ES2930009T3 - Methods for cancer and immunotherapy with glutamine analogues, comprising deoxynivalenol DON included - Google Patents

Abstract

The disclosure provides methods of treating cancer in a subject or preventing a relapse or reducing the incidence of relapse of cancer in a subject in remission, comprising administering to the subject: (a) a therapeutically effective amount of an immunotherapeutic agent, e.g. checkpoint blockade therapy, an adoptive cell therapy, marrow-infiltrating lymphocytes, an adenosine A2aR inhibitor, or an antibody; and (b) a compound having formula (I): and pharmaceutically acceptable salts thereof, wherein R1, R2, R2' and X are defined as set forth in the specification. Compounds having formula (I) are prodrugs that release glutamine analogues, eg 6-diazo-5-oxo-L-norleucine (DON).

Description

DESCRIPTION

Methods for cancer and immunotherapy with glutamine analogues, comprising deoxynivalenol DON included

BACKGROUND

[0001] Cells under certain conditions may undergo a metabolic shift from a metabolic profile that requires less activity of certain metabolic pathways to meet the energy demands of the cell to a metabolic profile that requires increased activity of those metabolic pathways or a increased activity of other metabolic pathways. to meet your energy demands. For example, cells under certain conditions can undergo a shift toward increased glycolysis and away from oxidative phosphorylation (OXPHOS). Although glycolysis provides less adenosine triphosphate (ATP) than oxidative phosphorylation, aerobic glycolysis has been proposed to allow the generation of the substrates necessary for the generation of amino acids, nucleic acids, and lipids, all of which are crucial for proliferation. (Vander Heiden et al. (2009) Science 324 (5930): 1029-1033). This use of glycolysis in the presence of oxygen was first described by Otto Warburg in cancer cells (Warburg (1956) Science 124 (3215):269-270) and was later found to be important in activated T cells (Warburg et al. al (1958) [Metabolism of leukocytes]. Zeitschriftfur Naturforschung. Teil B: Chemie, Biochemie, Biophysik, Biologie 13B (8): 515 516). These metabolically reprogrammed cells depend on increased activity of certain metabolic pathways, such as pathways involved in glutamine metabolism, glycolysis, and fatty acid synthesis. However, specific inhibitors of individual enzymes in these metabolic pathways alone have not been shown to be effective because multiple sites within each metabolic pathway are modulated as a cell's metabolism reprograms itself to meet the extraordinarily large energy demands of the body. abnormal, harmful or unhealthy state.

[0002] WO 2014/160071, in the name of St. Jude Children's Research Hospital, describes glutaminase inhibitors such as DON, azaserin or acivicin in combination with at least one proapoptotic Bcl-2 inhibitor in the treatment of cancer. WO 2014/148391, in the name of Johns Hopkins University, describes inhibitors of glutamine metabolism such as DON, acivicin or azaserin for use in the treatment of cancer. EP 0123170, in the name of Amiercan Cyanamid Co, describes a compound having a similar structure to DON for use in the treatment of leukemia and tumors. WO 2004/113363 describes derivatives of the DON transglutaminase inhibitor obtained by reaction of a protected N-alpha amino acid with 6-diazo-5-oxo-L-norleucine. Said derivatives do not include a diazo group. Alan Ramsay, in "Immune Checkpoint Blockade Immunotherapy to Activate Antitumor T-Cell Immunity," British Journal of Haematology, Vol. 162, no. 3, May 21, 2013, pages 313-325, describes the use of immunotherapy in the treatment of cancer and proposes its use in combination with additional targeted agents. This document does not suggest a combination with a glutaminase inhibitor.

RESUME

[0003] The practice of the present invention will typically employ, unless otherwise indicated, conventional techniques of cell biology, cell culture, molecular biology, transgenic biology, microbiology, recombinant nucleic acid (eg, DNA) technology, immunology and RNA interference. (iRNA) that are within the skill in the art. Non-limiting descriptions of some of these techniques are found in the following publications: Ausubel, F., et al., (eds.), Current Protocols in Molecular Biology, Current Protocols in Immunology, Current Protocols in Protein Science and Current Protocols in Cell Biology, all John Wiley & Sons, NY, December 2008 issue; Sambrook, Russell and Sambrook, Molecular Cloning. A Laboratory Manual, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 2001; Harlow, E. and Lane, D., Antibodies-A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 1988; Freshney, RI, "Culture of Animal Cells, A Manual of Basic Technique", 5th Ed., John Wiley & Sons, Hoboken, NJ, 2005. Unlimited information on therapeutic agents and human diseases is found in Goodman and Gilman's The Pharmacological Basis of Therapeutics, 11th Ed., McGraw Hill, 2005, Katzung, B. (ed.) Basic and Clinical Pharmacology, McGraw-Hill/Appleton & Lange 10th Ed. (2006) or 11th Edition (July 2009). Non-limiting information on genes and genetic disorders is found in McKusick, VA: Mendelian Inheritance in Man. A Catalog of Human Genes and Genetic Disorders. Baltimore: Johns Hopkins University Press, 1998 (12th Edition) or the most recent online database: Online Mendelian Inheritance in Man, OMIM™. McKusick-Nathans Institute for Genetic Medicine, Johns Hopkins University (Baltimore, Md.) and National Center for Biotechnology Information, National Library of Medicine (Bethesda, Md.), as of May 1, 2010, World Wide Web URL: http: / /www.ncbi.nlm.nih.gov/omim/ and Online Mendelian Inheritance in Animals (OMIA), a database of genes, inherited disorders, and traits in animal species (other than humans and mice), at

. . . . .

[0004] According to the invention there is provided a compound having Formula (I):

or a pharmaceutically acceptable salt thereof, in which:

X is -(CH2)n-;

n is 1;

R1 is selected from the group consisting of C1-6 alkyl and substituted C1-6 alkyl;

R2 is selected from the group consisting of an amino acid, an N-substituted amino acid, -C(=O)-Y-(CR3R4)m-NR5R6, and -C(=O)-O-(CR3R4)m-O-C(=O) -R10;

Y is -O- or a link;

R2' is selected from the group consisting of H, C1-C6 alkyl, and substituted C1-C6 alkyl; each R3 and R4 are independently H, C1-C6 alkyl, substituted C1-C6 alkyl, aryl, substituted aryl, -(CR3R4)m-NR5R6, or

m is an integer selected from the group consisting of 1,2, 3, 4, 5, 6, 7, and 8;

R5 and R6 are independently H or alkyl; Y

R10 is selected from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, monosaccharide, acylated monosaccharide, aryl, substituted aryl, heteroaryl, and substituted heteroaryl,

for use in the treatment of cancer, wherein the compound having Formula (I) is to be administered with immune checkpoint blockade therapy, adoptive cell therapy, or an adenosine A2aR inhibitor.

[0005] In particular embodiments, the immune checkpoint blockade therapy is selected from the group consisting of PD-1 antagonists, PD-L1 antagonists, CTLA-4 antagonists, LAG3 antagonists, B7 antagonists -H3 and combinations thereof.

[0006] In particular embodiments, the first immunotherapy and the second immunotherapy are administered to the subject in the absence of an anticancer therapy selected from the group consisting of: (i) chemotherapy; (ii) photodynamic therapy; (iii) proton therapy; (i.v.) radiotherapy; (v) surgery; and combinations thereof.

[0007] In particular embodiments, the cancer is a newly diagnosed, recurrent, and/or refractory cancer selected from the group consisting of cellnasopharyngeal cancer, synovial cancer, hepatocellular cancer, renal cancer, connective tissue cancer, melanoma, lung, bowel cancer, colon cancer, rectal cancer, colorectal cancer, brain cancer, throat cancer, oral cancer, liver cancer, bone cancer, pancreatic cancer, choriocarcinoma, gastrinoma, pheochromocytoma, prolactinoma, leukemia / T-cell lymphoma, neuroma, von Hippel-Lindau disease, Zollinger-Ellison syndrome, adrenal cancer, anal cancer, bile duct cancer, bladder cancer, ureteral cancer, brain cancer, oligodendroglioma, neuroblastoma, meningioma, spinal cord tumor, bone cancer, osteochondroma, chondrosarcoma, Ewing's sarcoma, cancer of unknown primary site, carcinoid, gastrointestinal tract carcinoid, fibro sarcoma, breast cancer, Paget's disease, cervical cancer, colorectal cancer, rectal cancer, esophageal cancer, gallbladder cancer, head cancer, eye cancer, neck cancer, kidney cancer, Wilms tumor , liver cancer, Kaposi's sarcoma, prostate cancer, lung cancer, testicular cancer, Hodgkin's disease, non-Hodgkin's lymphoma, oral cancer, skin cancer, mesothelioma, multiple myeloma, ovarian cancer, endocrine pancreatic cancer, glucagonoma , pancreatic cancer, parathyroid cancer, penile cancer, pituitary cancer, soft tissue sarcoma, retinoblastoma, small intestine cancer, stomach cancer, thymus cancer, thyroid cancer, trophoblastic cancer, hydatidiform mole, uterine cancer, endometrial cancer, vaginal cancer, vulvar cancer, acoustic neuroma, mycosis fungoides, insulinoma, carcinoid syndrome, somatostatinoma, gum cancer, heart cancer, lip cancer, meningeal cancer, mouth cancer, cancer Nerve cancer, palate cancer, parotid gland cancer, peritoneal cancer, pharyngeal cancer, pleural cancer, salivary gland cancer, tongue cancer, and tonsil cancer.

[0008] In unclaimed aspects, the subject matter described herein provides a method of preventing a relapse in a subject with cancer in remission, the method comprising administering to the subject a therapeutically effective amount of a metabolic reprogramming agent, wherein the metabolic reprogramming agent is selected from the group consisting of acivicin (L-(alpha S, 5S)-alpha-amino-3-chloro-4,5-dihydro-5-isoxazolacetic acid), azaserin, and 6-diazo-5- oxonorleucine (DON), and 5-diazo-4-oxo-L-norvaline (L-DONV) and prodrugs thereof.

[0009] Certain aspects of the presently disclosed object having been established above, which are addressed in whole or in part by the presently disclosed object, other aspects will become apparent as the description proceeds when taken in connection with the accompanying Examples and Drawings , as better described below.

BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Having thus described the object described herein in general terms, reference will now be made to the attached Figures, which are not necessarily drawn to scale, and in which:

FIG. 1 illustrates that metabolic reprogramming therapy with at least one metabolic reprogramming agent (eg, DON) markedly inhibits lymphoma growth in an EL4 mouse lymphoma model, suggesting that bone marrow-derived tumors they may be highly amenable to metabolic reprogramming therapy with at least one metabolic reprogramming agent (eg, DON);

FIG. 2 illustrates that metabolic reprogramming therapy with at least one metabolic reprogramming agent (eg, DON) has a modest effect in inhibiting the growth of melanoma, which is not a bone marrow-derived tumor;

FIG. 3 illustrates that metabolic reprogramming therapy with at least one metabolic reprogramming agent (eg, DON) conditions B16 melanoma to be eliminated by immunotherapy by inhibiting tumor-infiltrating regulatory T cells (Foxp3+);

FIG. 4 illustrates DON structures and DON-based prodrugs;

FIG. 5A and FIG. 5B illustrates that DON(1) inhibits glutamine metabolism and GBM tumor growth in vivo. FIG. 5A shows that compound 1 (0.8 mg/kg, i.p.) inhibited glutamine metabolism as evidenced by increased endogenous glutamine concentrations in flank GBM tumors 2 hours after administration relative to controls. treated with vehicle; * p < 0.05. FIG. 5B shows in efficacy studies, compared to baseline at day 0, vehicle-treated mice exhibited significant growth of GBM tumors on the flanks during the course of the experiment. In contrast, systemic administration of 1 (0.8 mg/kg, i.p., q.d. on days 1-6) caused a dramatic reduction in tumor size; ***p < 0.001,****p < 0.0001. Note the bold numbers following the terms "DON", "DON prodrugs", "DON-based prodrugs", refer to particular compounds described in Table 1 below.

FIG. 6 illustrates the in vivo brain and plasma pharmacokinetics of the compound DON(1) after oral administration of DON(1) and 5c in mice. 1 and 5c were dosed in mice at 0.8 mg/kg equivalent, by oral gavage, and plasma and brain concentrations of compound 1 were assessed by LC/MS. Oral administration of compound 1 and 5c exhibited similar plasma and brain pharmacokinetic profiles due to complete and rapid metabolism of 5c to 1 in mouse plasma;

FIG. 7 illustrates the in vivo pharmacokinetics of DON after i.v. of DON (1) and 5c in monkey plasma and CSF. 1 and 5c were dosed in two pig-tailed macaques at 1.6 mg/kg equivalent to 1 route i.v. and DON concentrations in plasma (0.25-6 h) and CSF (30 min) were assessed by LC/MS. Relative to 1.5c provided a substantially lower plasma DON concentration. The opposite was observed in CSF, where 5c delivered significantly higher CSF concentrations of DON, achieving a 10-fold higher CSF-to-plasma ratio at 30 minutes post-dose;

FIG. 8 illustrates that 25 (5 day dosing beginning on day 7) is superior to CB-839 (30 day dosing beginning on day 1) in the CT26 tumor model;

FIG. 9 illustrates that 25 (4 days from day 6) is superior to CB-839 (continuous dosing twice daily from day 1 prior to engraftment) in a CT26 tumor model.

FIG. 9 shows mice receiving 25 (1.9 mg/kg) daily on days 6-9 vs. BID glutaminase inhibitor on days 1-15;

FIG. 10 illustrates that 25 (daily on days 7-22) is superior to CB-839 (continuous dosing twice daily on days 1-29) in a 4T1 breast cancer model. Mice received 25 (1.0 mg.kg/d) daily for days 7-22 compared to BID glutaminase inhibitor for days 1-29;

FIG. 11 illustrates that dosing of 1 mg/kg followed by 0.3 mg/kg leads to a complete and long-lasting response in the MC38 tumor;

FIG. 12 illustrates that 25 provides a robust response and improved overall survival in multiple tumor models including, for example, CT26 colon cancer;

FIG. 13 illustrates that 25 provides a robust response and improved overall survival in multiple tumor models including, for example, 4T1 breast cancer;

FIG. 14 illustrates that 25-cured mice only immunologically reject tumors upon rechallenge, demonstrating that 25 monotherapy is immunotherapy;

FIG. 15 further illustrates that 25 monotherapy is immunotherapy;

FIG. 16 illustrates that inhibition of glutamine (eg, DON) reduces oxygen consumption and lactate production of tumor cells;

FIG. 17 illustrates that inhibition of glutamine (eg, DON) improved the CD8/Treg ratio in tumor and reduces hypoxia 6 in TILs;

FIG. 18 illustrates that 25 conditions the tumor to be eliminated by anti-PDI therapy in Model MC38, and in particular that 25 rescues anti-PDI failures;

FIG. 19 illustrates that even in the most difficult CT26 model, 25 response to anti-PDI therapy is improved;

FIG. 20 illustrates that inhibition of glutamine metabolism also potentiates the antitumor response to adenosine A2a receptor (A2aR) blockade;

FIG. 21 illustrates that inhibition of glutamine metabolism improves the efficacy of adoptive cell therapy (ACT) in a B16-OVA model;

FIG. 22A, FIG. 22B, and FIG. 23C illustrate the in vivo pharmacokinetics of DON after i.v. of DON (1) and 14 in monkey plasma and cerebrospinal fluid (CSF). 1 and 14 were dosed in two pig-tailed macaques at 1.6 mg/kg equivalent to 1 by i.v. administration. and DON concentrations in plasma (0.25-6 h) and CSF (30 min) were assessed by LC/MS. Relative to 1, 14 administered a substantially lower plasma concentration of DON. The opposite was observed in CSF, where 14 administered significantly higher CSF DON concentrations, achieving a 10-fold higher CSF-to-plasma ratio at 30 minutes post-dose;

FIG. 23 illustrates the species-specific plasma stability of (14); 14 is stable in the plasma of humans, pigs, dogs, and monkeys, but is rapidly metabolized in mice;

FIG. 24 illustrates exemplary structures of ^{dO n} and prodrugs based on DON 25, 9, 38 and 60; different N-amino acid promoters (eg, leucine, tryptophan) provide differential plasmas and microsomal stability;

FIG. 25A, FIG. 25B, FIG. 25C, and FIG. 25D illustrate the in vitro plasma stability of DON prodrugs 9, 25, 38 and 60. Metabolism occurs through removal of the N-protecting group; both ethyl and isopropyl esters are stable in pig and human plasma;

FIG. 26A, FIG. 26B, FIG. 26C, and FIG. 26D illustrates the in vitro hepatic microsomal stability of DON prodrugs 9, 25, 38 and 60; all prodrugs showed moderate-high stability in human and porcine microsomes;

FIG. 27A, FIG. 27B, FIG. 27C, FIG. 27D, FIG. 27E, FIG. 27F, FIG. 27G, FIG. 27H, FIG. 271, and FIG. 27J illustrate the results of ex vivo studies on human and pig whole blood from 9, 25, 38 and 60; DON prodrugs selectively deliver DON to PBMC in both humans and pigs versus plasma; Compared to DON, the PBMC/plasma ratio was increased 10-100+ fold;

FIG. 28A FIG. 28B, FIG. 28C, FIG. 28D and FIG. 28E illustrate the results of in vivo studies in pigs with DON prodrugs of 9, 25, 38 and 60; DON prodrugs selectively deliver DON to PBMCs versus plasma; Compared to DON, the PBMC/plasma ratio was increased 6-10 fold;

FIG. 29A, FIG. 29B, and F ⁱ G. 29C illustrate the plasma stability of the compound Methyl-POM 14 and its derivatives; FIG. 30 illustrate exemplary structures of N-acylalkyloxy DON-based prodrug analogs for intracellular targeting and brain penetration; adding steric bulk to the "bridge" could result in slower hydrolysis;

FIG. 31 illustrates that anti-PD1 monotherapy does not work in a 4T1 tumor model. 4T1 tumor cells (0.1 million) were injected into the mammary fat pads of 8-week-old female BALB/c mice. Anti-PD1 (5 mg/kg) was administered on days 3, 5, 8 and 11 and tumor volume was measured 2-3 times per week until tumors were evaluated for tumor-infiltrating cells. Group: a-PD1 (d3, 5, 8, 11 100 ug/mouse) = 5 mg/kg i.p.;

FIG. 32A and FIG. 32B illustrate that glutamine analog (6-diazo-5-oxo-L-norleucine (L-DON) derivative = 25) inhibits tumor growth. FIG. 32A shows 4T1 (breast cancer cell lines) after 30 days of inoculation (8 days without drug). FIG. 32B shows Group 1: vehicle PBS, Group 2: 251 mg/kg every day (day 5 to day 22). 4T1 tumor cells (0.1 million) were injected into the mammary fat pads of 8-week-old female BALB/c mice. Mice received vehicle (PBS) or 1 mg/kg daily from day 5 to day 22. Pictures were taken on day 30 after tumor inoculation. Tumor volume was measured 2-3 times per week until WT mice were sacrificed (when size reached 20 mM in length or necrosis occurred). Day 0: 100K 4T1 cells s.c. on the 4th breast pad. Day 5-22: Daily 25;

FIG. 33 illustrates that glutamine analogue (6-diazo-5-oxo-L-norleucine (L-DON) derivative = 25) inhibits tumor growth. 4T1-Luc tumor cells (0.1 million) were injected into the mammary fat pads of 8-week-old female BALB/c mice. Mice received vehicle (PBS) or 1 mg/kg daily from day 7. Anti-PD1 (5 mg/kg) was administered on days 5, 8, and 12. Mice bearing 4T1-luc tumors were given injected Luciferin to measure the luminescence of the tumor. IVIS images were taken on day 13; FIG. 34A and FIG 34B illustrate that glutamine analogue (6-diazo-5-oxo-L-norleucine (L-DON) derivative = 25) inhibits tumor growth. FIG. 34B shows tumor weight (mg) on day 21 of harvest. 4T1-Luc tumor cells (0.1 million) were injected into the mammary fat pads of 8-week-old female BALB/c mice. Mice received vehicle (PBS) or 1 mg/kg daily from day 7 to day 16. Anti-PDI (5 mg/kg) was administered on days 5, 8, 12, and 17. Tumor volume was measured. 2-3 times per week until tumors were assessed for tumor-infiltrating cells on day 21. Tumor weights were measured on day 21. On day 21, the tumor size of the PD1 group appears to have reduced, however, the result was not due to a reduction in the size of the tumor, but rather because one of the mice with large tumors died;

FIG. 35 illustrates reduced CDIIb+ cells and G-MDSC in 25 treated mice, demonstrating that metabolic reprogramming agents that decrease glutamine metabolic activity inhibit myeloid-derived suppressor cells. 4T1-Luc tumor Cells (0.1 million) were injected into the mammary fat pads of 8-week-old female BALB/c mice. Mice received vehicle (PBS) or 1 mg/kg daily from day 7 to day 16. Anti-PD1 (5 mg/kg) was administered on days 5, 8, 12, and 17.

Percentages of myeloid-derived suppressor cells (MDSCs) from circulating blood were monitored on days 6, 9, and 17 by flow cytometry with CD11b and Ly6C/Ly6G granulocytic MDSC (G-MDSC): CDllb+ Ly6g+ Ly6c lo monocytic MDSC (Mo- MDSC): CDllb+ Ly6G-Ly6c Hi;

FIG. 36 illustrates the increase in TNF alpha in 25 treated mice, demonstrating that metabolic reprogramming agents that decrease glutamine metabolic activity increase inflammatory tumor-associated macrophages (TAMs). 4T1-Luc tumor cells (0.2 million) were injected into the mammary fat pads of 8-week-old female BALB/c mice. Mice received vehicle (PBS) or 1 mg/kg daily from day 7 to day 16. Anti-PD1 (5 mg/kg) was administered on days 5, 8, 12, and 17. Tumors were evaluated for infiltrating tumor cells on day 21. Cells were plated and 200 ul Golgi plug was added to inhibit cytokine secretion overnight (without further stimulation). Tumor-associated macrophage markers: Live/CD45+/CD11b+/F4-80+/CD8- for flow cytometric analysis. Tumor-derived macrophages. Cells were incubated with Golgi plug without stimulation. TAM: Live/CD45+/CD11b+/F4-80+/CD8-;

FIG. 37 shows reduced fibrocytes in 25 treated mice, demonstrating that metabolic reprogramming agents that decrease glutamine metabolic activity inhibit bone marrow-derived fibrocytes that are thought to play a role in immunotherapy inhibition as well as in the generation of an extracellular matrix that surrounds tumors and inhibits chemotherapy. 4T1-Luc tumor cells (0.1 million) were injected into the mammary fat pads of 8-week-old female BALB/c mice. Mice received vehicle (PBS) or 1 mg/kg daily from day 7 to day 16. Anti-PDI (5 mg/kg) was administered on days 5, 8, 12, and 17. Tumors were evaluated for tumor-infiltrating cells on day 21. Fibrocyte markers: collagen I+CD11b+CD45+live were used for flow cytometric analysis.

DETAILED DESCRIPTION

[0011] The presently disclosed object will now be described in more detail hereinafter with reference to the attached figures, in which some, but not all, embodiments of the presently disclosed object are shown. Like numbers refer to like items throughout. The subject matter of this disclosure can be embodied in many different ways and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements. In fact, many modifications and other embodiments of the presently disclosed subject matter set forth herein will come to mind to one skilled in the art to which the presently disclosed subject matter pertains having the benefit of the teachings presented in the above descriptions and the associated figures. Therefore, it is to be understood that the subject matter described herein is not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims.

[0012] The object of the present description demonstrates that certain conditions, diseases and/or disorders involve metabolically reprogrammed cells whose activation, function, growth, proliferation and/or survival in an abnormal, harmful and/or unhealthy state depend on increased activity. of at least one, at least two, or at least three metabolic pathways selected from the group consisting of glutamine metabolism, glycolysis, and fatty acid synthesis. It should be appreciated that the abnormal, harmful and/or unhealthy state of the cell refers to its effect on or in relation to the subject whose cells are affected by the condition, disease or disorder rather than on or in relation to the cell itself. exhibiting an increased ability to thrive in the abnormal, harmful, and/or unhealthy state in a manner believed to be proportional to the increase in activity of at least one, at least two, or at least three metabolic pathways (e.g., metabolism glutamine, glycolysis and/or fatty acid synthesis).

[0013] The object of the present description has shown that some of said conditions, diseases and/or disorders, referred to herein as "metabolic reprogramming disorders", are amenable to treatment using at least one, at least two or at least three metabolic reprogramming agents that decrease the activity of at least one, at least two, or at least three metabolic pathways selected from the group consisting of glutamine metabolism, glycolysis, and fatty acid synthesis. In some cases, metabolic reprogramming disorders comprise conditions, diseases, or disorders involving aberrant and/or excessive glutamine metabolism, aberrant and/or excessive glycolysis, or aberrant and/or excessive fatty acid synthesis.

