known to those skilled in the art, and refers to a statistical range in which the probability of a given parameter being within a certain range is a specific value.

[0026] The terms “administered” and “administering” are used interchangeably herein to refer to modes of delivery, including but not limited to administering a treatment (e.g., arginine deficiency therapy or anticancer agents) by intravenous, intratumoral, intramuscular, intraperitoneal, intra-arterial, or subcutaneous administration.

[0027] Unless otherwise specified, the terms “treat,” “treating,” and “treatment” refer to actions that reduce the severity of a disease or disorder, or one or more of its symptoms, or that slow or delay the progression of a disease or disorder, which occur when a patient has a particular disease or disorder.

[0028] The terms “cancer” and “tumor” are used interchangeably in this disclosure and preferably refer to or describe a physiological condition in mammals that is typically characterized by uncontrolled cell growth. In this regard, cancer includes metastatic cancer and / or drug-resistant cancer. Examples of cancer include, but are not limited to, breast cancer, brain tumors, colorectal cancer, head and neck squamous cell carcinoma, hepatocellular carcinoma (HCC), leukemia, acute myeloid leukemia (AML), lymphoma, lung cancer, melanoma, mesothelioma, malignant pleural mesothelioma (MPM), neuroblastoma, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, sarcoma, and combinations thereof.

[0029] Unless otherwise indicated, the terms “patient” and “subject” are interchangeable in this disclosure and refer to animals, including humans, that are treatable by the methods of the present invention. The terms “patient” or “subject” refer to both males and females unless one sex is specifically referred to.

[0030] Although the numerical ranges and parameters describing the broad scope of the present invention are approximations, the numerical values ​​described in specific examples are described as precisely as possible. However, any numerical value inherently includes errors that inevitably result from the standard deviation appearing in each test measurement. Furthermore, in this specification, the term “approximately” generally means within 10%, 5%, 1%, or 0.5% of a given value or range. Alternatively, the term “approximately” means the average standard error that would be acceptable to those skilled in the art. Except in examples / executions, or unless explicitly indicated otherwise, all numerical ranges, quantities, values, and percentages of materials, times, temperatures, operating conditions, ratios of quantities, and similar items disclosed herein should be understood to be modified in all cases by the term “approximately.” Accordingly, unless otherwise stated, the numerical parameters described in this disclosure and the appended claims are approximations that may vary as desired. At a minimum, each numerical parameter should be interpreted in light of the number of significant figures stated and by applying customary rounding methods.

[0031] In this specification, the singular forms "a," "and," and "the" are used to refer to multiple objects unless the text clearly reads otherwise.

[0032] II. Description of the Invention (1) Predicting patients' responses to arginine deficiency therapy A diverse range of responses to cancer treatment has long been recognized, due to the heterogeneity in the biological characteristics of cancer, the variety of physiological functions, and the differences in patients' genetic profiles. Therefore, one objective of this disclosure is to provide molecular markers associated with the response of cancer patients to arginine deficiency therapy. According to embodiments of this disclosure, plasma arginine concentration is associated with the therapeutic response of in-patient tumors (e.g., HCC) to therapy based on the PEG form of ADI (ADI-PEG20, molecular weight 20,000, arginine deiminase conjugated to polyethylene glycol).

[0033] Accordingly, a first aspect of this disclosure provides a method for predicting whether a subject suffering from cancer (e.g., hepatocellular carcinoma) will exhibit a beneficial response to arginine deficiency therapy. This method is (a) A step of determining the plasma concentration of arginine in the subject, and (b) The step includes making a prediction based on the plasma arginine concentration determined in step (a), and it is determined that if the plasma arginine concentration is 84.2 or higher, the subject is showing a beneficial response to arginine deficiency therapy.

[0034] In step (a), the plasma concentration of arginine is measured. Suitable assays used to determine the plasma concentration of arginine include, but are not limited to, spectrophotometric analysis, capillary electrophoresis (CE), enzyme-linked immunosorbent assay (ELISA), high-performance liquid chromatography (HPLC), mass spectrometry (MS), Sakaguchi reaction, biosensors, and combinations thereof.

[0035] In step (b), a person skilled in the art and a clinician can make a prediction based on the plasma concentration of arginine. According to some embodiments of the present disclosure, if the plasma concentration of arginine is 84.2 μmol / L or greater (≧84.2 μmol / L, e.g., 84.2, 84.3, 84.4, 84.5, 84.6, 84.7, 84.8, 84.9, 85, 85.1, 85.2, 85.3, 85.4, 85.5, 85.6, 85.7, 85.8, 85.9, 86, 86.1, 86.2, 86.3, 86.4, 86.5, 86.6, 86.7, 86.8, 86.9, 87, 87.1, 87.2, 87.3, 87.4, 87.5, 87 0.6, 87.7, 87.8, 87.9, 88, 88.1, 88.2, 88.3, ​​88.4, 88.5, 88.6, 88.7, 88.8, 88.9, 89, 89.1, 89.2, 89.3, 89.4, 89.5, 89.6, 89.7, 89.8, 89.9, 90, 90.1, 90.2, 90.3, 90.4, 90.5, 90.6, 90.7, 90.8, 90.9, 91, 91.1, 91.2, 91.3, 91.4, 91.5, 91.6, 91.7, 91.8, 91.9, 92, 92.1, 92.2, 9 2.3, 92.4, 92.5, 92.6, 92.7, 92.8, 92.9, 93, 93.1, 93.2, 93.3, 93.4, 93.5, 93.6, 93.7, 93.8, 93.9, 94, 94.1, 94.2, 94.3, 94.4, 94.5, 94.6, 94.7, 94.8, 94.9, 95, 95.1, 95.2, 95.3, 95.4, 95.5, 95.6, 95.7, 95.8, 95.9, 96, 96.1, 96.2, 96.3, 96.4, 96.5, 96.6, 96.7, 96.8, 96. When the arginine deficiency level is 9, 97, 97.1, 97.2, 97.3, 97.4, 97.5, 97.6, 97.7, 97.8, 97.9, 98, 98.1, 98.2, 98.3, 98.4, 98.5, 98.6, 98.7, 98.8, 98.9, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8, 99.9, or 100 μmol / L or higher, it can be seen that the subject exhibited a beneficial response to arginine deficiency therapy, i.e., a positive response to arginine deficiency therapy.In some studies, beneficial responses were associated with overall survival, with subjects with plasma arginine concentrations of ≥84.2 μmol / L having longer overall survival after treatment compared to subjects with plasma arginine concentrations <84.2 μmol / L.

