Methods for predicting response to arginine deprivation therapy based on plasma arginine levels in cancer patients
12569488 ยท 2026-03-10
Assignee
Inventors
- Hung-Wen CHEN (Taipei City, TW)
- Shaw Tsen CHEN (Taipei City, TW)
- Hui-Fen LIU (Taipei City, TW)
- Chih-Ling KUO (Taipei City, TW)
- Chiung-Fang SHIU (Taipei City, TW)
Cpc classification
A61K31/519
HUMAN NECESSITIES
A61K31/513
HUMAN NECESSITIES
C12N9/78
CHEMISTRY; METALLURGY
International classification
A61K31/513
HUMAN NECESSITIES
A61K31/519
HUMAN NECESSITIES
A61K31/555
HUMAN NECESSITIES
A61K38/50
HUMAN NECESSITIES
A61K39/395
HUMAN NECESSITIES
A61K45/06
HUMAN NECESSITIES
A61P35/00
HUMAN NECESSITIES
C07K16/28
CHEMISTRY; METALLURGY
Abstract
A method for providing a prediction as to whether a subject with a cancer exhibits a beneficial response to arginine deprivation therapy including determining the plasma level of arginine in the subject, followed by administering to the subject an arginine deprivation therapy alone or in combination with an anti-cancer agent based on the determined plasma level of arginine. 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 a combination thereof.
Claims
1. A method of in vitro identifying the arginine threshold in biological samples of cancer subjects to predict whether the subjects respond to arginine deprivation therapy, comprising: providing a biological sample, the biological sample taken from a subject prior to be administrated an arginine deprivation agent; determining an arginine concentration, measuring the arginine concentration in the biological sample; calculating an overall survival, the overall survival statistics are assessed after the subjects undergo the arginine deprivation therapy; calculating an arginine cut-off point, inputting the arginine concentration and the overall survival of the subjects into the statistical method of maximally selected log-rank statistics to determine the arginine cut-off point; and calculating an arginine threshold, based on the arginine cut-off point to be the highest arginine value and gradually decrease the value of arginine and calculate the corresponding p-value for each arginine value, continuing this process until the p-value for the nth arginine value is higher than 0.05, then the (n1)th arginine value is the arginine threshold, wherein, when the arginine concentration of the subjects is greater than or equal to the arginine threshold, it means that the cancer subject responds well to the arginine deprivation therapy.
2. The method according to claim 1, wherein the results of the maximally selected log-rank statistics further comprising a statistical chart generated with levels of arginine plotted on the X-axis and the standardized log-rank statistics on the Y-axis.
3. The method according to claim 2, wherein the arginine cut-off point is determined through the following steps: identifying the highest value of the standardized log-rank statistics on the Y-axis of the statistical chart; and determining the highest point on the Y-axis and find the corresponding arginine value on the X-axis, wherein the level of arginine corresponding to this highest point is considered the arginine cut-off value.
4. The method according to claim 1, wherein the arginine threshold is analysis by the statistics of the log-rank test.
5. The method according to claim 1, wherein the arginine threshold is less than the arginine cut-off point among 5%10%.
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 a combination thereof.
7. The method according to claim 6, wherein the cancer is the hepatocellular carcinoma (HCC) and the arginine cut-off point is 84.2 mol/L.
8. The method according to claim 6, wherein the cancer is the hepatocellular carcinoma (HCC) and the arginine threshold is 79 mol/L.
9. The method according to claim 1, wherein the biological sample is plasma.
10. The method according to claim 1, wherein the arginine deprivation agent is selected from the group consisting of a recombinant arginine deiminase (rADI), a recombinant arginase (rArg), a recombinant arginine decarboxylase (rADC), a pegylated form of the rADI, the rArg, the rADC, difluoromethylornithine (DFMO) and a combination thereof.
11. The method according to claim 1, wherein the arginine deprivation agent is in further combination with an anti-cancer agent.
12. The method according to claim 11, wherein the anti-cancer agent is selected from the group consisting of FOLFOX, docetaxel, cisplatin, pemetrexed, pembrolizumab, and a combination thereof.
13. The method according to claim 12, wherein the arginine deprivation agent is pegylated form of the recombinant arginine deiminase (rADI) and the anti-cancer agent is cisplatin and the arginine cut-off point value is 122 mol/L.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
(1) The present description will be better understood from the following detailed description read in light of the accompanying drawings, where:
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DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
(14) The detailed description provided below in connection with the appended drawings is intended as a description of the present examples and is not intended to represent the only forms in which the present example may be constructed or utilized. The description sets forth the functions of the example and the sequence of steps for constructing and operating the example. However, the same or equivalent functions and sequences may be accomplished by different examples.
I. DEFINITIONS
(15) As used herein, the term hepatocellular carcinoma (or HCC for short) refers to a malignant tumor of hepatocellular origin. HCC is a type of liver cancer. HCC may undergo hemorrhage and necrosis because of a lack of fibrous stroma. According to the Barcelona Clinic Liver Cancer (BCLC) staging system updated in 2022, HCC can be classified into five stages, including (1) stage 0 (very early stage), which is defined as the presence of a single nodule being equal to or less than 2 cm without vascular invasion or extrahepatic spread; (2) stage A (early stage), which is defined as the presence of one nodule irrespective of size or as a multifocal HCC up to 3 nodules, without vascular invasion or extrahepatic spread; (3) stage B (intermediate stage), which is defined as the presence of multifocal HCC without vascular invasion or extrahepatic spread; (4) stage C (advanced stage), which is defined as patients having portal invention and/or extrahepatic spread; and (5) stage D (end stage or terminal stage), which is defined as patients having major cancer-related symptoms and/or impaired liver function. Based on the classification of the BCLC system, the term advanced hepatocellular carcinoma or advanced HCC refers to a locally advanced HCC or a metastatic HCC (i.e., HCC that has spread from liver to another part of the body). In general, advanced HCC is unresectable (i.e., it has spread to surrounding tissue and cannot be surgically removed), and is not amenable to cure by local modalities of treatment, such as radiotherapy.