[0014] As used herein, the term "excessive glutamine metabolism" means an increase in the amount of glutamine metabolic activity in a subject with a condition, disease, or disorder ( eg, metabolic reprogramming disorder). ) compared to the amount of glutamine metabolic activity in a subject without a similar disease or condition, such as an increase of approximately 100%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1,000%, or more. As used herein, the term "aberrant glutamine metabolism" means a change in the biologic activity of glutamine in a subject with a condition, disease, or disorder ( eg, metabolic reprogramming disorder) in compared to glutamine activity in a subject without a similar condition, disease, or disorder, such as increased glutamine utilization in the growth and/or proliferation of malignant, neoplastic, or other pathological cellular processes ( eg, immune disorders, neurodegenerative disorders, inflammatory disorders, etc.).

[0015] As used herein, the term "excessive glycolysis metabolism" means an increase in the amount of glycolytic metabolic activity in a subject with a condition, disease, or disorder (for example, a metabolic reprogramming disorder) compared to the amount of glycolytic metabolic activity in a subject without a similar disease or condition, as an increase of approximately 100%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1,000% or more. As used herein, the term "aberrant glycolytic metabolism" means a change in the biologic activity of glycolysis in a subject with a condition, disease, or disorder ( eg, metabolic reprogramming disorder) as compared to glycolytic activity in a subject without a similar disorder. condition, disease, or disorder, such as increased utilization of glucose in the growth and/or proliferation of malignant, neoplastic, or other pathological cellular processes ( eg, immune disorders, neurodegenerative disorders, inflammatory disorders, etc.).

[0016] As used herein, the term "excessive fatty acid synthesis" means an increase in the amount of fatty acid synthesis in a subject with a condition, disease, or disorder ( eg, an eating disorder). metabolic reprogramming) compared to the amount of fatty acid synthesis in a subject without a similar condition, disease, or disorder, as an increase of approximately 100%, 100%, 200%, 300%, 400%, 500%, 600% , 700%, 800%, 900%, 1,000% or more. As used herein, the term "aberrant fatty acid synthesis" means a change in the biological activity of fatty acid synthesis in a subject with a condition, disease, or disorder ( eg, a reprogramming disorder). metabolic) compared to fatty acid synthesis in a subject without a similar condition, disease, or disorder, such as increased utilization of fatty acids in the growth and/or proliferation of malignant, neoplastic, or other pathological cellular processes ( p. g., immune disorders, neurodegenerative disorders, inflammatory disorders, etc.).

[0017] As used herein, a "metabolically reprogrammed" cell refers to a cell in which the activity of at least one, at least two, or at least three metabolic pathways ( eg, metabolism of glutamine, glycolysis, and fatty acid synthesis) increases in response to the energetic and biosynthetic demands of cells placed on the cell in order for the cell to activate, function, grow, proliferate, and/or survive in the abnormal, damaging, and/or unhealthy. As used herein, a "metabolic reprogramming agent" refers to an agent that is capable of reversing the metabolic reprogramming of a cell of a cell whose activation, function, growth, proliferation, and/or survival in an abnormal manner. , harmful, and/or unhealthy state depends on increased activity of at least one, at least two, or at least three metabolic pathways (for example, glutamine metabolism, glycolysis, and fatty acid synthesis) at a cell that has a reduced ability or has lost its ability to thrive (eg, activate, function, grow, proliferate, and/or survive) in the abnormal, harmful, and/or unhealthy state. In some contexts, a "metabolic reprogramming agent" inhibits at least one, at least two, or all of aberrant and/or excessive glutamine metabolism, aberrant and/or excessive glycolysis, and aberrant and/or excessive fatty acid synthesis. .

[0018] The object described herein will now be described in more detail hereinafter with reference to the attached figures, in which some, but not all, embodiments of the inventions are shown. Like numbers refer to like items throughout. The subject matter of this disclosure can be embodied in many different ways and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements. In fact, many modifications and other embodiments of the presently disclosed subject matter set forth herein will come to mind to one skilled in the art to which the presently disclosed subject matter pertains having the benefit of the teachings presented in the above descriptions and the associated figures. Therefore, it is to be understood that the subject matter described herein is not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims.

I. TREATMENT METHODS USING METABOLIC REPROGRAMMING AGENTS

[0019] The presently disclosed subject matter describes a method for treating a subject having a condition, disease or disorder involving metabolically reprogrammed cells whose activation, function, growth, proliferation and/or survival depends on increased activity of at least one metabolic pathway selected from the group consisting of glutamine metabolism, glycolysis and fatty acid synthesis, the method comprising administering to the subject at least one metabolic reprogramming agent that decreases the activity of at least one metabolic pathway selected from the group consisting of it involves the metabolism of glutamine, glycolysis, and the synthesis of fatty acids in an effective amount to treat the condition, disease, or disorder.

[0020] The object of the present description describes a method for treating a subject having a condition, disease or disorder involving at least one of aberrant and/or excessive glutamine metabolism, aberrant and/or excessive glycolysis, or aberrant glycolysis and/or excessive fatty acid synthesis, the method comprising administering to the subject at least one metabolic reprogramming agent that decreases the activity of at least one metabolic pathway selected from the group consisting of glutamine metabolism, glycolysis, and synthesis of fatty acids in an amount effective to treat the condition, disease or disorder.

[0021] In general, the methods described herein result in a decrease in the severity of a condition, disease, or disorder ( eg, a metabolic reprogramming disorder) in a subject. The term "lessen" is intended to inhibit, suppress, attenuate, lessen, arrest, or stabilize a symptom of the condition, disease, or disorder.

As used herein, the terms "treat", "treated", "treatment", refer to reducing or ameliorating a disease or condition, and/or symptoms associated therewith. It will be appreciated that, although not excluded, treatment of a disease or condition does not require that the disorder, condition, or symptoms associated therewith be completely eliminated.

[0022] The method comprises administering to the subject at least two metabolic reprogramming agents that decrease the activity of at least two metabolic pathways selected from the group consisting of glutamine metabolism, glycolysis, and fatty acid synthesis in an effective amount to treat the condition, disease or disorder. The method comprises administering to the subject at least three metabolic reprogramming agents, each of which decreases the activity of a different metabolic pathway selected from the group consisting of glutamine metabolism, glycolysis, and fatty acid synthesis in an amount effective in treating the condition, disease, or disorder.

[0023] The terms "subject" and "patient" are used interchangeably in this document. The subject treated by the methods, uses, metabolic reprogramming agents, and compositions comprising those agents in their many embodiments described herein is desirably a human subject, although it is to be understood that the methods described herein are effective with respect to to all species of vertebrates, which are intended to be included in the term "subject". Accordingly, a "subject" may include a human subject for medical purposes, such as for treatment of an existing condition or disease or prophylactic treatment to prevent the occurrence of a condition or disease, or an animal subject for medical, veterinary, or development purposes. Suitable animal subjects include mammals including, but not limited to primates, eg, humans, monkeys, apes; cattle, eg, cattle, oxen; sheep, eg sheep; caprine, eg, goats; porcine, eg, pigs, pigs; equines, eg, horses, donkeys, zebras; felines, including wild and domestic cats; canines, including dogs; lagomorphs, including rabbits, hares; and rodents, including mice, rats. An animal may be a transgenic animal. T subject is a human including, but not limited to, fetal, neonatal, infant, juvenile, and adult subjects. Furthermore, a "subject" may include a patient afflicted or suspected of being afflicted with a condition or disease.

A. CANCER

[0024] Examples of malignant or cancer cells whose activation, function, growth, proliferation, and/or survival in an abnormal, harmful, or unhealthy state is dependent upon increased metabolic activity of at least one, at least two, or at least at least three metabolic pathways selected from the group consisting of glutamine metabolism, glycolysis, and fatty acid synthesis include, but are not limited to cMyc-dependent cancer cells, glutamine-dependent cancer cells, and combinations thereof. As used herein, a "glutamine-dependent cancer cell" is a cancer cell in which glutamine is an important fuel source for cellular energy in the cancer cell ( eg, hematopoietic tumors, hepatomas, Ehrilich's carcinoma (see Huber et al., "Uptake of glutamine antimetabolites 6-diazo-5-oxo-L-norleucine (DON) and acivicin in sensitive and resistant tumor cell lines," Int. J. Cancer. 1988; 41: 752-755)). As used herein, "cMyc-dependent cancer cells" refers to cancer cells that exhibit activation, overexpression, and/or amplification of c-Myc. In some contexts, a "Myc-dependent cancer" is a cancer in which c-Myc plays a role in increased glutamine metabolism in cancer cells, i.e., cMyc-dependent glutamine-addicted cancer cells Examples of Myc-dependent cancers include, without limitation, lymphoma, neuroblastoma, and small cell lung cancer.

[0025] Use of at least one metabolic reprogramming agent (eg, a metabolic reprogramming agent that decreases glutamine metabolism) as cancer maintenance therapy. As used herein, "cancer maintenance therapy" refers to a therapy administered to a cancer patient who is in cancer remission.

[0026] The presently disclosed subject matter provides a method of preventing a relapse in a subject with cancer in remission, the method comprising administering to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. As used herein, "remission" includes partial and complete remission and refers to a decrease or disappearance of signs and symptoms of cancer. "Partial remission" means that the cancer responded to treatment with the primary therapy, but at least a portion of the tumor and/or at least a portion of the cancer cells are still present in the subject, eg, at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 49% of a measurable tumor and/or measurable cancer cells are still present in the subject after therapy. "Complete remission" means that the subject shows no signs or symptoms of cancer, for example, after a health care provider has used the most accurate and up-to-date tests available to detect cancer and is unable to detect any signs or symptoms of cancer . It should be understood that cancer cells may still exist in a subject in complete remission at levels that are undetectable.

[0027] The metabolic reprogramming agent is administered to the subject after transplantation. As used herein, "post-transplant" refers to a subject who has recently received a cell, tissue, or organ transplant, including, for example, subjects receiving immunosuppressive agents to prevent or reduce risk and/or severity of transplant rejection. The metabolic reprogramming agent is administered to the subject after the chemotherapy. The metabolic reprogramming agent is administered to the subject after immunotherapy. The metabolic reprogramming agent is administered to the subject after photodynamic therapy. The metabolic reprogramming agent is administered to the subject after proton therapy. The metabolic reprogramming agent is administered to the subject after radiation therapy. The metabolic reprogramming agent is administered to the subject after surgery; and combinations thereof. The metabolic reprogramming agent is administered to the subject in two or more of post-transplant, post-chemotherapy, post-immunotherapy, post-photodynamic therapy, post-proton therapy, post-radiotherapy, post-surgery, and combinations thereof.

[0028] As used herein, a "cancer" in a subject refers to the presence of cells that possess characteristics typical of cancer-causing cells, eg, uncontrolled proliferation, loss of specialized functions, immortality, potential significant metastatic disease, significantly increased anti-apoptotic activity, rapid growth and proliferation rate, and some characteristic cellular morphology and markers. In some circumstances, the cancer cells will be in the form of a tumor; such cells may exist locally within an animal or circulate in the bloodstream as independent cells, eg, leukemic cells. A "tumor", as used herein, refers to all growth and proliferation of neoplastic cells, whether malignant or benign, and all precancerous and cancerous cells and tissues. A "solid tumor" as used herein is an abnormal mass of tissue that generally does not contain cysts or fluid areas. A solid tumor can be in the brain, colon, breasts, prostate, liver, kidneys, lungs, esophagus, head and neck, ovaries, cervix, stomach, colon, rectum, bladder, uterus, testicles, and pancreas, as non-limiting examples. . In some embodiments, the solid tumor regresses or its growth slows or stops after treating the solid tumor with the methods described herein. In other embodiments, the solid tumor is malignant. In some embodiments, the cancer comprises stage 0 cancer. In some embodiments, the cancer comprises Step I cancer. In some embodiments, the cancer comprises stage II cancer. In some embodiments, the cancer comprises stage III cancer. In some embodiments, the cancer comprises stage I.V. cancer. In some embodiments, the cancer is refractory and/or metastatic. For example, the cancer may be refractory to treatment with radiation therapy, chemotherapy, or immunotherapy monotherapy.

[0029] In particular embodiments, the cancer is a cancer of the central nervous system (CNS cancer). Some of the currently disclosed metabolic reprogramming agents and compositions are believed to be particularly useful in the treatment of CNS cancers and cancers of CNS origin. In particular, the data described in FIG. 22A, FIG. 22B, FIG. 22C, FIG. 29A, FIG. 29B, FIG. 29C and FIG. 30 demonstrate that certain metabolic reprogramming agents ( eg, glutamine analogue prodrugs, eg, DON prodrugs ^{) effectively target and deliver DON to the brain, eg, achieving a ratio of lCr to} plasma up to 10 times higher at 30 minutes post dosing. Accordingly, some of the currently described metabolic reprogramming agents are contemplated for use as cancer therapy ( eg, maintenance therapy), immunotherapy, and enhancement of immunotherapy, for the treatment of CNS cancers and cancers of the origin of the CNS.

[0030] Exemplary CNS cancers treatable with the methods, compositions, and agents described herein include, without limitation, gliomas, astrocytomas, oligodendrogliomas, ependymomas, mixed gliomas ( eg, oligoastrocytomas), meningiomas ( eg. , atypical, invasive, anaplastic, etc.), medulloblastomas, gangliogliomas, schwannomas (neuroliemmomas), craniopharyngiomas, chordomas, CNS non-Hodgkin lymphoma and pituitary tumors. In particular embodiments, the CNS cancer comprises glioblastoma multiforme (GBM).

[0031] In particular embodiments, the cancer is a cancer that is associated with transplantation and/or immunosuppression. It is well known that organ transplants ( eg, kidney, liver, heart, lung, etc.) in the United States carry a high risk of developing various types of cancer (see, eg, Engels et al. 2011). In some cases, the cancer risk is elevated for infection-related cancer due to immunosuppression, for example, due to drugs given to suppress the immune system and prevent transplant ( eg, organ) rejection. In some embodiments, the cancer associated with the transplant and/or immunosuppression is related to an infectious agent. Examples of cancers associated with transplantation and/or immunosuppression include, without limitation, anal cancer, Kaposi's sarcoma, kidney cancer, liver cancer, lung cancer, melanoma, non-Hodgkin lymphoma, and thyroid cancer. In particular embodiments, the subject is a child or elderly transplant recipient ( eg, liver, heart, kidney, etc.) who may or may not be infected with Epstein-Barr virus. In particular embodiments, the cancer is a cancer that is refractory to chemotherapy. In particular embodiments, the cancer is a cancer that is refractory to photodynamic therapy. In particular embodiments, the cancer is a cancer that is refractory to proton therapy.

[0032] In particular embodiments, the cancer is a cancer refractory to radiation therapy.

[0033] In particular embodiments, the cancer is a cancer refractory to surgery.

[0034] Cancer as used herein includes newly diagnosed or recurrent and/or refractory cancers, including, but not limited to, acute lymphoblastic leukemia, acute myelogenous leukemia, advanced soft tissue sarcoma, brain cancer, metastatic or aggressive breast cancer , breast carcinoma, bronchogenic carcinoma, choriocarcinoma, chronic myelocytic leukemia, colon carcinoma, colorectal carcinoma, Ewing's sarcoma, gastrointestinal tract carcinoma, glioma, glioblastoma multiforme, head and neck squamous cell carcinoma, carcinoma hepatocellular, Hodgkin's disease, intracranial ependymoblastoma, large intestine cancer, leukemia, liver cancer, lung carcinoma, Lewis lung carcinoma, lymphoma, malignant fibrous histiocytoma, breast tumor, melanoma, mesothelioma, neuroblastoma, osteosarcoma, ovarian cancer, pancreatic cancer, pontine tumor, premenopausal breast cancer, prostate cancer, rhabdomyosarcoma, reticulum cell sarcoma, sarcoma, small cell lung cancer, a solid tumor, stomach cancer, testicular cancer, and uterine carcinoma.

[0035] In particular embodiments, the cancer treated is a newly diagnosed, recurrent, and/or refractory cancer selected from the group consisting of cellnasopharyngeal cancer, synovial cancer, hepatocellular cancer, renal cancer, connective tissue cancer, melanoma, lung, bowel cancer, colon cancer, rectal cancer, colorectal cancer, brain cancer, throat cancer, oral cancer, liver cancer, bone cancer, pancreatic cancer, choriocarcinoma, gastrinoma, pheochromocytoma, prolactinoma, leukemia/ T-cell lymphoma, neuroma, von Hippel-Lindau disease, Zollinger-Ellison syndrome, adrenal cancer, anal cancer, bile duct cancer, bladder cancer, ureteral cancer, brain cancer, oligodendroglioma, neuroblastoma, meningioma, spinal cord tumor, bone cancer, osteochondroma, chondrosarcoma, Ewing's sarcoma, cancer of unknown primary site, carcinoid, carcinoid of the gastrointestinal tract, fibrosar coma, breast cancer, Paget's disease, cervical cancer, colorectal cancer, rectal cancer, esophageal cancer, gallbladder cancer, head cancer, eye cancer, neck cancer, kidney cancer, Wilms tumor , liver cancer, Kaposi's sarcoma, prostate cancer, lung cancer, testicular cancer, Hodgkin's disease, non-Hodgkin's lymphoma, oral cancer, skin cancer, mesothelioma, multiple myeloma, ovarian cancer, pancreatic cancer, endocrine cancer, glucagonoma, pancreatic cancer, parathyroid cancer, penile cancer, pituitary cancer, soft tissue sarcoma, retinoblastoma, small bowel cancer, stomach cancer, thymus cancer, thyroid cancer, trophoblastic cancer, hydatidiform mole, cancer of the uterus, endometrial cancer, vaginal cancer, vulvar cancer, acoustic neuroma, mycosis fungoides, insulinoma, carcinoid syndrome, somatostatinoma, gum cancer, heart cancer, lip cancer, meningeal cancer, mouth cancer, nerve cancer, palate cancer, parotid gland cancer, peritoneal cancer, pharyngeal cancer, pleural cancer, salivary gland cancer, tongue cancer, and tonsil cancer.

[0036] In some embodiments, the condition, disease, or disorder is lymphoma. Therefore, in one aspect, the object of the present description provides a method for the treatment of lymphoma in a subject in need thereof, the method comprising administering to the subject at least one metabolic reprogramming agent that decreases glutamine metabolism in a effective amount to treat the lymphoma in the subject.

[0037] In some embodiments, the condition, disease, or disorder is melanoma. Therefore, in one aspect, the object of the present description provides a method for the treatment of melanoma in a subject in need thereof, the method comprising administering to the subject at least one metabolic reprogramming agent that decreases glutamine metabolism in a effective amount to treat the melanoma in the subject.

[0038] In some embodiments, the methods include administering to the subject an effective amount of radiation therapy. In some embodiments, the methods include administering to the subject an effective amount of immunotherapy ( eg, a second immunotherapy). In some embodiments, the methods include administering to the subject an effective amount of photodynamic therapy. In some embodiments, the methods include administering to the subject an effective amount of proton therapy. In some embodiments, the methods include surgical resection of at least a portion of a tumor before, during, or after treatment with at least one, at least two, or at least three metabolic reprogramming agents and, optionally, at least one chemotherapeutic agent, immunotherapeutic. and/or radiotherapeutic agent.

[0039] In some embodiments, the condition, disease, or disorder is a newly diagnosed, recurrent, and/or refractory cancer selected from the group consisting of acute lymphoblastic leukemia, acute myelogenous leukemia, advanced soft tissue sarcoma, brain cancer, metastatic or aggressive breast cancer, breast carcinoma, bronchogenic carcinoma, choriocarcinoma, chronic myelocytic leukemia, colon carcinoma, colorectal carcinoma, Ewing's sarcoma, carcinoma of the gastrointestinal tract, glioma, glioblastoma multiforme, squamous cell carcinoma of the head and neck , hepatocellular carcinoma, Hodgkin's disease, intracranial ependymoblastoma, large intestine cancer, leukemia, liver cancer, lung carcinoma, Lewis lung carcinoma, lymphoma, malignant fibrous histiocytoma, breast tumor, melanoma, mesothelioma, neuroblastoma, osteosarcoma, cancer ovarian cancer, pancreatic cancer, pontine tumor, premenopausal breast cancer, prostate cancer a, rhabdomyosarcoma, reticulum cell sarcoma, sarcoma, small cell lung cancer, a solid tumor, stomach cancer, testicular cancer, and uterine carcinoma.

[0040] In some embodiments, the cancer is not acute lymphoblastic leukemia. In some embodiments, the cancer is not acute myelogenous leukemia. In some embodiments, the cancer is not an advanced soft tissue sarcoma. In some embodiments, the cancer is not brain cancer. In some embodiments, the cancer is not metastatic or aggressive breast cancer. In some embodiments, the cancer is not breast carcinoma. In some embodiments, the cancer is not a bronchogenic carcinoma. In some embodiments, the cancer is not choriocarcinoma. In some embodiments, the cancer is not chronic myelocytic leukemia. In some embodiments, the cancer is not colon carcinoma. In some embodiments, the cancer is not colorectal carcinoma. In some embodiments, the cancer is not Ewing's sarcoma. In some embodiments, the cancer is not a carcinoma of the gastrointestinal tract. In some embodiments, cancer is not a glioma. In some embodiments, the cancer is not glioblastoma multiforme. In some embodiments, the cancer is not head and neck squamous cell carcinoma. In some embodiments, the cancer is not hepatocellular carcinoma. In some embodiments, the cancer is not Hodgkin's disease. In some embodiments, the cancer is not an intracranial ependymoblastoma. In some embodiments, the cancer is not colon cancer. In some embodiments, the cancer is not leukemia. In some embodiments, the cancer is not liver cancer. In some embodiments, the cancer is not lung carcinoma. In some embodiments, the cancer is not a Lewis lung carcinoma. In some embodiments, the cancer is not a lymphoma. In some embodiments, the cancer is not a malignant fibrous histiocytoma. In some embodiments, the cancer is not a breast tumor. In some embodiments, the cancer is not melanoma. In some embodiments, the cancer is not mesothelioma. In some embodiments, the cancer is not a neuroblastoma. In some embodiments, the cancer is not osteosarcoma. In some embodiments, the cancer is not ovarian cancer. In some embodiments, the cancer is not pancreatic cancer. In some embodiments, the cancer is not a pontine tumor. In some embodiments, the cancer is not premenopausal breast cancer. In some embodiments, the cancer is not prostate cancer. In some embodiments, the cancer is not rhabdomyosarcoma. In some embodiments, the cancer is not a reticulum cell sarcoma. In some embodiments, the cancer is not a sarcoma. In some embodiments, the cancer is not small cell lung cancer. In some embodiments, the cancer is not a solid tumor. In some embodiments, the cancer is not stomach cancer. In some embodiments, the cancer is not testicular cancer. In some embodiments, the cancer is not uterine carcinoma.

B. IMMUNOTHERAPY

[0041] Unclaimed aspects of the presently disclosed subject matter involve the use of at least one, at least two, or at least three metabolic reprogramming agents, alone or optionally together in combination with additional immunotherapy ( eg, blockade of checkpoints, adoptive cell therapy (ACT), vaccines ( eg, tumor vaccines), passive immunotherapy antibodies, for cancer treatment

[0042] Consequently, in an unclaimed aspect, the object of the present description provides a method for treating a cancer in a subject in need thereof, the method comprising: (a) administering a therapeutically effective amount of a first immunotherapy to the subject , wherein the first immunotherapy is a metabolic reprogramming agent; and (b) optionally administering a therapeutically effective amount of a second immunotherapy to the subject. In particular embodiments described herein, the metabolic reprogramming agent is a glutamine antagonist. In particular embodiments described herein, the metabolic reprogramming agent is a glutamine analog that interferes with a glutamine metabolic pathway. In particular embodiments described herein, the metabolic reprogramming agent is selected from the group consisting of acivicin (L-(alpha S, 5S)-alpha-amino-3-chloro-4,5-dihydro-5-acid -isoxazolacetic), azaserine, and 6-diazo-5-oxonorleucine (DON), and 5-diazo-4-oxo-L-norvaline (L-DONV). In particular embodiments, the metabolic reprogramming agent is a prodrug of a glutamine analog that interferes with a glutamine metabolic pathway. In particular embodiments, at least one metabolic reprogramming agent is a prodrug of acivicin, azaserin, DON, and L-DONV.

[0043] In some aspects described herein, a prodrug of a glutamine antagonist, or a pharmaceutically acceptable salt or ester thereof has a structure of Formula (I):

where: X is selected from the group consisting of a bond, -O-, and -(CH2)n-, where n is an integer selected from the group consisting of 1,2, 3, 4, 5, 6 , 7 and 8; R1 is selected from the group consisting of H and a first prodrug-forming moiety capable of forming a salt or ester; and R2 is H or a second prodrug-forming moiety capable of forming an amide bond, carbamate bond, phosphoramidate bond, or phosphorodiamidate bond with the nitrogen adjacent to R2; R2' is selected from the group consisting of H, C1-C6 alkyl, substituted C1-C6 alkyl, or R2 and R2' together form a ring structure comprising -C(=O)-G-C(=O)-, where G is selected from the group consisting of C1-C8 alkylene, C1-C8 heteroalkylene, C5-C8 cycloalkylene, C6-C12 arylene, C5-C14 heteroarylene, C4-C10 bivalent heterocycle, each of which may be optionally substituted; or R1 and R2' together form a 4 to 6 membered heterocyclic ring comprising the oxygen atom adjacent to R1 and the nitrogen atom adjacent to R2'; with the proviso that the compound has at least one prodrug-forming moiety selected from the group consisting of the first and second prodrug-forming moieties.

