TW201408321A - Synergistic combination for treating cancer - Google Patents

Abstract

The present invention relates to a new approach for treating cancer by using a synergistic combination of an arginine degradation enzyme and an inhibitor of autophagy to deprive cancer cells of arginine. A pharmaceutical composition for treating cancers comprising the synergistic combination and a method for treating cancers comprising the synergistic combination are provided.

Description

Synergistic combination for treating cancer [related application]

The present application claims the benefit of U.S. Provisional Application No. 61/660,966, the entire disclosure of which is incorporated herein by reference.

The present invention relates to the treatment of cancer, in particular by the use of a combination of a arginine degrading enzyme and an autophagy inhibitor.

High amounts of arginase have been found in rat liver extracts and exhibit anti-tumor properties in in vitro assays (Burton et al., 1967, Can J. Biochem. 45, 289-297). In subsequent experiments, the anti-tumor properties of the enzyme were due to depletion of arginine and cell death in cancer cells that could not be repaired in low arginine-based medium (Storr and Burton, 1974, Br) .J. Cancer 30, 50-59). Arginine is an amino acid that cannot be substituted. In the absence of arginine, the synthesis of protein can cause disorder, and the repeated depletion of arginine can kill the tumor without causing tumor resistance. (Lamb et al., 2000, Experimental Cell Research 225, 238-249). The anti-tumor properties exhibited by arginine depletion in vitro have been confirmed in many recent studies (Scott et al., 2000, Br. J. Cancer 83, 800-810; Wheatley et al., 2000, Cellular Physiol. Biochem. 10). , 37-55).

There are three arginine degrading enzymes that can cause arginine depletion: arginase, arginine deiminase (ADI), and arginine decarboxylase. U.S. Patent No. 7,951,366 (Cheng et al., issued Dec. 23, 2003) discloses a human recombinant arginase and a pharmaceutical composition thereof having a sufficiently high amount of enzyme activity and stability Sexuality to maintain "sufficient arginine depletion" in patients. ADI-PEG20 is a polydiethanolated arginine deiminase used to reduce immunogenicity and improve bioavailability. It has been reported that ADI-PEG20 can cause arginine depletion, thus causing tumor cell death ( Feun and Savaraj, 2006, Expert Opin Investig Drugs 15, 815-22). However, metabolic targeting locked by ADI-PEG 20 typically results in stabilization of the tumor rather than a reduction in the absolute volume of the tumor (M.P. Kelly et al., 2012, Brit. J. Cancer 106, 324-332). U.S. Patent No. 7,323,167 (Clark, first published on June 16, 2005) discloses arginine deiminase modified with polyethylene glycol (PEG) and modified arginine A method of treating cancer by aminase. US Patent Application Publication No. 20030017146 (Tepic, published Jan. 23, 2003) discloses a medical composition for treating cancer comprising a arginolytic enzyme, in particular a biosynthesis of Escherichia coli The arginine decarboxylase and its modifications can be polydiethanolated.

In particular, U.S. Patent No. 6,261,557 (Tepic et al., issued July 17, 2001) discloses a medical composition and a method of treating cancer, wherein a arginolytic enzyme and a protein degradation inhibitor (such as insulin) are disclosed. Combined use to prevent the body from replenishing the depleted arginine from the body muscles because of the natural balance mechanism in the body. Although insulin can act as a protein degradation inhibitor, it also has profound physiological effects on the human body. If the patient's blood glucose concentration is not strictly maintained within the narrow normal range, it may have a fatal effect.

Therefore, there is still a need to develop a new method for treating cancer through arginine depletion to improve anticancer efficacy and safety.

The present invention relates to a novel method of treating cancer by using a synergistic combination of a arginine degrading enzyme and an autophagy inhibitor.

In one aspect, the invention provides a combination for treating cancer comprising a arginine degrading enzyme and an autophagy inhibitor to deplete arginine of a cancer cell, providing inhibition of cancer cell growth and/or even causing cancer cell death. Synergistic effect.