(16) The term arginine deprivation therapy refers to compounds or agents that remove the supply of arginine to cancers with disrupted urea cycle, thereby halting the growth of the cancers and induce cell death.
(17) The term FOLFOX refers to a chemotherapy made up of 5-fluorouracil (5-FU), folinic acid (leucovorin) and oxaliplatin. The term FOLFOX as used herein is not intended to be limited to any particular amounts of or dosing regimens for those components. Rather, as used herein, FOLFOX includes all combinations of those components in any amounts and dosing regimens. Based on the does and ways in which the three drugs are given, there are several different FOLFOX regimens known in the art, including FOLFOX-4, FOLFOX-6, modified FOLFOX-6 (mFOLFOX-6), and FOLFOX-7. According to some embodiments of the present disclosure, the FOLFOX is mFOLFOX-6.
(18) As used herein, the term survival, refers to the act or fact of living. The phrase overall survival (OS) refers to a prolongation in life expectancy as compared to naive or untreated individuals or patients.
(19) The term confidence interval (CI) as used herein has its plain meaning known to one of ordinary skill in the art, and refers to a statistical range with a specified probability that a given parameter lies within the range.
(20) The terms administered and administering are used interchangeably herein to refer a mode of delivery, including, without limitation, intravenously, intratumorally, intramuscularly, intraperitoneally, intraarterially, or subcutaneously administering a treatment (e.g., arginine deprivation therapy or anti-cancer agent).
(21) Unless otherwise indicated, the terms treat, treating and treatment contemplate an action that occurs while a patient is suffering from the specified disease or disorder, which reduces the severity of the disease or disorder, or one or more of its symptoms, or retards or slows the progression of the disease or disorder.
(22) The terms cancer and tumor are used alternatively in the present disclosure, and preferably refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth. Cancers in this respect include metastases cancers, and/or drug-resistant cancers. Examples of cancer include, but are not limited to, 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 a combination thereof.
(23) Unless otherwise indicated, the terms patient and subject are used interchangeably in the present disclosure, and refer to an animal including the human species that is treatable by the method of the present invention. The term patient or subject is intended to refer to both the male and female gender unless one gender is specifically indicated.
(24) Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in the respective testing measurements. Also, as used herein, the term about generally means within 10%, 5%, 1%, or 0.5% of a given value or range. Alternatively, the term about means within an acceptable standard error of the mean when considered by one of ordinary skill in the art. Other than in the operating/working examples, or unless otherwise expressly specified, all of the numerical ranges, amounts, values and percentages such as those for quantities of materials, durations of times, temperatures, operating conditions, ratios of amounts, and the likes thereof disclosed herein should be understood as modified in all instances by the term about. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the present disclosure and attached claims are approximations that can vary as desired. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.
(25) The singular forms a, and, and the are used herein to include plural referents unless the context clearly dictates otherwise.
II. DESCRIPTION OF THE INVENTION
(26) (1) Prediction of Cancer Patients' Responses to Arginine Deprivation Therapy
(27) Diversity of cancer treatment responses has long been recognized, largely owing to the underlying heterogeneities in cancer biology, variations in physiological function and distinctions in patients genetic profiles. Therefore, one objective of the present disclosure aims at providing a molecular marker associated with the responses of cancer patients to arginine deprivation therapy. According to the embodiments of the present disclosure, the plasma level of arginine is associated with the therapeutic response of tumors (e.g., HCC) in patients to a pegylated form of ADI (ADI-PEG 20, an arginine deiminase conjugated to polyethylene glycol with 20,000 molecular weight) based therapy.
(28) Accordingly, the first aspect of the present disclosure provides a method of making a prognosis as to whether a subject with a cancer (e.g., HCC) exhibits a beneficial response to arginine deprivation therapy. The method comprises, (a) determining the plasma level of arginine in the subject; and (b) making the prognosis based on the determined plasma level of arginine in step (a), wherein the plasma level of arginine being equal to or greater than 84.2 indicates that the subject exhibits a beneficial response to arginine deprivation therapy.
(29) In step (a), the plasma level of arginine is measured. Suitable assays used to determine the plasma level of arginine include, but are not limited to, spectrophotometric method, capillary electrophoresis (CE), enzyme-linked immunosorbent assay (ELISA), high performance liquid chromatography (HPLC), mass spectrometry (MS), Sakaguchi test, biosensors, and a combination thereof.
(30) Then, in step (b), a skilled artisan or a clinical practitioner may make the prognosis based on the plasma level of arginine. According to some embodiments of the present disclosure, the plasma level of arginine being equal to or greater than 84.2 mol/L (84.2 mol/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, 87.6, 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, 92.3, 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, 96.9, 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 mol/L, or higher) indicates that the subject exhibits a beneficial response to arginine deprivation therapy. i.e., responding to arginine deprivation therapy in a positive manner. According to certain working examples, the beneficial response is associated with overall survival, in which compared to the subject with plasma level of arginine 84.2 mol/L, the subject with plasma level of arginine 84.2 mol/L has a longer overall survival post-treatment.