[0036] In some embodiments of this disclosure, arginine deficiency therapy involves administering to a subject (e.g., a patient with HCC) an agent selected from the group consisting of DFMO, rADI, rArg, rADC, PEGylated rADI, PEGylated rArg, PEGylated rADC, and combinations thereof. In some exemplary embodiments, PEGylated rADI (ADI-PEG20) is administered weekly to a body surface area of ​​1 m². 2 In subjects administered approximately 18 mg of arginine per dose, the median overall survival for those with a plasma arginine concentration of ≥84.2 μmol was approximately 8.6 months (95% confidence interval (CI) ranging from 7.3 months to 10.5 months), while the median overall survival for subjects with a plasma arginine concentration of <84.2 μmol was approximately 5.7 months after treatment (95% confidence interval (CI) ranging from 4.9 months to 7.2 months).

[0037] Examples of cancer include, but are not limited to, breast cancer, brain tumors, colorectal cancer, head and neck squamous cell carcinoma, HCC, leukemia, AML, lymphoma, lung cancer, melanoma, mesothelioma, MPM, neuroblastoma, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, sarcoma, and combinations thereof.

[0038] In some embodiments of this disclosure, the cancer is HCC. In one particular embodiment, the cancer is advanced HCC.

[0039] (2) Predicting the patient's response to combination therapy According to some embodiments of the present disclosure, arginine deficiency therapy is combined with one or more additional therapies to improve its therapeutic effect. Accordingly, a second aspect of the present invention relates to a method for predicting whether a subject with cancer (e.g., HCC) will show a beneficial response to combination therapy (i.e., arginine deficiency therapy + additional therapy). This method includes (a) determining the plasma concentration of arginine in the subject, and (b) making a prediction based on the plasma concentration of arginine determined in step (a).

[0040] According to some embodiments of this disclosure, an arginine deficiency agent is combined with FOLFOX therapy. In these embodiments, the plasma concentration of arginine is 34 μmol / L or greater (≧34 μmol / L, e.g., 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72 When the plasma arginine concentration is 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 μmol / L or higher, subjects exhibit a beneficial response to combination therapy, i.e., a positive response to arginine deficiency agent + FOLFOX treatment. In some studies, the beneficial response was associated with overall survival, with subjects with plasma arginine concentrations of ≥34 μmol / L having a longer overall survival after treatment compared to subjects with plasma arginine concentrations of <34 μmol / L.

[0041] In some exemplary embodiments, PEGylated rADI (ADI-PEG20) is administered weekly to a body surface area of ​​1 m². 2In subjects administered approximately 36 mg of arginine per day, with FOLFOX treatment performed every other week, the median overall survival was approximately 9.5 months (95% CI range: 7.5 to 15.1 months) for subjects with a plasma arginine concentration of ≥34 μmol, and the median overall survival was approximately 4.3 months after treatment (95% CI range: 4.0 to 4.6 months) for subjects with a plasma arginine concentration <34 μmol.

[0042] According to some embodiments of this disclosure, an arginine deficiency agent is combined with pembrolizumab. In these embodiments, it is found that when the plasma concentration of arginine is 60.2 μmol / L or higher (≥60.2 μmol / L, e.g., 60.2, 60.3, 60.4, 60.5, 60.6, 60.7, 60.8, 60.9, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 μmol / L or higher), subjects exhibit a beneficial response to the combination therapy, i.e., a positive response to arginine deficiency agent + pembrolizumab treatment. In some studies, beneficial responses were associated with overall survival, with subjects with plasma arginine concentrations of ≥60.2 μmol / L having longer overall survival after treatment compared to subjects with plasma arginine concentrations <60.2 μmol / L.

[0043] In some exemplary embodiments, PEGylated rADI (ADI-PEG20) is administered weekly to a body surface area of ​​1 m². 2 The drug is administered at a dose of approximately 36 mg per dose, and pembrolizumab is administered at a dose of approximately 200 mg every three weeks.

[0044] According to some embodiments of this disclosure, an arginine deficiency agent is administered in combination with pemetrexed and cisplatin. In these embodiments, it is found that when the plasma concentration of arginine is 68.2 μmol / L or higher (≥68.2 μmol / L, e.g., 68.2, 68.3, 68.4, 68.5, 68.6, 68.7, 68.8, 68.9, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100 μmol / L or higher), subjects exhibit a beneficial response to the combination therapy, i.e., a positive response to arginine deficiency agent + pemetrexed and cisplatin treatment. In some studies, beneficial responses were associated with overall survival, with subjects with plasma arginine concentrations of ≥68.2 μmol / L having longer overall survival after treatment compared to subjects with plasma arginine concentrations <68.2 μmol / L.