[0044] As used herein, the term "amide bond" comprises a structure represented by the Formula:

wherein Rv is selected from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl, heterocyclyl, substituted heterocyclyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkylamine, substituted alkylamine , heteroaryl and substituted heteroaryl.

[0045] As used herein, the term "carbamate bond" encompasses a structure represented by the Formula:

where RW is selected from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl, heterocyclyl, heterocyclyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkylamine, substituted alkylamine, heteroaryl, and heteroaryl substituted.

[0046] As used herein, the term "phosphoramidate bond" comprises a structure represented by the Formula:

wherein R ^x and R ^x are each independently selected from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl, heterocyclyl, substituted heterocyclyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkylamine, substituted alkylamine, heteroaryl, and substituted heteroaryl.

[0047] As used herein, the term "phosphorodiamidate bond" comprises a structure represented by the Formula:

wherein Ry and Rz are each independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl, heterocyclyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, -(CR3R4)mZ, -( CR3R4)mQZ, aryl, substituted aryl, alkylamine, substituted alkylamine, heteroaryl, substituted heteroaryl, and

[0048] In some embodiments described herein, X is -CH2-, and n is 1.

[0049] In other embodiments described herein, X is -O-. In some embodiments described herein, the prodrug compound has both the first prodrug-forming moiety and the second prodrug-forming moiety. In some embodiments described herein, the glutamine analog is a glutamine antagonist, ie, the prodrug is a prodrug of a glutamine analog that antagonizes a glutamine pathway. Examples of glutamine antagonists include, without limitation, 6-diazo-5-oxo-norleucine (DON) and azaserine, and 5-diazo-4-oxo-L-norvaline (L-DONV).

[0050] In some embodiments described herein, the presently disclosed subject matter provides a prodrug of DON. In some embodiments, the DON prodrug has a structure of Formula (I). In some embodiments described herein, the presently disclosed subject matter provides a prodrug of L-DONV. In some embodiments, the L-DONV prodrug has a structure of Formula (I). In some embodiments described herein, the presently disclosed subject matter provides a prodrug of azaserin. In some embodiments, the azaserin prodrug has a structure of Formula (I).

[0051] In some embodiments described herein, R1 of Formula (I) comprises a PRO1 moiety of the prodrug-forming moiety which, together with a basic moiety and the terminal hydroxyl group, forms a salt.

[0052] In some embodiments described herein, R1 of Formula (I) comprises a PRO1 residue of the prodrug-forming moiety which, together with an alkyl group and the oxygen of a neighboring hydroxyl group, forms an ester.

[0053] In some embodiments described herein, R1 of Formula (I) comprises a PRO1 residue of the prodrug-forming moiety which, together with an alkyl group and the nitrogen found adjacent to the R2' group, forms a azlactone or an oxazolidone.

[0054] In some embodiments described herein, R1 of Formula (I) is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkenyl , substituted cycloalkenyl, tri(hydrocarbyl)ammonium and tetra(hydrocarbyl)ammonium. Preferred substituents of alkyl group, cycloalkyl group, alkenyl group, alkynyl group and cycloalkenyl group include alkyl, substituted alkyl, halo, arylamino, acyl, hydroxyl, aryloxy, alkoxy, alkylthio, arylthio, aralkyloxy, aralkylthio, carboxyl, alkoxycarbonyl, oxo and cycloalkyl.

[0055] In some embodiments described herein, R1 of Formula (I) is not H. In some embodiments, R1 of Formula (I) is not H when R2 and R2' are H. In some embodiments As an embodiment, R2 and R2' of Formula (I) are each H when and R1 is not H.

[0056] In some embodiments described herein, R1 of Formula (I) is selected from the group consisting of a C1-6 straight chain alkyl, an alkyl substituted C1-6 straight chain alkyl, a branched alkyl C1-6, a substituted branched C1-6 alkyl, tri(C1-C8 alkyl)ammonium, tetra(C1-C8 alkyl)ammonium, triphenylammonium, tri(hydroxyC1-C8alkyl)ammonium and tetra(hydroxy-C1-C8-alkyl )ammonium.

[0057] In some embodiments described herein, R1 of Formula (I) is selected from the group consisting of methyl, ethyl, isopropyl, cyclopentyl, cyclohexyl, trimethylammonium, triethylammonium, tri(hydroxyethyl)ammonium, tripropylammonium, and tri (hydroxypropyl)ammonium. In some embodiments described herein, R1 of Formula (I) is methyl. In some embodiments, R1 of Formula (I) is ethyl. In some embodiments, R1 of Formula (I) is isopropyl.

[0058] In some embodiments described herein, R2 of Formula (I) comprises a residue PRO2 of the second prodrug-forming moiety, which, together with a carbonyl, oxycarbonyl, or phosphonyl group and the adjoining NH nitrogen, forms an amide, a carbamate, phosphoramidate or phosphorodiamidate bond.

[0059] In some embodiments described herein, R2 of Formula (I) comprises a moiety selected from the group consisting of an amino acid, an N-substituted amino acid, a peptide, a substituted peptide, a monocyclic ring, a substituted monocyclic ring, a bicyclic ring, a substituted bicyclic ring, a purine nucleoside, a substituted purine nucleoside, a pyrimidine nucleoside, and a substituted pyrimidine nucleoside.

[0060] In some aspects described herein, a prodrug of a glutamine antagonist, or a pharmaceutically acceptable salt or ester thereof has a structure of Formula (I):

wherein: X is selected from the group consisting of a bond, -O-, and -(CH2V, where n is an integer selected from the group consisting of 1,2, 3, 4, 5, 6, 7, and 8; R1 is selected from the group consisting of H and a first prodrug-forming moiety capable of forming a salt or ester; and R2 is H or a second prodrug-forming moiety capable of forming an amide bond, a carbamate bond, a phosphoramidate bond or a phosphorodiamidate bond with the nitrogen adjacent to R2; R2' is selected from the group consisting of H, C1-C6 alkyl, substituted C1-C6 alkyl, or R2 and R2' together form a ring structure comprising -C (=O)-G-C(=O)-, where G is selected from the group consisting of C1-C8 alkylene, C1-C8 heteroalkylene, C5-C8 cycloalkylene, C6-C12 arylene, C5-C14 heteroarylene, C4- bivalent heterocycle C10, each of which may be optionally substituted; or R1 and R2' together form a 4 to 6 membered heterocyclic ring comprising the oxygen atom adya central to R1 and the nitrogen atom adjacent to R2'; with the proviso that the compound has at least one prodrug-forming moiety selected from the group consisting of the first and second prodrug-forming moieties.

[0061] As used herein, the term "amide bond" comprises a structure represented by the Formula:

where Rv is selected from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl, heterocyclyl, heterocyclyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkylamine, substituted alkylamine, heteroaryl, and substituted heteroaryl.

[0062] As used herein, the term "carbamate bond" comprises a structure represented by the Formula:

wherein Rw is selected from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl, heterocyclyl, substituted heterocyclyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkylamine, substituted alkylamine , heteroaryl and substituted heteroaryl.

[0063] As used herein, the term "phosphoramidate bond" comprises a structure represented by the Formula:

wherein Rx and Rx' are each independently selected from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, aralkyl, substituted aralkyl, heterocyclyl, substituted heterocyclyl, alkenyl, substituted alkenyl, cycloalkenyl, cycloalkenyl substituted, alkylamine, substituted alkylamine, heteroaryl, and substituted heteroaryl.

[0064] As used herein, the term "phosphorodiamidate bond" comprises a structure represented by the Formula:

wherein Ry and Rz are each independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl, heterocyclyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, -(CR3R4)m-Z, -( CR3R4)m-Q-Z, aryl, substituted aryl, alkylamine, substituted alkylamine, heteroaryl, substituted heteroaryl, and

[0065] In some embodiments described herein, X is -CH2-, and n is 1.

[0066] In other embodiments described herein, X is -O-. In some embodiments, the prodrug compound has both the first prodrug-forming moiety and the second prodrug-forming moiety. In some embodiments described herein, the glutamine analog is a glutamine antagonist, ie, the prodrug is a prodrug of a glutamine analog that antagonizes a glutamine pathway. Examples of glutamine antagonists include, without limitation, 6-diazo-5-oxo-norleucine (DON) and azaserine, and 5-diazo-4-oxo-L-norvaline (L-DONV).

[0067] In some embodiments described herein, the presently disclosed subject matter provides a prodrug of DON. In some embodiments described herein, the DON prodrug has a structure of Formula (I). In some embodiments described herein, the presently disclosed subject matter provides a prodrug of L-DONV. In some embodiments described herein, the L-DONV prodrug has a structure of Formula (I). In some embodiments, the subject of the present invention provides a prodrug of azaserin. In some embodiments described herein, the azaserin prodrug has a structure of Formula (I).

[0068] In some embodiments described herein, R1 of Formula (I) comprises a PRO1 residue of the prodrug-forming moiety which, together with a basic moiety and the terminal hydroxyl group, forms a salt.

[0069] In some embodiments described herein, R1 of Formula (I) comprises a PRO1 residue of the prodrug-forming moiety which, together with an alkyl group and the oxygen of a neighboring hydroxyl group, forms an ester.

[0070] In some embodiments described herein, R1 of Formula (I) comprises a PRO1 residue of the prodrug-forming moiety which, together with an alkyl group and the nitrogen adjacent to the R2' group, forms a azlactone or an oxazolidone.

[0071] In some embodiments described herein, R1 of Formula (I) is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkenyl , substituted cycloalkenyl, tri(hydrocarbyl)ammonium and tetra(hydrocarbyl)ammonium. Preferred substituents of alkyl group, cycloalkyl group, alkenyl group, alkynyl group and cycloalkenyl group include alkyl, substituted alkyl, halo, arylamino, acyl, hydroxyl, aryloxy, alkoxy, alkylthio, arylthio, aralkyloxy, aralkylthio, carboxyl, alkoxycarbonyl, oxo and cycloalkyl.

[0072] In some embodiments described herein, Ri of Formula (I) is not H. In some embodiments, R1 of Formula (I) is not H when R2 and R2' are H. In some embodiments As an embodiment, R2 and R2' of Formula (I) are each H when and R1 is not H.

[0073] In some embodiments described herein, R1 of Formula (I) is selected from the group consisting of a straight chain C1-6 alkyl, a straight chain alkyl substituted C1-6 straight chain alkyl, C1-6 branched, substituted branched C1-6 alkyl, tri(C1-C8-alkyl)ammonium, tetra(C1-C8-alkyl)ammonium, triphenylammonium, tri(hydroxy-C1-C8alkyl)ammonium, and tetra(hydroxy-C1 -C8-alkyl)ammonium.

[0074] In some embodiments described herein, R1 of Formula (I) is selected from the group consisting of methyl, ethyl, isopropyl, cyclopentyl, cyclohexyl, trimethylammonium, triethylammonium, tri(hydroxyethyl)ammonium, tripropylammonium, and tri (hydroxypropyl)ammonium. In some embodiments described herein, R1 of Formula (I) is methyl. In some embodiments, R1 of Formula (I) is ethyl. In some embodiments, R1 of Formula (I) is isopropyl.

[0075] In some embodiments described herein, R2 of Formula (I) comprises a residue PRO2 of the second prodrug-forming moiety, which, together with a carbonyl, oxycarbonyl, or phosphonyl group and the adjoining NH nitrogen, forms an amide, a carbamate, phosphoramidate or phosphorodiamidate bond.

[0076] In some embodiments described herein, R2 of Formula (I) comprises a moiety selected from the group consisting of an amino acid, an N-substituted amino acid, a peptide, a substituted peptide, a monocyclic ring, a substituted monocyclic ring, a bicyclic ring, a substituted bicyclic ring, a purine nucleoside, a substituted purine nucleoside, a pyrimidine nucleoside, and a substituted pyrimidine nucleoside.

[0077] In some embodiments described herein, R2 of Formula (I) is selected from the group consisting of H, alkyl, -C(=O)-Ar, -C(=O)-Y-( CRaR4)m-Ar, -C(=O)-Y-(CRaR4)m-NR5R6, -P(=O)(OR7)n(NHRg)o, -C(=O)-Y-(CRaR4)m -Ar-O-C(=O)-R8, -C(=O)-Y-(CRgR4)m-Ar-O-R8, -C(=O)-O-(CR3R4)m-O-C(=O)-Rm -C(=O)-O-Rg, -C(=O)-Y-(CR3R4)m-Ar-O-C(=O)-Ar and -C(=O)-Y-(CR3R4)m-Ar -NR5R6i where: Y is -O- or a bond; m is an integer selected from the group consisting of 0, 1,2, 3, 4, 5, 6, 7, and 8; each n and o is an integer from 0 to 2 as long as the sum of n and o is 2; R3 and R4 are independently H, C1-C6 alkyl or substituted C1-C6 alkyl, aryl or substituted aryl, -(CR3R4)m-NR5Ra, or

; each R5 and R6 is independently H, alkyl, -C(=O)-(CR3R4)m, -C(=O)-(NR5R6) or -C(=O)-(CR3R4)m-NR5R6; each R7 is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl, substituted heterocyclyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, -(CR3R4)mZ, -(CR3R4WQ-Z, in where Q is a monosaccharide, aryl, substituted aryl, heteroaryl, substituted heteroaryl, and where Z is

or where R7 together with the oxygen atom to which it is attached forms a purine or pyrimidine nucleoside; each Rg is independently selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclyl, substituted heterocyclyl, alkenyl, substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, -(CR3R4)mZ, aryl, substituted aryl, heteroaryl, substituted heteroaryl, and

, where Ri and X are as defined above, provided that Ri is not H; each R8 is independently alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, monosaccharide, acylated monosaccharide, aryl, substituted aryl, heteroaryl, substituted heteroaryl; each R10 is independently alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, monosaccharide, acylated monosaccharide, aryl, substituted aryl, heteroaryl, substituted heteroaryl; and Ar is aryl, substituted aryl, heteroaryl, or substituted heteroaryl. It should be appreciated that, in addition to substitutions on the amino group of Z, one or more R3, R4, R5 and/or R6 substitutions may be made on the 5- or 6-membered rings of Z.

[0078] The structures of representative DON prodrugs described herein are provided in Table 1.

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1. BLOCK CONTROL POINTS

[0079] Subject aspects of the present disclosure involve the use of metabolic reprogramming agents ( eg, ^{dO n} , DON prodrugs, etc.) in combination immunotherapy in conjunction with checkpoint blockade modulators, for example, to improve checkpoint blockade therapies for cancer treatment. In some aspects, the topic currently described involves the use of metabolic reprogramming agents ( eg, DON, DON prodrugs, etc.) in combination immunotherapy in conjunction with A2aR blockade.

[0080] Examples of immune checkpoint modulators for use herein include, but are not limited to small organic molecules ( eg, haptens) or small inorganic molecules; saccharides; oligosaccharides; polysaccharides; a biological macromolecule selected from the group consisting of peptides ( eg, aptides), proteins, peptide analogues, and derivatives; peptidomimetics; nucleic acids selected from the group consisting of miRNA, siRNA, shRNA, antisense nucleic acids, such as antisense RNA, ribozymes, and aptamers; an extract made from biological materials selected from the group consisting of bacteria, plants, fungi, animal cells and animal tissues; natural or synthetic compositions; and any combination thereof. Other examples of immune checkpoint modulators include orthosteric inhibitors, allosteric regulators, interfacial binders, and molecular analogs of substrates that act as competitive inhibitors.

[0081] Specific examples of immune checkpoint modulators include, without limitation, PD-1 antagonists, PD-L1 antagonists, CTLA-4 antagonists, Lag-3 antagonists, CD137 antagonists, KIR antagonists, Tim3 antagonists, Ox40 agonists, B7-H3 antagonists, and combinations thereof.

[0082] Exemplary CTLA-4 antagonists include, without limitation, ipilimumab, tremelimumab, and combinations thereof. Anti-CTLA-4 antibodies are currently in clinical trials for the treatment of melanoma.

[0083] Exemplary Lag-3 antagonists include, without limitation, BMS-986016 and IMP321.

[0084] Exemplary CD137 antagonists include, without limitation, CD137-specific antibody, peptide, small organic molecule, antisense oligonucleotide, siRNA, antisense expression vector, or recombinant virus. In some embodiments, the CD137-specific antibody is clone BBK-2 or clone 4B4-1, as described in WIPO International Application Publication No. WO200405513A2.

[0085] T-cell immunoglobulin and mucin domain 3 (TIM3) antagonists ( eg, anti-TIM3 antibodies) have been described for use as immunotherapy (see, eg, Ngiow et al. 2011). Exemplary Tim3 antagonists include, without limitation, anti-TIM3 monoclonal antibodies, for example, as described in the poster presentation by Jun et al. "Generation of anti-TIM-3 and anti-LAG-3 antagonistic monoclonal antibodies for potential novel immunotherapy combinations," available online at http://www.tesarobio.com/documents/2014AAcRposterLB266.pdf.

[0086] Ox40 agonists are described by Linch et al., "OX40 Agonists and Combination Immunotherapy: Putting the Pedal to the Metal" Front Oncol 5:34; 2015. Examples of Ox40 agonists include, without limitation, anti-Ox40 agonist antibodies. Other examples of Ox40 agonists include, without limitation, OX86 and Fc-OX40L.

[0087] Examples of B7-H3 antagonists include, without limitation, MGA271.

[0088] Exemplary PD-L1 antagonists include, without limitation, BMS-936559/MDX-1105, MEDI4736, MPDL3280A, MPDL3280A, MSB0010718C, and combinations thereof. PD-L1 antagonists are currently in clinical trials, for example, for the treatment of melanoma, non-small cell lung cancer, renal cell carcinoma, and ovarian cancer.

[0089] PD-1 antagonists have been reviewed (see, eg, Dolan and Gupta 2014). Examples of PD-1 antagonists for use herein include, without limitation, AMP-224, AMP-554, nivolumab, pembrolizumab, pidilizumab, and combinations thereof.

[0090] In some embodiments described herein, PD-1 antagonists comprise anti-PD-1 antibodies. Examples of anti-PD-1 antibodies include, without limitation, atezolizumab, nivolumab, pembrolizumab, pidilizumab, and combinations thereof. PD-1 antagonists are currently in clinical trials, for example, for the treatment of colorectal cancer, gastric cancer, melanoma, non-small cell lung cancer, ovarian cancer, pancreatic cancer, and renal cell carcinoma.

[0091] In particular embodiments described herein, the presently disclosed subject matter provides a method of treating an advanced solid tumor, the method comprising: (a) administering to the subject a therapeutically effective amount of BMS-936559; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0092] In particular embodiments described herein, the presently disclosed subject matter provides a method of treating an advanced solid tumor, the method comprising: (a) administering to the subject a therapeutically effective amount of MEDI4736; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0093] In particular embodiments described herein, the subject matter described herein provides a method of treating melanoma, the method comprising: (a) administering to the subject a therapeutically effective amount of MPDL3280A in combination with vemurafenib; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0094] In particular embodiments described herein, the subject matter described herein provides a method of treating melanoma, the method comprising: (a) administering to the subject a therapeutically effective amount of MEDI4736 in combination with dabrafenib and trametinib ; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In particular embodiments, the subject matter described herein provides a method of treating melanoma, the method comprising: (a) administering to the subject a therapeutically effective amount of MEDI4736 in combination with trametinib; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0095] In particular embodiments described herein, the presently disclosed subject matter provides a method of treating non-small cell lung cancer, the method comprising: (a) administering to the subject a therapeutically effective amount of MPDL3280A; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, MPDL3280A is administered in combination with erlotinib. In particular embodiments described herein, the object currently disclosed provides a method of treating non-small cell lung cancer, the method comprising: (a) administering to the subject a therapeutically effective amount of MEDI4736; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments, MEDI4736 is administered in combination with tremelimumab.

[0096] In particular embodiments described herein, the subject matter described herein provides a method of treating renal cell carcinoma, the method comprising: (a) administering to the subject a therapeutically effective amount of MPDL3280A; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, MPDL3280A is administered in combination with bevacizumab.

[0097] In particular embodiments described herein, the presently disclosed subject matter provides a method of treating a solid or hematological malignancy, the method comprising: (a) administering to the subject a therapeutically effective amount of MPDL3280A; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0098] In particular embodiments described herein, the subject matter described herein provides a method of treating a solid tumor, the method comprising: (a) administering to the subject a therapeutically effective amount of MPDL3280A; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, MPDL3280A is administered in combination with bevacizumab and/or chemotherapy. In some embodiments, MPDL3280A is administered in combination with cobimetinib.

[0099] In the particular embodiments described herein, the subject of the present disclosure provides a method of treating a solid tumor, the method comprising: (a) administering to the subject a therapeutically effective amount of MEDI4736; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, MEDI4736 is administered in combination with tremelimumab.

[0100] In particular embodiments described herein, the subject matter described herein provides a method of treating a solid tumor, the method comprising: (a) administering to the subject a therapeutically effective amount of MSB0010718C; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0101] In particular embodiments described herein, the subject matter described herein provides a method of treating advanced cancer, the method comprising: (a) administering to the subject a therapeutically effective amount of AMP-224; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0102] In particular embodiments described herein, the presently disclosed subject matter provides a method of treating an advanced solid tumor, the method comprising: (a) administering to the subject a therapeutically effective amount of nivolumab in combination with iliolumbar (anti-lumbar) -KIR); and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0103] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating castration-resistant prostate cancer, hepatocellular carcinoma, melanoma, non-small cell lung cancer, or renal cell carcinoma, the method comprising: (a) administering to the subject a therapeutically effective amount of nivolumab; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0104] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating colon cancer, gastric cancer, head and neck cancer, Hodgkin's lymphoma, melanoma, myeloma, myelodysplastic syndrome, non-invasive lymphoma. Hodgkin, non-small cell lung. cancer, solid tumors or triple negative breast cancer, the method comprising: (a) administering to the subject a therapeutically effective amount of pembrolizumab; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0105] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating gastric cancer, pancreatic cancer, small cell lung cancer, glioblastoma, or triple negative breast cancer, the method comprising: (a) administering by subjecting to an amount therapeutically effective nivolumab in combination with ipilimumab; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0106] In particular embodiments described herein, the subject matter described herein provides a method of treating a malignant glioma, the method comprising: (a) administering to the subject a therapeutically effective amount of pidilizumab; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0107] In particular embodiments described herein, the presently disclosed subject matter provides a method of treating pancreatic cancer, the method comprising: (a) administering to the subject a therapeutically effective amount of pidilizumab in combination with gemcitabine; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0108] In particular embodiments described herein, the subject matter described herein provides a method of treating renal cell carcinoma, the method comprising: (a) administering to the subject a therapeutically effective amount of pidilizumab in combination with a dendritic cell/RCC fusion cell vaccine; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

1. ADOPTIVE CELL THERAPY

[0109] Adoptive cell therapies (ACTs) are a useful approach to treating cancer. Adoptive cell transfer refers to the passive transfer of ex vivo cultured cells, often cells derived from the immune system, into a host with the goal of transferring immunologic functionality and transplant characteristics. The adoptive cell transfer can be autologous, as is common in adoptive T-cell therapies, or allogeneic. Adoptive transfer of autologous tumor-infiltrating lymphocytes (TILs) or genetically redirected peripheral blood mononuclear cells has been used to successfully treat patients with advanced solid tumors. such as melanoma as well as patients with haematological malignancies expressing CD19. Exemplary cell types for use in ACTs include, without limitation, T cells ( eg, CD8+ cells, CD4+ cells, etc.), NK cells, delta-gamma T cells, regulatory T cells, and peripheral blood mononuclear cells. . Such cells may be unmodified, as in LIT therapy, or genetically modified. One way to achieve the genetic targeting of T cells to specific tumor targets is the transfer of a T cell receptor with known specificity (TCR therapy) and with matching human leukocyte antigen (HLA, known as major histocompatibility complex in rodents). . Another way is to modify cells with artificial molecules such as chimeric antigen receptors (CARs), commonly known as CAR-T cell therapy. For example, single chain antibodies can be used and CARs can also incorporate costimulatory domains.

[0110] Aspects of the subject matter of the present disclosure involve the use of metabolic reprogramming agents ( eg, ^{dO n} , DON prodrugs, etc.) in combination immunotherapy together with adoptive cell therapy, for example, to improve adoptive cell therapy for the treatment of cancer.