(31) According to some embodiments of the present disclosure, the arginine deprivation therapy comprises administering to the subject (e.g., HCC patient) an agent selected from the group consisting of DFMO, rADI, rArg, rADC, pegylated rADI, pegylated rArg, pegylated rADC, and a combination thereof. In some exemplary embodiments, the pegylated rADI (ADI-PEG 20) is administered in an amount of about 18 mg/m.sup.2 body surface area weekly, in which the median overall survival of the subjects with plasma level of arginine 84.2 mol/L is about 8.6 months (with a 95% confidence interval (CI) ranging from 7.3 months to 10.5 months), and the median overall survival of the subjects with plasma level of arginine 84.2 mol/L is about 5.7 months (with a 95% CI ranging from 4.9 months to 7.2 months) post-treatment.
(32) Examples of the cancer include, but are not limited to, 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 a combination thereof.
(33) In some embodiments of the present disclosure, the cancer is HCC. According to one specific example, the cancer is advanced HCC.
(34) (2) Prediction of Cancer Patients' Responses to Combined Treatment
(35) According to some embodiments of the present disclosure, the arginine deprivation therapy is combined with one or more additional treatments so as to improve its therapeutic effect. Accordingly, the second aspect of the present disclosure pertains to a method of making a prognosis as to whether a subject with a cancer (e.g., HCC) exhibits a beneficial response to the combined treatment (i.e., arginine deprivation therapy plus additional treatment(s)). The method comprises the steps of, (a) determining the plasma level of arginine in the subject; and (b) making the prognosis based on the determined plasma level of arginine in step (a).
(36) According to some embodiments of the present disclosure, the arginine deprivation agent is combined with FOLFOX treatment. In these embodiments, the plasma level of arginine being equal to or greater than 34 mol/L (34 mol/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, 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 mol/L, or higher) indicates that the subject exhibits a beneficial response to the combined treatment, i.e., responding to the arginine deprivation therapy plus FOLFOX treatment in a positive manner. According to certain working examples, the beneficial response is associated with overall survival, in which compared to the subject with plasma level of arginine <34 mol/L, the subject with plasma level of arginine 34 mol/L has a longer overall survival post-treatment.
(37) In some exemplary embodiments, the pegylated rADI (ADI-PEG 20) is administered in an amount of about 36 mg/m.sup.2 body surface area weekly, and the FOLFOX treatment is administered biweekly, in which the median overall survival of the subjects with plasma level of arginine 34 mol/L is about 9.5 months (with a 95% CI ranging from 7.5 months to 15.1 months), and the median overall survival of the subjects with plasma level of arginine 34 mol/L is about 4.3 months (with a 95% CI ranging from 4.0 months to 4.6 months) post-treatment.
(38) According to some embodiments of the present disclosure, the arginine deprivation therapy is combined with pembrolizumab. In these embodiments, the plasma level of arginine being equal to or greater than 60.2 mol/L (60.2 mol/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 mol/L, or higher) indicates that the subject exhibits a beneficial response to the combined treatment, i.e., responding to the arginine deprivation therapy plus pembrolizumab treatment in a positive manner. According to certain working examples, the beneficial response is associated with overall survival, in which compared to the subject with plasma level of arginine <60.2 mol/L, the subject with plasma level of arginine 60.2 mol/L has a longer overall survival post-treatment.
(39) In some exemplary embodiments, the pegylated rADI (ADI-PEG 20) is administered in an amount of about 36 mg/m.sup.2 body surface area weekly, and pembrolizumab is administered in the amount of about 200 mg every three weeks.
(40) According to certain embodiments of the present disclosure, the arginine deprivation agent is administered in combination with pemetrexed and cisplatin. In these embodiments, the plasma level of arginine being equal to or greater than 68.2 mol/L (68.2 mol/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 mol/L, or higher) indicates that the subject exhibits a beneficial response to the combined treatment, i.e., responding to the arginine deprivation agent plus pemetrexed and cisplatin treatments in a positive manner. According to certain examples, the beneficial response is associated with overall survival, in which compared to the subject with plasma level of arginine <68.2 mol/L, the subject with plasma level of arginine 68.2 mol/L has a longer overall survival post-treatment.
(41) In some exemplary embodiments, the pegylated rADI (ADI-PEG 20) is administered in an amount of about 36 mg/m.sup.2 body surface area weekly, pemetrexed is administered in an amount of about 500 mg/m.sup.2 body surface area every three weeks, and cisplatin is administered in an amount of about 75 mg/m.sup.2 body surface area every three weeks. According to these embodiments, the median overall survival of the subjects with plasma level of arginine 68.2 mol/L is about 12.5 months (with a 95% CI ranging from 9.8 months to 14.2 months), and the median overall survival of the subjects with plasma level of arginine <68.2 mol/L is about 6.5 months (with a 95% CI ranging from 3.8 months to 8.8 months) post-treatment.