[0045] In some exemplary embodiments, PEGylated rADI (ADI-PEG20) is administered weekly to a body surface area of ​​1 m². 2 It is administered in doses of approximately 36 mg per 1 m² of body surface area every 3 weeks, and pemetrexed is administered every 3 weeks. 2 It is administered in doses of approximately 500 mg per 1 m² of body surface area every 3 weeks, and cisplatin is added every 3 weeks. 2 The drug is administered at a dose of approximately 75 mg per dose. The median overall survival for subjects with a plasma arginine concentration of ≥68.2 μmol was approximately 12.5 months (95% CI range: 9.8 months to 14.2 months), while the median overall survival for subjects with a plasma arginine concentration of <68.2 μmol was approximately 6.5 months after treatment (95% CI range: 3.8 months to 8.8 months).

[0046] According to some embodiments of this disclosure, an arginine deficiency agent is administered in combination with docetaxel. In these embodiments, it is found that when the plasma concentration of arginine is 97.5 μmol / L or higher (≥97.5 μmol / L, e.g., 97.5, 97.6, 97.7, 97.8, 97.9, 98, 98.1, 98.2, 98.3, 98.4, 98.5, 98.6, 98.7, 98.8, 98.9, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8, 99.9, or 100 μmol / L or higher), subjects exhibit a beneficial response to the combination therapy, i.e., a positive response to arginine deficiency agent + docetaxel treatment. In some studies, beneficial responses were associated with overall survival, with subjects with plasma arginine concentrations of ≥97.5 μmol / L having longer overall survival after treatment compared to subjects with plasma arginine concentrations <97.5 μmol / L.

[0047] In some exemplary embodiments, PEGylated rADI (ADI-PEG20) is administered weekly to a body surface area of ​​1 m². 2 It is administered in doses of approximately 36 mg per 1 m² of body surface area every 3 weeks, and docetaxel is added every 3 weeks. 2 The drug is administered at a dose of approximately 75 mg per dose. The median overall survival for subjects with a plasma arginine concentration of ≥97.5 μmol was approximately 40.7 months (95% CI range: 7.2 months to 40.7 months), while the median overall survival for subjects with a plasma arginine concentration of <97.5 μmol was approximately 14.6 months after treatment (95% CI range: 5.7 months to 16.0 months).

[0048] According to some embodiments of this disclosure, the arginine deficiency agent is administered in combination with cisplatin. In these embodiments, the plasma concentration of arginine is 122 μmol / L or greater (≧122 μmol / L, for example, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, When the plasma arginine concentration is 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, or 200 μmol / L or higher, subjects exhibit a beneficial response to combination therapy, i.e., a positive response to arginine deficiency agent + cisplatin treatment. In some studies, the beneficial response is associated with overall survival, with subjects with plasma arginine concentrations of ≥122 μmol / L having longer overall survival after treatment compared to subjects with plasma arginine concentrations of <122 μmol / L.

[0049] In some exemplary embodiments, PEGylated rADI (ADI-PEG20) is administered weekly to a body surface area of ​​1 m². 2 Administered in doses of approximately 36 mg per unit, cisplatin is administered weekly to a body surface area of ​​1 m² per treatment cycle. 2 The drug is administered at a dose of approximately 30 mg per week for 3 weeks (weeks 1-3), followed by a 1-week (week 4) rest period. The median overall survival for subjects with a plasma arginine concentration of ≥122 μmol was approximately 15.7 months (95% CI range: 8.9 months to 28.5 months), while the median overall survival for subjects with a plasma arginine concentration of <122 μmol was approximately 6.4 months after treatment (95% CI range: 3.4 months to 8.2 months).

[0050] As described above, cancer may be breast cancer, brain tumor, colorectal cancer, head and neck squamous cell carcinoma, HCC, leukemia, AML, lymphoma, lung cancer, melanoma, mesothelioma, MPM, neuroblastoma, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, sarcoma, and combinations thereof. In some embodiments, the cancer is HCC. In a particular embodiment, the cancer is advanced HCC.

[0051] (3) Methods of treating cancer Another aspect of the present disclosure relates to a method for treating cancer with arginine deficiency agents for arginine deficiency therapy (i.e., DFMO, rADI, rArg, rADC, PEGylated rADI, PEGylated rArg, PEGylated rADC, and combinations thereof) alone or in combination with one or more additional therapies. In some embodiments of the present disclosure, the method includes (a) determining the plasma concentration of arginine in a subject, and (b) performing a suitable treatment for the subject based on the plasma concentration of arginine determined in step (a).

[0052] According to some embodiments of this disclosure, when the plasma concentration of arginine is 34 μmol / L or higher, PEGylated rADI (ADI-PEG20) and FOLFOX therapy are administered and performed individually to subjects. In some exemplary embodiments, to improve or alleviate cancer-related symptoms, PEGylated rADI (ADI-PEG20) is administered weekly to a body surface area of ​​1 m². 2 The drug is administered at a dose of approximately 36 mg per dose, and FOLFOX treatment is performed every other week. As is understandable, those skilled in the art or clinicians may adjust the treatment regimen (including the dosage, schedule, and duration of ADI-PEG20 and FOLFOX treatment) to suit their actual needs.