In some embodiments of the invention, the combination may be in the form of a pharmaceutical composition comprising a arginine degrading enzyme and an autophagy inhibitor; the combination may also be in the form of two pharmaceutical compositions, each comprising a fine Amino acid degrading enzymes and autophagy inhibitors, which are administered simultaneously or sequentially.

In another aspect, the invention provides a method of treating cancer comprising administering to a subject in need thereof a sufficient amount of a arginine degrading enzyme to maintain arginine in the serum of the individual at an extremely low level, and at the same time Or sequentially administering an autophagy inhibitor to inhibit release of arginine by means of autophagy, thereby maintaining the cancer cell in a starved environment by inhibiting the growth of the cancer cell and/or causing the cancer cell to die. To provide synergistic effects in the treatment of cancer. In a specific embodiment of the invention, the autophagy inhibitor is an agent that inhibits intracellular arginine cycling by means of autophagy.

In particular, the present invention provides a method of treating cancer by using a arginine degrading enzyme (for example, arginine) and chloroquine or a pharmaceutically acceptable salt thereof (for example, chloroquine diphosphate) , a combination of chloroquine diphosphate) to treat cancer.

The present invention also provides a combination of a arginine degrading enzyme (for example, arginine) and chloroquine or a pharmaceutically acceptable salt thereof (for example, chloroquine diphosphate) for treating cancer. .

The invention will now be described more specifically with reference to the following description and specific embodiments. It should be noted that the following description of the preferred embodiments of the present invention is intended to be illustrative and not restrictive.

For the purpose of illustrating the invention, embodiments are illustrated in the present preferred embodiments. However, it should be understood that the invention is not limited to the preferred embodiments presented.

In the drawings: Figure 1 shows the results of inhibition rates of different cells measured in Example 3, (A) and (B) are Hep 3B cells, (C) is PC-3 cells, and (D) is U2OS. Cells, and (E) are COLO205 cells (A & A+Q, t<0.01 * t<0.001** t<0.0001 ***; Q & A+Q, t<0.01# t<0.001## t<0.0001 ###).

Figure 2 shows the results of the animal test in Example 4.

All technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs, unless otherwise defined.

As used herein, the singular forms "a", "the" Thus, for example, reference to "a sample" includes plural forms of the sample and equivalents thereof as known to those skilled in the art.

In recent years, a arginine depleted drug called arginine deiminase (ADI) The substance has progressed to the clinical trial stage. The Phase I/II report of the clinical trial indicated that the drug has moderate activity against liver cancer and melanoma and has been confirmed to cause depletion of arginine in serum, resulting in rapid death of cancer cells. Cancers treated with arginine depletion are dependent on exogenous arginine for growth, which is known as arginine auxotrophy. However, in a starvation environment, cancer cells are protected by autophagy to counteract the consequences of arginine depletion. To some extent, it may not be able to deplete serum by enzymes in cancer patients. Amino acid related. Accordingly, the present invention provides a novel cancer treatment method for treating cancer by utilizing a combination of serum arginine depletion and autophagy inhibition.

The present invention provides a combination for treating cancer comprising a arginine degrading enzyme and an autophagy inhibitor for depleting serotonin cancer cells to provide for inhibiting cancer cell growth and/or even causing cancer cell death. Synergistic effect.

According to the present invention, the combination may be in the form of a pharmaceutical composition comprising a arginine degrading enzyme and an autophagy inhibitor; or the composition may be in the form of two pharmaceutical compositions comprising arginine degrading enzyme and autophagy, respectively Inhibitors of action, which are administered simultaneously or sequentially.

