Metabolic Therapy of Cancer

Abstract

The objective of the present invention is to alter the normal metabolic environment of cancer cells by using an artificial diet in which the levels and ratios of lipids and specific amino acids are manipulated. Due to DNA alterations, cancer cells are unable to fully adapt to the new metabolic environment, which reduces their survival and their protection from the immune system. The present invention provides an artificial diet composition that induces surprising antineoplastic activity in an animal model of metastatic cancer. These activities exceed those observed in mice receiving pharmacological treatments used in cancer patients. The present invention demonstrates that the levels of methionine and leucine have a significant impact on the in vivo antineoplastic activity of the diet. In all active diets, methionine levels are below 0.6% and leucine levels are below 10% by total weight of the entire dry composition.

Description

[Technical Field]

[0001] The present invention relates to an artificial diet composition for use in the treatment of cancer, characterized in that it contains controlled amounts of specific amino acids present in free, salt, ester form and / or provided through an amino acid source such as a protein, and the artificial diet composition further contains other ingredients such as vitamins, minerals, choline, in addition to carbohydrates and lipids, and optionally an aqueous and / or pharmaceutical carrier.

[0002] The compositions of the present invention have been shown to be effective in treating cancer in subjects, such as mammals, as confirmed by experimental studies showing increased survival rates in mice treated with the artificial diet compositions. [Background technology]

[0003] Chemotherapy is the standard of care for patients with metastatic disease. Once the disease has spread and surgery and radiation therapy are no longer curative, chemotherapy becomes the primary form of treatment. Chemotherapy can prolong a patient's life and alleviate some disease-related symptoms. However, it usually does not cure the disease. The low effectiveness of existing anticancer drugs is reflected in the low survival rates of patients diagnosed with the most common metastatic cancers. The 5-year relative survival rates for patients with distant metastases are 5% for lung cancer, 31% for prostate cancer, 27% for breast cancer, 14% for colorectal cancer, 25% for melanoma, 12% for renal cancer, 29% for ovarian cancer, 16% for endometrial cancer, 17% for cervical cancer, 5% for bladder cancer, 5% for esophageal cancer, 2% for liver cancer, and 3% for pancreatic cancer (Reference 1). Many patients with metastases do not survive their disease, despite surviving five years after diagnosis.

[0004] Understanding why drug therapies typically fail is crucial for developing better therapies. When cancer cells are treated with specific concentrations of approved anticancer drugs and examined under a microscope, carnage is typically observed. Most cancer cells die in response to treatment. However, these same drugs fail to save the lives of cancer patients. The primary reason is that these drugs have limited selectivity for cancer cells. As a result of this narrow selectivity, patients cannot receive the drug dose required to kill all cancer cells; such doses also kill normal body cells, resulting in fatal outcomes. Instead, they receive a maximum tolerated dose, which is usually insufficient to reach the drug concentration required to eradicate those cancer cells. Surviving cancer cells continue to proliferate uncontrollably, ultimately resulting in a fatal outcome (Reference 2).

[0005] Some cancer cells exhibit drug resistance, leading to the failure of drug therapy (Refs. 3, 4). The most common cause of resistance is the expression of ATP-binding cassette (ABC) efflux transporters, which expel anticancer drugs from cells. These transporters are expressed in normal stem cells under physiological conditions; these cells must remain intact throughout the organism's lifetime, requiring strong defense mechanisms against environmental chemical insults. Recent evidence strongly suggests that cancer arises from normal stem cells (Refs. 5-7). After accumulating sufficient DNA alterations, normal stem cells give rise to cancer stem cells (CSCs) (Refs. 5-7), which continue to express ABC transporters (Refs. 8, 9). CSCs likely expel drugs via these transporters and resist therapy. This suggests that even with the development of more selective anticancer drugs, mechanisms that evolved to protect cells from chemical insults will continue to act as an obstacle to successful cancer treatment (Ref. 3).

[0006] Cancer drug therapies can also fail because most drugs preferentially target rapidly dividing cells. Quiescent and slowly growing cancer cells, such as CSCs, typically resist therapy. Furthermore, some quiescent and slowly growing cancer cells are located in tumor regions with poor vascularization. Because anticancer drugs are delivered to cells via the blood, tumor cells located in these regions are exposed to lower drug concentrations than normal cells (with an adequate blood supply). This factor reduces the already limited selectivity of existing anticancer drugs, contributing to therapy failure.

[0007] Improving outcomes for patients with metastases requires the development of therapies that are highly selective for cancer cells. Furthermore, these therapies should overcome drug resistance mechanisms of these cells. They should also be effective against non-dividing cancer cells and poorly vascularized tumor cells.

[0008] A major limitation of cancer chemotherapy is its low selectivity for cancer cells. With the discovery of CSCs, it has often been assumed that a major limitation of current treatments is their inability to kill CSCs (Ref. 10). Evidence is accumulating that chemotherapy is ineffective at killing CSCs. However, this does not mean that existing drugs can selectively kill the remaining cancer cells. As discussed elsewhere, the challenge with most cancers is not that a small number of cancer cells survive treatment, but rather that only a small number of cancer cells die in response to treatment (Ref. 11). Successful cancer therapy requires the development of therapies with high selectivity for all types of cancer cells.

[0009] The basis for developing selective anti-cancer therapies is similar to that for developing selective anti-infective therapies: the goal is to eliminate infectious agents or cancer cells without causing undue harm to the patient. The approach is to find key, exploitable differences between our cells and the infectious agent, or between our normal cells and cancer cells.

[0010] There are significant differences between normal cells and all types of cancer cells. Unlike normal cells, cancer cells have highly altered DNA. As described elsewhere (Ref. 12), most tumor cells resemble a bomb set off in their nuclei. In most tumor cells, most chromosomes are joined together, and entire chromosomes are gained or lost (Refs. 12, 13). The karyotypes of some cancer cells are strikingly different from those of normal cells; for example, several studies have reported tumor cells with more than 100 chromosomes (http: / / cgap.nci.nih.gov / chromosomes / Mitelman). Within the chromosomes, many tumors harbor thousands of DNA and epigenetic alterations (Refs. 14-16). A recent study using whole-genome sequences from 22,086 cancer samples showed that the mean and median number of genetic mutations (representing genes, which make up less than 2% of the total DNA) was 177 and 61, respectively (Ref. 16). It is actually remarkable that cells with so many DNA alterations can survive.

[0011] Current therapies do not fully exploit these significant differences between cancer and normal cells. New drugs are typically designed to target single DNA defects in malignant cells. For example, cancer cells commonly harbor mutations in genes encoding specific protein kinases. Because these proteins play a critical role in cancer cell proliferation, many of the drugs recently approved for cancer treatment are designed to inhibit specific kinases. However, exploiting these subtle differences between cancer and normal cells typically results in modest improvements in patient survival. Seventy-one recently approved anticancer drugs have been shown to result in a median survival of 2.1 months, translating into an estimated savings of $2.7 million per year of life, compared with an estimated $10,000 per month of treatment (Refs. 17-20). Current trends suggest that successful treatment of a particular cancer may require identifying drugs for each of the cancer's driving mutations. Given the complexity and diversity of the cancer genome, the clinical benefit of this approach may be limited (Refs. 16, 21, 22).

[0012] The key to developing highly selective anticancer therapies likely lies in finding ways to exploit the full set of DNA alterations in cancer cells. This can be achieved by creating a challenging cellular environment that only cells with intact DNA can overcome. Normal cells use their intact DNA to activate genetic and epigenetic programs to adapt to new conditions and survive. However, cancer cells may not be able to survive in a new environment. Activation of these adaptive programs may require the expression of genes that may be lost, mutated, or silenced in cancer cells. Some of these genes may be in chromosomes or chromosomal fragments lost during carcinogenesis. Others may be mutated or nonfunctional. Furthermore, activation of genetic programs may require the alteration of other programs that cancer cells may need to remain unchanged for survival. A challenging cellular environment can be created without drugs. Because surgery and radiation therapy cannot eliminate nonlocalized tumor cells, it is often assumed that drug therapy is the only possible way to successfully treat patients with metastases. By entering the bloodstream, drugs can potentially reach and kill any nonlocalized cancer cells. Cancer cells can be killed by administering cytotoxic agents, but they can also be killed by restricting the things they need to survive. While the results appear similar, targeting cancer cells without drugs may overcome many of their drug resistance mechanisms (e.g., no drugs are exported from cells via ABC transporters). Furthermore, the location of cancer cells in poorly vascularized tumor regions may not impair the effectiveness of restrictive therapy.

[0013] Selective killing of cancer cells through amino acid restriction is one approach taken to combat cancer. The state of the art has made several attempts to solve this problem by providing protein-free artificial diets in which the levels of certain amino acids are eliminated or restricted.

[0014] WO 2017 / 144877 describes a dietary product for use in the treatment of cancer, comprising a plurality of amino acids, including all essential amino acids, but excluding at least two non-essential amino acids selected from the group consisting of glycine, serine, cysteine, tyrosine, and arginine. Suitable combinations of non-essential amino acids not present in the dietary composition include glycine, serine, and cysteine; glycine, serine, and arginine; glycine, serine, and tyrosine; glycine, serine, arginine, and cysteine; glycine, serine, tyrosine, and cysteine; cysteine ​​and arginine; cysteine ​​and tyrosine; cysteine ​​and glycine; cysteine, tyrosine, and arginine; or glycine, serine, arginine, tyrosine, and cysteine. Additionally, the dietary product may further comprise methionine at a level of less than 25 mg / kg of subject body weight / day, or less than 20 mg / kg / day, or less than 18 mg / kg / day, or less than 16 mg / kg / day.