[0053] Alternatively, if the plasma concentration of arginine is less than 34 μmol / L, an alternative anticancer treatment may be administered to the subject for therapeutic purposes, preferably selected from the group consisting of surgery, chemotherapy, molecular targeted therapy, radiotherapy, hormone therapy, immunotherapy, and combinations thereof. Those skilled in the art or clinicians may select a suitable treatment for a cancer patient in accordance with clinical factors such as the patient's age, sex and physical condition, as well as the type and stage of cancer.

[0054] Drugs commonly used in chemotherapy include doxorubicin, adriamycin, bleomycin, actinomycin, dactinomycin, mutamycin, daunorubicin, epirubicin, idarubicin, mitoxantrone, mitomycin, epipodophyllotoxins, etoposide, teniposide, microtubule inhibitors, vinblastine, vincristine, vindesine, vinorelbine, taxanes, paclitaxel (Taxol), nitrogen mustard, chlorambucil, cyclophosphamide, estramustine, ifosfamide, mechloretamine, melphalan, aziridines, thiotepa, and alkylsulfonic acids. Examples include, but are not limited to, busulfan, nitrosoureas, carmustine, lomustine, streptozosin, platinum complexes, carboplatin, alkylating agents, altretamine, dacarbazine, procarbazine, temozolomide, methotrexate, fludarabine, mercaptopurine, thioguanine, cladribine, pentostatin, capecitabine, cytarabine, phloxuridine, fluorouracil, gemcitabine, hydroxyurea, camptothecin, irinotecan, busuphan, epothyron, azathioprine, halofudinone, sirolimus, everolimus, mitomycin, and topotecan.

[0055] Examples of drugs for molecular targeted therapy include, but are not limited to, trastuzumab or pertuzumab (antibodies specific to the tumor antigen HER-2 / neu), bevacizumab (antibody specific to vascular endothelial growth factor (VEGF)), ramucirumab (antibody specific to the VEGF receptor), nivolumab or semiplimab (antibodies specific to programmed cell death protein 1 (PD-1)), atezolizumab, avelumab or durvalumab (antibodies specific to the ligand for programmed cell death protein 1 (PD-L1)), ipilimumab (antibody specific to cytotoxic T lymphocyte-associated protein (CTLA-4)), and rituximab (antibody specific to CD20 on the surface of B cells).

[0056] Non-exclusive examples of immunomodulatory agents for immunotherapy include thalidomide, lenalidomide, pomalidomide, anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-CTLA-4 antibodies, interleukin (IL)-2, IL-6, IL-12, interferon-α (IFN-α), IFN-β, IFN-γ, granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), and cancer vaccines (e.g., human papillomavirus (HPV) vaccine and hepatitis B vaccine).

[0057] According to some embodiments of this disclosure, when the plasma concentration of arginine is 60.2 μmol / L or higher, PEGylated rADI (ADI-PEG20) and pembrolizumab are administered individually to subjects. In some exemplary embodiments, to improve or alleviate cancer-related symptoms, PEGylated rADI (ADI-PEG20) is administered weekly to a body surface area of ​​1 m². 2 The drug is administered at a dose of approximately 36 mg per dose, and pembrolizumab is administered at a dose of approximately 200 mg every three weeks. It should be understandable that a person skilled in the art or a clinician may adjust the treatment regimen (including the dosage, schedule, and duration of treatment with ADI-PEG20 and pembrolizumab) to suit the actual needs.

[0058] Alternatively, if the plasma concentration of arginine is less than 60.2 μmol / L, an alternative anticancer treatment is administered to the subject for therapeutic purposes. As described above, the alternative anticancer treatment is preferably selected from the group consisting of surgery, chemotherapy, molecular targeted therapy, radiotherapy, hormone therapy, immunotherapy, and combinations thereof. Those skilled in the art or clinicians may select a suitable treatment for a cancer patient in accordance with the patient's clinical factors.

[0059] According to some embodiments of this disclosure, when the plasma concentration of arginine is 68.2 μmol / L or higher, PEGylated rADI (ADI-PEG20), pemetrexed, and cisplatin are administered individually to the subject. In some exemplary embodiments, to improve or alleviate cancer-related symptoms, PEGylated rADI (ADI-PEG20) is administered weekly to a body surface area of ​​1 m². 2 It is administered in doses of approximately 36 mg per 1 m² of body surface area every 3 weeks, and pemetrexed is administered every 3 weeks. 2 It is administered in doses of approximately 500 mg per 1 m² of body surface area every 3 weeks, and cisplatin is added every 3 weeks. 2 It is administered in doses of approximately 75 mg per dose. Those skilled in the art or clinicians may adjust the treatment regimen (including the dosage, schedule, and duration of treatment with ADI-PEG20, pemetrexed, and cisplatin) to suit the actual needs.

[0060] Alternatively, if the plasma concentration of arginine is less than 68.2 μmol / L, an alternative anticancer treatment is administered to the subject for therapeutic purposes. The alternative anticancer treatment is preferably selected from the group consisting of surgery, chemotherapy, molecular targeted therapy, radiotherapy, hormone therapy, immunotherapy, and combinations thereof. Those skilled in the art or clinicians may select the appropriate treatment for a cancer patient in accordance with the patient's clinical factors.