[0111] Accordingly, in some embodiments described herein, the method of treating cancer further includes simultaneous or sequential administration of a therapeutically effective amount of the second immunotherapy to the subject, wherein the second immunotherapy is a adoptive cell therapy. In some embodiments described herein, the adoptive cell therapy is selected from the group consisting of T cells, CD8+ cells, CD4+ cells, NK cells, delta-gamma T cells, marrow-infiltrating lymphocytes (MIL), T cells regulatory and peripheral blood mononuclear cells. In some embodiments described herein, the adoptive cell therapy comprises a tumor infiltrating lymphocyte (TIL). In some embodiments described herein, the adoptive cell therapy comprises a T-cell receptor-modified lymphocyte. In some embodiments described herein, the adoptive cell therapy comprises a chimeric antigen receptor-modified lymphocyte. In some embodiments described herein, the adoptive cell therapy comprises a chimeric antigen receptor T (CAR-T) cell. In some embodiments described herein, the adoptive cell therapy comprises marrow infiltrating lymphocytes (MIL). In some embodiments described herein, the method of treating cancer further includes simultaneous or sequential administration of a therapeutically effective amount of the second immunotherapy to the subject, wherein the second immunotherapy is marrow-infiltrating lymphocytes (MILs). .

2. ADENOSINE A2AR BLOCKADE

[0112] Adenosine A2a receptor (A2aR) blockade has been reviewed and is reported to improve tumor vaccines, checkpoint blockade, and adoptive T-cell therapy (see, for example, Powell et al. 2015 ). Consequently, aspects of the topic currently described involve the use of metabolic reprogramming agents ( eg, DON, DON prodrugs, etc.) in combination immunotherapy in conjunction with adenosine A2a receptor (A2aR) blockade, for example , to improve A2aR blockade for the treatment of cancer. In some respects, the subject currently described involves the use of metabolic reprogramming agents ( eg, DON, DON prodrugs, etc.) in combination immunotherapy in conjunction with A2aR blockade, and optionally in combination with a third, fourth, or fifth immunotherapy Immunotherapy, such as tumor vaccinations, checkpoint blockade, and/or adoptive cell therapy.

[0113] In some embodiments described herein, the method of treating cancer further includes simultaneous or sequential administration of a therapeutically effective amount of the second immunotherapy to the subject, wherein the second immunotherapy is an adenosine blockade. A2aR. Exemplary A2aR inhibitors for use in blocking A2aR as immunotherapy include, without limitation, SCH58261, SYN115, ZM241365, and FSPTP.

[0114] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating a subject with a CD73-expressing tumor, the method comprising: (a) administering a therapeutically effective amount of SCH58261 to the subject ; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. Exemplary tumors that express CD73 include, without limitation, breast tumors ( eg, adenocarcinoma of the breast, metastatic breast cancer) and melanoma ( eg, metastatic). In some embodiments, the CD73-expressing tumor is a metastatic tumor and administration of SCH58261 suppresses metastases in the CD73-expressing tumor. In some embodiments described herein, the CD73-expressing tumor is melanoma and SCH58261 and the metabolic reprogramming agent are administered in combination with an anti-PD1 antibody to prolong survival and reduce the burden of metastatic melanoma. In some embodiments described herein, the CD73-expressing tumor is breast cancer and SCH58261 and the metabolic reprogramming agent are administered in combination with an anti-PD1 antibody to prolong survival and reduce the burden of breast cancer. metastatic. In some embodiments described herein, the CD73-expressing tumor is a breast cancer tumor and SCH58261 and the metabolic reprogramming agent are administered in combination with a chemotherapeutic agent ( eg, doxorubicin) to increase the breast cancer tumor sensitivity to chemotherapeutic agent.

[0115] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating a subject with a CD73-expressing tumor, the method comprising: (a) administering a therapeutically effective amount of SYN115 to the subject ; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, an anti-PD-1 antibody is administered in combination with SYN115 and the metabolic reprogramming agent.

[0116] In particular embodiments described herein, the presently disclosed subject matter provides a method for the treatment of melanoma, the method comprising: (a) administering a therapeutically effective amount of ZM241365 to the subject; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments, an anti-CTLA4 antibody is administered in combination with ZM241365 and the metabolic reprogramming agent.

[0117] In particular embodiments described herein, the presently disclosed subject matter provides a method for the treatment of bladder cancer, the method comprising: (a) administering a therapeutically effective amount of FSPTP to the subject; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments, FSPTP is administered by intratumoral injection.

3. KILLER CELL IMMUNOGLOBULIN TYPE RECEPTOR (KIR) BLOCKADE

[0118] Unclaimed aspects of the presently disclosed subject matter involve the use of metabolic reprogramming agents ( eg, DON, DON prodrugs, etc.) in combination immunotherapy in conjunction with killer cell-like immunoglobulin receptor (KIR) blockade ), for example, to enhance KIR blockade for cancer treatment. In some aspects, the topic currently described involves the use of metabolic reprogramming agents ( eg, DON, DON prodrugs, etc.) in combination immunotherapy in conjunction with KIR blockade, and optionally in combination with a third immunotherapy, a fourth immunotherapy and/or a fifth immunotherapy, such as tumor vaccines, adoptive cell therapy, A2aR blockade, and/or checkpoint blockade.

[0119] In some embodiments described herein, the method of treating cancer further includes simultaneous or sequential administration of a therapeutically effective amount of the second immunotherapy to the subject, wherein the second immunotherapy is a KIR blockade. .

[0120] Exemplary KIR inhibitors for use in KIR blockade as immunotherapy include, without limitation, IPH2102/BMS-986015 (lirilumab).

[0121] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating acute myeloid leukemia, the method comprising: (a) administering an amount therapeutically effective dose of lirilumab to the subject; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0122] In particular embodiments described herein, the presently disclosed subject matter provides a method of treating a solid tumor, the method comprising: (a) administering a therapeutically effective amount of lirilumab to the subject; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, the solid tumor is a melanoma tumor and lirilumab is administered in combination with nivolumab. In some embodiments described herein, the solid tumor is a non-small cell lung cancer tumor, and lirilumab is administered in combination with nivolumab. In some embodiments described herein, the solid tumor is a gastrointestinal tumor and lirilumab is administered in combination with nivolumab. In some embodiments described herein, the solid tumor is a squamous cell carcinoma of the head and neck tumor and lirilumab is administered in combination with nivolumab. In some embodiments described herein, the solid tumor is a hepatocellular carcinoma tumor and lirilumab is administered in combination with nivolumab.

[0123] In a particular embodiment described herein, the subject of the present disclosure provides a method of treating a hematologic tumor, the method comprising: (a) administering a therapeutically effective amount of lirilumab to the subject; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, the hematologic tumor is relapsed and/or refractory non-Hodgkin lymphoma and lirilumab is administered in combination with nivolumab. In some embodiments described herein, the hematologic tumor is relapsed and/or refractory Hodgkin lymphoma and lirilumab is administered in combination with nivolumab. In some embodiments described herein, the hematologic tumor is relapsed and/or refractory multiple myeloma and lirilumab is administered in combination with nivolumab. In some embodiments, the hematologic tumor is relapsed and/or refractory chromic myelogenous leukemia, and lirilumab is administered in combination with nivolumab.

[0124] In particular embodiments described herein, the subject matter described herein provides a method of treating relapsed and/or refractory multiple myeloma after autologous transplantation, the method comprising: (a) administering an amount therapeutically effective delivery of lirilumab to the subject optionally in combination with elotuzumab; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0125] In particular embodiments described herein, the subject matter described herein provides a method of treating relapsed and/or refractory acute myeloid leukemia, the method comprising: (a) administering a therapeutically effective amount of lirilumab to the subject optionally in combination with 5-azacitidine; and (b) sequentially or simultaneously administering to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

4. VACCINES

[0126] Vaccines stimulate the body's immune system to attack abnormal or malignant cells, such as cancer, resulting in a reduction of those cells. Vaccines against cancer or tumors often contain a tumor antigen in an immunogenic formulation that stimulates tumor antigen-specific helper cells, CTLs, and/or B cells. Examples of vaccine formulations include, but are not limited to, dendritic cells ( p eg, autologous dendritic cells pulsed with tumor cells or antigens), heterologous tumor cells transfected with immunostimulatory agents, such as GM-CSF, recombinant virus, and/or adjuvanted peptides or proteins, such as CpG.

[0127] Unclaimed aspects of the currently disclosed topic involve combination immunotherapy using metabolic reprogramming agents ( eg, that decrease glutamine metabolism ( eg, DON or a DON program) sequentially or simultaneously with vaccines, for example, for the treatment of cancer It is believed that when used as combination immunotherapy with vaccines, metabolic reprogramming agents may help slow or stop the growth of cancer cells, shrink the tumor, prevent recurrence cancer and/or kill cancer cells that have not been killed by other treatments.

[0128] Accordingly, in some embodiments described herein, the method of treating cancer further includes simultaneous or sequential administration of a therapeutically effective amount of the second immunotherapy to the subject, wherein the second immunotherapy is a vaccine ( eg, tumor vaccine).

[0129] Such exemplary vaccines include, without limitation, peptide vaccines, dendritic cell (DC) vaccines, EGFRvIII vaccines, mesotiline vaccine, G-VAX, listeria vaccines, and a dendritic cell/RCC fusion cell vaccine.

[0130] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating or preventing a human papillomavirus (HPV)-associated cancer in a subject in need thereof, the method comprising: (a) administering to the subject a therapeutically effective amount of a recombinant HPV vaccine ; and (b) simultaneously or sequentially administering a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0131] In some embodiments described herein, the subject has an oncogenic HPV type 6 infection and the rHPV vaccine comprises a vaccine against rHPV type 6. In some embodiments, the subject has an infection by oncogenic HPV type 11 and the rHPV vaccine comprises an HPV type 11 vaccine. In some embodiments described herein, the subject has an oncogenic HPV type 16 infection and the rHPV vaccine comprises an rHPV type 16 vaccine. In some embodiments described herein, the subject has an oncogenic HPV type 18 infection and the rHPV vaccine comprises an rHPV type 18 vaccine. In some embodiments described herein, the subject is a female and the cancer is selected from the group consisting of cervical, vaginal and vulvar cancer. In some embodiments described herein, the subject is a female between the ages of 9 and 26. In some embodiments, the subject is a female between the ages of 9 and 26 and the HPV-associated cancer is cervical cancer. In some embodiments described herein, the subject is a female between the ages of 9 and 26 and the HPV-associated cancer is vaginal cancer. In some embodiments described herein, the subject is a female between the ages of 9 and 26 and the HPV-associated cancer is vulvar cancer. In some embodiments, the subject is male or female and the HPV-associated cancer comprises anal cancer. In some embodiments described herein, the subject is a male or female between the ages of 9 and 26 and the HPV-associated cancer is anal cancer. In some embodiments described herein, the HPV vaccine comprises GARDASIL. In some embodiments described herein, the subject is a female between the ages of 10 and 25 and the HPV-associated cancer is cervical cancer. In some embodiments described herein, the HPV vaccine comprises CERVARIX. In some embodiments described herein, the HPV vaccine is administered intramuscularly by injection in the form of a suspension.

[0132] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating or preventing hepatitis B-associated cancer in a subject in need thereof, the method comprising: (a) administering to the subject a therapeutically effective amount of a hepatitis B vaccine; and (b) simultaneously or sequentially administering a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, the subject has a hepatitis B virus infection and the cancer is liver cancer.

6. PASSIVE IMMUNOTHERAPY

[0133] Unclaimed aspects of the currently disclosed topic involve combination immunotherapy using metabolic reprogramming agents ( eg, that decrease glutamine metabolism ( eg, DON or a DON prodrug) sequentially or simultaneous with antibodies passive immunotherapy for cancer treatment

[0134] Accordingly, in some embodiments described herein, the method of treating cancer further includes simultaneous or sequential administration of a therapeutically effective amount of the second immunotherapy to the subject, wherein the second immunotherapy is a passive immunotherapy antibody. As used herein, "passive immunotherapy antibody" refers to monoclonal antibodies directed to specific antigens on the surface of cancer cells to provide immunity against cancer without actively stimulating a patient's immune system.

[0135] Examples of passive immunotherapy antibodies for use herein include, without limitation, bevacizumab ( eg, Av ASTIN), cetuximab ( eg, Er B iTUX), rituximab ( eg, RITUXAN) , trastuzumab ( eg, HERCEPTIN), alemtuzumab ( eg, CAMPATH), ibritumomab tiuxetan ( eg, ZEVa L iN), panitumumab ( eg, VEc T iBIX).

[0136] In particular embodiments described herein, the subject matter described herein provides a method for treating a subject with metastatic colorectal cancer, the method comprising: (a) administering a therapeutically effective amount of bevacizumab to the subject ; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, bevacizumab is administered as first-line or second-line treatment in combination with 5-fluorouracil-based intravenous chemotherapy. In some embodiments described herein, bevacizumab is administered as second-line treatment in combination with fluoropyrimidine-based chemotherapy (combined with irinotecan or oxaliplatin) after cancer progression after first-line treatment including bevacizumab.

[0137] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating a subject with non-small cell lung cancer, the method comprising: (a) administering a therapeutically effective amount of bevacizumab to the subject; and (b) administer sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, non-small cell lung cancer comprises advanced non-squamous non-small cell lung cancer and bevacizumab is administered to subjects who have not received chemotherapy for their advanced disease in combination with carboplatin and paclitaxel.

[0138] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating a subject with plantin-resistant ovarian cancer, the method comprising: (a) administering a therapeutically effective amount of bevacizumab to the subject; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, the subject has not received more than two prior chemotherapy treatments and bevacizumab is used to treat plantin-resistant recurrent epithelial ovarian, fallopian tube, or primary peritoneal cancer in the subject in combination with paclitaxel, pegylated liposomal doxorubicin, or topotecan.

[0139] In particular embodiments described herein, the subject matter described herein provides a method for treating a subject with advanced cervical cancer, the method comprising: (a) administering a therapeutically effective amount of bevacizumab to the subject; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, the subject has persistent, recurrent, or metastatic cervical cancer and bevacizumab is administered in combination with paclitaxel and cisplatin. In some embodiments described herein, the subject has persistent, recurrent, or metastatic cervical cancer and bevacizumab is administered in combination with paclitaxel and topotecan.

[0140] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating a subject with metastatic renal cell carcinoma, the method comprising: (a) administering a therapeutically effective amount of bevacizumab to the subject ; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, the subject has metastatic kidney cancer and bevacizumab is administered in combination with alpha interferon.

[0141] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating a subject with recurrent glioblastoma, the method comprising: (a) administering a therapeutically effective amount of bevacizumab to the subject; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, the subject with recurrent glioblastoma is an adult.

[0142] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating a subject with head and neck cancer, the method comprising: (a) administering a therapeutically effective amount of cetuximab to the subject ; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, the subject has locally or regionally advanced head and neck squamous cell carcinoma and cetuximab is administered in combination with radiation therapy. In some embodiments described herein, the subject has recurrent locoregional disease or metastatic squamous cell carcinoma of the head and neck and cetuximab is administered in combination with 5-FU platinum-based therapy. In some embodiments described herein, the subject has recurrent or metastatic head and neck squamous cell carcinoma and has not responded to prior platinum-based therapy.

[0143] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating a subject with metastatic colorectal cancer, the method comprising: (a) administering a therapeutically effective amount of cetuximab to the subject; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, the metastatic colorectal cancer comprises wild-type KRAS epidermal growth factor receptor (EGFR)-expressing metastatic colorectal cancer, and cetuximab is administered in combination with FOLFIRI (irinotecan, 5-fluorouracil and lucovorin). In some embodiments described herein, the metastatic colorectal cancer comprises metastatic colorectal cancer that expresses wild-type KRAS epidermal growth factor receptor (EGFR), the subject is refractory to irinotecan-based chemotherapy, and is administered cetuximab. in combination with irinotecan. In some embodiments described herein, the metastatic colorectal cancer comprises metastatic colorectal cancer that expresses wild-type KRAS epidermal growth factor receptor (EGFR), and the subject has not responded to oxaliplatin- and irinotecan-based chemotherapy. . In some embodiments described herein, the metastatic colorectal cancer comprises metastatic colorectal cancer that expresses the wild-type epidermal growth factor receptor (EGFR) KRAS, and the subject is intelerant to irinotecan.

[0144] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating a subject with non-Hodgkin lymphoma, the method comprising: (a) administering a therapeutically effective amount of rituximab to the subject; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0145] In some embodiments described herein, the subject has relapsed or refractory CD20-positive low-grade or follicular non-Hodgkin's lymphoma. In some embodiment, the subject has a newly diagnosed cD20-positive non-Hodgkin's lymphoma. In some embodiments described herein, the subject has low-grade or follicular CD20-positive non-Hodgkin lymphoma and responded to initial treatment with CVP chemotherapy (cyclophosphamide, vincristine, and prednisone). In some embodiments described herein, the subject has CD20-positive diffuse large B-cell non-Hodgkin lymphoma and rituximab is administered in combination with CHOP chemotherapy (cyclophosphamide, doxorubicin hydrochloride, vincristine, and prednisolone). In some embodiments described herein, the subject has newly diagnosed or recurrent CD20-positive chronic lymphocytic leukemia and rituximab is administered in combination with CF chemotherapy (fludarabine and cyclophosphamide).

[0146] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating a subject with early stage breast cancer that is human epidermal growth factor receptor 2 (HER2+) positive. the method comprising: (a) administering a therapeutically effective amount of alemtuzumab to the subject; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0147] In some embodiments described herein, alemtuzumab is administered to the subject as part of a treatment cycle comprising doxorubicin, cyclophosphamide, and paclitaxel or docetaxel. In some embodiments described herein, the subject is administered alemtuzumab with docetaxel and carboplatin. In some embodiments described herein, alemtuxumab is administered to the subject after treatment with an anthracycline-based therapy. In some embodiments described herein, the HER2+ breast cancer has spread to the subject's lymph nodes. In some embodiments described herein, the HER2+ breast cancer has not spread to the subject's lymph nodes and the cancer is estrogen receptor/progesterone receptor (ER/PR) negative or has a high-risk feature selected from the group consisting of tumor size > 2 MC, age <35 years, or tumor grade 2 or 3.

[0148] In particular embodiments described herein, the subject of the present disclosure provides a method for treating a subject with B-cell chronic lymphocytic leukemia (B-CLL), the method comprising: (a) administering a therapeutically effective amount of alemtuzumab to the subject; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0149] In particular embodiments described herein, the subject matter described herein provides a method for treating a subject with low-grade or follicular B-cell non-Hodgkin lymphoma (NHL) who has relapsed during or after of treatment with other anticancer drugs or newly diagnosed follicular NHL following a response to initial anticancer therapy, the method comprising: (a) administering a therapeutically effective amount of ibritumomab tiuxetan to the subject; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism. In some embodiments described herein, administration of ibritumomab tiuxetan comprises an intravenous injection comprising two infusions of rituximab ( eg, to reduce the number of B cells in the subject's blood) and one injection of Yttrium -90 ibritumomab tiuxetan.

[0150] In particular embodiments described herein, the presently disclosed subject matter provides a method for treating a subject with wild-type KRAS (exon 2 at codons 12 or 13) metastatic colorectal cancer, the method comprising: (a) administering a therapeutically effective amount of panitumumab to the subject as first-line therapy in combination with folinic acid, fluorouracil, and oxaliplatin; and (b) administering sequentially or simultaneously to the subject a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

[0151] In particular embodiments described herein, the subject matter described herein provides a method for treating a subject with wild-type KRAS (exon 2 at codons 12 or 13) metastatic colorectal cancer, comprising the method: (a) administering a therapeutically effective amount of panitumumab after disease progression after prior treatment with chemotherapy containing fluoropyrimidine, oxaliptin and irinotecan; and (b) administering sequentially or simultaneously a therapeutically effective amount of a metabolic reprogramming agent that decreases glutamine metabolism.

7. COMBINATION IMMUNOTHERAPY

[0152] Unclaimed aspects of the presently disclosed subject matter involve the use of metabolic reprogramming agents ( eg, DON, DON prodrugs, etc.) in combination immunotherapy in conjunction with a second, third, fourth, and/or fifth immunotherapy, for example, to enhance immunotherapy for the treatment of cancer.

[0153] In some embodiments described herein, the method of treating cancer further includes simultaneous or sequential administration of a therapeutically effective amount of a third immunotherapy to the subject, wherein the third immunotherapy is selected from the group that consists of checkpoint blockade, adoptive cell therapy, CAR-T cell therapy, marrow-infiltrating lymphocytes, A2aR blockade, KIR blockade, vaccines ( eg, tumor vaccines), passive immunotherapy antibodies, and combinations thereof.

[0154] In some embodiments described herein, the method of treating cancer further includes simultaneous or sequential administration of a therapeutically effective amount of a third and/or fourth immunotherapy to the subject, wherein the third and/or o Fourth immunotherapy is selected from the group consisting of checkpoint blockade, adoptive cell therapy, c AR-T cell therapy, marrow-infiltrating lymphocytes, A2aR blockade, KIR blockade, vaccines ( eg, tumor vaccines) , passive immunotherapy antibodies and combinations thereof.

[0155] In some embodiments described herein, the method of treating cancer further includes simultaneous or sequential administration of a therapeutically effective amount of a third, fourth, and/or fifth immunotherapy to the subject, wherein the third, fourth and/or fifth immunotherapy is selected from the group consisting of checkpoint blockade, adoptive cell therapy, CAR-T cell therapy, marrow-infiltrating lymphocytes, A2aR blockade, KIR blockade, vaccines ( eg. , tumor vaccines), passive immunotherapy antibodies and combinations thereof.

8. ADJUVANT FOR CANCER THERAPY

[0156] Unclaimed aspects of the presently disclosed subject matter involve the use of metabolic reprogramming agents ( eg, DON or a DON prodrug) as an adjuvant to cancer therapy, eg, for the treatment or prevention of cancer. cancer.

[0157] Accordingly, in some embodiments described herein, the method of treating cancer further includes simultaneous or sequential administration to the subject of a therapeutically effective amount of an anticancer therapy selected from the group consisting of: (i) chemotherapy; (ii) photodynamic therapy; (iii) proton therapy; (i.v.) radiotherapy; (v) surgery; and combinations thereof.

[0158] In some embodiments described herein, the first immunotherapy and the second immunotherapy, if given, are administered to the subject in the absence of chemotherapy. In some embodiments described herein, the first immunotherapy and the second immunotherapy, if administered, are administered to the subject in the absence of photodynamic therapy. In some embodiments described herein, the first immunotherapy and the second immunotherapy, if administered, are administered to the subject in the absence of proton therapy. In some embodiments described herein, the first immunotherapy and the second immunotherapy, if administered, are administered to the subject in the absence of radiation therapy. In some embodiments described herein, the first immunotherapy and the second immunotherapy, if administered, are administered to the subject in the absence of surgery.

II. METABOLIC REPROGRAMMING AGENTS

[0159] The subject matter of the present disclosure describes the use of various agents in connection with the methods, uses, and compositions described herein. Some of the methods and compositions described herein relate to the metabolic reprogramming of cells using at least one metabolic reprogramming agent described herein to treat conditions, diseases or disorders involving metabolically reprogrammed cells whose activation, function, growth, proliferation and/or survival is dependent on increased activity of at least one, at least two, or at least three metabolic pathways selected from the group consisting of glutamine metabolism, glycolysis, and fatty acid synthesis. Aspects of the methods and compositions described herein relate to the use of at least one metabolic reprogramming agent described herein to treat conditions, diseases, or disorders that involve aberrant and/or excessive amounts of at least one, at least two or at least three metabolic. pathways selected from the group consisting of aberrant and/or excessive glutamine metabolism, aberrant and/or excessive glycolysis, or aberrant and/or excessive fatty acid synthesis.

[0160] As used herein, "metabolic reprogramming agent" generally refers to an agent that modulates the metabolic activity of at least one metabolic pathway in a cell, for example, to alter activation, function, growth, cell proliferation and/or survival. As used herein, "modular" means broadly to cause or facilitate a qualitative or quantitative change, alteration, or modification in a molecule, process, pathway, or phenomenon of interest. Without limitation, said change may be an increase, a decrease, a change in binding characteristics, or a change in the relative strength or activity of different components or branches of the process, pathway, or phenomenon. The term "modulator" refers in general terms to any molecule or agent that causes or facilitates a qualitative or quantitative change, alteration, or modification in a process, pathway, or phenomenon of interest. As used herein, the term "modulator" encompasses both inhibitors and activators of a metabolic pathway or target. For example, "modulator" comprises inhibitors and activators of the expression and/or activity of a component involved in glutamine metabolism, a component involved in glycolysis, and/or a component involved in fatty acid metabolism ( p. g., fatty acid synthesis or fatty acid oxidation).

[0161] As used herein, the phrase "modulation of a metabolic pathway" refers to modulation of the activity of at least one component of the metabolic pathway. It is contemplated herein that the modulator of the metabolic pathway may be, for example, a receptor ligand ( eg, small molecule, antibody, siRNA), a ligand sequestrant ( eg, antibody, an binding protein), a modulator of phosphorylation of a component of the pathway, or a combination of such modulators. One skilled in the art can readily test an agent to determine whether it modulates a metabolic pathway by assessing, for example, the phosphorylation status of a receptor or the expression or synthesis of downstream proteins or enzymes controlled by the pathway in cultured cells and comparing the results. to cells not treated with a modulator. A modulator is determined to be a metabolic pathway modulator if the level of receptor phosphorylation or expression or synthesis of downstream proteins or enzymes in a cell culture is reduced by at least 20% compared to the level of receptor phosphorylation or expression or synthesis of downstream proteins or enzymes in cells grown in the absence of the modulator; preferably the level of phosphorylation or expression or synthesis of downstream proteins or enzymes is altered by at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 99% in the presence of a metabolic pathway modulator.