[0015] WO 2017 / 053328 describes a method for treating cancer by suppressing cancer using nutritional therapy by identifying the nutritional weaknesses of cancer cells and providing a subject with a diet that deprives cancer cells of the nutrients necessary for cancer growth and proliferation. The invention also describes that such nutritional therapy can be used to enhance the effectiveness of current cancer treatments. In one embodiment of the invention described in this patent application, the amino acid-containing supplement does not contain cysteine ​​or cystine, thereby reducing the patient's daily intake of the amino acid by 70-100%.

[0016] US2013 / 0330419 refers to a dietary composition for tumor-bearing patients comprising a depleted or reduced amino acid concentration of at least one amino acid selected from the group consisting of arginine, glutamine, methionine, asparagine, phenylalanine, histidine, glycine, tryptophan, leucine, threonine, valine, cystine, isoleucine, lysine, aspartic acid, and tyrosine, with a reduction of at least 50% from normal consumption. In particular, the invention contemplates a dietary composition useful for treating breast cancer comprising a depleted or reduced amino acid concentration of at least one of Arg, Gln, Asn, Phe, and His.

[0017] When the tumor is associated with prostate cancer, the depleted or decreased amino acid concentrations are Gln, Gly, Trp, Arg, Leu, His, and Met. When the tumor is associated with lung cancer, the depleted or decreased amino acid concentrations are His, Gln, Asn, Cys, Leu, Met, and Trp. When the tumor is associated with colorectal cancer, the depleted or decreased amino acid concentrations are Thr, Gly, Met, Cys, Phe, Tyr, Trp, Asn, and Val. When the tumor is associated with head and neck cancer, the depleted or decreased amino acid concentrations are Met, Cys, Tyr, Leu, and Asp.

[0018] EP1572093 discloses a method for preventing various conditions, particularly conditions associated with cancer and cancer treatment, including metastasis, by administering glutamine or a pharmaceutically acceptable salt thereof. In a preferred embodiment, the patent discloses co-administration of glutamine with an effective amount of a carbohydrate, such as a sugar.

[0019] Despite the efforts made in this field, there remains a need in the art to provide alternative artificial dietary compositions that are effective in treating cancer and / or that exhibit enhanced anti-cancer activity relative to pharmacological treatments used in patients with cancer.

[0020] Thus, the present invention addresses the problem of providing an effective anti-cancer artificial diet composition, as well as methods of using the same. Summary of the Invention

[0021] The following disclosure is presented as an illustration of the general principles of the present invention and is in no way meant to limit the inventive concepts contained herein.

[0022] All terms defined herein are to be given their broadest possible interpretation, including any implied meaning.

[0023] It should be noted that, as recited herein, the singular forms "a," "an," and "the" include plural referents unless otherwise indicated. Additionally, the terms "comprises" and "comprising," as used herein, specify that certain features are present in an embodiment, but the phrase should not be interpreted to exclude the presence or addition of additional steps, operations, features, and / or components.

[0024] As used herein, the term "dry composition" includes all ingredients in the artificial diet composition of the present invention except water, e.g., amino acid mixture, protein, carbohydrates, lipids, choline, vitamins and minerals.

[0025] The terms "subject" or "patient" are used interchangeably herein and refer to a vertebrate, preferably a mammal, including, but not limited to, a human.

[0026] As used herein, the terms "treat," "treatment," and the like are used herein, but are not limited to, to mean obtaining a desired pharmacological and / or physiological effect. The effect may be prophylactic, in that it completely or partially prevents a disorder or sign or symptom thereof, and / or may be therapeutic, in that it ameliorates the symptoms of a disease or infection, or partially or completely cures a disorder and / or adverse effects resulting from a disorder.

[0027] As used herein, a given viscosity characteristic is provided by measurement by methods known to those skilled in the art, including the use of various types of viscometers and rheometers.

[0028] The object of the present invention is an artificial dietary composition for use in the treatment and / or prevention of cancer, comprising: Based on the total weight of the dry ingredient composition, 4-40% amino acid mixture, 0-25% fat, 40-95% carbohydrates, 1-5% vitamin and mineral mixture, and 0-1% choline Including, 1. An artificial diet composition for use, characterized in that leucine is present in the composition as part of the mixture of amino acids in an amount of ≦10% by weight, based on the total weight of the dry ingredient composition, and methionine is present in the composition as part of the mixture of amino acids in an amount of ≦0.6% by weight, based on the total weight of the dry ingredient composition.

[0029] A further object of the present invention is an artificial diet composition for the use according to the preceding paragraph, characterized in that leucine is present in an amount of 0.5 to 6% by weight and methionine is present in an amount of 0.1 to 0.6% by weight, based on the total weight of said dry ingredient composition.

[0030] A further object of the invention is an artificial diet composition for use according to any of the preceding paragraphs, characterized in that said mixture of amino acids is a mixture of essential and non-essential amino acids selected from the group consisting of leucine, isoleucine, valine, methionine, lysine, phenylalanine, tryptophan, threonine, histidine, asparagine, alanine, arginine, aspartic acid, cysteine / cystine, glutamic acid, glutamine, proline, glycine, tyrosine, serine and mixtures thereof.

[0031] A further object of the invention is an artificial diet composition for use according to any one of the preceding paragraphs, characterized in that said amino acids are in free form, in salt form, in ester form and / or in the form of peptides, polypeptides or proteins.

[0032] A further object of the invention is an artificial diet composition for use according to any one of the preceding paragraphs, characterized in that the amino acids present in said composition are a combination of amino acids in free form and as proteins.

[0033] A further object of the invention is an artificial diet composition for the use according to the preceding paragraph, characterized in that said protein is casein.

[0034] A further object of the invention is an artificial diet composition for use according to any one of the preceding paragraphs, characterized in that said lipid component is present in an amount of 0 to 14% by weight relative to the total weight of the dry composition.

[0035] A further object of the invention is an artificial feed composition for the use according to the previous paragraph, characterized in that said lipid component is selected from any edible vegetable or animal oil selected from rapeseed oil, sunflower oil, corn oil, soybean oil, linseed oil, rice oil, safflower oil, olive oil, coconut oil, cottonseed oil, fish oil, and mixtures thereof.

[0036] A further object of the invention is an artificial feed composition for use according to the previous paragraph, characterized in that said lipid component or components are selected from the group consisting of olive oil, coconut oil, salmon oil, corn oil, soybean oil, canola oil, rapeseed oil, sunflower oil, linseed oil, rice oil, safflower oil, cottonseed oil, palm oil, castor seed oil, peanut oil, wheat oil, pumpkin seed oil, poppy seed oil, hemp oil, pomegranate seed oil, cod oil, herring oil, whale oil, seal oil, margarine, butter, lard, tallow, and mixtures thereof.

[0037] A further object of the invention is an artificial diet composition for use according to any one of the preceding paragraphs, characterized in that said carbohydrates may be selected from the group consisting of sucrose, cellulose, starch and mixtures thereof.

[0038] A further object of the invention is an artificial diet composition for the use according to any one of the preceding paragraphs, characterized in that it is in a form suitable for oral administration.

[0039] A further object of the present invention is an artificial diet composition for the use described in the previous paragraph, characterized in that it is in solid, semi-solid or liquid form and is selected from dry powders, shakes, liquid concentrates, ready-to-drink beverages, chilled or shelf-stable beverages, soups, pastes, purees, nutritional bars.

[0040] A further object of the present invention is an artificial diet composition for use according to any one of the preceding paragraphs, characterized in that the treatment of cancer includes renal cancer, lung cancer, colon cancer, breast cancer, melanoma, ovarian cancer, prostate cancer, pancreatic cancer, liver cancer, endometrial cancer, cervical cancer, bladder cancer, esophageal cancer, gastric cancer, head and neck cancer, leukemia, lymphoma, non-melanoma skin cancer, sarcoma, central nervous system cancer, testicular cancer, thyroid cancer and cancer of unknown primary site.

[0041] A further object of the invention is an artificial diet composition for use according to any one of the preceding paragraphs, characterized in that said treatment comprises a treatment cycle of 2 to 12 weeks with 4 to 6 daily doses.

[0042] A further object of the invention is an artificial feed composition for use according to any one of the preceding paragraphs, characterized in that it additionally comprises water or an aqueous carrier.

[0043] A further object of the present invention is a pharmaceutical composition for use in the treatment of cancer, characterized in that it comprises an artificial diet composition according to any one of the preceding paragraphs and a pharmaceutically acceptable carrier or vehicle.

[0044] A further object of the present invention is the pharmaceutical composition as defined in the preceding paragraph for use in the treatment of cancer, characterized in that it comprises renal cancer, lung cancer, colon cancer, breast cancer, melanoma, ovarian cancer, prostate cancer, pancreatic cancer, liver cancer, endometrial cancer, cervical cancer, bladder cancer, esophageal cancer, gastric cancer, head and neck cancer, leukemia, lymphoma, non-melanoma skin cancer, sarcoma, central nervous system cancer, testicular cancer, thyroid cancer and cancer of unknown primary site.

[0045] It is a further object of the present invention to provide a method for treating rheumatoid arthritis in patients with rheumatoid arthritis, wherein the treatment is selected from the group consisting of alkylating agents (e.g. cisplatin, carboplatin, oxaliplatin, cyclophosphamide, hydroxyurea, etc.), antimetabolites (e.g. fluorouracil, capecitabine, gemcitabine, methotrexate, cytarabine, etc.), mitotic inhibitors (e.g. paclitaxel, docetaxel, cabacitaxel, vincristine, vinblastine, vinorelbine, vindesine, etc.), topoisomerase inhibitors (e.g. etoposide, teniposide, irinotecan, topodecane, doxorubicin, epirubicin, etc.), hormonal therapy (e.g. antiestrogens such as tamoxifen, aromatase inhibitors such as anastrozole, LHRH agonists such as goserelin, antiandrogen drugs such as abiraterone and flutamide, etc.). and the like), immunotherapy (e.g., anti-PD1 such as nivolumab, anti-PDL1 such as avelumab, anti-CTLA4 such as ipilimumab, cytokines such as interleukin-2 and interferons), targeted therapy (e.g., anti-VEGF agents such as bevacizumab, anti-VEGFR such as sunitinib and sorafenib, anti-EGFR such as cetuximab or erlotinib, anti-HER2 such as trastuzumab, anti-PARP such as olaparib, anti-BRAF such as vemurafenib), and any other anti-cancer agent, as well as mixtures thereof.