Furthermore, the present invention provides a method of treating cancer comprising administering to a subject in need thereof a arginine degrading enzyme in an amount sufficient to maintain arginine in the serum of the individual at an extremely low level, and simultaneously or sequentially Inhibiting the administration of an autophagy inhibitor in an amount sufficient to inhibit release of arginine by means of autophagy, thereby maintaining the cancer cell in a starved environment by inhibiting the growth of the cancer cell and/or causing the cancer cell to die, To provide synergistic effects in the treatment of cancer. The normal concentration of human serum arginine is 50 to 150 μM. A sufficient reduction in serum arginine content to maintain cancer cells in a starvation environment is expected to provide synergistic efficacy in cancer treatment. The reduction in serum arginine content used herein means reducing the amount or concentration of serum arginine to below normal The amount, for example, is about 70% or less of the normal amount, about 60% or less, about 50% or less, about 40% or less, about 30% or less.

As used herein, the term "arginine degrading enzyme" refers to an enzyme used to degrade or decompose arginine, including but not limited to arginase, arginine deiminase, and arginine decarboxylase, which may An enzyme that is naturally occurring or recombined, derived from humans, animals, plants, fungi, or bacteria (eg, Bacillus caldovelox ). The arginine degrading enzyme may optimally be PEGylated to achieve the desired biological and pharmacological activity.

As used herein, the term "arginase" refers to a manganese metalloenzyme containing a metal-activated hydroxide ion, which is a key nucleophile in metalloenzymes. Catalytic hydrolysis or hydration reaction.

In a particular embodiment of the invention, the arginine degrading enzyme may be a arginine deiminase, which may optimally be PEGylated to achieve the desired biological and pharmacological activity. For example, the arginine degrading enzyme can be a modified arginine deiminase (ADI-PEG20), a polyethylene glycol (PEG) modified arginine deiminase, such as U.S. Patent No. 7,323,167. The disclosure is hereby incorporated by reference. The arginine degrading enzyme may also be a arginine degrading enzyme biosynthesized by E. coli , as disclosed in U.S. Patent Application Publication No. 20030017146, which is incorporated herein by reference. In another aspect, the arginine degrading enzyme can be a human recombinant arginase, as disclosed in U.S. Patent No. 7,951,366, the disclosure of which is incorporated herein by reference.

In a specific embodiment, the PEG used to carry out the polydiethanolation according to the invention has a range from 200 to 20,000 daltons, in particular from 400 to 10,000 daltons, more specifically from 800 to Molecular weight of 8,000 Daltons. Examples of PEG include, but are not limited to PEG 200, PEG 300, PEG 400, PEG 600, PEG 1,000, PEG 2,000, PEG 3,000, PEG 4,000, PEG 5,000, PEG 6,000, PEG 8,000 or PEG 20,000.

As used herein, the term "autophagy" or "autophagy" refers to a process of catabolism that involves the degradation of components contained in cells by lysosomal means. Autophagy is a major mechanism by which starved cells redistribute nutrients from unnecessary procedures into more desirable programs.

As used herein, the term "autophagy inhibitor" refers to an agent for inhibiting the recycling of cellular nutrients caused by autophagy. In particular, the recycling of cell nutrients is caused by cancer cells. The exhaustion of the share. In one embodiment of the invention, the autophagy inhibitor is an agent for inhibiting cellular arginine recycling by autophagy. The autophagy inhibitor can be, but is not limited to, hydroxychloroquine, chloroquine diphosphate, lucanthone, bafilomycin A1, and succinyl- 1 (Spautin-1), KU-55933, Go 6976, 3-Methyladenine, N-Acetyl-L-cysteine, L- L-Asparagine, E-64d, Wortmannin, Thapsigargin, LY-294, 002 hydrogen chloride (LY-294, 002 hydrochloride), or DBeQ.

As used herein, the term "cancer" refers to a group of diseases in which a group of cells exhibit uncontrolled growth, invasion (invasion and destruction of adjacent tissues), and sometimes metastasis (through lymph or blood to other locations in the body). According to the present invention, cancers to be treated include, but are not limited to, breast cancer, lung cancer, prostate cancer, ovarian cancer, liver cancer (for example, hepatocellular carcinoma), colon cancer, kidney cancer, rectal cancer, skin cancer (for example, melanoma). ), osteosarcoma and mesothelioma.