[0162] The terms "decrease", "reduce", "reduce", "decrease" or "inhibit" are all used herein generally to mean a decrease by a statistically significant amount. However, for the avoidance of doubt, "reduced", "reduced", "decreased" or "inhibited" means a decrease of at least 10% compared to a reference level, for example, a decrease of at least 20% , or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least around 90%, where the decrease is less than 100%. In one embodiment, the decrease includes a 100% decrease (eg, missing level compared to a reference sample), or any decrease between 10 and 100% compared to a reference level.

[0163] The terms "increased", "increase", "improve" or "activate" are all used here to generally mean an increase by a statically significant amount; for the avoidance of doubt, the terms "increased", "increase", "improve" or "activate" mean an increase of at least 10% compared to a baseline level, for example, an increase of at least 20% , or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or more up to and including an increase of 100% or any increase between 10 and 100% compared to a baseline level, or at least about 2-fold, or at least about 3-fold, or at least about 4-fold, or at least about a 5-fold increase, or at least about a 10-fold increase, or any 2- to 10-fold increase or more compared to a baseline level.

[0164] Certain methods, compositions, and agents contemplated herein modulate an immune response. In settings of decreased immune response ( eg, inhibition of an immunosuppressive pathway, eg, via checkpoint blockade, A2aR blockade, KIR blockade, etc.), the methods, compositions, and agents contemplated herein can decrease the immune response by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or up to 100% compared to a baseline level ( eg, an objective measure of immune response prior to employing the method, composition, and/or agent). In contexts of augmentation of an immune response ( eg, activation of a T-cell costimulatory signal), the methods, compositions, and agents contemplated herein can augment the immune response by at least about 10%. %, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or as much as 100%, at least about 2 times, or at least about 3 times, or at least about 4 times times, or at least about a 5-fold or at least a 10-fold increase, or any 2- to 10-fold increase or more compared to a baseline level ( eg, an objective measure of immune response before employ the method, composition and/or agent).

[0165] Certain methods, compositions, and agents contemplated herein modulate the growth, proliferation, metastasis, and invasion of malignant cells ( eg, cancer cells). In the contexts of inhibiting the growth, proliferation, metastasis, invasion, and/or survival of malignant cells (eg, cancer cells), the methods, compositions, and agents contemplated herein may decrease the growth, proliferation, metastasis, invasion, and/or or survival of malignant cells ( eg, cancer cells) by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or up to 100% in comparison with a reference level (eg, an objective measure of malignant cell growth, proliferation, metastasis, invasion, and/or survival prior to employing the method, composition, and/or agent).

[0166] The term "statistically significant" or "significantly" refers to statistical significance and generally means two standard deviations (2SD) below normal, or lower, concentration of the marker. The term refers to statistical evidence that a difference exists. It is defined as the probability of making the decision to reject the null hypothesis when the null hypothesis is actually true. The decision is often made using the p-value.

[0167] As used more particularly herein in some contexts, "modula", "modulating" and "modulating" are used interchangeably and refer to any or a combination of a decrease in glutamine metabolism, a decrease in glycolysis and a decrease in the synthesis of fatty acids. In other contexts, "modula," "modulator," and "modulation" are used interchangeably and refer to any or a combination of increased glutamine metabolism, increased glycolysis, and increased fatty acid synthesis. . In certain contexts, "modulates", "modulator" and "modulation" are used interchangeably and refer to any or a combination of an increase in oxidative phosphorylation.

[0168] Glutamine (2-amino-4-carbamoylbutanoic acid) is used by the cell for both bioenergetic and biosynthetic needs. Glutamine can be used as an amino acid for protein synthesis, as a carbon source, or as the main nitrogen donor for multiple essential biosynthetic reactions in the cell. Once taken up by the cell, mitochondrial glutaminase converts much of the glutamine to glutamate. Both glutamine and glutamate contribute to anabolic metabolism; glutamine supplies nitrogen for the synthesis of nucleotides and hexosamine, while glutamate is the nitrogen donor for the synthesis of many non-essential amino acids. Glutamate can be used to support NADPH production or be converted to the metabolic intermediates pyruvate and a-ketoglutaramate. As used herein, the term "glutamine metabolism" or "glutamine metabolic activity" refers to the chemical reactions, enzymes, and pathways that involve glutamine. As used herein, the term "glutamine metabolic pathway" is a biochemical pathway involving glutamine.

[0169] As one skilled in the art can imagine, a metabolic reprogramming agent can modulate any of the chemical reactions, enzymes and/or pathways that involve glutamine. In some embodiments, at least one metabolic reprogramming agent can modulate chemical reactions, enzymes, and/or pathways that do not directly involve glutamine, such as the conversion of pyruvate to acetyl CoA or the citric acid cycle, but indirectly affect either of chemical reactions, enzymes and/or pathways involving glutamine. Certain methods, compositions, and metabolic reprogramming agents contemplated herein reduce glutamine metabolism in cells. In the context of decreasing glutamine metabolism in cells, the methods, compositions, and agents contemplated herein can decrease glutamine metabolism in cells by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or as much as 100% compared to a reference level ( eg, an objective measure of glutamine metabolic activity prior to employing the method , the composition and/or the agent).

[0170] In some embodiments described herein, at least one metabolic reprogramming agent is a glutamine antagonist (ie, an agent that reduces glutamine metabolism). As used herein, the term "glutamine antagonist" refers to an agent that blocks or interferes with the synthesis or use of glutamine in a cell, and preferably in a cell that is part of a living organism. When the glutamine antagonist is said to interfere with glutamine synthesis, it is meant that the antagonist acts to reduce the amount or rate of glutamine synthesis to less than the amount or rate that would be experienced in the absence of the glutamine antagonist. When the glutamine antagonist is said to interfere with glutamine use, it is meant that the antagonist acts to inhibit or block a metabolic pathway downstream of glutamine, i.e., a pathway in which glutamine acts as a precursor. of one or more glutamine compounds, or that the antagonist acts to deplete glutamine in a cell or organism by reacting glutamine to form a non-glutamine product, or by reversibly or irreversibly binding glutamine to reduce its availability .

[0171] In some embodiments described herein, at least one metabolic reprogramming agent of the presently disclosed subject matter may be a glutamine analog that interferes with a glutamine metabolic pathway, an antagonist that inhibits glutamine synthesis, a glutamine-depleting enzyme, a compound that reacts with glutamine under intracellular conditions to form a non-glutamine product, an antagonist that inhibits glutamine uptake by cells, an agent that inhibits glutamine oxidation, an agent that that inhibits a glutamine transporter, an agent that inhibits glutaminolysis (a series of biochemical reactions by which glutamine is lysed to glutamate, aspartate, carbon dioxide, pyruvate, lactate, alanine, and/or citrate), or a glutamine compound glutamine binding which reduces the biological availability of glutamine. It should be recognized that a compound that is a useful metabolic reprogramming agent may have two or more of these characteristics. For example, a compound that is a glutamine analog that interferes with a glutamine metabolic pathway could also act as an antagonist that inhibits glutamine synthesis.

[0172] In some embodiments described herein, at least one metabolic reprogramming agent may be an antagonist that inhibits glutamine synthesis. Examples of compounds having this activity include glutamine synthase inhibitors (EC 6.3.1.2), such as L-methionine-DL-sulfoximine and phosphinothricin; glutamate synthase inhibitors (EC 1,4,1,13); amidophosphoribosyltransferase inhibitors (EC 2,4,2,14); and inhibitors glutamate dehydrogenase; and mixtures of any two or more of these.

[0173] In some embodiments described herein, at least one metabolic reprogramming agent may be a glutamine-depleting enzyme. Examples of such enzymes include carbamoyl-phosphate synthase (EC6.3,5,5), glutamine-pyruvate transaminase (EC 2,6,1,15), glutamine-tRNA ligase (EC6,1,1,18), glutaminase (EC3,5,1,2), D-glutaminase (EC3,5,1,35), glutamine N-acyltransferase (EC2,3,1,68), glutaminase-asparaginase (particularly Pseudomonas glutaminase-asparaginase 7a and Acinatobacter sp.), and mixtures of two or more of these.

[0174] In some embodiments described herein, at least one metabolic reprogramming agent may be a compound that reacts with glutamine under intracellular conditions to form a non-glutamine product. An example of a compound having this property is phenylbutyrate (see Darmaun et al., Phenylbutyrate-induces glutamine depletion in human: effect on leucine metabolism, pp. E801-E807, in Glutamine Depletion and Protein Catabolism, Am. Physiol. Soc. (1998)). Another example of a glutamine antagonist having this feature is phenylacetate (see US Patent No. 6,362,226).

[0175] In some embodiments described herein, at least one metabolic reprogramming agent may be an antagonist that inhibits glutamine uptake by cells. Examples of compounds having this property include alpha-methylaminoisobutyric acid (inhibits the plasma membrane glutamine transporter GynT; See, Varoqui et al., J. Biol. Chem., 275(6):4049-4054 (2000), wortmannin and LY-294002 (inhibits the hepatic glutamine transporter; See, Pawlik et al., Am. J. Physiol. Gastrointest. Liver Physiol., 278:G532-G541 (2000)).

[0176] In some embodiments described herein, at least one metabolic reprogramming agent may be a glutamine-binding compound that reduces the biological availability of glutamine.

[0177] In some embodiments described herein, at least one metabolic reprogramming agent may be a glutamine analog that interferes with a glutamine metabolic pathway (eg. Examples of compounds that may act in this manner include acivicin ( L-(alpha S,5S)-alpha-amino-3-chloro-4,5-dihydro-5-isoxazolacetic acid), DON (6-diazo-5-oxo-L-norleucine), azaserine, azotomycin, chloroketone ( L-2-amino-4-oxo-5-chloropentanoic acid), N3-(4-methoxyfumaroyl)-L-2,3-diaminopropanoic acid (FMDP) (inactivates glucosamine-6-phosphate synthase (EC 2.6, 1,16), See, Zgódka et al., Microbiology, 147:1955 -1959 (2001)), (3S,4R)-3,4-dimethyl-L-glutamine, (3S,4R)-3 acid ,4-dimethyl-L-pyroglutamic acid (See, Acevedo et al., Tetrahedron., 57:6353-6359 (2001)), 1,5-N,N'-disubstituted-2-(substituted benzenesulfonyl) glutamamides (See, Srikanth et al., Bioorganic and Medicinal Chemistry, (2002)), or a mixture of two or more of these.In some embodiments, at least a metabolic reprogramming agent is selected from the group consisting of acivicin (L-(alpha S, 5S)-alpha-amino-3-chloro-4,5-dihydro-5-isoxazolacetic acid), azaserin, 6-diazo-5 -oxo-norleucine (DON) and 5-diazo-4-oxo-L-norvaline (L-DONV).

[0178] In some embodiments described herein, at least one metabolic reprogramming agent is a prodrug of a glutamine analog that interferes with a glutamine metabolic pathway ( eg, decreases glutamine metabolism/activity). metabolism of glutamine). In some embodiments, at least one metabolic reprogramming agent is a prodrug of acivicin (L-(alpha S, 5S)-alpha-amino-3-chloro-4,5-dihydro-5-isoxazolacetic acid), azaserin, 6-diazo-5-oxo-norleucine (DON) and 5-diazo-4-oxo-L-norvaline (L-DONV). Illustrative suitable prodrugs of acivicin, azaserin, DON and L-DONV can be found in "Prodrugs of Glutamine Analogs" (Attorney Docket No. 111232-00403, filed at the same time, now published as WO 2017/023774 A1).

[0179] Glycolysis is the metabolic pathway that converts glucose to pyruvate with the simultaneous production of ATP. Pyruvate is a metabolic intermediate that can then enter the tricarboxylic acid (TCA) cycle within mitochondria to produce NADH and FADH2. The first step of glycolysis is the phosphorylation of glucose by hexokinase to form glucose 6-phosphate.

[0180] In some embodiments described herein, at least one metabolic reprogramming agent can modulate any of the chemical reactions and/or enzymes involved in glycolysis. In some embodiments described herein, at least one metabolic reprogramming agent can modulate chemical reactions, enzymes, and/or pathways that do not directly involve glycolysis, but indirectly affect any of the chemical reactions, enzymes, and/or pathways that involve glycolysis. Certain methods, compositions, and metabolic reprogramming agents contemplated herein reduce glycolysis in cells. In the context of decreasing glycolysis in cells, the methods, compositions, and agents contemplated herein can decrease glycolysis in cells by at least 10%, 20%, 30%, 40%, 50 %, 60%, 70%, 80%, 90%, or as much as 100% compared to a reference level ( eg, an objective measure of glycolytic metabolic activity prior to use of the method, composition, and/or or the agent). As used herein, the term "glycolytic metabolic activity" refers to chemical reactions and enzymes that involve the glycolysis pathway.

[0181] In some embodiments described herein, at least one metabolic reprogramming agent of the presently disclosed subject may be an agent that interferes with glycolysis or a related pathway. that affects glycolysis; an agent that inhibits the synthesis of pyruvate and/or one of the intermediate products of glycolysis; an agent that inhibits one or more of the enzymes involved in glycolysis, such as hexokinase, phosphoglucose isomerase, phosphofructokinase, fructose bisphosphate aldolase, triophosphate isomerase, glyceraldehyde phosphate dehydrogenase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, and/or pyruvate kinase; an agent that depletes glucose-6-phosphate, one of the rate-limiting products of glycolysis; an agent that inhibits the uptake and/or transport of glucose across the plasma membrane by cells; or a glucose binding compound that reduces the biological availability of glucose. It should be recognized that a compound that is a useful metabolic reprogramming agent may have two or more of these characteristics.

[0182] In some embodiments described herein, at least one metabolic reprogramming agent interferes or inhibits hexokinase expression and/or activity. Examples of hexokinase inhibitors include, but are not limited to 2-deoxyglucose (2-DG), 3-bromopyruvate (3-BrPA), lonidamine (LND), sodium fluoride, and potassium fluoride. In some embodiments, at least one metabolic reprogramming agent is 2-deoxy-D-glucose (2-DG).

[0183] Fatty acid synthesis is the process in the cell that creates fatty acids from acetyl-CoA and malonyl-CoA precursors. Fatty acid oxidation is the process by which fatty acid molecules are broken down in mitochondria to generate acetyl-CoA, which enters the citric acid cycle, and NADH and FADH2, which are used in the transport chain of electrons. The enzyme AMP-activated protein kinase (AMPK) plays a role in cellular energy homeostasis and is a stimulator of fatty acid oxidation.

[0184] In some embodiments described herein, at least one metabolic reprogramming agent can modulate any of the chemical reactions and/or enzymes involved in fatty acid synthesis and/or fatty acid oxidation. In some embodiments described herein, at least one metabolic reprogramming agent can modulate chemical reactions, enzymes, and/or pathways that do not directly involve fatty acid synthesis and/or fatty acid oxidation, but indirectly affect either of chemical reactions, enzymes and/or pathways that involve the synthesis of fatty acids and/or the oxidation of fatty acids.

[0185] Certain methods, compositions, and metabolic reprogramming agents contemplated herein reduce fatty acid synthesis and/or increase fatty acid oxidation in cells. In the context of decreasing fatty acid synthesis and/or increasing fatty acid oxidation in cells, the methods, compositions, and agents contemplated herein may decrease fatty acid synthesis and/or increase fatty acid oxidation. in cells by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or as much as 100% compared to a reference level ( eg. ., an objective measure of fat synthesis prior to employing the method, composition, and/or agent).

[0186] In some embodiments described herein, at least one metabolic reprogramming agent of the presently disclosed subject matter may be an agent that interferes with fatty acid synthesis and/or fatty acid oxidation or a related pathway that affects fatty acid synthesis and/or fatty acid oxidation. acid oxidation; an agent that increases the oxidation of fatty acids; an agent that increases one or more of the fatty acid oxidation products; an agent that increases the expression and/or activity of one or more of the enzymes involved in the oxidation of fatty acids, such as acyl CoA dehydrogenase, enoyl CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase and p-ketothiolase; an agent that increases the expression and/or activity of AMP-activated protein kinase (AMPK); an agent that increases the uptake and/or transfer of activated fatty acids across the mitochondrial membrane; and an agent that increases the expression and/or activity of enzymes involved in the uptake and/or transfer of activated fatty acids across the mitochondrial membrane. It should be recognized that a compound that is a useful metabolic reprogramming agent may have two or more of these characteristics. In some embodiments, at least one metabolic reprogramming agent is an activator of 5' AMP-activated protein kinase (AMPK) activity.

[0187] At least one metabolic reprogramming agent that is an activator of AMPK activity can be an agent that increases AMP concentrations in the cell; an AMP analogue, such as 5-amino-4-imidazolecarboxamide ribotide (ZMP); an agent that increases AMPK phosphorylation, such as an agent that increases the expression and/or activity of a kinase that can phosphorylate AMPK; and an agent that is an AMPK allosteric modulator, such as one that can modify AMPK to make it a better substrate for a kinase that can phosphorylate AMPK.

[0188] In some embodiments described herein, at least one metabolic reprogramming agent is metformin.

[0189] It should be appreciated that modulation of glutamine metabolism, glycolysis and fatty acid metabolism may result in modulation of one or more genes or gene expression products or biosynthesis or degradation of one or more enzymes.

[0190] The term "expression" means the process by which information from a gene or nucleic acid ( eg, DNA) is used in the synthesis of gene products ( eg, mRNA, RNA and/or proteins) and includes, but is not limited to, one or more of the steps of replication, transcription, and translation. Expression steps that can be modulated by agents contemplated herein may include, for example, transcription, splicing, translation, and post-translational modification of a protein. Those of skill in the art will appreciate that the method for modulating any particular protein may depend on the type of protein ( eg, protein kinase, transcriptional regulator, enzyme, etc.), its function ( eg, transcriptional regulation, catalysis, phosphorylation, signal transduction, etc.), and their subcellular location (eg, extracellular space, cytoplasm, nucleus, membrane, etc.). Those skilled in the art will readily appreciate the appropriate agents to use for modulation depending on the particular context ( eg, protein type, biological function, subcellular localization, composition, method of use, mode of inhibition, etc.). For example, an agent can be used to inhibit the enzymatic activity of an enzyme ( eg, at least one metabolic reprogramming agent that inhibits glutaminase-catalyzed glutaminolysis ( eg, a glutamine antagonist), by least one metabolic reprogramming agent that inhibits glycolysis catalyzed in part by hexokinase ( eg, 2-DG), etc.), inhibits the phosphorylation level or activity of a protein kinase, inhibits transcriptional activation, or a signaling pathway.

[0191] The metabolic reprogramming agents, chemotherapeutic agents, cytotoxic agents, immunotherapeutic agents, immunosuppressive agents, radiotherapeutic agents, anti-inflammatory agents, and neuroprotective agents described herein can be any type of agent. Exemplary types of agents that can be used as such agents in the methods, compositions, and uses described herein include small inorganic or organic molecules; saccharides; oligosaccharides; polysaccharides; a biological macromolecule selected from the group consisting of peptides, proteins, peptide analogues, and derivatives; peptidomimetics; nucleic acids selected from the group consisting of siRNA, shRNA, antisense RNA, ribozymes, dendrimers, and aptamers; an extract made from biological materials selected from the group consisting of bacteria, plants, fungi, animal cells and animal tissues; natural or synthetic compositions; microcarrier or nanocarrier consisting of one or more polymers, proteins, nucleic acids, lips, or metals; and any combination thereof.

[0192] As used herein, the term "small molecule" can refer to agents that are "natural product-like", however, the term "small molecule" is not limited to "natural product-like" agents. . Rather, a small molecule is typically characterized as containing multiple carbon-carbon bonds and having a molecular weight of less than 5,000 Daltons (5 kD), preferably less than 3 kD, even more preferably less than 2 kD, and most preferably less of 1kD. In some cases it is preferred that a small molecule have a molecular weight of 700 Daltons or less.

[0193] As used herein, an "RNA interference molecule" refers to an agent that interferes or inhibits the expression of a target gene or genomic sequence by RNA interference (RNAi). Such Rn interference agents include, but are not limited to, nucleic acid molecules including RNA molecules that are homologous to the target gene or genomic sequence, or a fragment thereof, short interference RNA (siRNA), short hairpin RNA or small hairpin (shRNA), microRNA (miRNA) and small molecules that interfere or inhibit the expression of a target gene by RNA interference (RNAi).

[0194] The term "polynucleotide" is used herein interchangeably with "nucleic acid" to indicate a polymer of nucleosides. Typically, a polynucleotide is composed of nucleosides naturally found in DNA or RNA ( eg, adenosine, thymidine, guanosine, cytidine, uridine, deoxyadenosine, deoxythymidine, deoxyguanosine, and deoxycytidine) linked by phosphodiester bonds. However, the term encompasses molecules comprising nucleosides or nucleoside analogs containing chemically or biologically modified bases, modified backbones, etc., whether or not they are found in natural nucleic acids, and such molecules may be preferred for certain applications. When this application refers to a polynucleotide, it is understood that both DNA and RNA are provided and, in each case, both single-stranded and double-stranded forms (and complements of each single-stranded molecule). Polynucleotide sequence, as used herein, can refer to the polynucleotide material itself and/or to the sequence information ( eg, the sequence of letters used as abbreviations for bases) that biochemically characterizes a specific nucleic acid. direction 'to 3' unless otherwise stated.

[0195] Nucleic acid molecules that modulate metabolic pathways or targets described herein can, in some embodiments, be inserted into vectors and used as gene therapy vectors. Gene therapy vectors can be administered to a subject by, for example, intravenous injection, local administration (see US Patent No. 5,328,470), or by stereotactic injection (see, for example, Chen et al. Proc Natl Acad Sci USA 91:3054-3057, 1994). The gene therapy vector pharmaceutical preparation may include the gene therapy vector in an acceptable diluent, or may comprise a slow release matrix into which the gene delivery vehicle is embedded. If the complete gene delivery vector can be produced intact from recombinant cells, eg, retroviral vectors, the pharmaceutical preparation can include one or more cells that produce the gene delivery system.

[0196] The term "polypeptide" as used herein refers to a polymer of amino acids. The terms "protein" and "polypeptide" are used interchangeably in this document. A peptide is a relatively short polypeptide, typically between about 2 and 60 amino acids in length. The polypeptides used herein typically contain amino acids, such as the 20 L-amino acids most commonly found in proteins. However, other amino acids and/or amino acid analogues known in the art may be used. One or more of the amino acids in a polypeptide can be modified, for example, by the addition of a chemical entity, such as a carbohydrate group, a phosphate group, a fatty acid group, a linker for conjugation, functionalization, etc. polypeptide having a non-polypeptide moiety covalently or non-covalently associated therewith is still considered a "polypeptide". Exemplary modifications include glycosylation and palmitoylation. Polypeptides can be purified from natural sources, produced using recombinant DNA technology, synthesized by chemical means, such as conventional solid phase peptide synthesis, etc. itself and/or sequence information ( eg, the sequence of letters or three-letter codes used as abbreviations for amino acid names) that biochemically characterizes a polypeptide. A polypeptide sequence presented herein is presented in an N-terminal to C-terminal direction unless otherwise indicated.

[0197] The term "identity" as used herein refers to the degree to which the sequence of two or more nucleic acids or polypeptides is the same. The percent identity between a sequence of interest and a second sequence over an evaluation window, for example, over the length of the sequence of interest, can be calculated by aligning the sequences, determining the number of residues (nucleotides or amino acids) within the window of evaluation found against an identical residue that allows for the introduction of gaps to maximize identity, dividing by the total number of residues from the sequence of interest or the second sequence (whichever is greater) that lie within the window, and multiplying by 100. When calculating the number of identical residues required to achieve a particular percent identity, fractions should be rounded to the nearest whole number. Percent identity can be calculated using a variety of computer programs known in the art. For example, computer programs such as BLAST2, BLASTN, BLASTP, Gapped BLAST, etc., generate alignments and provide percent identity between sequences of interest. The Karlin and Altschul algorithm (Karlin and Altschul, Proc. Natl. Acad. Sci. USA 87:22264-2268, 1990) modified as in Karlin and Altschul, Proc. Natl. Acad. Sci. USA 90:5873-5877, 1993 is incorporated into the NBLAST and XBLAST programs of Altschul et al. (Altschul et al., J. MoT Biol. 215:403-410, 1990). To obtain gapped alignments for comparison purposes, Gapped BLAST is used as described in Altschul et al. (Altschul et al. Nucleic Acids Res. 25: 3389-3402, 1997). When using the BLAST and Gapped BLAST programs, the default parameters of the respective programs can be used. A PAM250 or BLOSUM62 matrix can be used. Software for performing BLAST analysis is publicly available through the National Center for Biotechnology Information (NCBI). Check the website that has URL www.ncbi.nlm.nih.gov for these programs. In a specific embodiment, percent identity is calculated using BLAST2 with default parameters provided by NCBI.