[0046] A further object of the invention is a pharmaceutical composition as defined in the previous paragraph, characterized in that said co-administration comprises sequential, combined or simultaneous administration of the active ingredients.

[0047] A further object of the present invention is a pharmaceutical composition as defined in the previous paragraph, characterized in that the treatment comprises administering to a subject in need thereof an effective amount of said pharmaceutical composition during surgery and / or radiotherapy treatment. [Brief explanation of the drawings]

[0048] [Figure 1] Anticancer activity of dietary P2, sunitinib, and anti-PD1 in mice with renal cancer (intraperitoneal model). [Figure 2] Anticancer activity of dietary P10 and capecitabine in colon cancer-bearing mice (intraperitoneal model). [Figure 3] Anticancer activity of dietary P6 and capecitabine in colon cancer-bearing mice (intravenous model). [Figure 4] Anticancer activity of dietary P6, dietary P8, cisplatin, and capecitabine in mice with triple-negative breast cancer (intravenous model). [Figure 5] Cell viability after treatment with dietary M0, M1, M2, M3, 5-FU, cisplatin, doxorubicin, and paclitaxel. DETAILED DESCRIPTION OF THE INVENTION

[0049] Detailed Description of the Invention All cancer cells acquire DNA modifications and develop in a relatively constant metabolic environment. In this normal environment, cancer cells can grow uncontrollably, evade the immune system, and generally resist most types of systemic therapy. The purpose of this invention is to change the normal metabolic environment of cancer cells by using an artificial diet in which the levels and ratios of specific amino acids and lipids are manipulated. Normal cells use their normal DNA to adapt to the new environment and resist therapy. Due to these DNA modifications, cancer cells may not be able to fully adapt to the new metabolic environment and may therefore die.

[0050] Limiting amino acids can lead to cancer cell death. Simply put, protein synthesis is necessary for cell survival. Proteins are constantly degraded and replaced with new proteins, ensuring a constant supply of functional proteins (References 23, 24). Protein synthesis in humans requires adequate levels of the 20 standard amino acids (AAs). Insufficient supply of just one of them for a long enough period compromises protein synthesis and leads to cell death. Many protein-constituting AAs are also required for other cellular processes. All cancer cells, including CSCs, non-dividing cancer cells, or any type of resistant cancer cell, will die if they do not receive adequate levels of any protein-constituting AA.

[0051] AA restriction can result in the selective death of cancer cells. Human cells cannot synthesize nine of the 20 protein-building AAs, called essential AAs (EAAs), which must be obtained from the diet. The remaining AAs, called non-essential AAs (NEAAs), can be synthesized from glucose and several essential and non-essential AAs. NEAA biosynthesis requires various enzymes that catalyze several reactions and pathways. Some genes encoding these enzymes may not be functional in cancer cells; they may be mutated, silenced, or located on missing chromosomes. However, because dietary proteins provide each of the 20 AAs required for protein synthesis, these DNA modifications do not jeopardize the survival of cancer cells. This can be altered by an artificial diet in which the levels of specific NEAAs are temporarily restricted. Cancer cells with defects in the synthesis of a specific AA will not survive the restriction of this AA, while normal cells will. Amino acid manipulation can also be lethal to cancer cells that lack mutations in genes involved in NEAA synthesis. Carcinogenesis is an evolutionary process in which normal cells acquire multiple DNA modifications. However, not all of them provide a survival advantage. Because many DNA modifications are incompatible with cell survival under specific environmental conditions, cells can only acquire those modifications that allow them to survive in their existing environment. It is important to recognize that carcinogenesis occurs in environments in which the levels and ratios of the 20 proteinogenic AA remain relatively constant. This is primarily because virtually all food proteins contain each of the 20 proteinogenic AA, and standard diets typically provide AA in relatively constant ratios. However, this invention proposes modifying the environment in which cancer cells evolved with an artificial diet in which the levels of specific AA are manipulated. This new environment can cause their death, as DNA modifications that provide a survival advantage under certain environmental conditions may be lethal under others. Scott et al. observed that more than 90% of human cancer cells derived from a wide range of tumors and established cell lines died in vitro after arginine deprivation, while normal cells survived (Reference 25).It is unlikely that all susceptible cancer cells had mutations in genes involved in the synthesis of the NEAA arginine. Arginine deprivation likely caused cells to activate various genetic adaptive programs that function in normal cells but not in cancer cells. The accumulation of DNA alterations in cancer cells during carcinogenesis likely inactivated genetic programs necessary for survival and adaptation to the new environment created by arginine deprivation.

[0052] The present authors have surprisingly found that for an amino acid engineered diet to be successful, it is essential to control the intake of specific amino acids, and further to control the amounts of these specific amino acids to be ingested, so that the daily intake of specific amino acid combinations is modified to specific amounts, in relation to the increased survival rate of patients receiving such a diet compared to patients not receiving such a dietary composition.In contrast to known amino acid restricted diets of the prior art, in which groups of amino acids are deprived to specific levels or alternatively completely suppressed, the present authors have found that the antitumor activity of an amino acid engineered diet depends on the interaction of controlled amounts of specific groups of amino acids.

[0053] Furthermore, the activity of amino acid-engineered diets may also depend on the levels of other dietary components, such as lipids. Lipids are involved in multiple cellular processes important for tumor development and disease progression. For example, they are essential for the synthesis of new cancer cell membranes, provide substrates for energy generation, and are the starting point for the biosynthesis of cellular mediators highly implicated in cancer, such as prostaglandin E2 (PGE2). PGE2 promotes angiogenesis, activates cancer stem cell division, blocks type 1 interferon-dependent innate immune responses, induces macrophage reprogramming to the M2 subtype, and promotes cancer cachexia (26, 27).

[0054] Malnutrition is a common secondary diagnosis in cancer patients. Even patients who are dieting can become malnourished due to specific biochemical and metabolic changes associated with cancer. These metabolic changes impair nutritional status and contribute to cancer-associated malnutrition, anorexia, and cachexia. At least 50% of cancer patients are cachectic (Ref. 28). A recent review indicates that cachexia is even more prevalent among patients with advanced cancer (Ref. 29).

[0055] Cachexia, derived from the Greek word meaning "ill-condition," is characterized by anorexia (loss of appetite), weight loss, muscle wasting, and chronic nausea. Other noted effects include altered body composition, altered carbohydrate, protein, and lipid metabolism, and depression. Metabolic changes associated with cancer result in the preferential depletion of lean body mass as a source of calories. Thus, cachexia differs from simple starvation, in which the body metabolizes fat stores and preserves lean body mass.

[0056] Anorexia, loss of appetite, and loss of food intake are present in 50% of newly diagnosed cancer patients. Early satiety, altered taste and smell, food aversions, nausea, and vomiting contribute to anorexia.

[0057] Anorexia-cachexia syndrome is a major cause of morbidity and mortality in cancer patients. Characterized by progressive nutritional changes, weakness, and wasting, anorexia-cachexia syndrome is often long-term, debilitating, and potentially life-threatening. Therefore, amino acid-engineered diets for cancer treatment should not only impede cancer cell proliferation but also create a metabolic environment to avoid or ameliorate anorexia-cachexia syndrome.

[0058] Thus, the present invention relates to an artificial dietary composition for use in the treatment of cancer, characterized in that it contains controlled amounts of specific amino acids, present in free, salt, ester form and / or provided through an amino acid source such as a protein, together with additional ingredients such as a mixture of vitamins and minerals, in addition to carbohydrates and lipids, and optionally a dietary carrier and / or pharmaceutical vehicle.

[0059] Thus, a first embodiment of the present invention is an artificial diet composition comprising approximately 4-40% by weight of amino acids calculated on the total weight of the dry composition, the content of essential amino acids present in the composition being 2-25% by weight.

[0060] The essential amino acids present in the composition include leucine, isoleucine, valine, methionine, lysine, phenylalanine, tryptophan, threonine, histidine, and mixtures thereof. In certain embodiments of the present invention, the essential amino acids leucine and methionine are present in the composition in controlled amounts.

[0061] Non-essential amino acids that may be present in the composition are selected from the group consisting of asparagine, alanine, arginine, aspartic acid, cysteine / cystine, glutamic acid, glutamine, proline, glycine, tyrosine, serine, and mixtures thereof.

[0062] Other amino acids such as betaine, taurine, etc. may also form part of the composition, as well as amino acid metabolites and / or precursors thereof.

[0063] In a preferred embodiment, the composition according to the invention comprises ≦10% by weight of leucine and ≦0.6% by weight of methionine relative to the total weight of the dry composition.

[0064] More preferably, the composition according to the invention comprises leucine in an amount of 0.5 to 6% by weight and methionine in an amount of 0.1 to 0.6% by weight relative to the total weight of the dry composition.

[0065] In an even more preferred embodiment of the present invention, glutamine is present in an amount of 0-10% by weight and cysteine / cystine is present in an amount of 0-0.5% by weight, relative to the total weight of the dry composition, in addition to leucine in an amount of 0.5-6% by weight and methionine in an amount of 0.1-0.6% by weight.