As used herein, an individual in need of treatment in accordance with the present invention includes humans and non-human mammals. Non-human mammals include, but are not limited to, pet animals such as cats, dogs, and the like, as well as livestock animals such as cattle, horses, sheep, goats, pigs, and the like.

An "effective amount" or similar vocabulary means an amount of the active agent or composition sufficient to achieve the desired therapeutic, prophylactic, and/or biological effect in one body, for example, reducing the amount of the arginine serum, provided Synergistic effect on inhibiting cancer cell growth or causing cancer cell death. The exact effective amount for a particular application may vary, for example, depending on the type or dosage of the agent or composition, and the weight, age, and health of the individual to be treated. The effective amount to be used in a human subject can be determined based on animal models and clinical trials.

In one aspect, the invention provides a method of treating cancer by treating a cancer by using a combination of a arginine degrading enzyme and chloroquine or a pharmaceutically acceptable salt thereof.

As used herein, the term "pharmaceutically acceptable salts" refers to a salt that is not deleterious to the individual to be treated, and which maintains the biological effectiveness and properties of the active compound. Salts may also be derived from the following pharmacologically or physiologically acceptable inorganic and organic acids: hydrochloric acid, cyanuric acid, sulfuric acid, nitric acid, fumaric acid, phosphoric acid, diphosphoric acid, succinic acid, tartaric acid, acetic acid, citric acid, A Sulfonic acid, formic acid, benzoic acid, and malonic acid, but are not limited thereto. The salts may also be derived from the following pharmacologically or physiologically acceptable inorganic and organic bases: alkali metal (e.g., sodium), alkaline earth metal (e.g., magnesium), and ammonium salts, but are not limited thereto. Examples of pharmaceutically acceptable salts of chloroquine include chloroquine phosphate, chloroquine diphosphate or chloroquine sulphate.

In addition, combinations of the invention are also suggested for use in conjunction with chemotherapeutic drugs or target agents for treating cancer to produce synergistic effects.

In the present invention, any chemotherapeutic drug or a target agent for treating cancer can be used. Chemotherapeutic drugs include, but are not limited to, small molecule drugs such as 13-cis-Retinoic Acid, 2-Chlorodeoxyadenosine, 5-Azacitidine (5- Azacitidine), 5-fluorourouracil, 6-Mercaptopurine, 6-Thioguanine, Actinomycin-D, doxorubicin (Azacitidine) Adriamycin), Amethopterin, cytarabine (Ara-C), bleomycin, camptothecin, cisplatin, daunorubicin, etoposide ( Etoposide), Gefitinib, Gemcitabine, Hydroxyurea, Methotrexate, Paclitaxel, Sorafenib, Tamoxifen , Taxol, Temozolomide, Thalidomide, Vinblastine or VP-16; monoclonal antibodies or proteins, for example, Alemtuzumab, Rituximab, trastuzumab, Nimotuzumab, Cetuximab Monoclonal antibody (cetuximab), Bevacizumab antibody, granulocyte mononuclear globulin-stimulated growth factor (GM-CSF), interferon alpha (IFN-alpha), interleukin-2 (IL-2), Interleukin-11 (IL-11); or DNA demethylase inhibitor (DNA demethylase inhibitor).

The pharmaceutical composition comprises a therapeutically effective amount of the synergistic combination depending on the mode of administration and the condition of the individual to be treated, including age, weight, symptoms, therapeutic effect, route of administration, and time of treatment.

The pharmaceutical compositions of this invention may be administered by any suitable route including, but not limited to, parenteral or oral administration. For example, the pharmaceutical composition can be administered intravenously, intraperitoneally , intramuscular administration, intraventricular administration, nasal administration, in vitro administration, or oral administration. The pharmaceutical compositions for parenteral administration can be prepared in the form of solutions, suspensions, emulsions, and solid injectable compositions which are dissolved or suspended in a solvent immediately before use. The injection can be prepared by dissolving, suspending, or emulsifying one or more active ingredients in a diluent. Examples of such diluents are distilled water for injection, physiological saline, vegetable oil, alcohol, and combinations thereof. Further, the injection may contain an adjuvant, a stabilizer, a solubilizer, a suspending agent, an emulsifier, a soothing agent, a buffering agent, a preservative, and the like. The injection is sterilized in the final formulation step or is prepared in a sterile process.