(42) According to certain embodiments of the present disclosure, the arginine deprivation agent is administered in combination with docetaxel. In these embodiments, the plasma level of arginine being equal to or greater than 97.5 mol/L (97.5 mol/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 mol/L, or higher) indicates that the subject exhibits a beneficial response to the combined treatment, i.e., responding to the arginine deprivation therapy plus docetaxel treatment in a positive manner. According to certain working examples, the beneficial response is associated with overall survival, in which compared to the subject with plasma level of arginine <97.5 mol/L, the subject with plasma level of arginine 97.5 mol/L has a longer overall survival post-treatment.
(43) In some exemplary embodiments, the pegylated rADI (ADI-PEG 20) is administered in an amount of about 36 mg/m.sup.2 body surface area weekly, and docetaxel is administered in an amount of about 75 mg/m.sup.2 body surface area every three weeks. According to these embodiments, the median overall survival of the subjects with plasma level of arginine 97.5 mol/L is about 40.7 months (with a 95% CI ranging from 7.2 months to 40.7 months), and the median overall survival of the subjects with plasma level of arginine <97.5 mol/L is about 14.6 months (with a 95% CI ranging from 5.7 months to 16.0 months) post-treatment.
(44) According to some embodiments of the present disclosure, the arginine deprivation agent is administered in combination with cisplatin. In these embodiments, the plasma level of arginine being equal to or greater than 122 mol/L (122 mol/L; e.g., 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, 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 mol/L, or higher) indicates that the subject exhibits a beneficial response to the combined treatment, i.e., responding to the arginine deprivation agent plus cisplatin treatment in a positive manner. According to certain examples, the beneficial response is associated with overall survival, in which compared to the subject with plasma level of arginine <122 mol/L, the subject with plasma level of arginine 122 mol/L has a longer overall survival post-treatment.
(45) In some exemplary embodiments, the pegylated rADI (ADI-PEG 20) is administered in an amount of about 36 mg/m.sup.2 body surface area weekly, and cisplatin is administered in an amount of about 30 mg/m.sup.2 body surface area weekly for three weeks (week 1 to week 3) followed by one week (week 4) of rest period in one treatment cycle. According to these embodiments, the median overall survival of the subjects with plasma level of arginine 122 mol/L is about 15.7 months (with a 95% CI ranging from 8.9 months to 28.5 months), and the median overall survival of the subjects with plasma level of arginine <122 mol/L is about 6.4 months (with a 95% CI ranging from 3.4 months to 8.2 months) post-treatment.
(46) As described above, the 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, or a combination thereof. According to some working examples, the cancer is HCC. In one specific example, the cancer is advanced HCC.
(47) (3) Method of Treating Cancers
(48) Another aspect of the present disclosure pertains to a method of treating cancers via arginine deprivation agent alone (i.e., the administration of DFMO, rADI, rArg, rADC, pegylated rADI, pegylated rArg, pegylated rADC, or a combination thereof) for arginine deprivation therapy, or in combination with one or more additional treatments. According to some embodiments of the present disclosure, the method comprises the steps of, (a) determining the plasma level of arginine in the subject; and (b) administering to the subject a suitable treatment based on the determined plasma level of arginine in step (a).
(49) According to some embodiments of the present disclosure, in the case when the plasma level of arginine is equal to or greater than 34 mol/L, then the pegylated rADI (ADI-PEG 20) and FOLFOX treatment are independently administered to the subject. In some exemplary embodiments, the pegylated rADI (ADI-PEG 20) is administered in an amount of about 36 mg/m.sup.2 body surface area weekly, and the FOLFOX treatment is administered biweekly, so as to ameliorate or alleviate the symptoms associated with cancers. As could be appreciated, a skilled artisan or a clinical practitioner may adjust the treatment regimen (including the dosages, schedule and duration of ADI-PEG 20 and FOLFOX treatments) in accordance with practical requirements.
(50) Alternatively, in the case when the plasma level of arginine is less than 34 mol/L, then an alternative anti-cancer treatment is administered to the subject for therapeutic purpose, wherein the alternative anti-cancer treatment is preferably selected from the group consisting of surgery, chemotherapy, targeted therapy, radiotherapy, hormone therapy, immunotherapy, and a combination thereof. A skilled artisan or a clinical practitioner may choose a suitable treatment for the cancer patient in accordance with clinical factors such as age, sex and physical conditions of the patient, and the type and stage of the cancer.
(51) The agents commonly used in chemotherapy include, but are not limited to, doxorubicin, adriamycin, bleomycin, actinomycin, dactinomycin, mutamycin, daunorubicin, epirubicin, idarubicin, mitoxantrone, mitomycin, epipodophyllotoxins, etoposide, teniposide, antimicrotubule agent, vinblastine, vincristine, vindesine, vinorelbine, taxane, paclitaxel (taxol), nitrogen mustard, chlorambucil, cyclophosphamide, estramustine, ifosfamide, mechlorethamine, melphalan, aziridines, thiotepa alkyl sulfonate, busulfan, nitrosoureas, carmustine, lomustine, streptozocin, platinum complex, carboplatin, alkylator, altretamine, dacarbazine, procarbazine, temozolamide, methotrexate, fludarabine, mercaptopurine, thiogaunine, cladribine, pentostatin, capecitabine, cytarabine, floxuridine, fluorouracil, gemcitabine, hydroxyurea, camptothecin, irinotecan, busufane, epothilone, azathioprine, halofuginone, sirolimus, everolimus, mytomycin, and topotecan.