[0061] According to some embodiments of this disclosure, when the plasma concentration of arginine is 84.2 μmol / L or higher, PEGylated rADI (ADI-PEG20) is administered to a subject alone without any additional treatment. In some exemplary embodiments, to improve or alleviate cancer-related symptoms, PEGylated rADI (ADI-PEG20) is administered weekly to a body surface area of ​​1 m². 2 It is administered in doses of approximately 18 mg per dose. Those skilled in the art or clinicians may adjust the treatment regimen (including the dosage, schedule, and duration of ADI-PEG20 treatment) to suit their actual needs.

[0062] In contrast, if the plasma concentration of arginine is less than 84.2 μmol / L, alternative anticancer therapies are administered to the subject for therapeutic purposes. These alternative anticancer therapies are preferably selected from the group consisting of surgery, chemotherapy, molecular targeted therapy, radiotherapy, hormone therapy, immunotherapy, and combinations thereof. Those skilled in the art or clinicians may select the appropriate treatment for cancer patients in accordance with the patient's clinical factors.

[0063] According to some embodiments of this disclosure, when the plasma concentration of arginine is 97.5 μmol / L or higher, PEGylated rADI (ADI-PEG20) and docetaxel are administered individually to the subject. In some exemplary embodiments, to improve or alleviate cancer-related symptoms, PEGylated rADI (ADI-PEG20) is administered weekly to a body surface area of ​​1 m². 2 It is administered in doses of approximately 36 mg per 1 m² of body surface area every 3 weeks, and docetaxel is added every 3 weeks. 2 It is administered in doses of approximately 75 mg per dose. Those skilled in the art or clinicians may adjust the treatment regimen (including the dosage, schedule, and duration of treatment with ADI-PEG20 and docetaxel) to suit their actual needs.

[0064] Alternatively, if the plasma concentration of arginine is less than 97.5 μmol / L, an alternative anticancer treatment is administered to the subject for therapeutic purposes. The alternative anticancer treatment is preferably selected from the group consisting of surgery, chemotherapy, molecular targeted therapy, radiotherapy, hormone therapy, immunotherapy, and combinations thereof. Those skilled in the art or clinicians may select the appropriate treatment for a cancer patient in accordance with the patient's clinical factors.

[0065] According to some embodiments of this disclosure, when the plasma concentration of arginine is 122 μmol / L or higher, PEGylated rADI (ADI-PEG20) and cisplatin are administered individually to the subject. In some exemplary embodiments, to improve or alleviate cancer-related symptoms, PEGylated rADI (ADI-PEG20) is administered weekly to a body surface area of ​​1 m². 2 Administered in doses of approximately 36 mg per unit, cisplatin is administered weekly to a body surface area of ​​1 m² per treatment cycle. 2 Approximately 30 mg is administered per week for 3 weeks (weeks 1-3), followed by a 1-week (week 4) rest period. Those skilled in the art or clinicians may adjust the treatment regimen (including the dosage, schedule, and duration of ADI-PEG20 and cisplatin treatment) to suit actual needs.

[0066] In contrast, if the plasma concentration of arginine is less than 122 μmol / L, alternative anticancer therapies are administered to the subject for therapeutic purposes. These alternative anticancer therapies are preferably selected from the group consisting of surgery, chemotherapy, molecular targeted therapy, radiotherapy, hormone therapy, immunotherapy, and combinations thereof. Those skilled in the art or clinicians may select the appropriate treatment for cancer patients in accordance with the patient's clinical factors.

[0067] Cancers treatable by this method include, but are not limited to, breast cancer, brain tumors, colorectal cancer, head and neck squamous cell carcinoma, HCC, leukemia, AML, lymphoma, lung cancer, melanoma, mesothelioma, MPM, neuroblastoma, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, and sarcoma. In some exemplary embodiments, the cancer is HCC. In a particular embodiment, the cancer is advanced HCC.

[0068] The subjects are mammals such as humans, mice, rats, guinea pigs, monkeys, sheep, goats, cats, dogs, horses, or chimpanzees. Preferably, the subjects are humans.

[0069] The present invention will now be described more specifically with reference to the following embodiments, which are provided for illustrative purposes rather than limitation. They belong to those that are commonly used, but other procedures, methodologies, or techniques known to those skilled in the art may be used instead. [Examples]

[0070] Raw materials and methods Patient registration This study used plasma samples from patients receiving arginine deficiency therapy. Patients enrolled in this study (including cohorts 1-6) were diagnosed with advanced stages of HCC and participated in the ADI-PEG20 clinical trial in Taiwan. Samples and clinicopathological parameters, de-linked to personal information, were used for post-hoc analysis.

[0071] Cohort 1, consisting of 422 patients, received ADI-PEG20 monotherapy, with each patient receiving 18 mg / m² of ADI-PEG20 weekly, unless disease progression, an unacceptable adverse event occurred, or other discontinuation criteria were met. 2 It was administered intramuscularly at the specified dosage.

[0072] Cohort 2, consisting of 31 patients, received ADI-PEG20 along with docetaxel, with each patient receiving 36 mg / m² of ADI-PEG20 weekly, unless disease progression, unacceptable adverse events occurred, or other discontinuation criteria were met. 2 The dosage is administered intramuscularly, and docetaxel is added every three weeks at a dose of 75 mg / m². 2 It was administered intravenously at the specified dosage.