The pharmaceutical composition can also be administered orally, wherein the composition can be in solid or liquid form. Solid compositions include lozenges, pills, capsules, dispersible powders, granules, and the like. In another aspect, liquid compositions for oral administration include pharmaceutically acceptable aqueous solutions, suspensions, emulsions, syrups, elixirs, and the like. In the composition, one or more of the active compounds may be dissolved, suspended or emulsified in a conventional diluent (for example, purified water, alcohol or a mixture thereof, etc.).

The present invention will now be described more specifically by reference to the preferred embodiments thereof,

Example 1: Performance and purification of arginase

E. coli containing pDEST17-rArg1 (rat arginase I) plastid was used to produce recombinant arginase (rArg), pArg was collected from pig liver, L-arginase (L-Arg) Collected from bovine liver (A3233 SIGMA).

The rat arginase I gene line was derived from rat liver cDNA by PCR amplification and colonized into the bacterial expression vector pDEST17. The resulting quality system was used to perform the performance of recombinant arginase.

Escherichia coli, Arctic Express (DE3) strain, which contains pDEST17-rArg1 plastids, was grown in Teriffic liquid medium containing ampicillin (100 g/ml). After culturing at 37 ° C for 8 hours, 1 mM isopropyl-β-D-thiogalactoside (IPTG) and 0.1 mM manganese chloride (MnCl ₂ ) were added to the liquid medium, and the culture temperature was lowered to 20 °C. After the Escherichia coli was cultured overnight at 20 ° C, the bacteria were collected by centrifugation. The obtained bacterial mass was centrifuged in a 20 mM Tris/HCl pH 7.5, 1% Triton X-114 solution with repeated ultrasonic shock for 3 times and then centrifuged. After the last centrifugation, the protein pellet was washed several times with PBS solution, dispensed, and stored at -80 °C.

In addition to obtaining arginase from rat liver, arginase was also obtained from pig liver. The frozen pig liver was homogenized in a 20 mM Tris/HCl pH 7.5 solution and centrifuged at 20,000 g for 30 minutes to obtain a clear lysate. The supernatant was taken to a glass beaker and manganese chloride (MnCl ₂ ) was added to a final concentration of 10 mM. After heating the protein solution at 60 ° C for 30 minutes, it was rapidly placed on ice for cooling, followed by centrifugation at 20,000 g for 30 minutes. Ammonium sulfate was added to 70% saturation to precipitate the protein in the supernatant, and the precipitated protein was resuspended in a 20 mM Tris/HCl pH 7.5 solution, followed by concentration of the protein with an Amicon Ultra-15 centrifugal filter device. The crude arginase extract is then dispensed and stored at a temperature of -80 ° C, and the arginase activity is determined by urea measurement as known in the art.

Example 2: Polydiethanolation of recombinant and purified porcine arginase

First, the arginase was polydiethanolated with mPEG-SPA (sigma 85969 PEG5000). Conjugation of mPEG-SPA to recombinant arginase or purified porcine arginase (herein referred to as Arg) in PBS buffer (pH 7.4) with a protein:PEG molecular weight ratio of 1:20 . The sample was stirred at 25 ° C for 3 hours until the PEG molecule reacted sufficiently with rArg (recombinant arginase). Continue to remove the polydiethanol mixture with a centrifugal filter (Amicon Ultra-15) The free form of mPEG-SPA in the complex. This protein concentration was measured by BCA reagent (pierce). The degree of rArg polydiethanolation is determined by SDS-PAGE.