[0198] Generally, at least one metabolic reprogramming agent described herein can be used in combination with an additional therapeutic agent ( eg, a pharmaceutically active agent, eg, a drug approved by a regulatory agency). . The therapeutic agent may act synergistically with the agent described herein, or they may independently serve their purpose. effects. The description contemplates any therapeutic agent that one of ordinary skill in the art would use in connection with a method, use, or composition described herein. Examples of therapeutic agents contemplated for use in the presently disclosed methods, uses, and compositions in combination with metabolic reprogramming agents include, but are not limited to, chemotherapeutic agents/chemotherapy, immunotherapeutic agents/immunotherapy, immunosuppressive agents, anti-inflammatory agents, neuroprotective agents, neuroregenerative agents, neurotrophic factors, radiotherapeutic agents/radiotherapy, proton therapy, photodynamic therapy, and stem and progenitor cells used to replace and/or repair endogenous populations of abnormal, harmful, or unhealthy cells.

[0199] Chemotherapy and chemotherapeutic agent are used herein synonymously. A "chemotherapeutic agent" is used to connote a compound or composition that is administered in the treatment of cancer. Chemotherapeutic agents contemplated for use in combination with at least one metabolic reprogramming agent, at least two metabolic reprogramming agents, or at least three metabolic reprogramming agents described herein include, but are not limited to alkylating agents, such as thiotepa and cyclophosphamide; alkyl sulfonates, such as busulfan, improsulfan, and piposulfan; aziridines, such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolomelamine; nitrogen mustards, such as chlorambucil, chlornaphazine, colophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembiquine, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas, such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics, such as aclacinomycin, actinomycin, autramycin, azaserin, bleomycin, cactinomycin, calicheamicin, carabicin, cadomycin, carzinophyllin, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucin, doxorubicin, epirubicin, esorubicin, idarubicin, marcelomycin, mitomycins, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, chelamycin, rhodorubicin, streptongrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites, such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues, such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues, such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues, such as ancitabine, azacitidine, 6-azauridine, carmofur, cytosine arabinoside, dideoxyuridine, doxifluridine, enocitabine, floxuridine, 5-FU; androgens, such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; antiadrenals, such as aminoglutethimide, mitotane, trilostane; folic acid replenishers, such as folinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisanthrene; edatraxate; defamine; demecolcine; Diaziquone; elformitin; elliptinium acetate; ethoglycide; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; freak; pyrarubicin; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK; razoxane; sizofuran; spirogermanium; tenuazonic acid; triazicuone; 2,2',2"trichlorotriethylamine;urethane;vindesine;dacarbazine;mannomustine;mitobronitol;mitolactol;pipobroman;gacytosine; arabinoside (Ara-C); taxoids, eg, paclitaxel and docetaxel; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine ; platinum analogues, such as cisplatin and carboplatin; vinblastine; platinum; etoposide; ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT11; topoisomerase inhibitor RFS 2,000; difluoromethylornithine, retinoic acid, esperamicins, capecitabine, and pharmaceutically acceptable salts, acids, or derivatives of any of the foregoing Chemotherapeutic agents also include antihormonal agents that act to regulate or inhibit hormonal action on tumors, such as antiestrogens, including, for example , tamoxifen, raloxifene, 4(5)-imidazoles that inhibit aromatase, 4-hydroxytamoxifen, trioxifen, keoxifene, LY117 018, onapristone and toremifene (Fareston); and anti-androgens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptable salts, acids or derivatives of any of the foregoing.

[0200] In some embodiments described herein, the chemotherapeutic agent is a topoisomerase inhibitor. Topoisomerase inhibitors are chemotherapy agents that interfere with the action of a topoisomerase enzyme ( eg, topoisomerase I or II). Topoisomerase inhibitors include, but are not limited to, doxorubicin HCl, daunorubicin citrate, mitoxantrone HCl, actinomycin D, etoposide, topotecan HCl, teniposide, and irinotecan, as well as pharmaceutically acceptable salts, acids, or derivatives of any of these.

[0201] In some embodiments described herein, the chemotherapeutic agent is an antimetabolite. An antimetabolite is a chemical substance similar in structure to a metabolite required for normal biochemical reactions, but different enough to interfere with one or more normal cell functions, such as cell division. Antimetabolites include, but are not limited to, gemcitabine, fluorouracil, capecitabine, methotrexate sodium, ralitrexed, pemetrexed, tegafur, cytosine arabinoside, thioguanine, 5-azacitidine, 6-mercaptopurine, azathioprine, 6-thioguanine, pentostatin, fludarabine phosphate, and cladribine , as well as pharmaceutically acceptable salts, acids or derivatives of any of these.

[0202] In certain embodiments described herein, the chemotherapeutic agent is an antimitotic agent including, but not limited to, tubulin-binding agents. In some embodiments, the agent is a taxane. In certain embodiments described herein, the agent is paclitaxel or docetaxel, or a pharmaceutically acceptable salt, acid, or derivative of paclitaxel or docetaxel. In certain alternate embodiments, the antimitotic agent comprises a vinca alkaloid, such as vincristine, binblastine, vinorelbine, or vindesine, or pharmaceutically acceptable salts, acids, or derivatives thereof.

[0203] As used herein, the term "immunotherapeutic agent" refers to a molecule that can aid in the treatment of a disease by inducing, potentiating, or suppressing an immune response in a cell, tissue, organ, or subject. Examples of immunotherapeutic agents contemplated for use in combination with at least one metabolic reprogramming agent, at least two metabolic reprogramming agents, or at least three metabolic reprogramming agents described herein include, but are not limited to, dotted molecules. immune control cells ( eg, antibodies to immune checkpoint proteins), interleukins ( eg, IL-2, IL-7, IL-12, IL-15), cytokines ( eg, interferons , G-CSF, imiquimod), chemokines ( eg, CCL3, CCL26, CXCL7), vaccines ( eg, peptide vaccines, dendritic cell (DC) vaccines), EGFRvIII vaccines, mesotilin vaccine, G-VAX , listeria vaccines) and adoptive T cell therapy including chimeric antigen receptor T cells (CAR T cells).

[0204] As used herein, "immunosuppressive agent" means an agent that can be used in immunotherapy to reduce or prevent an immune response in a cell, organ, tissue, or subject. Examples of immunosuppressive agents contemplated for use in combination with at least one metabolic reprogramming agent, at least two metabolic reprogramming agents, or at least three metabolic reprogramming agents include, without limitation, corticosteroids, calcineurin inhibitors, antiproliferative agents, agonists of SIP receptor, kinase inhibitors, monoclonal antilymphocyte antibodies, and polyclonal antilymphocyte antibodies. Non-limiting examples of corticosteroids include prednisone (Deltasone® and Orasone®) and methylprednisolone (SoluMedrol®). Non-limiting examples of calcineurin inhibitors include cyclosporine (cyclosporine A, SangCya, Sandimmune ® , Neoral ® , Gengraf ® ), ISA, Tx247, ABT-281, ASM 981, and tacrolimus (Prograf ® , FK506). Non-limiting examples of antiproliferative agents include mycophenolate mofetil (CellCept®), azathioprene (Imuran®), and sirolimus (Rapamune®). Non-limiting examples of SIP receptor agonists include FTY 720 or analogs thereof. Non-limiting examples of kinase inhibitors include mTOR kinase inhibitors, which are compounds, proteins, or antibodies that target, decrease, or inhibit the activity and/or function of members of the serine/threonine mTOR family. These include, without limitation, CCI-779, ABT578, SAR543, Rapamycin, and derivatives or analogs thereof, including 40-O-(2-hydroxyethyl)-rapamycin, Rapalogs, including AP23573, AP23464, AP23675, and AP23841 from Ariad, Everolimus (CERTICAN, RAD001), biolimus 7, biolimus 9, and sirolimus (RAPAMUNE). Kinase inhibitors also include protein kinase C inhibitors, including the compounds described in PCT publications WO 2005/097108 and WO 2005/068455. Non-limiting examples of antilymphocyte monoclonal antibodies include Muromonab-CD3 (Orthoclone OKT3®), Interleukin-2 Receptor Antagonist (Basiliximab, Simulect®), and Daclizumab (Zenapax®). Non-limiting examples of polyclonal antilymphocyte antibodies include equine antithymocyte globulin (Atgam®) and rabbit antithymocyte globulin (RATG, Thymoglobulin®). Other immunosuppressants include, without limitation, SERP-1, a serine protease inhibitor produced by malignant rabbit fibroma virus (MRV) and myxoma virus (MYX), described in US Patent Publication No. 2004/0029801.

[0205] Immunosuppressive agents can be classified according to their specific molecular mode of action. The four main categories of immunosuppressive drugs currently used in the treatment of patients with organ transplants are as follows. Calcineurin inhibitors inhibit T cell activation, thus preventing T cells from attacking the transplanted organ. Azathioprines disrupt the synthesis of DNA and RNA, as well as the process of cell division. Monoclonal antibodies inhibit the binding of interleukin-2, which in turn slows the production of T cells in the patient's immune system. Corticosteroids suppress the inflammation associated with transplant rejection.

[0206] Immunosuppressants can also be classified according to the specific organ being transplanted. Basiliximab (Simulect) is also used in combination with other drugs such as cyclosporine and corticosteroids in kidney transplants. IL-2 blockers, including Novartis Simulect, FK506 or CyA, MMF, prednisone, or rapamycin are also used in kidney transplants. Daclizumab (Zenapax) is also used in combination with other drugs such as cyclosporine and corticosteroids in kidney transplants. Similar drugs are used in heart transplants, but antilymphocyte globulin (ALG) is often used instead of Simulect. Muromonab CD3 (Orthoclone OKT3) is used together with cyclosporine in kidney, liver, and heart transplants. Tacrolimus (Prograf) is used in liver and kidney transplants. He is being studied for bone marrow, heart, pancreas, pancreatic islet, and small intestine transplantation.

[0207] As used herein, "photodynamic therapy," also known as photoradiation therapy, phototherapy, and photochemotherapy, refers to a treatment that uses photosensitizing agents in combination with light to kill cancer cells. Photosensitizing agents kill cancer cells by activating light.

[0208] As used herein, "proton therapy", also known as proton beam therapy, refers to a treatment that uses a beam of protons to irradiate and kill cancer cells.

[0209] As used herein, "anti-inflammatory agent" refers to an agent that can be used to prevent or reduce an inflammatory response or inflammation in a cell, tissue, organ, or subject. Examples of anti-inflammatory agents contemplated for use in combination with at least one metabolic reprogramming agent, at least two metabolic reprogramming agents, or at least three metabolic reprogramming agents include, without limitation, steroidal anti-inflammatory agents, a nonsteroidal anti-inflammatory agent, or a combination thereof. In some embodiments, anti-inflammatory agents include clobetasol, alclofenac, alclomethasone dipropionate, algestone acetonide, alpha amylase, amcinafal, amcinafide, amfenac sodium, amiprilose hydrochloride, anakinra, anirolac, anitrazafen, apazone, balsalazide disodium, bendazac, benoxaprofen , benzydamine hydrochloride, bromelains, broperamole, budesonide, carprofen, cycloprofen, cintazone, cliprofen, clobetasol propionate, clobetasone butyrate, clopirac, cloticasone propionate, cormethasone acetate, cortodoxone, deflazacort, desonide, desoximetasone, dexamethasone, dexamethasone acetate , dexamethasone dipropionate, diclofenac potassium, diclofenac sodium, diflorasone diacetate, diluminidone sodium, diflunisal, difluprednate, diphthalone, dimethyl sulfoxide, drocinonide, endrysone, enlimomab, enolicam sodium, epirizol, etodolac, etofenamate, felbinac, fenamol, fenbufenacfen , fenclorac, fendosal, fenpipalone, fentiazac, flazalone, fluazacort, flufenamic acid, flumizole, flunisolide acetate, flunixin, flunixin meglumine, fluocortin butyl, fluorometholone acetate, fluquazone, flurbiprofen, fluretofen, fluticasone propionate, furaprofen, furobufen, halcinonide, halobetasol propionate, halopredone acetate, ibufenac, ibuprofen, ibuprofen aluminum, ibuprofen piconol, ilonidap, indomethacin, indomethacin sodium, indoprofen, indoxol, intrazole, isoflupredone acetate, isoxepac, isoxicam, ketoprofen, lofemizol hydrochloride, lomoxicam, loteprednol etabonate, meclofenamate sodium, meclofenamic acid, dibutyrate mechlorisone, mefenamic acid, mesalamine, meseclazone, methylprednisolone suleptanate, momiflumate, nabumetone, naproxen, naproxen sodium, naproxol, nimazine, olsaline sodium, orgotein, orpanoxin, oxaprozin, oxyphenbutazone, paraniline hydrochloride, pentosan polysul sodium, fenbutazone glycerate sodium , pirfenidone, piroxicam, piroxicam cinnamate, piroxy cam olamine, pirprofen, prednazate, prifelone, prodolic acid, proquazone, proxazole, proxazole citrate, rimexolone, romazarit, salcolex, salnacedin, salsalate, sanguinarine chloride, seclazone, sermethacin, sudoxicam, sulindac, suprofen, talmethacin, talniflumate, thallosalate, tebufelone, tenidap, tenidap sodium, tenoxicam, tesicam, tesimide, tetridamine, thiopinac, tixocortol pivalate, tolmetin, tolmetin sodium, triclonide, triflumidate, zidomethacin, zomepirac sodium, aspirin (acetylsalicylic acid), salicylic acid, corticosteroids, glucocorticoids, tacrolimus , pimecorlimus, prodrugs thereof, codrugs thereof, and combinations thereof. The anti-inflammatory agent can also be a biological inhibitor of proinflammatory signaling molecules including antibodies against such biological inflammatory signaling molecules.

[0210] Examples of neuroprotective agents include, without limitation, L-dopa, dopamine agonists ( eg, apomorphine, bromocriptine, pergolide, ropinirole, pramipexole, or cabergoline), adenosine A2a antagonists (Shah et al., Curr Opinion Drug Discov Dev 13:466-80 (2010)); serotonin receptor agonists; continuous release levodopa (Sinemet CR.RTM., MSD, Israel); continuous administration of duodenal levodopa (Duodopa.RTM., Abbott, UK); catechol-O-methyltransferase (COMT) inhibitors ( eg, Stalevo.RTM., Novartis Pharma, USA; entacapone (Comtan.RTM., Novartis Pharma, USA)); tolcapone; coenzyme Q10 and/or MAO-B inhibitors ( eg selegiline or rasagiline). Additional neuroprotective agents are described, for example, in Hart et al., Mov. disorder. 24: 647-54 (2009).

[0211] As used herein, a "radiotherapeutic agent" refers to agents conventionally adopted in the therapeutic field of cancer treatment and includes photons that have sufficient energy for the ionization of chemical bonds, such as alpha (a), beta (p), and gamma ( ^y ) rays from radioactive nuclei, as well as X. The radiation can be high LET (linear energy transfer) or low LET. LET is the energy transferred per unit length of the distance. A high LET is said to be heavily ionizing radiation and a low LET is said to be poorly ionizing radiation. Representative examples of high LET are neutrons and alpha particles. Representative examples of low LET are X-rays and gamma rays. Low-LET radiation, which includes X-rays ^and y-rays, is most commonly used for radiation therapy of cancer patients. Radiation can be used for external radiation therapy, which is usually given on an outpatient basis, or for internal radiation therapy, which uses radiation that is placed very close to or inside the tumor. For internal radiation therapy, the radiation source is usually sealed in a small holder called an implant. Implants can be in the form of thin wires, plastic tubes called catheters, tapes, capsules, or seeds. The implant is placed directly into the body. Internal radiation therapy may require hospitalization. The ionizing radiation source is provided as a unit dose of radiation and is preferably an X-ray tube as it offers many advantages, such as convenient adjustable dosage where the source can be easily turned on and off, minimal disposal problems. A unit dose of radiation is usually measured in gray (Gy). The source of ionizing radiation may also comprise a radioisotope, such as a solid source of radioisotopes ( eg, wire, strip, pellet, seed, bead), or a balloon filled with liquid radioisotopes. In the latter case, the balloon has been specially configured to prevent leakage of the radioisotopic material from the balloon into the body or into the bloodstream. Still further, the source of ionizing radiation may comprise a receptacle in the catheter body for receiving radioisotopic materials as pellets or liquids. The radioisotope material can be selected to emit a, p, and y. Generally, a and p radiations are preferred as they can be rapidly absorbed by surrounding tissue and will not penetrate substantially beyond the lumen wall of the body being treated. Consequently, incidental irradiation of the heart and other organs adjacent to the treatment region can be substantially eliminated. The total number of units provided will be an amount determined to be therapeutically effective by one skilled in treatment using ionizing radiation. This amount will vary with the subject and the type of malignancy or neoplasm being treated. The amount can vary, but a patient may receive a dose of around 30 to 75 Gy for several weeks.

[0212] Examples of radiotherapeutic agents contemplated for use in combination with at least one metabolic reprogramming agent, at least two metabolic reprogramming agents, or at least three metabolic reprogramming agents include factors that cause DNA damage, such as lightning ^and , X-rays and/or targeted delivery of radioisotopes to tumor cells. Other forms of DNA damaging factors such as microwaves and UV radiation are also contemplated. Dosage ranges for X-rays from daily doses of 50 to 200 roentgens for prolonged periods (3 to 4 weeks), to single doses of 2,000 to 6,000 roentgens. Dose ranges for radioisotopes vary widely and depend on the half-life of the isotope, the strength and type of radiation emitted, and absorption by the target cell. In some embodiments, the radiotherapeutic agent is selected from the group consisting of 47Sc, 67Cu, 90Y, 109Pd, 123I, 125I, 131I, 186Re, 188Re, 199Au, 211At, 212Pb, 212B, 32P and 33P, 71Ge, 77As, The PB, 105rh, 111ag, 119sb, 121Sn, 131cs, 143pr, 161TB, 177lu, 191os, 193npt, 197h, 43K, 43K, 52FE, 57CO, 67CU, 67GA, 68GA, 77BR, 81RR/81MKR, 87MSR, 99MTC, 113MIN, 113MIN 113Min, 127Cs, 129Cs, 132I, 197Hg, 203Pb, and 206Bi, as described in US Patent No. 8,946,168.

[0213] In some contexts, an agent described herein may be administered with an antigen ( eg, to induce an immune response). In some embodiments, an adjuvant may be used in combination with the antigen.

[0214] An agent described herein can also be used in combination with an imaging agent. An agent ( eg, a metabolic reprogramming agent) can be linked to imaging agents for imaging and diagnosis of various diseased organs, tissues or cell types. The agent can be labeled or conjugated to a fluorophore or radiotracer for use as an imaging agent. Many suitable imaging agents are known in the art, as are methods for their binding to agents ( eg, binding of an imaging agent to proteins or peptides using metal chelate complexes, radioisotopes, fluorescent labels). or enzymes whose presence can be detected using a colorimetric). markers (such as, but not limited to, urease, alkaline phosphatase, hydrogen peroxidase (horseradish), and glucose oxidase). An agent can also be dual-labeled with a radioisotope to combine imaging via nuclear approaches and become a single cyclic structure and optimized for binding affinity and pharmacokinetics. Such agents can be administered by any number of methods known to those skilled in the art, including, but not limited to, oral administration, inhalation, subcutaneous (sub-q), intravenous (IV), intraperitoneal (LP), intramuscular (IM), injection intrathecal or intratumoral injection. The methods, compositions, and uses described herein can be used alone or in combination with other techniques to diagnose access and control and direct therapy of metabolic reprogramming disorders. In some contexts, the imaging agent can be used to detect and/or monitor tumors or sites of metastasis in a subject. For example, an agent ( eg, a metabolic reprogramming agent) can be administered in vivo and monitored using an appropriate marker. Examples of methods for detecting and/or monitoring a labeled agent with an imaging agent in vivo include scintigraphy, positron emission tomography (PET), single photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), X-rays, computer-assisted x-ray tomography (CT), near-infrared spectroscopy, and ultrasound. These techniques provide information on the detection of neoplastic involvement, particularly of inaccessible nodes in subjects with diseases malignant Knowledge about the size of the node and the filling of the nodes can also be instructive. For example, agents or compositions targeted to lymph nodes in screening applications will contain suitable imaging or contrast agents, such as ferromagnetic materials such as iron oxide, perfluorochemicals such as perfluorooctyl bromide, or gamma-emitting radiotracers. such as technetium-99m, indium-Ill, gallium-67, thallium-201, iodine-131, 125 or 123, positron-emitting radiotracers, such as fluorine-18, or those produced by neutron activation, such as samarium -153.

[0215] Imaging agents for use herein include radioisotopes and dyes. Any conventional method under radioactive labeling that is suitable for labeling isotopes for use in vivo will generally be suitable for labeling detection agents in accordance with the disclosure. Internal screening procedures include intraoperative, intravascular, or endoscopic techniques, including laparoscopic techniques, both surgically invasive and non-invasive. For example, when a lymph node is detected, a high signal-to-background ratio should be achieved. The therapy also requires a high absolute accumulation of the therapeutic agent in the lymph node, as well as a reasonably long duration of uptake and binding.

[0216] Radioisotopes suitable for the methods of the disclosure include: Actinium-225, Astatin-211, Iodine-123, Iodine-125, Iodine-126, Iodine-131, Iodine-133, Bismuth-212, Bromine-77, Indium-111, Indium-113m, Gallium-67, Gallium-68, Ruthenium-95, Ruthenium-97, Ruthenium-103, Ruthenium-105, Mercury-107, Mercury-203, Rhenium-186, Rhenium-188, Tellurium- 121m, Tellurium-122m, Tellurium-125m, Thulium-165, Thulium-167, Thulium-168, Technetium-99m, Fluorine-18, Silver-111, Platinum-197, Palladium-109, Copper-67, Phosphorus-32, Phosphorus-33, Yttrium-90, Scandium-47, Samarium-153, Lutetium-177, Rhodium-105, Praseodymium-142, Praseodymium-143, Terbium-161, Holmium-166, Gold-199, Cobalt-57, Cobalt- 58, Chromium-51, Iron-59, Selenium-75, Thallium-201 and Ytterbium-169. The most preferred radioisotope for use in the object of the present invention is Technetium-99m. Preferably, the radioisotope will emit a particle or beam in the range of 10 to 7000 keV, more preferably in the range of 50 to 1500 keV and most preferably in the range of 80 to 250 keV.

[0217] Preferred isotopes for external imaging include: iodine-123, iodine-131, indium-111, gallium-67, ruthenium-97, technetium-99m, cobalt-57, cobalt-58, chromium-51, iron-59, selenium-75, thallium-201 and ytterbium-169. Technetium-99m is the most preferred radioisotope for external imaging in disclosure.

[0218] The most preferred isotopes for internal detection include: iodine-125, iodine-123, iodine-131, indium-111, technetium-99m and gallium-67. Technetium-99m is the preferred isotope for internal detection.

III. USES OF METABOLIC REPROGRAMMING AGENTS

[0219] The object of the present description contemplates the use of at least one, at least two or at least three metabolic reprogramming agents that decrease the activity of at least one metabolic pathway selected from the group consisting of glutamine metabolism, glycolysis and fatty acid synthesis, alone or optionally together with one or more additional therapeutic agents described herein. Accordingly, an unclaimed aspect of the presently disclosed subject matter involves the use of at least one metabolic reprogramming agent that decreases the activity of at least one metabolic pathway selected from the group consisting of glutamine metabolism, glycolysis, and metabolism. fatty acid synthesis to treat a condition, disease or disorder involving (i) metabolically reprogrammed cells whose activation, function, growth, proliferation and/or survival is dependent on increased activity of at least one metabolic pathway selected from the group consisting of glutamine metabolism, glycolysis and fatty acid synthesis or (ii) at least one of aberrant and/or excessive glutamine metabolism, aberrant and/or excessive glycolysis, or aberrant and/or excessive fatty acid synthesis.

[0220] In some embodiments described herein, the presently disclosed object involves the use of at least two metabolic reprogramming agents. In some embodiments, the object of the present invention involves the use of at least three metabolic reprogramming agents.

[0221] In some aspects described herein, the presently disclosed subject involves the use of at least one metabolic reprogramming agent that decreases glutamine metabolism as an immunotherapy to treat cancer. In other aspects described herein, the presently disclosed subject matter involves the use of at least one metabolic reprogramming agent that decreases glutamine metabolism as an immunotherapy in combination with an additional immunotherapy to treat cancer. Examples of additional immunotherapy contemplated for use in combination with at least one metabolic reprogramming agent include, without limitation, checkpoint blockade, adoptive cell therapy, CAR-T cell therapy, marrow-infiltrating lymphocytes, A2aR blockade, blockade KIR, vaccines (eg, tumor vaccines), passive immunotherapy antibodies, and combinations thereof.

[0222] In one aspect described herein, the presently disclosed subject matter involves the use of an effective amount of at least one metabolic reprogramming agent that decreases glutamine metabolism to treat lymphoma in a subject in need thereof.

[0223] In one aspect described herein, the presently disclosed subject matter involves the use of an effective amount of at least one metabolic reprogramming agent that decreases glutamine metabolism to treat glutamine metabolism. melanoma in a subject who needs it.