(52) Exemplary agents for targeted therapy include, but are not limited to, trastuzumab or pertuzumab (an antibody specific to tumor antigen HER-2/neu); bevacizumab (an antibody specific to vascular endothelial growth factor (VEGF)); ramucirumab (an antibody specific to VEGF receptor); nivolumab or cemiplimab (an antibody specific to programmed cell death protein 1 (PD-1)); atezolizumab, avelumab or durvalumab (an antibody specific to the ligand of programmed cell death protein 1 (PD-L1). Ipilimumab (an antibody specific to cytotoxic T-lymphocyte-associated protein 4 (CTLA-4)), rituximab (an antibody specific to CD20 on B cells), and etc.
(53) Non-limiting examples of immunomodulatory agents for immunotherapy include, thalidomide, lenolidomide, pomalidomide, anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, interleukin (IL)-2, IL-6, IL-12, interferon-alpha (IFN-), IFN-, IFN-, granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), and cancer vaccine (e.g., human papillomavirus (HPV) vaccine, and hepatitis B vaccine).
(54) According to some embodiments of the present disclosure, in the case when the plasma level of arginine is equal to or greater than 60.2 mol/L, then the pegylated rADI (ADI-PEG 20) and pembrolizumab are independently administered to the subject. In certain exemplary embodiments, the pegylated rADI (ADI-PEG 20) is administered in an amount of about 36 mg/m.sup.2 body surface area weekly, and pembrolizumab is administered in the amount of about 200 mg every three weeks, so as to ameliorate or alleviate the symptoms associated with cancers. As could be appreciated, a skilled artisan or a clinical practitioner may adjust the treatment regimen (including the dosages, schedule and duration of ADI-PEG 20 and pembrolizumab treatments) in accordance with practical requirements.
(55) Alternatively, in the case when the plasma level of arginine is less than 60.2 mol/L, then an alternative anti-cancer treatment is administered to the subject for therapeutic purpose. As described above, the alternative anti-cancer treatment is preferably selected from the group consisting of surgery, chemotherapy, targeted therapy, radiotherapy, hormone therapy, immunotherapy, and a combination thereof. A skilled artisan or a clinical practitioner may choose a suitable treatment for the cancer patient in accordance with clinical factors of the patient.
(56) According to some embodiments of the present disclosure, in the case when the plasma level of arginine is equal to or greater than 68.2 mol/L, then the pegylated rADI (ADI-PEG 20), pemetrexed and cisplatin are independently administered to the subject. In certain exemplary embodiments, the pegylated rADI (ADI-PEG 20) is administered in an amount of about 36 mg/m.sup.2 body surface area weekly, pemetrexed is administered in an amount of about 500 mg/m.sup.2 body surface area every three weeks, and cisplatin is administered in an amount of about 75 mg/m.sup.2 body surface area every three weeks, so as to ameliorate or alleviate the symptoms associated with cancers. A skilled artisan or a clinical practitioner may adjust the treatment regimen (including the dosages, schedule and duration of ADI-PEG 20, pemetrexed and cisplatin treatments) in accordance with practical requirements.
(57) Alternatively, in the case when the plasma level of arginine is less than 68.2 mol/L, then an alternative anti-cancer treatment is administered to the subject for therapeutic purpose. The alternative anti-cancer treatment is preferably selected from the group consisting of surgery, chemotherapy, targeted therapy, radiotherapy, hormone therapy, immunotherapy, and a combination thereof. A skilled artisan or a clinical practitioner may choose a suitable treatment for the cancer patient in accordance with clinical factors of the patient.
(58) According to certain embodiments of the present disclosure, in the case when the plasma level of arginine is equal to or greater than 84.2 mol/L, then the pegylated rADI (ADI-PEG 20) is administered to the subject alone, without combining with any additional treatments. In certain exemplary embodiments, the pegylated rADI (ADI-PEG 20) is administered in an amount of about 18 mg/m.sup.2 body surface area weekly, so as to ameliorate or alleviate the symptoms associated with cancers. A skilled artisan or a clinical practitioner may adjust the treatment regimen (including the dosage, schedule and duration of ADI-PEG 20 treatment) in accordance with practical requirements.
(59) By contrast, in the case when the plasma level of arginine is less than 84.2 mol/L, then an alternative anti-cancer treatment is administered to the subject for therapeutic purpose. The alternative anti-cancer treatment is preferably selected from the group consisting of surgery, chemotherapy, targeted therapy, radiotherapy, hormone therapy, immunotherapy, and a combination thereof. A skilled artisan or a clinical practitioner may choose a suitable treatment for the cancer patient in accordance with clinical factors of the patient.
(60) According to certain embodiments of the present disclosure, in the case when the plasma level of arginine is equal to or greater than 97.5 mol/L, then the pegylated rADI (ADI-PEG 20) and docetaxel are independently administered to the subject. In some exemplary embodiments, the pegylated rADI (ADI-PEG 20) is administered in an amount of about 36 mg/m.sup.2 body surface area weekly, and docetaxel is administered in an amount of about 75 mg/m.sup.2 body surface area every three weeks, so as to ameliorate or alleviate the symptoms associated with cancers. A skilled artisan or a clinical practitioner may adjust the treatment regimen (including the dosages, schedule and duration of ADI-PEG 20 and docetaxel treatments) in accordance with practical requirements.
(61) Alternatively, in the case when the plasma level of arginine is less than 97.5 mol/L, then an alternative anti-cancer treatment is administered to the subject for therapeutic purpose. The alternative anti-cancer treatment is preferably selected from the group consisting of surgery, chemotherapy, targeted therapy, radiotherapy, hormone therapy, immunotherapy, and a combination thereof. A skilled artisan or a clinical practitioner may choose a suitable treatment for the cancer patient in accordance with clinical factors of the patient.