The protein sample was mixed with sample buffer and allowed to stand at 99 ° C for 10 minutes to prepare for 8% SDS-PAGE. Next, the boiled sample is loaded into sample wells in the gel and electrophoresed. When the protein molecular weight marker reaches the bottom of the glass plate, SDS-PAGE electrophoresis is stopped. Next, the colloid was stained with Coomassie Blue for 30 minutes, followed by de-staining in a solution containing 40% methanol and 10% glacial acetic acid.

Example 3: Cell culture and cell proliferation analysis (MTS assay)

Cells (Hep3B cells, COLO205 cells, PC-3 cells, U2OS cells) were MEM, DMEM, F-12 or McCoy's containing 10% (v/v) fetal bovine serum and 100 units/mL penicillin/streptomycin, respectively. 5A medium was maintained.

The cell line was seeded in a flat-bottom 96-well culture dish at a density of 2 x 10 ³ cells per well per 100 μl of medium and cultured for 24 hours. Next, L-Arg-PEG5000 (14 μl, 0-1 U/ml) and chloroquine diphosphate (CQ, 6 μl, 0-10 U/ml) were added to each well. The plate was further incubated for 72 hours. Next, it will contain 24 μl of MTS/PMS solution [(333 μg/ml MTS, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-() A mixture of 4-sulfophenyl)-2H-tetrazolium, internal salt, Promega G111A) and 25 μM PMS (methylthiophene, Sigma P9625) was added to each well, and the plate was incubated for an additional 2 hours. The cell proliferation was determined by measurement of the absorbance at a wavelength of 490 nm. The inhibition rate (%) is calculated by the following equation:

Figure 1 (A) - Figure 1 (E) shows the inhibition of different cells. Thus, the combination of arginine degrading enzyme and autophagy inhibitor exhibited a synergistic effect in inhibiting cancer cell proliferation regardless of whether arginine degrading enzyme alone or autophagy inhibitor was used alone.

Example 4: Animal test

Analysis of arginase activity

The release rate of urea released from L-arginine by arginase was measured using a commercially available arginine active colorimetric assay kit (BioAssay Systems, Hayward, CA). One unit of arginase is defined as the amount of enzyme that converts 1 μmole of L-arginine to ornithine and urea per minute at pH 9.5 and 37 °C.

Mice and xenograft models evaluated for pharmacodynamics (PD): BALB/c nude mice used in this study were from BioLASCO Taiwan Co., Ltd., all mice At the beginning of the trial, the 6-week-old mother was used. The mice live in a plastic cage with a cushion at room temperature and free access to water and food during a 12-hour light/dark cycle. These mice were used for pharmacodynamic (PD) assessment of rArg-peg 5,000 and rArg-peg 5,000 plus chloroquine diphosphate (CQ).

Tumor cells were implanted into BALB/c nude mice on day 0 of the experiment. Tumor cell lines were previously treated in the following steps. COLO205 cells and PC-3 cell lines were collected from tissue culture flasks, trypsinized and resuspended in PBS or PBS containing 50% matrigel (BD) for xenografting. About 1×10 ⁵ COLO205 cells or 1×10 ⁶ PC-3 cells were injected subcutaneously (sc) into the right axilla of each mouse. About two weeks later, the starting solid tumor system was firmly established in mice. This xenograft is then used to perform pharmacodynamic (PD) assessment.

The mice were then randomized into the following four groups: group 1, PBS (control group); group 2, CQ group (35 mg/kg); group 3, Arg-peg 5,000 group (250 U per mouse) And Group 4, Arg-peg 5,000 (250 U/per mouse) plus CQ (35 mg/kg). The combination of arginase or arginase with CQ was administered intraperitoneally (i.p.) to mice in 4 doses every 6 or 7 days. The CQ line was administered 24 hours after Arg-peg 5,000 administration. PBS was used as a control test.