[0224] In some embodiments described herein, a use described herein further comprises the use of an effective amount of at least one metabolic reprogramming agent that decreases glycolysis. In some embodiments, a use described herein further comprises the use of an effective amount of at least one metabolic reprogramming agent that enhances fatty acid oxidation.

I.V. PHARMACEUTICAL COMPOSITIONS INCLUDING METABOLIC REPROGRAMMING AGENTS

[0225] The presently disclosed subject matter also contemplates pharmaceutical compositions comprising one or more metabolic reprogramming agents for the treatment of certain conditions, diseases and/or disorders involving metabolically reprogrammed cells. In some embodiments described herein, currently disclosed methods comprise the use of currently disclosed metabolic reprogramming agents for the manufacture of a medicament for the treatment of certain conditions, diseases and/or disorders involving metabolically reprogrammed cells. The disclosure contemplates various pharmaceutical compositions that comprise at least one, at least two and/or at least three metabolic reprogramming agents.

[0226] Consequently, in a non-claimed aspect, the object currently disclosed provides a pharmaceutical composition comprising an effective amount of at least one, at least two or at least three metabolic reprogramming agents that decrease the activity of at least one pathway metabolic process selected from the group consisting of glutamine metabolism, glycolysis, and fatty acid synthesis, and a pharmaceutically acceptable carrier, diluent, or excipient.

[0227] In some unclaimed aspects, the currently disclosed object provides a pharmaceutical composition comprising at least one metabolic reprogramming agent that decreases glutamine metabolism as an immunotherapy to treat cancer, and a pharmaceutically acceptable carrier, diluent or excipient. It should be appreciated that additional forms of immunotherapy are contemplated for use in combination with the pharmaceutical composition comprising at least one metabolic reprogramming agent, such as checkpoint blockade, adoptive cell therapy, CAR-T cell therapy, infiltrating lymphocytes of marrow, A2aR blockade, KIR blockade, vaccines ( eg, tumor vaccines), passive immunotherapy antibodies, and combinations thereof.

[0228] In some embodiments described herein, the metabolic reprogramming composition comprises one or more additional therapeutic agents described herein. In general, the compositions described herein ( eg, comprising at least one metabolic reprogramming agent) can be administered to a subject for therapy by any suitable route of administration, including oral, nasal, transmucosal, ocular, rectal, intravaginal, parenteral, including intramuscular, subcutaneous, intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intraarticular, intrasternal, intrasynovial, intrahepatic, intralesional, intracranial, intraperitoneal, intranasal, intraocular, intratumoral, intracisternally, topically injections , as a powder, ointment, or drops (including eye drops), including by the buccal and sublingual routes, transdermal, via inhalation aerosol, or other modes of administration known in the art.

[0229] The phrases "systemic administration", "systemically administered", "peripheral administration" and "peripherally administered", as used herein, mean the administration of compositions comprising at least one metabolic reprogramming agent, of way they enter the patient's system and are therefore subject to metabolism and other similar processes, eg subcutaneous administration.

[0230] The phrases "parenteral administration" and "parenterally administered" as used herein mean modes of administration other than enteral and topical administration, generally by injection, and include, without limitation, intravenous, intramuscular, intrathecal, intracapsular, intraorbital, intraocular, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0231] The pharmaceutical compositions described herein can be manufactured in a manner known in the art, for example, by means of conventional mixing, dissolving, granulating, dragee-making, levitation, emulsifying, encapsulating, entrapment or freeze-drying processes.

[0232] In some embodiments described herein, currently described pharmaceutical compositions can be delivered via refillable or biodegradable devices. For example, a variety of slow-release polymeric devices for controlled drug delivery, including proteinaceous biopharmaceuticals, have been developed and tested in vivo . Suitable examples of sustained release preparations include semipermeable polymeric matrices in the form of shaped articles, eg films or microcapsules. Sustained-release matrices include polyesters, hydrogels, polylactides (US Patent No. 3,773,919; EP 58,481), copolymers of L-glutamic acid and gamma ethyl-L-glutamate (Sidman et al., Biopolymers 22:547, 1983), poly(2-hydroxyethyl-methacrylate) (Langer et al. (1981) J. Biomed. Mater. Res. 15:167; Langer (1982), Chem. Tech. 12:98), ethylene vinyl acetate (Langer et al (1981) J. Biomed. Mater. Res. 15:167), or acid poly-D-()-3-hydroxybutyric acid (EP 133,988A). Sustained release compositions also include liposome-entrapped compositions comprising at least one metabolic reprogramming agent that can be prepared by methods known in the art (Epstein et al. (1985) Proc. Natl. Acad. Sci. USA 82 :3688, Hwang et al (1980) Proc. Natl. Acad. Sci. USA 77:4030, US Patent Nos. 4,485,045 and 4,544,545 and EP 102,324A). Typically, liposomes are of the small unilamellar type (about 200-800 angstroms) in which the lipid content is greater than about 30 mol% cholesterol, the selected ratio being adjusted for optimal therapy. Such materials may comprise an implant, eg, for the sustained release of the presently disclosed compositions, which, in some embodiments, may be implanted at a particular predetermined target site.

[0233] In another embodiment described herein, the currently disclosed pharmaceutical compositions may comprise PEGylated therapeutic agents ( eg, PEGylated antibodies). PEGylation is a well-established and validated approach for the modification of a variety of antibodies, proteins, and peptides and involves the attachment of polyethylene glycol (PEG) at specific sites on the antibodies, proteins, and peptides (Chapman (2002) Adv. Drug Deliv. Rev. 54:531-545). Some effects of PEGylation include: (a) markedly improved in vivo circulating half-lives due to evasion of renal clearance as a result of the polymer increasing the apparent size of the molecule above the glomerular filtration limit and/or through evasion of cell cleaning mechanisms; (b) improved pharmacokinetics; (c) improved solubility: PEG has been found to be soluble in many different solvents, from water to many organic solvents, such as toluene, methylene chloride, ethanol, and acetone; (d) PEGylated antibody fragments can be concentrated to 200 mg/mL, and the ability to do so opens formulation and dosing options, such as subcutaneous administration of a high dose of protein; this is in contrast to many other therapeutic antibodies that are typically administered intravenously; (e) improved proteolytic resistance of the conjugated protein (Cunningham-Rundles TE.al. (1992) J. Immunol. Meth. 152:177-190); (f) improved bioavailability through reduced losses at subcutaneous injection sites; (g) reduced toxicity has been observed; For agents where toxicity is related to peak plasma level, a flatter pharmacokinetic profile achieved by subcutaneous administration of PEGylated protein is advantageous; proteins that elicit an immune response that has toxic consequences may also benefit as a result of PEGylation; and (h) improved thermal and mechanical stability of the PEGylated molecule.

[0234] Pharmaceutical compositions for parenteral administration include aqueous solutions of compositions comprising at least one metabolic reprogramming agent. For injection, the pharmaceutical compositions described herein can be formulated in aqueous solutions, for example, in some embodiments, in physiologically compatible buffers, such as Hank's solution, Ringer's solution, or physiologically buffered saline. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. In addition, composition suspensions include fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of compositions comprising at least one metabolic reprogramming agent to allow the preparation of highly concentrated solutions.

[0235] For nasal or transmucosal administration generally, penetrants appropriate for the particular barrier to be penetrated are used in the formulation. Such penetrants are generally known in the art.

[0236] Additional ingredients may be added to compositions for topical administration, provided that such ingredients are pharmaceutically acceptable and are not detrimental to epithelial cells or their function. In addition, such additional ingredients should not negatively affect the epithelial penetrating efficacy of the composition and should not cause deterioration of the stability of the composition. For example, fragrances, opacifiers, antioxidants, gelling agents, stabilizers, surfactants, emollients, colorants, preservatives, buffering agents may be present. The pH of the topical composition described herein can be adjusted to a physiologically acceptable range of about 6.0 to about 9.0 by the addition of buffering agents so that the composition is physiologically compatible with the skin of the subject.

[0237] Regardless of the route of administration selected, the presently disclosed compositions are formulated into pharmaceutically acceptable dosage forms, as described herein or by other conventional methods known to those skilled in the art.

[0238] In general, the "effective amount" or "therapeutically effective amount" of an active agent or drug delivery device refers to the amount necessary to elicit the desired biological response. As will be appreciated by those skilled in this art, the effective amount of an agent or device can vary depending on factors such as the desired biological endpoint, the agent to be administered, the composition of the encapsulating matrix, the target tissue.

[0239] The term "combination" is used in its broadest sense and means that a subject is administered at least two agents. More particularly, the term "in combination" refers to the concomitant administration of two (or more) active agents for the treatment of, for example, a single disease state. As used herein, the active agents can be combined and administered in a single dosage form, they can be administered as separate dosage forms at the same time, or they can be administered as separate dosage forms that are administered alternately or sequentially on the same day or on separate days. In one embodiment of the subject of the present invention, the active agents are combined and administered in a single dosage form. In another embodiment, the active agents are administered in separate dosage forms ( eg, where it is desirable to vary the amount of one, but not the other). The single dosage form may include additional active agents for treatment of the disease state.

[0240] In addition, the currently described compositions can be administered alone or in combination with adjuvants that improve the stability of the agents, facilitate the administration of pharmaceutical compositions containing them in certain embodiments, provide greater dissolution or dispersion, increase the activity, provide an adjuvant therapy, which includes other active ingredients. Advantageously, such combination therapies use lower doses than conventional therapies, thus avoiding the potential toxicity and adverse side effects that occur when these agents are used as monotherapies.

[0241] The timing of administration of at least one metabolic reprogramming agent can be varied as long as the beneficial effects of the combination of these agents are achieved. Accordingly, the phrase "in combination with" refers to the administration of at least one metabolic reprogramming agent, at least two metabolic reprogramming agents, or at least three metabolic reprogramming agents, and optionally additional agents simultaneously, sequentially. or a combination thereof. Thus, a subject administered a combination of at least one, at least two, or at least three metabolic reprogramming agents and optionally additional agents may receive at least one metabolic reprogramming agent, at least two metabolic reprogramming agents. and at least three metabolic reprogramming agents. agents, and optionally additional agents at the same time (i.e., simultaneously) or at different times (i.e., sequentially, in any order, on the same day or on different days), provided that the effect of the combination of all agents be accomplished in the matter.

[0242] When administered sequentially, the agents can be administered within 1, 5, 10, 30, 60, 120, 180, 240 minutes or more of each other. In other embodiments, the sequentially administered agents can be administered within 1,2, 3, 4, 5, 10, 15, 20, or more days of each other. When the agents are administered simultaneously, they may be administered to the subject as separate pharmaceutical compositions, each comprising at least one metabolic reprogramming agent, at least two metabolic reprogramming agents, or at least three metabolic reprogramming agents, and optionally additional agents, or they can be administered to a subject as a single pharmaceutical composition comprising all of the agents.

[0243] When administered in combination, the effective concentration of each of the agents to elicit a particular biological response may be less than the effective concentration of each agent when administered alone, allowing for a reduction in dose of one or more. more than agents. relative to the dose that would be required if the agent were administered as a single agent. The effects of multiple agents can, but need not be, additive or synergistic. Agents can be administered multiple times.

[0244] In some embodiments, when administered in combination, the two or more agents may have a synergistic effect. As used herein, the terms "synergistically", "synergistically", "synergistically" and their derivatives, as in a "synergistic effect" or a "synergistic combination" or a "synergistic composition" refer to circumstances in which that the biological activity of a combination of an agent and at least one additional therapeutic agent is greater than the sum of the biological activities of the respective agents when administered individually.

[0245] Synergy can be expressed in terms of a "Synergy Index (SI)", which can generally be determined by the method described by FC Kull et al. Applied Microbiology 9, 538 (1961), from the relationship determined by:

QaQA QbQB = Synergy Index (SI)

where:

Qa is the concentration of component A, acting alone, that produced an end point relative to component A;

Qa is the concentration of component A, in a mixture, that produced an end point;

Qb is the concentration of a component B, acting alone, that produced an endpoint relative to component B; Y

Qb is the concentration of component B, in a mixture, that produced an end point.

[0246] Generally, when the sum of Qa/QA and Qb/QB is greater than one, antagonism is indicated. When the sum is equal to one, additivity is indicated. When the sum is less than one, synergism is demonstrated. The lower the SI, the greater the synergy displayed by that particular blend. Therefore, a "synergistic combination" has activity in excess of what might be expected based on the observed activities of the individual components when used alone. In addition, a "synergistically effective amount" of a component refers to the amount of the component necessary to cause a synergistic effect, for example, in another therapeutic agent present in the composition.

[0247] In another aspect, the presently disclosed subject matter provides a pharmaceutical composition that includes at least one metabolic reprogramming agent, at least two metabolic reprogramming agents, at least three metabolic reprogramming agents, and optionally additional agents, alone or in combination. combination with one or more additional therapeutic agents mixed with a pharmaceutically acceptable excipient.

[0248] More particularly, the object of the present description provides a pharmaceutical composition comprising at least one metabolic reprogramming agent, at least two metabolic reprogramming agents, at least three metabolic reprogramming agents and, optionally, additional agents and a vehicle pharmaceutically acceptable.

[0249] In therapeutic and/or diagnostic applications, the compounds of the disclosure may be formulated for a variety of modes of administration, including systemic and topical or localized administration. Techniques and formulations can generally be found in Remington: The Science and Practice of Pharmacy (20th ed.) Lippincott, Williams, and Wilkins (2,000).

[0250] The use of inert pharmaceutically acceptable vehicles to formulate the compounds described herein for the practice of the disclosure in doses suitable for systemic administration is within the scope of the disclosure. With proper choice of vehicle and proper manufacturing practice, the compositions of the present disclosure, particularly those formulated as solutions, can be administered parenterally, such as by intravenous injection. The compounds can be readily formulated using pharmaceutically acceptable carriers well known in the art in doses suitable for oral administration. Such vehicles allow the compounds of the disclosure to be formulated as tablets, pills, capsules, liquids, gels, syrups, slurries, suspensions, for oral ingestion by a subject ( eg, a patient) to be treated.

[0251] For nasal or inhalation administration, the agents of the disclosure may also be formulated by methods known to those skilled in the art and may include, for example, but are not limited to examples of solubilizing, diluting or dispersing substances, such as Saline solution; preservatives, such as benzyl alcohol; absorption promoters; and fluorocarbons.

[0252] Pharmaceutical compositions suitable for use herein include compositions in which the active ingredients are contained in an amount effective to achieve their intended purpose. Determination of effective amounts is within the ability of those skilled in the art, especially in light of the detailed description provided herein. Generally, the compounds according to the disclosure are effective over a wide dosage range. For example, in the treatment of adult humans, doses of 0.01 to 1,000 mg, 0.5 to 100 mg, 1 to 50 mg per day, and 5 to 40 mg per day are examples of doses that may be used. A non-limiting dosage is 10 to 30 mg per day. The exact dosage will depend on the route of administration, the manner in which the compound is administered, the subject to be treated, the body weight of the subject to be treated, and the preference and experience of the treating physician.

[0253] In addition to the active ingredients, these pharmaceutical compositions may contain suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into pharmaceutically usable preparations. Preparations formulated for oral administration may be in the form of tablets, dragees, capsules or solutions.

[0254] The term "instructing" a patient as used herein means providing instructions for the therapy, medication, treatment, applicable treatment regimens, by any means, but preferably in writing. The instruction may be in the form of a prescription for a course of treatment, or it may be in the form of package inserts or other written promotional material. Accordingly, aspects of the subject matter described herein include instructing a patient to receive a treatment method or use an agent to treat a metabolic reprogramming disorder described herein.

[0255] The term "promotion" as used herein means offering, advertising, selling, or describing a particular drug, drug combination, or treatment modality, by any means, including writing, such as in the form of package inserts. Promotion in this document refers to promotion of a metabolic reprogramming agent for an indication, when such promotion is cleared by the Food and Drug Administration (FDA) because it has been shown to be associated with statistically significant therapeutic efficacy and safety. acceptable in a population of subjects. In some embodiments, the promotion is not authorized by the Food and Drug Administration (FDA) (or other health regulatory agency, such as the European Medicines Agency (EMA), and the promotion is for a use not indicated in Accordingly, aspects of this disclosed subject matter include promotion of a method of treatment or use described in this document.

V. General definitions

[0256] Although specific terms are used herein, they are used only in a generic and descriptive sense and not for the purpose of limitation. Unless otherwise defined, all technical terms and scientific terms used herein have the same meaning as is commonly understood by those skilled in the art to which this topic described at this time pertains.

[0257] While the following terms in relation to compounds of Formula (I) are believed to be well understood by one skilled in the art, the following definitions are set forth to facilitate explanation of the subject matter currently disclosed. These definitions are intended to supplement and illustrate, not exclude, definitions that would be apparent to one skilled in the art upon review of this disclosure.

[0258] The terms substituted, whether preceded by the term "optionally" or not, and substituent, as used herein, refer to the ability, as will be appreciated by one skilled in the art, to exchange one functional group for another functional group in a molecule, as long as the valence of all atoms is maintained. When more than one position in any given structure can be substituted with more than one substituent selected from a specific group, the substituent may be the same or different at each position. Substituents may also be substituted ( eg, a substituent on the aryl group may have another substituent, such as another aryl group, that is substituted at one or more positions).

[0259] When substituent groups or linking groups are specified by their conventional chemical Formulas, written from left to right, they equally encompass chemically identical substituents that would result from writing the structure from right to left, for example, -CH2O- is equivalent to -OCH2-; -C(=O)O- is equivalent to -O-C(=O)-; -O-C(=O)NR- is equivalent to -NRC(=O)O-.

[0260] When the term "independently selected" is used, the substituents referred to ( eg, R groups, such as the R1, R2 groups, or variables, such as "m" and "n"), they may be identical or different. For example, both R1 and R2 can be substituted alkyl, or R1 can be hydrogen and R2 can be substituted alkyl.

[0261] The terms "a" or "an", when used in reference to a group of substituents in this document, mean at least one. For example, when a compound is substituted with "an" alkyl or aryl, the compound is optionally substituted with at least one alkyl and/or at least one aryl. Furthermore, when a moiety is substituted with an R substituent, the group may be referred to as "R-substituted". When a moiety is substituted with R, the moiety is substituted with at least one R substituent and each R substituent is optionally different.

[0262] A named "R" or group will generally have the structure that is recognized in the art as corresponding to a group having that name, unless otherwise specified herein. For purposes of illustration, certain representative "R" groups as set forth above are defined below.

[0263] Descriptions of the compounds herein are limited by chemical bonding principles known to those skilled in the art. Accordingly, where a group can be substituted by one or more of a number of substituents, such substitutions are selected to comply with chemical bonding principles and to give compounds that are not intrinsically unstable and/or would be known to those of ordinary skill in the art. the technique are likely to be unstable under environmental conditions, such as aqueous, neutral, and various known physiological conditions. For example, a heterocycloalkyl or heteroaryl is attached to the remainder of the molecule via a ring heteroatom in accordance with chemical bonding principles known to those skilled in the art, thus avoiding intrinsically unstable compounds.

[0264] Unless otherwise explicitly defined, a "substituent group", as used herein, includes a functional group selected from one or more of the following moieties, which are defined herein: The term hydrocarbon , as used herein, refers to any chemical group consisting of hydrogen and carbon. The hydrocarbon may be substituted or unsubstituted. As one skilled in this art will know, all valences must be satisfied when making any substitution. The hydrocarbon can be unsaturated, saturated, branched, unbranched, cyclic, polycyclic, or heterocyclic. Illustrative hydrocarbons are defined herein below and include, for example, methyl, ethyl, N-propyl, isopropyl, cyclopropyl, allyl, vinyl, N-butyl, tert-butyl, ethynyl, cyclohexyl.

[0265] The term "alkyl", by itself or as part of another substituent, means, unless otherwise indicated, a straight chain (i.e. unbranched) or branched acyclic hydrocarbon group, or a combination of the same, which is fully saturated and may include divalent and multivalent groups, having the designated number of carbon atoms (i.e., C1-C10 means one to ten carbons, including 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 carbons). In particular embodiments, the term "alkyl" refers to C1-20 inclusive, including 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 carbons, linear (i.e., "straight chain"), branched, saturated hydrocarbon radicals derived from a hydrocarbon moiety containing between one and twenty carbon atoms by removal of a single carbon atom hydrogen.

[0266] Representative saturated hydrocarbon groups include, but are not limited to, methyl, ethyl, N-propyl, isopropyl, N-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, isopentyl, neopentyl, n -hexyl, sec-hexyl, n-heptyl, n-octyl, n-decyl, n-undecyl, dodecyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, and homologues and isomers thereof.

[0267] "Branched" refers to an alkyl group in which a lower alkyl group, such as methyl, ethyl, or propyl, is attached to a linear alkyl chain. "Lower alkyl" refers to an alkyl group having from 1 to about 8 carbon atoms (i.e., a C-i-s alkyl), for example, 1,2, 3, 4, 5, 6, 7, or 8 carbon atoms. . "Higher alkyl" refers to an alkyl group having 10 to 20 carbon atoms, for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms. In certain embodiments, "alkyl" refers in particular to C1-8 straight chain alkyls. In other embodiments, "alkyl" refers in particular to C1-8 branched chain alkyls.

[0268] Alkyl groups may be optionally substituted (a "substituted alkyl") with one or more alkyl group substituents, which may be the same or different. The term "alkyl group substituent" includes, but is not limited to, alkyl, substituted alkyl, halo, arylamino, acyl, hydroxyl, aryloxy, alkoxy, alkylthio, arylthio, aralkyloxy, aralkylthio, carboxyl, alkoxycarbonyl, oxo, and cycloalkyl. Optionally one or more oxygen, sulfur, or substituted or unsubstituted nitrogen atoms may be inserted along the alkyl chain, wherein the nitrogen substituent is hydrogen, lower alkyl (also referred to herein as "alkylaminoalkyl"), or aryl.

[0269] Therefore, As used herein, the term "substituted alkyl" includes alkyl groups, as defined herein, in which one or more atoms or functional groups of the alkyl group are replaced with another atom or functional group, including, for example, alkyl, substituted alkyl, halogen, aryl, substituted aryl, alkoxy, hydroxy, nitro, amino, alkylamino, dialkylamino, sulfate, and mercapto.

[0270] The term "heteroalkyl", alone or in combination with another term, means, unless otherwise indicated, a stable straight or branched chain hydrocarbon group, or combinations thereof, consisting of at least one atom carbon and at least one heteroatom selected from the group consisting of O, N, P, Si, and S, and wherein the nitrogen, phosphorous, and sulfur atoms may be optionally oxidized and the nitrogen heteroatom may be optionally quaternized. The heteroatom(s) O, N, P and S and Si can be placed at any position within the heteroalkyl group or at the position where the alkyl group is attached to the rest of the molecule. Examples include, but are not limited to -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH25 -S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CHO-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, -CH=CH-N (CH3)-CH3, O-CH3, -O-CH2-CH3 and -CN. Up to two or three heteroatoms may be consecutive, such as -CH2-NH-OCH3 and -CH2-O-Si(CH3)3.

[0271] As described above, heteroalkyl groups, as used herein, include those groups that are attached to the rest of the molecule through a heteroatom, such as -C(O)NR', -NR'R", - OR', -SR, -S(O)R, and/or -S(O2)R' When "heteroalkyl" is mentioned, followed by listings of specific heteroalkyl groups, such as -NR'R, it is understood that the terms heteroalkyl and -NR'R" are neither redundant nor mutually exclusive. Rather, specific heteroalkyl groups are listed to add clarity. Therefore, the term "heteroalkyl" should not be construed herein as excluding specific heteroalkyl groups, such as -NR'R".

[0272] "Cyclic" and "cycloalkyl" refer to a non-aromatic mono or multicyclic ring system of about 3 to about 10 carbon atoms, for example, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. The cycloalkyl group can optionally be partially unsaturated. The cycloalkyl group can also be optionally substituted with an alkyl group substituent as defined herein, oxo, and/or alkylene. One or more substituted or unsubstituted oxygen, sulfur, or nitrogen atoms may optionally be inserted along the cyclic alkyl chain, wherein the nitrogen substituent is hydrogen, unsubstituted alkyl, substituted alkyl, aryl, or substituted aryl, providing thus a heterocyclic group. Representative monocyclic cycloalkyl rings include cyclopentyl, cyclohexyl, and cycloheptyl. Multicyclic cycloalkyl rings include adamantyl, octahydronaphthyl, decalin, camphor, camphane, and noradamantyl, and fused ring systems, such as dihydro- and tetrahydronaphthalene.

[0273] The term "cycloalkylalkyl", as used herein, refers to a cycloalkyl group as defined herein above, which is attached to the parent molecular moiety through an alkyl group, also as defined previously. Examples of cycloalkylalkyl groups include cyclopropylmethyl and cyclopentylethyl.

[0274] The terms "cycloheteroalkyl" or "heterocycloalkyl" refer to a non-aromatic ring system, unsaturated or partially unsaturated ring system, such as a 3 to 10-membered substituted or unsubstituted cycloalkyl ring system, including one or more heteroatoms, which may be the same or different, and are selected from the group consisting of nitrogen (N), oxygen (O), sulfur (S), phosphorus (P), and silicon (Si), and may optionally include one or more more double bonds.