(62) According to certain embodiments of the present disclosure, in the case when the plasma level of arginine is equal to or greater than 122 mol/L, then the pegylated rADI (ADI-PEG 20) and cisplatin are independently administered to the subject. In some exemplary embodiments, the pegylated rADI (ADI-PEG 20) is administered in an amount of about 36 mg/m.sup.2 body surface area weekly, and cisplatin is administered in an amount of about 30 mg/m.sup.2 body surface area weekly for three weeks (week 1 to week 3) followed by one week (week 4) of rest period in one treatment cycle, so as to ameliorate or alleviate the symptoms associated with cancers. A skilled artisan or a clinical practitioner may adjust the treatment regimen (including the dosages, schedule and duration of ADI-PEG 20 and cisplatin treatments) in accordance with practical requirements.
(63) By contrast, in the case when the plasma level of arginine is less than 122 mol/L, then an alternative anti-cancer treatment is administered to the subject for therapeutic purpose. The alternative anti-cancer treatment is preferably selected from the group consisting of surgery, chemotherapy, targeted therapy, radiotherapy, hormone therapy, immunotherapy, and a combination thereof. A skilled artisan or a clinical practitioner may choose a suitable treatment for the cancer patient in accordance with clinical factors of the patient.
(64) The cancers treatable with the present method include, but are not limited to, 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, and sarcoma. According to some exemplary embodiments, the cancer is HCC. In one specific example, the cancer is advanced HCC.
(65) The subject is a mammal, such as a human, a mouse, a rat, a guinea pig, a monkey, a sheep, a goat, a cat, a dog, a horse, or a chimpanzee. Preferably, the subject is a human.
(66) The present invention will now be described more specifically with reference to the following embodiments, which are provided for the purpose of demonstration rather than limitation. While they are typically of those that might be used, other procedures, methodologies, or techniques known to those skilled in the art may alternatively be used.
EXAMPLES
Materials and Methods
(67) Patient Enrollment
(68) The plasma samples from patients receiving arginine deprivation therapy were used in this study. Patients (including cohort-1 to cohort-6) enrolled in the study were diagnosed with advanced stage of HCC and had joined the clinical trial of ADI-PEG 20 in Taiwan. The de-linked samples and clinicopathological parameters were used for post hoc analysis.
(69) The cohort-1, containing 422 patients, received ADI-PEG 20 monotherapy, in which ADI-PEG 20 was intramuscularly administered to each patient at a dose of 18 mg/m.sup.2 weekly, until disease progression or unacceptable adverse event occurred or other withdrawal criteria met.
(70) The cohort-2, containing 31 patients, received ADI-PEG 20 in combination with docetaxel, in which ADI-PEG 20 was intramuscularly administered to each patient at a dose of 36 mg/m.sup.2 weekly, and docetaxel was intravenously administered at a dose of 75 mg/m.sup.2 every three week, until disease progression or unacceptable adverse event occurred or other withdrawal criteria met.
(71) The cohort-3, containing 78 patients, received ADI-PEG 20 in combination with cisplatin, in which ADI-PEG 20 was intramuscularly administered to each patient at a dose of 36 mg/m.sup.2 weekly, and cisplatin was intravenously administered to each patient at a dose of 30 mg/m.sup.2 weekly for three weeks (week 1 to week 3) followed by one week (week 4) of rest period in one treatment cycle, until disease progression or unacceptable adverse event occurred or other withdrawal criteria met.
(72) The cohort-4, containing 39 patients, received ADI-PEG 20 in combination with modified FOLFOX6 regimen (mFOLFOX6 regimen, composed of 85 mg/m.sup.2 oxaliplatin, 400 mg/m.sup.2 bolus of 5-FU, and 400 mg/m.sup.2 leucovorin on the first day, followed by 2,400 mg/m.sup.2 of 5-FU as a continuous infusion in 2 days), in which ADI-PEG 20 was intramuscularly administered to each patient at a dose of 36 mg/m.sup.2 weekly, and mFOLFOX6 regimen was intravenously administered to each patient every two weeks, until disease progression or unacceptable adverse event occurred or other withdrawal criteria met.
(73) The cohort-5, containing 111 patients, received ADI-PEG 20 in combination with pemetrexed and cisplatin, in which ADI-PEG 20 was intramuscularly administered to each patient at a dose of 36 mg/m.sup.2 weekly, pemetrexed was intravenously administered to each patient at a dose of 500 mg/m.sup.2 every three weeks, and cisplatin was intravenously administered to each patient at a dose of 75 mg/m.sup.2 every three weeks, until disease progression or unacceptable adverse event occurred or other withdrawal criteria met.
(74) The cohort-6, containing 27 patients, received ADI-PEG 20 in combination with pembrolizumab, in which ADI-PEG 20 was intramuscularly administered to each patient at a dose of 36 mg/m.sup.2 weekly, and pembrolizumab was intravenously administered to each patient at a dose of 200 mg every three weeks, until disease progression or unacceptable adverse event occurred or other withdrawal criteria met.