[0073] Cohort 3, consisting of 78 patients, received ADI-PEG20 along with cisplatin, with each patient receiving 36 mg / m² of ADI-PEG20 weekly, unless disease progression, unacceptable adverse events occurred, or other discontinuation criteria were met. 2 The drug is administered intramuscularly at the specified dose, with each patient receiving cisplatin at 30 mg / m² per week during one treatment cycle. 2 The drug was administered at this dosage for 3 weeks (weeks 1-3), followed by a 1-week (week 4) drug-free period.

[0074] Cohort 4, which included 39 patients, received a modified FOLFOX6 regimen of ADI-PEG20 (oxaliplatin 85 mg / m² on day 1). 2 , 5-FU bolus 400 mg / m² 2 , and leucovorin 400 mg / m² 2 After administering [the drug], 2400 mg / m² 2 In combination with the mFOLFOX6 regimen (5-FU infusion over two consecutive days), ADI-PEG20 is administered to each patient at a dose of 36 mg / m² weekly, unless disease progression, unacceptable adverse events occur, or other discontinuation criteria are met. 2 The drug was administered intramuscularly at the specified dose, and the mFOLFOX6 regimen was administered intravenously to each patient every other week.

[0075] Cohort 5, consisting of 111 patients, received ADI-PEG20 along with pemetrexed and cisplatin, with each patient receiving 36 mg / m² of ADI-PEG20 weekly, unless disease progression, an unacceptable adverse event occurred, or other discontinuation criteria were met. 2 The drug is administered intramuscularly at the specified dose, with each patient receiving pemetrexed at 500 mg / m² every three weeks. 2 The following dosage is administered intravenously: each patient receives 75 mg / m² of cisplatin every three weeks. 2 Administer intravenously at the specified dosage.

[0076] Cohort 6, consisting of 27 patients, received ADI-PEG20 in combination with pembrolizumab, with each patient receiving 36 mg / m² of ADI-PEG20 weekly, unless disease progression, an unacceptable adverse event occurred, or other discontinuation criteria were met. 2 The drug was administered intramuscularly at the specified dose, and pembrolizumab was administered intravenously at a dose of 200 mg every three weeks to each patient.

[0077] Evaluation of treatment outcomes Overall survival (OS) for patients in Cohorts 1 and 5 was calculated from the date of enrollment for randomization to the date of death, regardless of cause, or the date of loss of follow-up. For patients in Cohorts 2, 3, 4, and 6, OS was calculated as the time from the first administration of the study's treatment to death from any cause, and if a subject was alive or lost follow-up, the data was terminated at the date of the last contact.

[0078] statistical analysis The best truncation point for arginine plasma concentration related to survival outcomes was determined using the maximum selection log-rank statistic. The Kaplan-Meier method was used to estimate the median OS of patients. A p-value < 0.05 obtained by the log-rank test was considered statistically significant. Statistical analysis was performed using software.

[0079] Example 1: Correlation of plasma arginine concentration with overall survival in patients with advanced HCC receiving arginine deficiency therapy alone or in combination with additional treatments. To investigate whether plasma arginine levels correlate with overall survival (OS) in cancer patients, we performed maximum-selection log-rank statistics. The analysis results are shown in Figures 1-12 and summarized in Tables 1-3.

[0080] [Table 1]

[0081] Table 1 Correlation of arginine plasma concentration with overall survival (OS) in cancer patients receiving specific treatments. [1] The maximum value is the selected log-rank statistic. [2] Moon [3] Positive limit estimate of the Kaplan-Meier method [4] The p-values ​​used to compare treatment groups are based on the log-rank test.

[0082] [Table 2]

[0083] Table 2 Correlation of arginine plasma concentration with overall survival (OS) in cancer patients receiving specific treatments [1] The maximum value is the selected log-rank statistic. [2] Moon [3] Positive limit estimate of the Kaplan-Meier method [4] The p-values ​​used to compare treatment groups are based on the log-rank test.

[0084] Example 2 Procedure for determining arginine cleavage values ​​(e.g., in HCC) In the first step, the arginine truncation value is determined. HCC patients agree to participate in Polaris' HCC clinical trial, and Polaris collects the patients' plasma before the initial ADI-PEG20 treatment. Subsequently, pharmacokinetics are evaluated by measuring peripheral blood concentrations of arginine and citrulline using LCMS. In this study, a total of 633 patients were enrolled (422 in the ADI-PEG20 group and 211 in the placebo group). Subsequently, once the number of deaths reaches the sample size requirement at the beginning of the study design, the clinical database is locked. Furthermore, a statistician calculates the overall survival for each patient. The arginine truncation value is determined using the maximum selection log-rank statistic (the statistician inputs each patient's arginine value and OS into the statistical model).

[0085] In the second step, the effectiveness of arginine cleavage levels is demonstrated. 422 HCC patients treated with ADI-PEG20 are divided into two groups: a high-arginine group and a low-arginine group. Overall survival is then aggregated using the Kaplan-Meier positive limit estimate. Point estimates with 95% confidence intervals (25th, 50th, and 75th percentiles) are provided for each treatment group. Survival estimates are also presented graphically for each treatment group. Furthermore, treatments are compared using a stratified log-rank test.