[0066] Preferred amounts of leucine in the composition are 0.5%, 2.5%, 5%, 6% and 10% by weight based on the total weight of the dry composition.

[0067] Preferred amounts of methionine in the composition are 0.17% by weight; 0.5% by weight and 0.6% by weight relative to the total weight of the dry composition.

[0068] Preferred amounts of glutamine in the composition are 1.3%, 5% and 6% by weight based on the total weight of the dry composition.

[0069] Preferred amounts of cysteine / cystine in the composition are 0%, 0.2% and 0.5% by weight based on the total weight of the dry composition.

[0070] Suitably, the amino acids present in the dietary compositions of the present invention may be in the form of amino acids in free form, salts, esters, and / or sources of amino acids such as polypeptides, peptides, proteins, amino acid metabolites and / or precursors.

[0071] The protein present in the composition as a source of amino acids can be selected from the group consisting of casein and any other protein that provides the amino acid levels described in the present invention, such as collagen, albumin, globulin, ovalbumin, zein, fibroin, keratin, gelatin, gluten, whey protein, egg protein, pea protein, hemp protein, soy protein, and plant and animal protein isolates.In a preferred embodiment, the composition of the present invention comprises casein as an amino acid source, alone or in combination with additional amino acids in free form, salts or esters.

[0072] Such amino acids in free form, salts or esters are commercially available from, for example, Applichem, Acros Organics, ProFoods, BulkSupplements.com, Blackburn Distributions, Myprotein, and the like.

[0073] In addition to the amino acids discussed in the previous paragraph, the compositions of the present invention comprise one or more lipid components in an amount of 0-25 wt. % based on the total weight of the dry composition. In a preferred embodiment of the present invention, the one or more lipid components are present in an amount of 0-14 wt. % based on the total weight of the dry composition. A lipid component suitable for the purposes of the compositions of the present invention may be selected from any edible vegetable or animal oil, such as rapeseed oil, sunflower oil, corn oil, soybean oil, linseed oil, rice oil, safflower oil, olive oil, coconut oil, cottonseed oil, fish oil, etc. In a preferred embodiment of the present invention, the lipid component is selected from the group consisting of olive oil, coconut oil, salmon oil, and mixtures thereof.

[0074] Such lipid components are commercially available, for example, from local markets.

[0075] In addition to the amino acids and lipids defined in any of the preceding paragraphs, the compositions of the present invention comprise one or more carbohydrates in an amount of 40 to 95% by weight, based on the total weight of the dry composition. The carbohydrate portion of the composition can be provided by any suitable carbohydrate source, such as starch, dextrin, glycogen, glucose, fructose, sugars, monosaccharides, disaccharides, oligosaccharides, polysaccharides, and mixtures thereof. Preferred carbohydrates according to the present invention can be selected from the group consisting of sucrose, cellulose, and starch, and mixtures thereof.

[0076] Such carbohydrates are commercially available from, for example, local markets, ProFoods, BulkSupplements.com, Blackburn Distributions, Myprotein, and the like.

[0077] The composition of the present invention comprises, in addition to the amino acids, lipids and carbohydrates defined in any of the preceding paragraphs, a mixture of vitamins and minerals in an amount of 1 to 5% by weight relative to the total weight of the dry composition.

[0078] Suitable mixtures of vitamins and minerals for use in the compositions of the present invention include calcium carbonate, monopotassium phosphate, potassium citrate, sodium chloride, potassium sulfate, magnesium oxide, iron citrate, zinc carbonate, manganese carbonate, copper carbonate, potassium iodate, sodium selenate, ammonium paramolybdate-tetrahydrate, sodium metasilicate-nopehydrate, potassium chromium sulfate-dodecahydrate, lithium chloride, boric acid, sodium fluoride, nickel hydroxide, ammonium meta-vanadium hydroxide, ammonium iodate, sodium ... The active ingredient may be selected from the group consisting of Calcium pantothenate, Thiamine hydrochloride, Riboflavin, Pyridoxine hydrochloride, Nicotinic acid, D-Calcium pantothenate, Folic acid, D-Biotin, Cyanocobalamin (Vitamin B12), Retinyl palmitate (Vitamin A) premix (250,000 IU / g), DL-α-tocopherol acetate (250 IU / g), Cholecalciferol (Vitamin D3, 400,000 IU / g), Menaquinone (Vitamin K2) and mixtures thereof.

[0079] The composition further comprises choline in free form, a salt, an ester such as choline chloride or choline bitartrate in an amount of 0-1% by weight, preferably 0.25% by weight, relative to the total weight of the dry composition.

[0080] Such vitamin and mineral mixtures are commercially available from, for example, Fisher Bioreagents, MP biomedical, and the like.

[0081] The dietary composition of the present invention may further contain optional ingredients selected from the group consisting of stabilizers, preservatives, emulsifiers, and flavorings, which are commonly known from "Formulation Engineering of Foods", Wiley-Blackwell, August 2013 and "Handbook of Food Chemistry", Springer, 2015.

[0082] The dietary products of the present invention are suitable for oral administration and therefore present in any form, from liquid to solid, with the corresponding consistency and viscosity to meet swallowing needs, such as clear liquid, soft, or fully solid diets. Suitable forms include powders, shakes, liquid concentrates, ready-to-drink beverages, chilled or shelf-stable beverages, soups, pastes, purees, nutritional bars, etc.

[0083] In a further embodiment of the present invention, the compositions of the present invention may optionally contain, in addition to any of the above-mentioned ingredients, water or an aqueous carrier to form the artificial diet of the present invention.

[0084] Water or any aqueous carrier can be mixed with the components of the composition listed in the previous paragraph, as needed, to form the artificial diet composition of the present invention with a desired consistency. The artificial diet composition of the present invention exists in forms of different viscosities and viscosities depending on the amount of water or aqueous carrier added to the composition to address a subject's health condition that impairs oral intake of the composition, such as dysphagia. The different viscosities and viscosities range from liquid to semi-solid and solid forms, with viscosities ranging from 0.001 to 0.05 Pa·s for thin liquids, 0.051 to 0.35 Pa·s for nectar-like liquids, 0.351 to 1.75 Pa·s for honey-like liquids, and 1.751 Pa·s or greater for spoon-thick liquids.

[0085] Alternatively, the compositions of the present invention do not contain water or an aqueous carrier, such that the compositions are in a solid to semi-solid dry form.

[0086] The solid or semi-solid dry composition can be consumed immediately. Optionally, such a composition can be mixed with water before consumption to reach the desired volume of the final artificial diet in liquid form.

[0087] A further embodiment of the present invention also provides pharmaceutical compositions comprising the dietary composition of the present invention as defined in any of the preceding paragraphs and claims, together with a pharmaceutically acceptable carrier, excipient, or diluent. These pharmaceutical compositions are formulated to be administered to patients orally, enterally, rectally, vaginally, parenterally, intrapulmonary, sublingually, pulmonary, and / or intranasally. Thus, the present invention is preferably directed to pharmaceutical formulations in the form of solid or liquid; such formulations are in the form of tablets, capsules, gel tabs, lozenges, orally dissolving strips, syrups, oral suspensions, emulsions, granules, sprinkles, and pellets. The preparation of any solid or liquid pharmaceutical dosage, including the dietary composition of the present invention, is well known and common practice to those skilled in the art. Pharmaceutical carriers, excipients, and diluents for preparing the pharmaceutical compositions are well known to those skilled in the art; see Remington, The Science and Practice of Pharmacy, 21st Edition; Lippincott Williams & Wilkins, and Handbook of Pharmaceutical Excipients, 5th Edition, Rowe et al.

[0088] The artificial diet composition of the present invention provides all of a subject's daily nutritional needs and is ingested as a food substitute. To meet a subject's daily nutritional needs, the artificial diet composition of the present invention can be ingested once, twice, or multiple times as needed to complete the required daily caloric intake. In a preferred embodiment, the artificial diet is ingested in 4 to 6 daily doses.

[0089] As shown in the experimental section of the description, a daily intake of the artificial diet composition of the present invention provides enhanced anti-cancer effects when compared with treatment with conventional drugs such as sunitinib, anti-PD1, capecitabine, or cisplatin. Therefore, the artificial diet composition of the present invention is suitable for use as the sole active agent in cancer treatment. Alternatively, the artificial diet composition or pharmaceutical composition containing the artificial diet of the present invention is suitable for use in cancer treatment in a co-administration regimen with other anti-cancer drugs, where such treatment regimen includes sequential, combined, or simultaneous administration of the active ingredients. Anticancer drugs contemplated for use in combination with the artificial diet composition or pharmaceutical composition comprising the artificial diet of the present invention include alkylating agents (e.g., cisplatin, carboplatin, oxaliplatin, cyclophosphamide, hydroxyurea, etc.), antimetabolites (e.g., fluorouracil, capecitabine, gemcitabine, methotrexate, cytarabine, etc.), mitotic inhibitors (e.g., paclitaxel, docetaxel, cabacitaxel, vincristine, vinblastine, vinorelbine, vindesine, etc.), topoisomerase inhibitors (e.g., etoposide, teniposide, irinotecan, topodecane, doxorubicin, epirubicin, etc.), hormone therapy (e.g., antiestrogens such as tamoxifen, aromatase inhibitors such as anastrozole, etc.), and the like. inhibitors, LHRH agonists such as goserelin, antiandrogens such as abiraterone and flutamide, corticosteroids such as dexamethasone and prednisone), immunotherapy (e.g. anti-PDL1 such as nivolumab, anti-PDL1 such as avelumab, anti-CTLA4 such as ipilimumab, cytokines such as interleukin-2 and interferons, etc.), targeted therapy (e.g. anti-VEGF agents such as bevacizumab, anti-VEGFR such as sunitinib and sorafenib, anti-EGFR such as cetuximab or erlotinib, anti-HER2 such as trastuzumab, anti-PARP such as olaparib, anti-BRAF such as vemurafenib, etc.), and any other anti-cancer agent, and mixtures thereof.