(75) Therapeutic Outcomes Evaluation
(76) The duration of overall survival (OS) of patients in cohort-1 and cohort-5 was calculated from the date of patients included for randomization to the date of death, regardless of any causes, or the date of losing follow-up. The duration of OS of patients in cohort-2, cohort-3, cohort-4 and cohort-6 was calculated as the time from the first dose of study treatment until death from any case; in the case when a subject is alive or lost to follow up, they were censored at last date of contact.
(77) Statistical Analysis
(78) The optimal cut-off point of plasma arginine level for survival outcomes was determined using maximally selected log-rank statistics. Kaplan-Meier method was employed to estimate the median OS of patients. A p value <0.05 determined by log-rank test was considered statistically significant. Statistical analysis was conducted using software.
Example 1 Correlation of Plasma Arginine Level with OS in Advanced HCC Patients Receiving Arginine Deprivation Therapy Alone or in Combination with Additional Treatment
(79) To investigate whether the plasma level of arginine was correlated with OS in cancer patients, maximally selected log-rank statistics was performed. The analytic results were respectively depicted in
(80) TABLE-US-00001 TABLE 1 Correlation of plasma arginine level with OS of cancer patients receiving specified treatments Sample cut- Arginine # of # of Median OS.sup.[2] Cohort Treatment Indication Size off.sup.[1] (umo/L) Event Censored (95% CI).sup.[3] P-value.sup.[4] cohort-1 ADI-PEG 20 (18 mg/m.sup.2) HCC 184 84.2 >=84.2 143 41 8.6 (7.3, 10.5) 0.0057 238 <=84.2 202 36 5.7 (4.9. 7.2) cohort-5 ADI-PEG 20 Malignant Pleural 72 68.2 >=68.2 58 14 12.5 (9.8, 14.2) 0.0011 (36 mg/m.sup.2) + Mesothelioma Pemetrexed (MPM) (500 mg/m.sup.2) + Cisplatin 39 <68.2 36 3 6.5 (3.8, 8.8) (75 mg/m.sup.2) .sup.[1]Maximally Selected log-rank, statistics. .sup.[2]Months. .sup.[3]Kaplan-Meier product-limit estimates. .sup.[4]P-value comparing the treatment groups is based on the log-rank test.
(81) TABLE-US-00002 TABLE 2 Correlation of plasma arginine level with OS of cancer patients receiving specified treatments Sample cut- Arginine # of # of Median OS.sup.[2] Protocol No. Treatment Indication Size off.sup.[1] (umo/L) Event Censored (95% CI).sup.[3] P-value.sup.[4] cohort-2 ADI-PEG 20 (36 mg/m.sup.2) + Advanced Solid Tumors with 12 97.5 >=97.5 5 7 40.7 (7.2, 40,7) 0.0117 Docetaxel(75 mg/m.sup.2) Emphasis on Castration Resistant Prostate Cancer (CRPC) and Advanced Non- Small Cell Lung Cancer 19 <97.5 14 5 14.6 (5.7, 16.0) (NSCLC) cohort-3 ADI-PEG 20 (36 mg/m2) + HCC with coexistent biliary 23 122 >=122 16 7 15.7 (8.9. 28.5) 0.0006 Cisplatin (30 mg/m.sup.2) tract carcinoma (BTC) of BTC Only Cutaneous Melanoma 55 <122 47 6.4 (3.4. 8.2) Ovarian Carcinoma Uveal Melanoma cohort-4 ADI-PEG 20 (36 mg/m.sup.2) + Gastro-esophageal Cancer 37 34 >=34 30 7 9.5 ( 7.5, 15.1) 0.0083 FOLFOX Colorectal Cancer Hepatocellular Carcinoma 2 <34 2 0 4.3 (4.0. 4.6) cohort-6 ADI-PEG 20 36 mg/m.sup.2 + Advanced Solid Cancers 10 60.2 >=60.2 2 8 (2.5,) 0.0119 Pembrolizumab 200 mg 17 <60.2 13 4 6.6 (1.8. 7.8) .sup.[1]Maximally Selected log-rank statistics. .sup.[2]Months. .sup.[3]Kaplan-Meier product-limit estimates. .sup.[4]P-value comparing the treatment groups is based on the log-rank test.
Example 2. The Procedure of Determining the Arginine Cut-Off Point (e.g. HCC)
(82) First stepto determine the cut-off value of arginine. HCC patients agree to join the Polaris HCC clinical study, and Polaris collect patients' plasma before first treatment of ADI-PEG 20. Then, pharmacodynamics will be assessed by measurement of peripheral blood levels of arginine and citrulline by LCMS. Total of 633 patients (422 for ADI-PEG 20 group and 211 for placebo group) were enrolled in the study. Then, clinical database is locked once the number of deaths reach the requirement of sample size calculation at the beginning of study design. Further, the statistician calculates each patient's overall survival. The Arginine cut-off point is determined using the Maximally Selected log-rank statistics (statistician inputs each patient's arginine value and OS into the statistical model).
(83) Second stepto validate the efficiency of arginine cut-off point. 422 HCC patient who were treated as ADI-PEG 20 are categorized into two groups: high arginine group and low arginine group. Then, the overall survival is summarized using the Kaplan-Meier product-limit estimates. Point estimates (25th, 50th, and 75th percentiles) along with 95% confidence intervals will be provided by treatment group. Survival estimates will also be shown graphically for each treatment group. Further, treatments will be compared using a stratified log-rank test.