[0086] In one embodiment, a method for in vitro identifying the arginine threshold in a subject's sample to predict whether a cancer subject will respond to arginine deficiency therapy includes: providing a sample taken from the subject before the administration of the arginine deficiency agent; determining the arginine concentration by measuring the arginine concentration in the sample; calculating overall survival by evaluating the overall survival statistic after the subject has received arginine deficiency therapy; (1) identifying the highest value of the standardized log-rank statistic on the Y-axis of a statistical graph; (2) finding the highest point on the Y-axis and finding the corresponding arginine value on the X-axis; and (3) determining the arginine concentration corresponding to this highest point as the arginine truncation value (for example) The arginine cleavage value is determined by the following steps (see Figure 1): first, the arginine cleavage value is 84.2 μmol / L in Cohort 1; second, the arginine cleavage value is taken as the maximum arginine value; third, the arginine value is gradually decreased, and the corresponding p-value is calculated for each arginine value; this process is continued until the p-value for the nth arginine value is greater than 0.05; and then the arginine value corresponding to the (n-1)th arginine value is taken as the arginine threshold (see Figure 3). When a subject's arginine concentration is above the arginine threshold, the cancer subject is considered to be responding well to arginine deficiency therapy.

[0087] Based on the results above, the 422 patients in Cohort 1 received 18 mg / m². 2The patients were treated with ADI-PEG20, and the estimated arginine cleavage value for overall survival was set at 84.2 μmol / L. The maximum log-rank statistic recorded was M = 2.8792 (Figure 1). The difference in overall survival between the two groups, defined by the arginine cleavage value of 84.2 μmol / L, is shown in the figure. The high-arginine group (arginine ≥ 84.2 μmol / L) had a median OS of 8.6 months (95% CI: 7.3, 10.5), while the low-arginine group (arginine < 84.2 μmol / L) had a median OS of 5.7 months (95% CI: 4.9, 7.2), indicating that the high-arginine group survived longer in Cohort 1 (p = 0.0057) (Figure 2 and Table 1).

[0088] According to the HCC trial of ADI-PEG20 monotherapy, the arginine cleavage value was 84.2 μmol / L based on maximum selection rank statistics. Patients with arginine concentrations ≥84.2 μmol / L may survive longer than patients with arginine concentrations <84.2 μmol / L.

[0089] To enroll more patients, predictor variable analysis was used to determine the arginine threshold. Based on an arginine truncation value of 84.2 μmol / L, the highest arginine value was assumed, and then the arginine value was gradually decreased to 83 μmol / L, 82 μmol / L, 81 μmol / L, 80 μmol / L, and 79 μmol / L. In Table 3, the corresponding significant *p value for each arginine value is less than 0.05. However, at an arginine value of 78 μmol / L, the *p value becomes greater than 0.05 for the first time. Therefore, 78 μmol / L is defined as the nth arginine value. The immediately preceding (n-1)th arginine value is 79 μmol / L, which also serves as the arginine threshold. Survival time is longer in the group with arginine values ​​above the threshold than in the low arginine group, reaching statistical significance. Statistical results indicate that the arginine threshold is, in most cases, 5% to 10% smaller than the arginine truncation value (Table 3).

[0090] [Table 3]

[0091] Table 3. Tracking values ​​for Cohort 1 A p-value less than 0.05 is considered statistically significant.

[0092] Based on the results above, the 31 patients in Cohort 2 received 36 mg / m². 2 ADI-PEG20+ 75mg / m² 2 Patients were treated with docetaxel, and the estimated arginine cleavage value for overall survival was set at 97.5 μmol / L. The maximum log-rank statistic recorded was M = 3.0559 (Figure 3). The difference in overall survival between the two groups, defined by the arginine cleavage value of 97.5 μmol / L, is shown in the figure. The high-arginine group (arginine ≥ 97.5 μmol / L) had a median OS of 40.7 months (95% CI: 7.2, 40.7), while the low-arginine group (arginine < 97.5 μmol / L) had a median OS of 14.6 months (95% CI: 5.7, 16.0), indicating that the high-arginine group survived longer in Cohort 2 (p = 0.0117) (Figure 4 and Table 2).

[0093] Based on the results above, the 78 patients in Cohort 3 received 36 mg / m². 2 ADI-PEG20+30mg / m² 2 The patients were treated with cisplatin, and the estimated arginine cleavage value for overall survival was set at 122 μmol / L. The maximum log-rank statistic recorded was M = 3.6943 (Figure 5). The difference in overall survival between the two groups, defined by the arginine cleavage value of 122 μmol / L, is shown in the figure. The high-arginine group (arginine ≥ 122 μmol / L) had a median OS of 15.7 months (95% CI: 8.9, 28.5), while the low-arginine group (arginine < 122 μmol / L) had a median OS of 6.4 months (95% CI: 3.4, 8.2), indicating that the high-arginine group survived longer in Cohort 3 (p = 0.0006) (Figure 6 and Table 2).

[0094] Based on the results above, the 39 patients in Cohort 4 received 36 mg / m². 2 The patients were treated with ADI-PEG20 + FOLFOX, and the estimated arginine cleavage value for overall survival was set at 34 μmol / L. The maximum log-rank statistic recorded was M = 1.6007 (Figure 7). The difference in overall survival between the two groups, defined by the arginine cleavage value of 34 μmol / L, is shown in the figure. The high-arginine group (arginine ≥ 34 μmol / L) had a median OS of 9.5 months (95% CI: 7.5, 15.1), while the low-arginine group (arginine < 34 μmol / L) had a median OS of 4.3 months (95% CI: 4.0, 4.6), indicating that the high-arginine group survived longer in Cohort 4 (p = 0.00083) (Figure 8 and Table 2).