[0090] The dietary composition of the present invention may be taken once or several times for approximately 2 to 12 weeks, depending on the progression of the disease. After 2 to 12 weeks of treatment with the metabolic diet, the patient returns to a normal diet for a predetermined period, such as 1, 2, or 3 weeks. The daily dosage intended to provide the desired therapeutic effect is one that provides the necessary caloric needs of a human. Note that the term "calorie" is commonly used as an abbreviation for "kilocalories" (Kcal), and a person's caloric needs depend on age, sex, height, weight, and physical activity. The calories (Kcal) provided by the metabolic diet can be estimated by considering that 1 gram of carbohydrate provides 4.1 calories (Kcal), 1 gram of protein (or amino acids) provides 4.1 calories, 1 gram of fat (lipids) provides 8.8 calories, and 1 gram of fiber provides 1.9 calories. For example, the calorie requirement for a person with the following characteristics (male, 50 years old, 175 cm tall, 75 kg, little or no exercise) is approximately 1900 calories (https: / / www.calculator.net / bmr-calculator.html). If this person is treated with diet P9, he or she must consume approximately 500 g of dry composition daily (500 g of this diet provides approximately 1900 calories). This total daily amount can be divided into 4-6 doses per day, for example, 100 g at 8:00, 100 g at 12:00, 100 g at 16:00, 100 g at 20:00, and 100 g at 24:00. Note that during treatment, the patient should only consume these amounts of dry composition (which can be prepared with different amounts of water) and drink only water.

[0091] [Cancer treatment:] The present invention provides artificial dietary and / or pharmaceutical compositions for use in methods of treating cancer.

[0092] The present invention also provides a method of treating cancer in a subject in need thereof, comprising administering to the subject in need thereof an effective amount of a dietary composition and / or pharmaceutical composition of the present invention.

[0093] Treatment of cancer according to the present invention includes any type of cancer, including kidney cancer, lung cancer, colon cancer, breast cancer, melanoma, ovarian cancer, prostate cancer, pancreatic cancer, liver cancer, endometrial cancer, cervical cancer, bladder cancer, esophageal cancer, gastric cancer, head and neck cancer, leukemia, lymphoma, non-melanoma skin cancer, sarcoma, central nervous system cancer, testicular cancer, thyroid cancer, cancer of unknown primary site, and the like.

[0094] Methods for preparing the compositions of the present invention The diets were prepared by mixing all the solid ingredients shown in the table according to the amount of lipid component used in each case at ambient temperature according to well-known methods until they formed a well-blended dry powder in a semi-solid to solid state.

[0095] Mineral mixture (Harlan Laboratories, AIN-93M-MX) comprised 3.5% of the dry diet. 100 g of dry diet contained 1.25% calcium carbonate, 0.875% monopotassium phosphate, 0.098% potassium citrate, 0.259% sodium chloride, 0.163% potassium sulfate, 0.085% magnesium oxide, 0.021% iron citrate, 0.0058% zinc carbonate, 0.0022% manganese carbonate, 0.0011% copper carbonate, 0.000035% potassium iodate, 0.000035% sodium selenate, and 0.000035% potassium iodate. It contained 0.000028% ammonium paramolybdate tetrahydrate, 0.0051% sodium metasilicate nonahydrate, 0.00095% potassium chromium sulfate dodecahydrate, 0.0000595% lithium chloride, 0.000284% boric acid, 0.00022% sodium fluoride, 0.00011% nickel hydroxide carbonate, 0.000021% ammonium metavanadate, and 0.73% sucrose.

[0096] The vitamin mixture (AIN Vitamin Mix 76) constituted 1% of the dry diet. 100 g of dry diet contained the following: thiamine hydrochloride (0.6 mg), riboflavin (0.6 mg), pyridoxine hydrochloride (0.7 mg), nicotinic acid (3 mg), D-calcium pantothenate (1.6 mg), folic acid (0.2 mg), D-biotin (0.02 mg), cyanocobalamin (vitamin B12) (0.001 mg), retinyl palmitate (vitamin A) premix (250,000 IU / g) (1.6 mg), DL-α-tocopheryl acetate (250 IU / g) (20 mg), cholecalciferol (vitamin D3, 400,000 IU / g) (0.25 mg), menaquinone (vitamin K2) (0.005 mg), and sucrose (972.9 mg).

[0097] Casein was obtained from Acros Organics. Amino acids were obtained from different sources, including Applichem, Acros Organics, and Myprotein. Choline (bitartrate) was obtained from Acros Organics, and olive oil, coconut oil, and sucrose were obtained from the local market. Corn starch and cellulose were obtained from Farmusal (a local pharmacy).

[0098] The preferred compositions (Tables 1 and 2) were obtained according to the method described in the previous paragraph.

[0099] [Table 1]

[0100] Table 1: Preferred composition. Typical amounts (g) of amino acids in 100 g and 6 g of casein used (shown in brackets): glutamine + glutamic acid: 21.7 (1.302), leucine: 9 (0.54), methionine: 2.9 (0.174), phenylalanine: 4.8 (0.288), histidine: 2.6 (0.156), lysine: 7.5 (0.45), threonine: 4.1 (0.246), isoleucine: 4.3 (0.258), Valine: 5.3 (0.318), tryptophan: 1.2 (0.072), cysteine / cystine: 0.7 (0.042), arginine: 3.4 (0.204), glycine: 1.7 (0.102), serine: 5.7 (0.342), tyrosine: 5.2 (0.312), alanine: 2.9 (0.174), aspartate + asparagine: 6.9 (0.414), proline: 10.1 (0.606).

[0101] [Table 2]

[0102] Table 2: Preferred compositions The present invention will be described in more detail below with reference to specific examples, but these examples are intended to illustrate the present invention in more detail and are not intended to limit the scope of the present invention. [Example]

[0103] [Experimental conditions] Cell lines and cell culture conditions The A549 cell line (human non-small cell lung carcinoma) was purchased from the European Collection of Authenticated Cell Cultures (ECACC). 786-O (renal carcinoma), MDA-MB-231 (triple-negative breast carcinoma), LLc1 (mouse lung carcinoma), Renca (mouse renal carcinoma), 4T1 (mouse breast carcinoma), CT26WT (mouse colorectal carcinoma), and B16-F10 (mouse melanoma) were obtained from the American Type Culture Collection (ATCC). A64-CLS (submaxillary gland adenoma), AN3Ca (endometrial adenocarcinoma), BT-474 (breast cancer; luminal B (ER+; PR+; Her-2+), Calu-1 (squamous cell lung carcinoma), HNO97 (tongue cancer), MeWo (melanoma; BRAF WT), NIH:OVCAR-3 (ovarian cancer), Sk-Br-3 (breast cancer; HER-2+), Sk-OV-3 (ovarian cancer), T24 (bladder cancer), T-47D (breast cancer; luminal A (ER+; PR+; Her-2-)), and HaCaT cell lines (skin normal) were purchased from Cell Lines Service (CLS). UACC-62 (melanoma; BRAF mut) was obtained from the National Cancer Institute (Rockville, MD). CAPAN-1 (pancreatic cancer), HepG2 (human hepatocellular carcinoma), HT29 (colorectal cancer), and PC3 (human prostate cancer) were generously provided by Dr. Helleday (Karolinska Institute, Sweden). GAMG (glioblastoma) was kindly provided by Dr. Ayala (University of Seville, Spain). A549, A64-CLS, AN3Ca, B16-F10, BT-474, GAMG, HaCaT, HepG2, HNO97, HT29, LLc1, MDA-MB-231, MeWo, Sk-Br-3, Sk-OV-3, and T24 were cultured in Dulbecco's modified Eagle's medium (DMEM) high glucose medium. 4T1, 786-O, Calu-1, CAPAN-1, CT26WT, NIH:OVCAR-3, PC-3, Renca, T-47D, and UACC-62 were cultured in RPMI medium. 1640. All media were supplemented with 100 U / mL penicillin, 100 μg / mL streptomycin, and 10% fetal bovine serum.All cells were kept at 37°C in a humidified atmosphere containing 5% CO Cell culture reagents were purchased from Biowest or Thermo Fisher Scientific.

[0104] [Cell viability analysis] Exponentially growing cells were seeded into 96-well plates and grown for 24 hours. Cells were then exposed to artificial medium or several concentrations of anticancer drugs for 7 days. Cells were then allowed to recover in their corresponding standard medium (drug-free) for 3 days. Cell viability was then estimated using the resazurin assay. This assay is a redox-based colorimetric method based on the ability of viable cells to reduce the blue reagent resazurin to a pink product. The number of viable cells is directly proportional to the amount of end product formed. After the treatment and recovery period, the medium was removed, and 150 μL of resazurin solution (20 μg / mL in medium) was added to each well for 5–7 hours (depending on the cell line). The optical density of each well was measured at 540 nm and 620 nm using a multiwell plate spectrophotometer reader. Results were expressed as the percentage of cell viability relative to untreated cells grown in their standard medium. Data from at least three independent experiments using artificial medium were averaged and presented as the mean ± standard error of the mean (SEM). Data for anticancer drugs were averaged from duplicate wells per experiment (data consistent with what the authors routinely obtain in their laboratory with these drugs).