(84) In one embodiment, a method of in vitro identifying the arginine threshold in biological samples of cancer subjects to predict whether the subjects respond to arginine deprivation therapy, comprising: providing a biological sample, the biological sample taken from a subject prior to be administrated an arginine deprivation agent; determining an arginine concentration, measuring the arginine concentration in the biological sample; calculating an overall survival, the overall survival statistics are assessed after the subjects undergo the arginine deprivation therapy: the arginine cut-off point is determined through the following steps: (1) Identifying the highest value of the standardized log-rank statistics on the Y-axis of the statistical chart; (2) Determining the highest point on the Y-axis and find the corresponding arginine value on the X-axis; and (3) The level of arginine corresponding to this highest point is considered the arginine cut-off value. (for example, arginine cut-off point is 84.2 mol/L in cohort-1, and please refer to
(85) According to the results, 422 patients in cohort-1 were treated with ADI-PEG 20 18 mg/m.sup.2, with the estimated arginine cut-off point for overall survival set at 84.2 mol/L. The maximum log-rank statistic recorded was M=2.8792 (
(86) According to prior ADI-PEG 20 monotherapy Phase HCC study, the cut-off value of arginine was 84.2 mol/L based on the maximally selected rank statistics. Patients with arginine levels 84.2 mol/L may survive longer than those with arginine levels <84.2 mol/L.
(87) To enroll more patients, the prognostic variable analysis was used to determine the arginine threshold. Based on the arginine cut-off point of 84.2 mol/L, the highest arginine value is considered, followed by a gradual decrease in arginine values as follows: 83 mol/L, 82 mol/L, 81 mol/L, 80 mol/L, and 79 mol/L. In Table 3, the significant corresponding *P-value of each arginine is less than 0.05. However, when the arginine is 78 mol/L, and for the first time, the *P-value is higher than 0.05. Therefore, the 78 mol/L is defined as the nth arginine. The previous one (n1)th arginine, which is 79 mol/L, serves as the arginine threshold. The survival of higher then the arginine threshold is longer than low arginine, reaching statistical significance. According to the statistical result mostly the arginine threshold is less than the arginine cut-off point among 5%10% (Table 3).
(88) TABLE-US-00003 TABLE 3 The cut-off value of cohort-1 Cut-off Judgment point of of nth Arginine # of # of Median OS arginine (mol/L) Level N event censor (95% CI) P-value value 78 >=78 215 172 43 8.17 (6.9, 9.4) 0.0671 nth <78 207 173 34 6.17 (4.93, 7.63) 79 >=79 211 168 43 8.17 (6.9, 9.53) 0.0414* (n-1)th <79 211 177 34 5.9 (4.9, 7.43) 80 >=80 208 165 43 8.3 (7.13, 9.57) 0.0223* <80 214 180 34 5.7 (4.9, 7.4) 81 >=81 203 161 42 8.3 (7.13, 9.93) 0.0383* <81 219 184 35 5.7 (4.9, 7.4) 82 >=82 199 158 41 8.3 (7.13, 9.93) 0.0342* <82 223 187 36 5.7 (4.93, 7.4) 83 >=83 196 155 41 8.47 (7.3, 10.0) 0.0211* <83 226 190 36 5.7 (4.93, 7.33) 84.2 >=84.2 184 143 41 8.57 (7.33, 10.47) 0.0057* arginine cut-off <84.2 238 202 36 5.7 (4.9, 7.3) point *P-value of less than 0.05 is considered significant.
(89) According to the results, 31 patients in cohort-2 were treated with ADI-PEG 20 36 mg/m.sup.2+Docetaxel 75 mg/m.sup.2, with the estimated arginine cut-off point for overall survival set at 97.5 mol/L. The maximum log-rank statistic recorded was M=3.0599 (
(90) According to the results, 78 patients in cohort-3 were treated with ADI-PEG 20 36 mg/m.sup.2+Cisplatin 30 mg/m.sup.2, with the estimated arginine cut-off point for overall survival set at 122 mo/L. The maximum log-rank statistic recorded was M=3.6943 (
(91) According to the results, 39 patients in cohort-4 were treated with ADI-PEG 20 36 mg/m.sup.2+FOFLOX, with the estimated arginine cut-off point for overall survival set at 34 mo/L. The maximum log-rank statistic recorded was M=1.6007 (
(92) According to the results, 111 patients in cohort-5 were treated with ADI-PEG 20 36 mg/m.sup.2+Pemetrexed 500 mg/m.sup.2+Cisplatin 75 mg/m.sup.2, with the estimated arginine cut-off point for overall survival set at 68.2 mol/L. The maximum log-rank statistic recorded was M=3.043 (
(93) According to the results, 27 patients were treated with ADI-PEG 20 36 mg/m.sup.2+Pembrolizumab 200 mg, with the estimated arginine cut-off point for overall survival set at 60.2 mol/L. The maximum log-rank statistic recorded was M=3.0899 (
(94) Taken together, the plasma level of arginine could predict the outcome (i.e., OS) of arginine deprivation therapy alone or in combination with different anti-cancer treatments, including FOLFOX, docetaxel, cisplatin, pemetrexed and pembrolizumab.
(95) It will be understood that the above description of embodiments is given by way of example only and that various modifications may be made by those with ordinary skill in the art. The above specification, examples and data provide a complete description of the structure and use of exemplary embodiments of the invention. Although various embodiments of the invention have been described above with a certain degree of particularity, or with reference to one or more individual embodiments, those with ordinary skill in the art could make numerous alterations to the disclosed embodiments without departing from the spirit or scope of the present disclosure.