[0095] Based on the results above, the 111 patients in Cohort 5 received 36 mg / m². 2 ADI-PEG20+ 500mg / m² 2 Pemetrexed + 75 mg / m² 2 The patients were treated with cisplatin, and the estimated arginine cleavage value for overall survival was set at 68.2 μmol / L. The maximum log-rank statistic recorded was M = 3.043 (Figure 9). The difference in overall survival between the two groups, defined by the arginine cleavage value of 68.2 μmol / L, is shown in the figure. The high-arginine group (arginine ≥ 68.2 μmol / L) had a median OS of 12.5 months (95% CI: 9.8, 14.2), while the low-arginine group (arginine < 68.2 μmol / L) had a median OS of 6.5 months (95% CI: 3.8, 8.8). This indicates that the high-arginine group survived longer in Cohort 5 (p = 0.0011) (Figure 10 and Table 1).

[0096] Based on the results above, 27 patients received 36 mg / m². 2The subjects were treated with ADI-PEG 20 + 200 mg of pembrolizumab, and the estimated arginine cleavage value for overall survival was set at 60.2 μmol / L. The maximum log-rank statistic recorded was M = 3.0899 (Figure 11). The difference in overall survival between the two groups, defined by the arginine cleavage value of 60.2 μmol / L, is shown in the figure. Due to a high censoring rate, the median OS could not be estimated as the majority of subjects are still alive. The low-arginine group (arginine < 60.2 μmol / L) had a median OS of 6.6 months (95% CI: 1.8, 7.8), indicating that the high-arginine group survived longer in Cohort 6 (p = 0.0119) (Figure 12 and Table 2).

[0097] In summary, plasma arginine concentrations may allow us to predict the outcome (i.e., overall survival) of arginine deficiency therapy, either alone or in combination with various anticancer therapies including FOLFOX, docetaxel, cisplatin, pemetrexed, and pembrolizumab.

[0098] The above description of embodiments is merely illustrative, and it should be understood that various modifications can be made by those skilled in the art. The above specifications, examples, and data fully describe the structure and use of exemplary embodiments of the present invention. Although various embodiments of the present invention have been described in a certain degree of specificity, or by reference to one or more individual embodiments, those skilled in the art will be able to make numerous changes to the disclosed embodiments without departing from the spirit or scope of this disclosure.

Claims

1. A method for in vitro determining the arginine threshold in a sample of a cancer subject in order to predict whether the subject will respond to arginine deficiency therapy, Provide samples taken from subjects before the administration of arginine deficiency agents. The arginine concentration in the aforementioned sample is measured to determine the arginine concentration. The overall survival period is calculated by evaluating the overall survival statistics after the subject receives the arginine deficiency therapy. The arginine cleavage value is calculated by inputting the arginine concentration and overall survival time of the subject into the statistical method of maximum selection log-rank statistics and determining the arginine cleavage value, and The arginine threshold is determined by setting the aforementioned arginine cleavage value as the highest arginine value, gradually decreasing the arginine value, calculating the corresponding p-value for each arginine value, continuing this process until the p-value for the nth arginine value becomes greater than 0.05, and then setting the (n-1)th arginine value as the arginine threshold. Includes, A method for determining the arginine threshold in vitro, wherein if the arginine concentration of the subject is above the arginine threshold, the cancer subject responds well to the arginine deficiency therapy.

2. The method according to claim 1, wherein the result of the maximum selection log-rank statistic, further including a statistical graph, is generated by the concentration of arginine plotted on the X-axis and the standardized log-rank statistic on the Y-axis.

3. The aforementioned arginine cleavage value is (1) A step of identifying the highest value of the standardized log-rank statistic on the Y-axis of the statistical graph, (2) The steps of finding the highest point on the Y axis and finding the corresponding arginine concentration on the X axis, (3) The step of setting the arginine concentration corresponding to the highest point as the arginine cutoff value. The method according to claim 2, as determined by [the specified method].

4. The method according to claim 1, wherein the arginine threshold is the result of analysis using the statistic of the log-rank test.

5. The method according to claim 1, wherein the arginine threshold is 5% to 10% smaller than the arginine cleavage value.

6. The method according to claim 1, wherein the cancer is selected from the group consisting of breast cancer, brain tumor, colorectal cancer, head and neck squamous cell carcinoma, hepatocellular carcinoma (HCC), leukemia, acute myeloid leukemia (AML), lymphoma, lung cancer, melanoma, mesothelioma, malignant pleural mesothelioma (MPM), neuroblastoma, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, sarcoma, and combinations thereof.

7. The method according to claim 6, wherein the cancer is hepatocellular carcinoma (HCC) and the arginine cleavage value is 84.2 μmol / L.

8. The method according to claim 6, wherein the cancer is hepatocellular carcinoma (HCC) and the arginine threshold is 79 μmol / L.

9. The method according to claim 1, wherein the sample is plasma.

10. The method according to claim 1, wherein the arginine deficiency agent is selected from the group consisting of recombinant arginine deiminase (rADI), recombinant arginase (rArg), recombinant arginine decarboxylase (rADC), the PEG form of the rADI, the rArg, the rADC, difluoromethylornithine (DFMO), and combinations thereof.

11. The method according to claim 1, wherein the arginine deficiency agent further comprises an anticancer agent.

12. The method according to claim 11, wherein the anticancer agent is selected from the group consisting of FOLFOX, docetaxel, cisplatin, pemetrexed, pembrolizumab, and combinations thereof.

13. The method according to claim 12, wherein the arginine deficiency agent is the PEG form of recombinant arginine deiminase (rADI), the anticancer agent is cisplatin, and the arginine cleavage value is 122 μmol / L.

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