[0105] [Mice and experimental in vivo conditions] All mice were purchased from Janvier Labs® (France). Male BALB / cAnNRJ mice were used for the renal cancer model (Renca cells, intraperitoneal model). Female BALB / cAnNRJ mice were used for the colon cancer model (CT26WT cells, intraperitoneal and intravenous models) and triple-negative breast cancer model (4T1 cells, intravenous model). Female C57BL / 6J mice were used for the lung cancer model (LLc1 cells, intravenous model) and melanoma model (B16-F10 cells, intravenous model). All mice were aged 12 weeks or older at the start of the experiment. For the 4T1 breast cancer model and the Renca renal cancer model (injected cells number 100,000), treatment began 8 days after injection of the cancer cells. For the Renca cancer model and the LLc1 lung cancer model (injected cells number 150,000), treatment began 7 days after injection. For the CT26WT colon cancer model (intraperitoneal and intravenous) and the B16-F10 melanoma model, treatment began 4 days after injection.

[0106] Mouse cells (passages 5-7) were cultured in 75 cm2 flasks. When the cells were approximately 60-70% confluent, the medium was removed and the cells were washed twice with sterile PBS. The cells were then incubated with trypsin / EDTA solution at 37°C for 2-3 minutes to allow the cells to have a rounded shape but not detach. Next, the trypsin / EDTA solution was aspirated, and the cells were resuspended in 5 mL of sterile PBS. The cell suspension was pipetted up and down to disrupt any cell aggregates, after which 10% FBS-supplemented medium was added. The working cell suspension (5 x 10 cells depending on the cancer model) was then added. 5 ~25×10 6A working cell suspension (1000 cells / mL) was prepared. This suspension was centrifuged at room temperature (250 g for 5 minutes). The medium was removed, and the cells were resuspended in warmed 2.5% FBS-supplemented medium. The working cell suspension was dispensed into 2 mL tubes and kept at 37 °C in a humidified atmosphere containing 5% CO2 until use. A few minutes before injection, the tubes were centrifuged at 300 g for 3 minutes at 4 °C, the medium was removed, and the cells were resuspended in sterile PBS. The cells were counted again. Finally, 0.2 mL of the working cell suspension was loaded into a 1 mL syringe (insulin type with a 29-G × 1 / 2" needle) and injected into the peritoneal cavity or tail vein of a mouse. After inoculating the last mouse, the cells in the last tube were placed in a flask and monitored under a microscope for several weeks to ensure that all mice were inoculated with viable cells.

[0107] One day before the start of treatment, mice were housed in individual cages to avoid cannibalism. Treatment began 4, 7, or 8 days (depending on the model) after cancer cell injection. Most treatments using artificial diets continued for at least 4 weeks. Artificial diet treatment consisted of simply replacing their normal diet with an artificial diet containing specific AA and lipid levels. Sunitinib and capecitabine were administered daily in the diet. Cisplatin and anti-PD1 antibodies were injected intraperitoneally. Animals were monitored daily and weighed regularly (at least three times weekly). If signs of disease progression were evident, mice were euthanized by cervical dislocation; these signs (e.g., excessive weight gain or loss, decreased mobility and curiosity, respiratory distress, and / or visible or palpable tumors larger than 15–20 mm) indicated a low likelihood of survival for an additional 48 hours. Postmortem examinations were performed to confirm the cause of death and observe the extent of lesions. Necropsy confirmed the presence of tumors in all euthanized mice.

[0108] [Food preparation] The diets were prepared by first mixing all of the solid ingredients shown in the table until they formed a well-blended dry powder. Oil (if present in the composition) was added to the mixture, and then sufficient water was added in small increments to create a soft dough. The dough was air-dried for approximately 2 hours, manually pelleted (approximately 5 g / pellet), air-dried for an additional 2 hours, and stored until use.

[0109] Mineral mixture (Harlan Laboratories, AIN-93M-MX) comprised 3.5% of the dry diet. 100 g of dry diet contained 1.25% calcium carbonate, 0.875% monopotassium phosphate, 0.098% potassium citrate, 0.259% sodium chloride, 0.163% potassium sulfate, 0.085% magnesium oxide, 0.021% iron citrate, 0.0058% zinc carbonate, 0.0022% manganese carbonate, 0.0011% copper carbonate, 0.000035% potassium iodate, 0.000035% sodium selenate, and 0.000035% potassium iodate. It contained 0.000028% ammonium paramolybdate tetrahydrate, 0.0051% sodium metasilicate nonahydrate, 0.00095% potassium chromium sulfate dodecahydrate, 0.0000595% lithium chloride, 0.000284% boric acid, 0.00022% sodium fluoride, 0.00011% nickel hydroxide carbonate, 0.000021% ammonium metavanadate, and 0.73% sucrose.

[0110] The vitamin mixture (AIN Vitamin Mixture 76, Fisher Bio-Reagent) constituted 1% of the dry diet; 100 g of dry diet contained thiamine hydrochloride (0.6 mg), riboflavin (0.6 mg), pyridoxine hydrochloride (0.7 mg), nicotinic acid (3 mg), D-calcium pantothenate (1.6 mg), folic acid (0.2 mg), D-biotin (0.02 mg), cyanocobalamin (vitamin B12) (0.001 mg), retinyl palmitate (vitamin A) premix (250,000 IU / g) (1.6 mg), DL-α-tocopherol acetate (250 IU / g) (20 mg), cholecalciferol (vitamin D3, 400,000 IU / g) (0.25 mg), menaquinone (vitamin K2) (0.005 mg), and sucrose (972.9 mg).

[0111] Casein was obtained from Acros organic (27607; bovine). Typical amounts (g) of amino acids in 100 g and 6 g of casein used in the experiments (shown in parentheses) were: glutamine + glutamic acid: 21.7 (1.302), leucine: 9 (0.54), methionine: 2.9 (0.174), phenylalanine: 4.8 (0.288), histidine: 2.6 (0.156), lysine: 7.5 (0.45), threonine: 4.1 (0.246), isoleucine: 4.3 (0.25). 8), valine: 5.3 (0.318), tryptophan: 1.2 (0.072), cysteine / cystine: 0.7 (0.042), arginine: 3.4 (0.204), glycine: 1.7 (0.102), serine: 5.7 (0.342), tyrosine: 5.2 (0.312), alanine: 2.9 (0.174), aspartic acid + asparagine: 6.9 (0.414), proline: 10.1 (0.606). Amino acids were obtained from different sources, including Applichem, Acros Organics, and Myprotein. Choline (bitartrate) was obtained from Acros Organics, and olive oil, coconut oil, and sucrose were obtained from local markets. Salmon oil was obtained from Petspurest. Corn starch and cellulose were obtained from Farmusal (a local pharmacy).

[0112] [Pharmaceuticals] Sunitinib malate (462640010, Acros Organics) was mixed into the food. Mice were fed a standard diet supplemented with sunitinib (350 mg / kg diet) for 28 days. A 25 g mouse typically consumed an average of 4.5 g of food per day, resulting in a dose of approximately 60 mg / kg / day. Regular diet was powdered and mixed with sunitinib. Sufficient water was then added to form a soft dough, which was air-dried for approximately 2 hours, manually pelleted (approximately 5 g / pellet), and stored until use. Capecitabine (500 mg / pill, 707278.2, Normon) was also mixed into the food (following the process described for sunitinib). Mice were fed a standard diet supplemented with capecitabine (2500 mg / kg diet) for 7 days, followed by 7 days of drug-free normal food. Mice underwent 2 to 3 cycles depending on their health status. A 25g mouse typically consumes an average of 4.5g of food per day, resulting in a dose of approximately 450mg / kg / day. Cisplatin (1mg / mL, 659219.9, Cisplatin Pharmacia, Pfizer) was administered intraperitoneally once weekly for 4 weeks. Mice received a dose of 5mg / kg at each dose. Anti-PD-1 (anti-mouse PD-1 (CD279), clone RMP1-14, BE0146, Bioxcell) was administered intraperitoneally every 4 days for a total of 4 doses. Mice received 250µg at each dose. Anti-PD-1 was diluted in pH 7.0 buffer (InVivoPure, IP0070, Bioxcell). In in vitro experiments, we also used the following anticancer drugs: doxorubicin (50 mg powder for solution, 958314.9, Farmiblastina, Pfizer), 5-fluorouracil (F6627, Sigma), and paclitaxel (66997, TEVA, 6 mg / ml).

[0113] [In vivo activity results] Table 3 shows the results from one experiment in mice with renal cancer treated with several diets. In most diets, one amino acid was added or removed from diet P2. Previous amino acid-restricted diets (e.g., WO2017 / 144877) have shown that the activity of a diet depends on the removal of one or several amino acids, such as serine and glycine. This experiment clearly demonstrates that serine removal is not required for activity; indeed, mice treated with a serine-containing diet (P13) live longer than mice treated with the same diet (P2) without serine. Previous data also showed that the removal of both serine and glycine is important for activity; however, the results shown in Table 3 indicate that the removal of both amino acids (diet P14) is worse than a diet containing both serine and glycine (P13) or a diet containing glycine but not serine (P2). In addition, this experiment also shows that increasing the level of lipids can decrease the activity of the composition (diet P21 is worse than diet P2), and decreasing the level of lipids can increase the activity of the diet (diet P20 is better than diet P2). Thus, in contrast to known amino acid-restricted diets of the prior art, in which groups of amino acids are deprived to specific levels or completely suppressed, these results show that the antitumor activity of amino acid-restricted diets depends on the interaction of controlled amounts of specific groups of amino acids, and also on their interaction with other dietary components, such as lipids.