Tumor size was measured with an electronic vernier caliper and calculated using the following equation: tumor volume = (width) ² x length / 2

The percentage increase in tumor volume was calculated using the following formula: (tumor volume per measurement) - (the tumor volume of the first injection of Arg-PEG 5000) / (the tumor volume of the first injection of Arg-PEG 5000)

The inhibition rate (%) is calculated by the following equation:

Figure 2 shows the results of this animal test: the combination of arginine degrading enzymes and autophagy inhibitors provides a synergistic effect in the treatment of cancer.

It is believed that one skilled in the art to which the invention pertains can apply the invention to the broadest scope of the invention. Accordingly, the description and claims of the invention are intended to be

Claims (15)

A combination for the manufacture of a medicament for the treatment of cancer, wherein the combination comprises a arginine degrading enzyme and an autophagy inhibitor to remove arginine from cancer cells, to provide inhibition of cancer cell growth, and/or to cause cancer cell death Synergistic effect. The use of the first aspect of the patent application, wherein the medicament is in the form of a pharmaceutical composition comprising the arginine degrading enzyme, or in the form of two separate pharmaceutical compositions comprising arginine degrading enzyme and The phagocytosis inhibitor is administered simultaneously or sequentially. The use of the first aspect of the invention, wherein the arginine degrading enzyme is selected from the group consisting of arginase, arginine deiminase, arginine decarboxylase, and any combination thereof. The use of the first aspect of the patent application, wherein the arginine degrading enzyme is PEGylated. The use of the first aspect of the invention, wherein the autophagy inhibitor is selected from the group consisting of hydroxychloroquine, chloroquine diphosphate, lucanthone, bafilomycin A1. (Bafilomycin A1), Spautin-1, KU-55933, Go 6976, 3-Methyladenine, N-Acetyl-L-cysteine (N-Acetyl- L-cysteine), L-Asparagine, E-64d, Wortmannin, Thapsigargin, LY-294, 002 Hydrogen Chloride (LY-294, 002 hydrochloride), DBeQ and any A group of combinations. The use of the first aspect of the patent application, wherein the cancer is breast cancer, lung cancer, prostate cancer, ovarian cancer, liver cancer, colon cancer, kidney cancer, rectal cancer, skin cancer, osteosarcoma or mesothelioma. The use of the sixth aspect of the patent application, wherein the liver cancer is hepatocellular carcinoma. The use of the sixth aspect of the patent application, wherein the skin cancer is melanoma. For example, the application of the scope of claim 1 can be combined with a chemotherapy drug or a target drug for cancer. The use of the ninth aspect of the patent application, wherein the chemotherapeutic drug is 13-cis-Retinoic Acid, 2-Chlorodeoxyadenosine, 5-Azacitidine (5-Azacitidine), 5-Fluorouracil, 6-Mercaptopurine, 6-Thioguanine, Actinomycin-D, A Adriamycin, Amethopterin, cytarabine (Ara-C), bleomycin, camptothecin, cisplatin, daunorubicin, relying on Etoposide, Gefitinib, Gemcitabine, Hydroxyurea, Methotrexate, Paclitaxel, Sorafenib, Tamoxifen (Tamoxifen), Taxol, Temozolomide, Thalidomide, Vinblastine or VP-16. The use of claim 9, wherein the target agent is a monoclonal antibody or protein. The use of the scope of claim 11 wherein the target agent is Alemtuzumab, Rituximab, Trastuzumab, Ninotex Antibody (Nimotuzumab), Cetuximab antibody (cetuximab), Bevacizumab antibody, granulocyte mononuclear globulin-stimulated growth factor (GM-CSF), interferon alpha, interleukin-2, or Interleukin-11. A use in combination for the manufacture of a medicament for the treatment of cancer, wherein the combination comprises a arginine degrading enzyme and chloroquine or a pharmaceutically acceptable salt thereof. A combination for the manufacture of a medicament for treating cancer, wherein the combination comprises arginase And chloroquine diphosphate. A pharmaceutical composition for treating cancer comprising a arginase degrading enzyme and chloroquine or a pharmaceutically acceptable salt thereof.