[0114] [Table 3]

[0115] Table 3: Survival of mice bearing renal cancer treated with several metabolic diets. Male BALB / cAnNRJ mice bearing renal cell carcinoma were treated with anti-PD1 antibody (250 μg administered intraperitoneally on days 8, 12, 16, and 20), treated with one of the following diets: P2, P3, P13, P14, P15, P16, P17, P18, P19, P20, and P21 (normal diet was replaced by one of these diets for 28 days), or left untreated (control, normal diet). Treatment began 8 days after intraperitoneal injection of 100,000 Renca cancer cells. At least three mice were included in each group. Mice were euthanized by cervical dislocation if signs of disease progression were evident; these signs (e.g., excessive weight gain or loss, decreased mobility and agility, and / or visible or palpable tumors larger than 15-20 mm) indicated a low likelihood of survival for an additional 48 hours. Postmortem examination confirmed the presence of tumors in all euthanized mice. Anti-PD1 antibody (nivolumab) is the first-line treatment for patients with metastatic renal cancer.

[0116] In all active diets described in this invention, the level of the amino acid methionine is 0.6% or less. The results shown in Table 4 indicate that the amount of methionine must not exceed 0.6% to achieve anti-cancer activity in cancer-bearing mice. Diets P23 and P24, containing 0.67% methionine (0.5% pure methionine + 0.17% methionine contained in 6 g of casein), did not have anti-tumor activity. However, diet P22 (containing only methionine provided by casein, i.e., 0.17%) showed significant anti-tumor activity, even higher than that observed in mice treated with sunitinib (a first-line treatment for patients with metastatic renal cancer). Previous amino acid diets (WO2017 / 053328) indicated that cysteine / cystine levels should be removed or reduced to achieve anti-cancer activity. However, this experiment shows that the cysteine / cystine-rich diet P22 (0.5042%; 0.5 pure cystine + 0.042 of the cysteine ​​in 6 g casein) was highly active, whereas diet P11 containing low amounts of cysteine ​​(only 0.042 in 6 g casein) had low antitumor activity (P22 and P11 differ only in the amount of cysteine / cystine).

[0117] [Table 4]

[0118] Table 4: Survival of mice bearing renal cancer treated with several metabolic diets. Male BALB / cAnNRJ mice bearing renal cell carcinoma were treated with sunitinib (60 mg / kg / day, orally for 28 days), one of the following diets: P22, P23, P24, P11, or P9 (normal diet was replaced by one of these diets for 28 days), or left untreated (control, normal diet). Treatment began 7 days after intraperitoneal injection of 150,000 Renca cancer cells. Mice surviving 28 days of treatment (with diet and sunitinib) were placed on a normal diet for 10 days and then returned to treatment (diet or sunitinib) for an additional 21 days (until day 66 after cancer cell inoculation). At least four mice were included in each group. Mice were euthanized by cervical dislocation if signs of disease progression were evident; these signs (e.g., excessive weight gain or loss, decreased mobility and agility, and / or visible or palpable tumors larger than 15-20 mm) indicated a low likelihood of survival for an additional 48 hours. Postmortem examination confirmed the presence of tumors in all euthanized mice. Sunitinib is a first-line treatment for patients with metastatic renal cancer.

[0119] In all active diets described in this invention, the level of the amino acid leucine is 10% or less. Several independent experiments have shown that controlling the level of leucine is important for increasing antitumor activity in mice with renal cancer. In the experiment whose results are shown in Table 4, diet P11 (containing only leucine provided by casein, i.e., 0.54%) showed moderate antitumor activity (mice survived 2.5 days longer than untreated treated mice), while diet P9 (containing 3.04% leucine) (pure leucine 2.5 + leucine 0.54 contained in 6 g of casein) showed significant antitumor effect (mice survived 23.75 days longer than untreated treated mice; one mouse is still alive and shows no signs of disease). The anticancer activity of this diet (P9) was higher than that observed in mice treated with sunitinib (first-line treatment for metastatic renal cancer patients), i.e., 17.25 days.

[0120] The effect of controlling leucine levels on dietary anticancer activity has also been observed in other cancers in vivo. For example, in mice bearing colon cancer, diet P10 (containing 3.04% leucine; 2.5% pure leucine + 0.54% of the leucine contained in 6 g of casein) was significantly more active than diet P11 (containing only leucine provided by casein, i.e., 0.54%). Briefly, BALB / cAnNRJ mice bearing colon cancer were treated with diet P10 (6 weeks), diet P11 (6 weeks), capecitabine (450 mg / kg / day, 7 days of treatment + 7 days of rest, orally administered) until excessive toxicity or death, or left untreated (control). At least three mice were included in each group. Treatment began 4 days after intraperitoneal injection of 100,000 CT26.WT cancer cells. Mice were euthanized by cervical dislocation if signs of disease progression were evident; these signs (e.g., excessive weight gain or loss, decreased mobility and agility, and / or visible or palpable tumors larger than 15–20 mm) indicated a low likelihood of survival for an additional 48 hours. Postmortem examination confirmed the presence of tumors in all euthanized mice. Mice treated with capecitabine (first-line treatment for patients with metastatic colon cancer) survived 4.5 days longer than untreated mice, mice treated with diet P11 (0.54% leucine) survived 2.3 days longer than untreated mice, and mice treated with diet P10 (3.04% leucine) survived >44.1 days longer than untreated mice (two of the seven mice used in this group survived and showed no signs of disease).

[0121] As mentioned above, cachexia and weight loss are common and significant problems in cancer patients. Because many of the diets described in this invention are low in protein (and / or amino acids) and lipids, one might think that these diets promote cachexia and weight loss. In the experiment shown in Table 4, two mice survived long enough to compare their weights when fed diet P22 (6.5% protein and 1% fat) and a normal diet (21% protein and 7% fat). At the start of diet P22 (day 7 after cancer cell inoculation), the weights of the two mice were 29.2 g and 30.6 g. After 28 days (day 35) on diet P22 (low in protein and lipid), the weights were 28.0 g and 33.4 g, respectively. The mice were returned to a normal diet (rich in protein and lipid), and 10 days later (day 45), their weights had decreased to 24.2 g and 27.0 g. The P22 diet was then resumed, and 10 days later (day 55), the body weights increased to 28.0 g and 30.4 g. The mice died on days 63 and 83 (P22 treatment was terminated on day 66). These results clearly demonstrate that protein and lipid restriction does not necessarily promote cachexia. In fact, many of the diets described in this invention induced anti-cancer effects without significantly affecting the animal's body weight and without inducing any obvious toxic effects.

[0122] Table 5 shows the survival improvement achieved with various metabolic diets in mice with several types of cancer relative to untreated mice (controls), as well as the survival improvement achieved with drugs used in cancer patients.

[0123] [Table 5] JPEG0007727286000006.jpg197169

[0124] Table 5: Anticancer activity of various diets and several anticancer drugs in mice bearing various types of cancer. Diet compositions (P1-P24) are shown in Tables 1 and 2. Experimental conditions are described in the "Mice and Experimental In Vivo Conditions" section. The > symbol indicates that one or several treated mice are still alive. The * symbol indicates that the results are shown in Figures 1-4.

[0125] Anticancer activity of dietary P2, sunitinib, and anti-PD1 in renal cancer-bearing mice. Figure 1 shows the survival of male BALB / cAnNRJ mice bearing renal cell carcinoma treated with sunitinib (60 mg / kg / day, orally administered for 28 days), anti-PD1 antibody (250 μg administered intraperitoneally on days 8, 12, 16, and 20), diet P2 (28 days; normal diet was replaced with P2 diet), or left untreated (control, normal diet). Treatment began 8 days after intraperitoneal injection of 100,000 Renca cancer cells. Mice were euthanized by cervical dislocation if signs of disease progression were evident; these signs (e.g., excessive weight gain or loss, decreased mobility and aggression, and / or visible or palpable tumors larger than 15–20 mm) indicated a low likelihood of survival for an additional 48 hours. Postmortem examination confirmed the presence of tumors in all euthanized mice. Data were averaged from at least two independent experiments, and the mean survival was 35.2 days (n = 16) in the control group, 62.2 days (n = 7) in the sunitinib group, 41.9 days (n = 9) in the anti-PD1 group, and >112.0 days (n = 31) in the dietary P2 group. Some mice treated with dietary P2 survived and showed no signs of disease. Sunitinib and the anti-PD1 antibody (nivolumab) are first-line treatments for patients with metastatic renal cancer.

[0126] Anticancer activity of dietary P10 and capecitabine in colon cancer-bearing mice (intraperitoneal model) Figure 2 shows the survival of female BALB / cAnNRJ mice bearing colon cancer treated with capecitabine (450 mg / kg / day, 7 days of treatment plus 7 days of rest, orally administered until excessive toxicity or death), diet P10 (6 weeks), or left untreated (control). Treatment began 4 days after intraperitoneal injection of 100,000 CT26.WT cancer cells. Mice were euthanized by cervical dislocation if signs of disease progression were evident; these signs (e.g., excessive weight gain or loss, decreased mobility and aggression, and / or visible or palpable tumors larger than 15–20 mm) indicated a low likelihood of survival for an additional 48 hours. Postmortem examination confirmed the presence of tumors in all euthanized mice. Data were averaged from at least two independent experiments, with mean survival of 24.6 days (n=7) in the control group, 27.7 days (n=7) in the capecitabine group, and >68.7 days (n=7) in the diet P10 group. Two mice treated with diet P10 are alive and without any signs of disease. Capecitabine (a prodrug of 5-fluorouracil) is a first-line treatment for patients with metastatic colon cancer.

[0127] Anticancer activity of dietary P6 and capecitabine in colon cancer-bearing mice (intravenous model) Figure 3 shows the survival of female BALB / cAnNRJ mice bearing colon cancer that were treated with capecitabine (450 mg / kg / day, 7 days of treatment + 7 days of rest, orally administered until excessive toxicity or death), diet P6 (treated for 28 days), or left untreated (control). Treatment began 4 days after tail vein injection of 100,000 CT26.WT cancer cells. Mice were euthanized by cervical dislocation if signs of disease progression were evident; these signs (e.g., respiratory distress, excessive weight gain or loss, decreased mobility and aggression, and / or visible or palpable tumors larger than 15-20 mm) indicated a low likelihood of survival for an additional 48 hours. Postmortem examination confirmed the presence of tumors, primarily in the lungs, in all euthanized mice. Median survival was 33.3 days (n=3) in the control group, 36.7 days (n=3) in the capecitabine group, and 78 days (n=3) in the diet P6 group.

[0128] Anticancer activity of dietary P6, dietary P8, cisplatin, and capecitabine in mice bearing triple-negative breast cancer (intravenous model). Figure 4 shows the survival of female BALB / cAnNRJ mice bearing triple-negative breast cancer that were treated with capecitabine (450 mg / kg / day, 7 days of treatment + 7 days of rest, orally administered until excessive toxicity or death), cisplatin (5 mg / kg weekly for 4 weeks, intraperitoneally injected), diet P6 (28 days), diet P8 (28 days), or left untreated (control). Treatment began 8 days after tail vein injection of 100,000 4T1 cancer cells. Mice were euthanized by cervical dislocation if signs of disease progression were evident; these signs (e.g., respiratory distress, excessive weight gain or loss, decreased mobility and aggression, and / or visible or palpable tumors larger than 15–20 mm) indicated a low likelihood of survival for an additional 48 hours. Postmortem examination confirmed the presence of tumors, primarily in the lungs, in all euthanized mice. The median survival was 26.1 days (n = 8) in the control group, 24.2 days (n = 10) in the capecitabine group (two mice died early due to excessive drug toxicity), 33.6 days (n = 5) in the cisplatin group, 43.8 days (n = 6) in the diet P6 group, and 60.1 days (n = 9) in the diet P8 group. One mouse treated with diet P8 is still alive (it developed signs of disease twice, but these disappeared within an additional 4 weeks on diet P8). Capecitabine (a prodrug of 5-fluorouracil) is the first-line treatment for patients with triple-negative breast cancer. Cisplatin is a drug widely used for many types of cancer.

[0129] [In vitro anticancer activity of amino acid-limited compositions] Cell culture media M1 (equivalent to diet P1), M2 (equivalent to diet P2), and M3 (equivalent to diet P3) were prepared and their cytotoxicity and selectivity toward cancer cells were evaluated. Medium M0, containing all amino acids, was also prepared and tested under the same experimental conditions. The medium composition is shown in Table 6. These media were tested on human normal skin cells (HaCaT), which have a high proliferation rate, and 20 types of human cancer cells representing the most common types of cancer. Four commonly used anticancer drugs (representing major types of chemotherapy) were also tested on 21 human cell lines to compare the selective anticancer activity of amino acid-limited media with that of standard anticancer drugs. The data in Figure 5 show that normal cells exposed to amino acid-limited media grew relatively well (cell viability was 78–98%), while most types of cancer cells were highly affected (cell viability was low) when cultured in these media. However, none of the anticancer drugs at any dose induced a significant decrease in cancer cell viability without reducing the viability of normal cells. These results indicate that amino acid-limited media is more selective for cancer cells than standard anti-cancer drugs used in patients.

[0130] [Table 6]

[0131] Table 6. Composition of cell culture media M0, M1, M2 and M3.

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Claims

1. 1. An artificial dietary composition for use in the treatment and / or prevention of cancer, comprising: Based on the total weight of the dry ingredient composition, 0.1 to 0.6% methionine, 0.5-6% leucine, 0-5% fat, 4-40% amino acid mixture, 40-95% carbohydrates, 1-5% vitamin and mineral mixture, and 0-1% choline, An artificial diet composition for use, comprising:

2. 2. An artificial diet composition for use according to claim 1, characterized in that the mixture of amino acids is a mixture of essential and non-essential amino acids selected from the group consisting of leucine, isoleucine, valine, methionine, lysine, phenylalanine, tryptophan, threonine, histidine, asparagine, alanine, arginine, aspartic acid, cysteine / cystine, glutamic acid, glutamine, proline, glycine, tyrosine, serine and mixtures thereof.

3. 3. The artificial diet composition for use according to claim 1 or 2, wherein glutamine is further present in said artificial diet composition as part of a mixture of amino acids in an amount of 1.3-10.0% by weight based on the total weight of said dry ingredient composition.

4. An artificial diet composition for use according to any one of claims 1 to 3, characterized in that glycine or glycine and serine are further present in said artificial diet composition.

5. 5. An artificial diet composition for use according to any one of claims 1 to 4, characterized in that the amino acids are in free form, in salt form, in ester form and / or in the form of peptides, polypeptides or proteins.

6. 6. An artificial diet composition for use according to any one of claims 1 to 5, characterized in that the amino acids present in said artificial diet composition are a combination of amino acids in free form and as proteins.

7. 7. An artificial diet composition for use according to claim 6, characterized in that said protein is casein.

8. 2. An artificial feed composition for use according to claim 1, characterized in that the lipid component is selected from any edible vegetable or animal oil selected from olive oil, coconut oil, salmon oil, corn oil, soybean oil, canola oil, rapeseed oil, sunflower oil, linseed oil, rice oil, safflower oil, cottonseed oil, palm oil, castor seed oil, peanut oil, wheat oil, pumpkin seed oil, poppy seed oil, hemp oil, pomegranate seed oil, cod oil, herring oil, whale oil, seal oil, margarine, butter, lard, tallow, and mixtures thereof.

9. 9. An artificial diet composition for use according to claim 1 or 8, characterized in that the one or more lipid components are selected from the group consisting of olive oil, coconut oil and salmon oil.

10. An artificial diet composition for use according to any one of claims 1 to 9, characterized in that the carbohydrates may be selected from the group consisting of sucrose, cellulose, starch and mixtures thereof.

11. An artificial diet composition for use according to any one of claims 1 to 10, characterized in that it is in a form suitable for oral administration.

12. 12. An artificial diet composition for use according to claim 11, characterized in that it is in solid, semi-solid or liquid form and is selected from dry powders, shakes, liquid concentrates, ready-to-drink beverages, chilled or shelf-stable beverages, soups, pastes, purees, nutritional bars.

13. 13. An artificial diet composition for use according to any one of claims 1 to 12, characterized in that the treatment of cancer includes renal cancer, lung cancer, colon cancer, breast cancer, melanoma, ovarian cancer, prostate cancer, pancreatic cancer, liver cancer, endometrial cancer, cervical cancer, bladder cancer, esophageal cancer, gastric cancer, head and neck cancer, leukemia, lymphoma, non-melanoma skin cancer, sarcoma, central nervous system cancer, testicular cancer, thyroid cancer and cancer of unknown primary site.

14. 14. An artificial diet composition for use according to any one of claims 1 to 13, wherein said treatment comprises ingesting said artificial diet composition for use as a replacement for food to provide the daily nutritional requirements of the subject.

15. An artificial diet composition for use according to any one of claims 1 to 14, characterized in that the treatment comprises a treatment cycle of 2 to 12 weeks with 4 to 6 daily doses.

16. 16. An artificial feed composition for use according to any one of claims 1 to 15, characterized in that it further comprises water or an aqueous carrier.

17. A pharmaceutical composition for use in the treatment of cancer, characterized in that it comprises an artificial diet composition for use according to any one of claims 1 to 16 and a pharmaceutically acceptable carrier or vehicle.

18. 18. The pharmaceutical composition for use according to claim 17, characterized in that the treatment of cancer includes renal cancer, lung cancer, colon cancer, breast cancer, melanoma, ovarian cancer, prostate cancer, pancreatic cancer, liver cancer, endometrial cancer, cervical cancer, bladder cancer, esophageal cancer, gastric cancer, head and neck cancer, leukemia, lymphoma, non-melanoma skin cancer, sarcoma, central nervous system cancer, testicular cancer, thyroid cancer and cancer of unknown primary site.

19. Treatment may include alkylating agents (e.g., cisplatin, carboplatin, oxaliplatin, cyclophosphamide, hydroxyurea, etc.), antimetabolites (e.g., fluorouracil, capecitabine, gemcitabine, methotrexate, cytarabine, etc.), mitotic inhibitors (e.g., paclitaxel, docetaxel, cabacitaxel, vincristine, vinblastine, vinorelbine, vindesine, etc.), topoisomerase inhibitors (e.g., etoposide, teniposide, irinotecan, topodecane, doxorubicin, epirubicin, etc.), hormone therapy (e.g., antiestrogens such as tamoxifen, aromatase inhibitors such as anastrozole, LHRH agonists such as goserelin, antiandrogens such as abiraterone and flutamide, dexamethasone, thiazolin ...

19. The pharmaceutical composition for use according to claim 17 or 18, characterized in that it comprises the co-administration of said pharmaceutical composition with any type of drug therapy, including cytotoxic chemotherapeutic agents such as anti-PD1 (e.g., anti-PD-L1 such as nivolumab, anti-PDL1 such as avelumab, anti-CTLA4 such as ipilimumab, cytokines such as interleukin-2 and interferons, etc.), targeted therapy (e.g., anti-VEGF agents such as bevacizumab, anti-VEGFR such as sunitinib and sorafenib, anti-EGFR such as cetuximab or erlotinib, anti-HER2 such as trastuzumab, anti-PARP such as olaparib, anti-BRAF such as vemurafenib, etc.), and any other anti-cancer agent, and mixtures thereof.

20. 20. The pharmaceutical composition for use according to claim 19, characterized in that said co-administration comprises sequential, combined or simultaneous administration of the active ingredients.

21. 21. The pharmaceutical composition for use according to any one of claims 17 to 20, characterized in that the treatment comprises administering to a subject in need thereof an effective amount of said pharmaceutical composition during surgery and / or radiotherapy treatment.

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