[0275] The cycloheteroalkyl ring may optionally be fused or otherwise attached to other cycloheteroalkyl rings and/or non-aromatic hydrocarbon rings. Heterocyclic rings include those having one to three heteroatoms independently selected from oxygen, sulfur, and nitrogen, wherein the nitrogen and sulfur heteroatoms may be optionally oxidized and the nitrogen heteroatom optionally quaternized. In certain embodiments, the term "heterocyclic" refers to a 5-, 6-, or 7-membered non-aromatic ring or polycyclic group in which at least one ring atom is a heteroatom selected from O, S, and N (where the nitrogen and sulfur heteroatoms may be optionally oxidized), which includes, among others, a bi- or tricyclic group, comprising fused six-membered rings having one to three heteroatoms independently selected from oxygen, sulfur, and nitrogen, where (i) each 5-membered ring has 0 to 2 double bonds, each 6-membered ring has 0 to 2 double bonds, with each 7-membered ring having 0 to 3 double bonds, (ii) the nitrogen and sulfur heteroatoms can be optionally oxidized, (iii) the nitrogen heteroatom can optionally be quaternized, and (iv) any of the heterocyclic rings above can be fused to an aryl or heteroaryl ring. Representative cycloheteroalkyl ring systems include, but are not limited to, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperidyl, piperazinyl, indolinyl, quinuclidinyl, morpholinyl, thiomorpholinyl, thiadiazinanyl, tetrahydrofuranyl.

[0276] The terms "cycloalkyl" and "heterocycloalkyl", by themselves or in combination with other terms, represent, unless otherwise indicated, cyclic versions of "alkyl" and "heteroalkyl", respectively. Also, for heterocycloalkyl, a heteroatom can occupy the position where the heterocycle is attached to the rest of the molecule. Examples of cycloalkyl include, but are not limited to, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl. Examples of heterocycloalkyl include, but are not limited to, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran -3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl. The terms "cycloalkylene" and "heterocycloalkylene" refer to the divalent derivatives of cycloalkyl and heterocycloalkyl, respectively.

[0277] An unsaturated alkyl group is one that has one or more double or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3- -propynyl, 3-butynyl and the higher homologues and isomers. Alkyl groups that are limited to hydrocarbon groups are called "homoalkyl."

[0278] More particularly, the term "alkenyl" as used herein refers to a monovalent group derived from a C1-20 linear or branched hydrocarbon moiety inclusively having at least one carbon-carbon double bond by removal of a single hydrogen molecule. Alkenyl groups include, for example, ethenyl (ie, vinyl), propenyl, butenyl, 1-methyl-2-buten-1-yl, pentenyl, hexenyl, octenyl, allenyl, and butadienyl.

[0279] The term "cycloalkenyl", as used herein, refers to a cyclic hydrocarbon containing at least one carbon-carbon double bond. Examples of cycloalkenyl groups include cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadiene, cyclohexenyl, 1,3-cyclohexadiene, cycloheptenyl, cycloheptatrienyl, and cyclooctenyl.

[0280] The term "alkynyl", as used herein, refers to a monovalent group derived from a straight or branched C1-20 hydrocarbon of a designated number of carbon atoms containing at least one carbon-carbon triple bond. carbon. Examples of "alkynyl" include ethynyl, 2-propynyl (propargyl), 1-propynyl, pentynyl, hexynyl and heptynyl groups.

[0281] The term "alkylene" by itself or as part of another substituent refers to a linear or branched bivalent aliphatic hydrocarbon group derived from an alkyl group having from 1 to about 20 carbon atoms, eg, 1, 2 , 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms. The alkylene group can be linear, branched or cyclic. The alkylene group can also be optionally unsaturated and/or substituted with one or more "alkyl group substituents". Optionally one or more substituted or unsubstituted oxygen, sulfur or nitrogen atoms (also referred to herein as "alkylaminoalkyl") may be inserted along the alkylene group, wherein the nitrogen substituent is alkyl as described above. . Examples of alkylene groups include methylene (-CH2-); ethylene (-CH2-CH2-); propylene (-(CH2)3-); cyclohexylene (-C6H10-); -Ch=CH-CH=CH-; -CH=CH-CH2-; -CH2CH2CH2CH2-, -CH2CH=CHCH2-, -CH2CsCCH2-, -CH2CH2CH(CH2CH2CH3)CH2-, -(CH2)q-N(R)-(CH2)r-, where each q and r is independently an integer from 0 to about 20, for example, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, and R is hydrogen or lower alkyl; methylenedioxy (-O-CH2-O-); and ethylenedioxy (-O-(CH2)2-O-). An alkylene group may have 2 to 3 carbon atoms and may additionally have 6 to 20 carbons. Typically, an alkyl (or alkylene) group will have from 1 to 24 carbon atoms, with those groups having 10 carbon atoms or fewer being some embodiments of the present disclosure. A "lower alkyl" or "lower alkylene" is a shorter chain alkyl or alkylene group, generally having eight or fewer carbon atoms.

[0282] The term "heteroalkylene" by itself or as part of another substituent means a divalent group derived from heteroalkyl, as exemplified, but not limited to -CH2-CH2-S-CH2-CH2- and -CH2-S -CH2-CH2-NH-CH2-. For heteroalkylene groups, heteroatoms can also occupy one or both ends of the chain ( eg, alkyleneoxo, alkylenedioxo, alkyleneamino, alkylenediamino). Furthermore, for alkylene and heteroalkylene linking groups, the direction in which the Linking Group Formula is written does not imply any orientation of the linking group. For example, the Formula -C(O)OR'- represents both -C(O)OR'- and -R'OC(O)-.

[0283] The term "aryl" means, unless otherwise indicated, an aromatic hydrocarbon substituent which may be a single ring or multiple rings (such as 1 to 3 rings), which are fused together or covalently bonded. The term "heteroaryl" refers to aryl groups (or rings) containing one to four heteroatoms (in each separate ring in the case of multiple rings) selected from N, O, and S, where the nitrogen and sulfur atoms are optionally present. oxidized and the nitrogen atom(s) are optionally quaternized. A group heteroaryl can be attached to the rest of the molecule through a carbon or a heteroatom. Non-limiting examples of aryl and heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3- furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5- isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl and 6-quinolyl. The substituents for each of the aryl and heteroaryl ring systems noted above are selected from the group of acceptable substituents described below. The terms "arylene" and "heteroarylene" refer to the divalent forms of aryl and heteroaryl, respectively.

[0284] For brevity, the term "aryl" when used in combination with other terms ( eg, aryloxy, arylthioxy, arylalkyl) includes aryl and heteroaryl rings as defined above. Thus, the terms "arylalkyl" and "heteroarylalkyl" include those groups in which an aryl or heteroaryl group is attached to an alkyl group ( eg, benzyl, phenethyl, pyridylmethyl, furylmethyl), including those alkyl groups in which that a carbon atom ( eg, a methylene group) has been replaced, for example, by an oxygen atom ( eg, phenoxymethyl, 2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl). However, the term "haloaryl", as used herein, is intended to cover only aryls substituted with one or more halogens.

[0285] When a heteroalkyl, heterocycloalkyl, or heteroaryl includes a specific number of members (eg, "3 to 7 membered"), the term "member" refers to a carbon or heteroatom.

[0286] Furthermore, a structure generally represented by the formula:

as used herein refers to a ring structure, for example, but not limited to 3 carbon, 4 carbon, 5 carbon, 6 carbon, a 7 carbon aliphatic and/or aromatic cyclic compound, including a saturated ring structure , a partially saturated ring structure and an unsaturated ring structure, comprising a substituent R group, in which the R group may be present or absent, and when present, one or more R groups may each be substituted by one or more carbon atoms available from the ring structure. The presence or absence of the R group and the number of R groups is determined by the value of the variable "n", which is an integer that generally has a value ranging from 0 to the number of ring carbon atoms available. for substitution. Each R group, if there is more than one, is substituted on an available carbon of the ring structure instead of another R group. For example, the structure above where n is 0 to 2 would comprise compound groups including, but not limited to:

[0287] A dashed line representing a bond in a cyclic ring structure indicates that the bond may be present or absent in the ring. That is, a dashed line representing a bond in a cyclic ring structure indicates that the ring structure is selected from the group consisting of a saturated ring structure, a partially saturated ring structure, and an unsaturated ring structure.

[0288] The symbol (vvwwwv) indicates the point of attachment of a moiety to the rest of the molecule.

[0289] When a named atom of an aromatic ring or heterocyclic aromatic ring is defined as "absent", the named atom is replaced by a direct bond.

[0290] Each of the above terms (for example, "alkyl", "heteroalkyl", "cycloalkyl and "heterocycloalkyl", "aryl", "heteroaryl", "phosphonate" and "sulfonate", as well as their divalent derivatives) means to include both substituted and unsubstituted forms of the indicated group.Optional substituents for each type of group are given below.

[0291] Substituents for monovalent and divalent alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl (including groups often referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) may be one or more of a variety of groups selected from, but not limited to: -OR', =O , =NR', =N- or R', -NR'R", -SR', -halogen, -SiR'R"R-, -OC(O)R', -C(O)R', - CO2R', -C(O)NR'R", -OC(O)NR'R", --S(O)R', -S(O)2R', -S(O)2NR'R", -NRSO2R', -CN and -NO2 in a number from zero to (2m'+1), where m' is the total number of carbon atoms in those groups. Each of R', R", R'"andR"" can independently refer to hydrogen, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl ( e.g., aryl substituted with 1-3 halogen), groups substituted or unsubstituted alkyl, alkoxy, or thioalkoxy, or arylalkyl groups.As used herein, an "alkoxy" group is an alkyl attached to the res of the molecule through a divalent oxygen. When a compound of the disclosure includes more than one R group, for example, each of the R groups is independently selected as is each R', R", R'" and R"" group when more than one of these groups. When R' and R" are attached to the same nitrogen atom, they can combine with the nitrogen atom to form a 4-, 5-, 6-, or 7-membered ring. For example, -NR'R" means to include, but not limited to 1-pyrrolidinyl and 4-morpholinyl. From the above discussion of substituents, one skilled in the art will understand that the term "alkyl" includes groups that include carbon atoms attached to groups other than hydrogen groups, such as haloalkyl ( eg, -CF3 and -CH2CF3) and acyl (for example, -C(O)CH3, -C(O)CF3, -C(O)CH2OCH3).

[0292] Similar to the substituents described above for alkyl groups, examples of substituents for aryl and heteroaryl groups (as well as their divalent derivatives) vary and are selected, for example, from: halogen, -OR', - NR'R", -SR', -SiR'R"R-, -OC(O)R', -C(O)R', -CO2R', -C(O)NR'R", -OC( O)NR'R”, -NR"C(O)R', -NR'-C(O)NR"R-, -NR"C(O)OR', -NRC(NR'R"R-) =NR'm, -NR-C(NR'R")=NR'm -S(O)R', -S(O)2R', -S(O)2NR'R", -NRSO2R', - CN and -NO2, -R', -N3, -CH(Ph)2, fluoro(Ci-C4)alcoxo and fluoro(Ci-C4)alkyl, in a number ranging from zero to the total number of open valences in the aromatic ring system; and where R', R", R'" and R"" can be independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl and heteroaryl When a compound of the disclosure includes more than one R group, po For example, each of the R groups is independently selected as each of the R', R", R- and R"" groups when more than one of these groups is present.

[0293] Two of the substituents on adjacent aryl or heteroaryl ring atoms may optionally form a ring of formula -T-C(O)-(CRR')q-U-, where T and U are independently -NR-, - O-, -CRR'- or a single bond, and q is an integer from 0 to 3. Alternatively, two of the substituents on adjacent aryl or heteroaryl ring atoms may optionally be replaced with a substituent of the formula -A-(CH2 )r-B-, where A and B are independently -CRR'-, -O-, -NR-, -S-, -S(O)-, -S(O)2-, -S(O)2NR'- or a single bond, and r is an integer from 1 to 4.

[0294] One of the single bonds of the new ring thus formed can optionally be replaced with a double bond. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may be optionally replaced with a substituent of the formula -(CRR')s-X'-(C"R-)d-, where s and d are independently integers of 0 to 3, and X' is -O-, -NR'-, -S-, -S(O)-, -S(O)2-, or -S(O)2NR'-. R', R" and R'" may be independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl.

[0295] As used herein, the term "acyl" refers to an organic acid group in which the -OH of the carboxyl group has been replaced with another substituent and has the general Formula RC(=O)- , wherein R is an alkyl, alkenyl, alkynyl, aryl, carbocyclic, heterocyclic, or aromatic heterocyclic group as defined herein). As such, the term "acyl" specifically includes arylacyl groups, such as a 2-(furan-2-yl)acetyl)- and 2-phenylacetyl group. Specific examples of acyl groups include acetyl and benzoyl. Acyl groups are also intended to include amides, -RC(=O)NR', esters, -RC(=O)OR', ketones, -RC(=O)R', and aldehydes, -RC(=O)H.

[0296] The terms "alkoxy" or "alkoxy" are used interchangeably in this document and refer to a saturated (i.e. alkyl-O-) or unsaturated (i.e. alkenyl-O- and alkynyl-O-) group. attached to the parent molecular moiety through an oxygen atom, where the terms "alkyl", "alkenyl" and "alkynyl" are as described above and can include C1 straight, branched or cyclic saturated or unsaturated oxohydrocarbon chains -20 inclusive, including, for example, methoxy, ethoxy, propoxy, isopropoxy, N-butoxy, sec-butoxy, tert-butoxy, and N-pentoxy, neopentoxy, N-hexoxy.

[0297] The term "alkoxyalkyl", as used herein, refers to an alkyl-O-alkyl ether, for example, a methoxyethyl or ethoxymethyl group.

[0298] "Aryloxy" refers to an aryl-O- group in which the aryl group is as described above, including substituted aryl. The term "aryloxy", as used herein, can refer to phenyloxy or hexyloxy, and alkyl-substituted, substituted alkyl, halo, or alkoxy-substituted phenyloxy or hexyloxy.

[0299] "Aralkyl" refers to an aryl-alkyl group in which aryl and alkyl are as described above, and include substituted aryl and substituted alkyl. Exemplary aralkyl groups include benzyl, phenylethyl, and naphthylmethyl.

[0300] "Aralkyloxy" refers to an aralkyl-O- group in which the aralkyl group is as previously described. An example of an aralkyloxy group is benzyloxy, ie, C6H5-CH2-O-. An aralkyloxy group may be optionally substituted.

[0301] "Alkoxycarbonyl" refers to an alkyl-O-C(=O)- group. Exemplary alkoxycarbonyl groups include methoxycarbonyl, ethoxycarbonyl, butyloxycarbonyl, and ferrt-butyloxycarbonyl.

[0302] "Aryloxycarbonyl" refers to an aryl-O-C(=O)- group. Exemplary aryloxycarbonyl groups include phenoxy- and naphthoxy-carbonyl.

[0303] "Aralkoxycarbonyl" refers to an aralkyl-O-C(=O)- group. An example of an aralkoxycarbonyl group is benzyloxycarbonyl.

[0304] "Carbamoyl" refers to an amide group of Formula -C(=O)NH2. "Alkylcarbamoyl" refers to a group R'RN-C(=O)- in which one of R and R' is hydrogen and the other of R and R' is alkyl and/or substituted alkyl as described above. "Dialkylcarbamoyl" refers to a group R'RN-C(=O)- in which R and R' are each independently alkyl and/or substituted alkyl as previously described.

[0305] The term carbonyldioxy, as used herein, refers to a carbonate group of the formula -O-C(=O)-OR.

[0306] "Acyloxy" refers to an acyl-O- group where acyl is as described above.

[0307] The term "amino" refers to the group -NH2 and also refers to a nitrogen-containing group as known in the art derived from ammonia by substitution of one or more hydrogen radicals with organic radicals. For example, the terms "acylamino" and "alkylamino" refer to specific N-substituted organic radicals with acyl and alkyl substituent groups, respectively.

[0308] An "aminoalkyl", as used herein, refers to an amino group covalently attached to an alkylene linker. More particularly, the terms alkylamino, dialkylamino, and trialkylamino as used herein refer to one, two, or three, respectively, alkyl groups, as previously defined, attached to the parent molecular moiety through a nitrogen atom. The term "alkylamino" refers to a group having the structure -NHR' where R' is an alkyl group, as defined above; while the term "dialkylamino" refers to a group having the structure -NR'R", where R' and R" are each independently selected from the group consisting of alkyl groups. The term "trialkylamino" refers to a group having the structure -NR'R"R-, where R', R" and R'" are each independently selected from the group consisting of alkyl groups. In addition, R', R" and/or R'" together may optionally be -(CH2)k- where k is an integer from 2 to 6. Examples include, but are not limited to methylamino, dimethylamino, ethylamino, diethylamino, diethylaminocarbonyl, methylethylamino, isopropylamino, piperidino, trimethylamino and propylamino.

[0309] The amino group is -NR'R", where R' and R" are typically selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, aryl substituted or unsubstituted, or substituted or unsubstituted heteroaryl.

[0310] The terms alkylthioether and thioalkoxy refer to a saturated (ie, alkyl-S-) or unsaturated (ie, alkenyl-S- and alkynyl-S-) group attached to the parent molecular moiety through a carbon atom. sulfur. Examples of thioalkoxy moieties include, but are not limited to, methylthio, ethylthio, propylthio, isopropylthio, n-butylthio.

[0311] "Acylamino" refers to an acyl-NH- group where acyl is as described above. "Aroylamino" refers to an aroyl-NH- group where aroyl is as previously described.

[0312] The term "carbonyl" refers to the group -C(=O)-, and may include an aldehyde group represented by the general Formula RC(=O)H.

[0313] The term "carboxyl" refers to the group -COOH. Such groups are also referred to herein as a "carboxylic acid" moiety.

[0314] The terms "halo", "halide" or "halogen", as used herein, refer to fluoro, chloro, bromo and iodo groups. Additionally, terms such as "haloalkyl" include monohaloalkyl and polyhaloalkyl. For example, the term "halo(C1-C4)alkyl" includes, but is not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl.

[0315] The term "hydroxyl" refers to the group -OH.

[0316] The term "hydroxyalkyl" refers to an alkyl group substituted with a -OH group.

[0317] The term "mercapto" refers to the -SH group.

[0318] The term "oxo", as used herein, means an oxygen atom having a double bond with a carbon atom or with another element.

[0319] The term "nitro" refers to the group -NO2.

[0320] The term "thio" refers to a compound described herein above in which a carbon or oxygen atom is replaced by a sulfur atom.

[0321] The term "sulfate" refers to the group -SO4.

[0322] The term thiohydroxyl or thiol, as used herein, refers to a group of the formula -SH.

[0323] More particularly, the term "sulfide" refers to a compound having a group of formula -SR.

[0324] The term "sulfone" refers to a compound having a sulfonyl group -S(O2)R.

[0325] The term "sulfoxide" refers to a compound having a sulfinyl group -S(O)R.

[0326] The term "ureido" refers to a urea group of the formula -NH-CO-NH2.

[0327] Throughout the specification and claims, a given chemical formula or name shall encompass all tautomers, congeners and optical and stereoisomers, as well as racemic mixtures where such isomers and mixtures exist.

[0328] Certain compounds of the present disclosure may possess asymmetric carbon atoms (optical or chiral centers) or double bonds; enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisometric forms that can be defined, in terms of absolute stereochemistry, as (R) or (S) or, as D or L for individual amino acids and isomers are included within the scope of the present disclosure. The compounds of the present disclosure do not include those known in the art to be too unstable to synthesize and/or isolate. The present disclosure is intended to include compounds in racemic, scalemic, and optically pure forms. The optically active (R) and (S), or D and L isomers can be prepared using chiral synthons or chiral reagents, or can be resolved using conventional techniques. When the compounds described herein contain olephenic bonds or other centers of geometric asymmetry, and unless otherwise specified, the compounds are intended to include both E and Z geometric isomers.

[0329] Unless otherwise indicated, the structures depicted herein also include all stereochemical forms of the structure; that is, the R and S configurations for each asymmetric center. Therefore, individual stereochemical isomers, as well as enantiomeric and diastereomeric mixtures of the present compounds are within the scope of the disclosure.

[0330] It will be apparent to one skilled in the art that certain compounds of this disclosure may exist in tautomeric forms, all such tautomeric forms of the compounds being within the scope of the disclosure. The term "tautomer" as used herein refers to one of two or more structural isomers that exist in equilibrium and are readily converted from one isomeric form to another.

[0331] Unless otherwise indicated, the structures depicted herein also include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures with the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by a 13C- or 14C-enriched carbon are within the scope of this disclosure.

[0332] The compounds of the present disclosure may also contain unnatural atomic isotope ratios on one or more of the atoms constituting such compounds. For example, compounds may be radiolabeled with radioactive isotopes, such as tritium (3H), iodine-125 (125I), or carbon-14 (14C). All isotopic variations of the compounds of the present disclosure, whether radioactive or not, are included within the scope of the present disclosure.

[0333] The compounds of the present disclosure may exist as salts. The present description includes such salts. Examples of applicable salt forms include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, maleates, acetates, citrates, fumarates, tartrates (for example, (+)-tartrates, (-)-tartrates or mixtures thereof, including mixtures racemics, succinates, benzoates, and salts with amino acids, such as glutamic acid.These salts can be prepared by methods known to those skilled in the art.Also include base addition salts, such as sodium, potassium, calcium, ammonium salts , organic amino or magnesium, or similar salts.When the compounds of the present description contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of said compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent or by ion exchange Among the acceptable acid addition salts Derivatives of inorganic acids such as hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenetic are included. hydrogenphosphoric, sulfuric, monohydrogensulfuric, hydroiodic or phosphorous acids, as well as the salts of derived organic acids such as acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p- tolylsulfonic, citric, tartaric, methanesulfonic. Also included are amino acid salts, such as arginate, and organic acid salts, such as glucuronic or galacturonic acids. Certain specific compounds of the present disclosure contain both basic and acidic functionalities that allow the compounds to be converted to acid or base addition salts.

[0334] The neutral forms of the compounds can be regenerated by contacting the salt with a base or an acid and isolating the parent compound in conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents.

[0335] Certain compounds of the present disclosure may exist in unsolvated forms, as well as solvated forms, including hydrated forms. In general, solvated forms are equivalent to unsolved forms and are included within the scope of the present disclosure. Certain compounds of the present disclosure can exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by this disclosure and are intended to be within the scope of this disclosure.

[0336] In addition to salt forms, the present disclosure provides compounds that are in prodrug form. Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of the present disclosure. Furthermore, prodrugs can be converted to the compounds of the present disclosure by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to the compounds of the present disclosure when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

[0337] Following long-standing patent law convention, the terms "a", "an", "the" and "she" refer to "one or more" when used in this application, including the claims . Thus, for example, a reference to "a subject" includes a plurality of subjects, unless the context clearly indicates otherwise (eg, a plurality of subjects), and so on.

[0338] Throughout this specification and the claims, the terms "comprising", "comprising" and "comprising" are used in a non-exclusive sense, except where the context otherwise requires. Similarly, the term "include" and its grammatical variants are intended to be non-limiting, so that the recitation of items in a list does not exclude other similar items that may be substituted for or added to the listed items.

[0339] For the purposes of this specification and attached claims, unless otherwise indicated, all numbers expressing quantities, sizes, dimensions, proportions, shapes, formulations, parameters, percentages, amounts, characteristics, and other numerical values used in the specification and claims, they shall be understood to be modified in any case by the term "envelope" even though the term "envelope" does not expressly appear with the value, amount or range. Accordingly, unless otherwise indicated, the numerical parameters set forth in the following specification and appended claims are not and need not be exact, but may be approximate and/or larger or smaller as desired, reflecting tolerances, conversion factors , rounding, measurement error, and other factors known to those skilled in the art depending on the desired properties that are intended to be obtained by means of the object of the present invention. For example, the term "approximately", when referring to a value, can mean that it encompasses variations of, in some embodiments ±100%, in some embodiments ±50%, in some embodiments ±20%, in some embodiments ±10%, in some embodiments ±5%, in some embodiments ±1%, in some embodiments ±0.5%, and in some embodiments ±0.1% of the amount specified, as such variations are appropriate for carrying out the described methods or employing the disclosed compositions.

[0340] Furthermore, the term "over" when used in relation to one or more numbers or numerical ranges, should be understood to refer to all of those numbers, including all numbers in a range and modifies that range by extending the limits by above and below the set numerical values. Reciting number ranges by endpoints includes all numbers, for example whole integers, including their fractions, included within that range (for example, reciting 1 to 5 includes 1, 2, 3, 4, and 5, so as fractions of it, for example, 1.5, 2.25, 3.75, 4.1) and any range within that range.

EXAMPLES

[0341] The following examples have been included to provide guidance to one skilled in the art in practicing representative embodiments of the currently disclosed subject matter. In light of the present disclosure and the general level of skill in the art, those skilled in the art can appreciate that the following examples are intended to be illustrative only and that numerous changes, modifications, and alterations may be employed without departing from the scope of the presently disclosed subject matter. The synthetic descriptions and specific examples that follow are for educational purposes only. illustrative and should not be construed as limiting in any way to making compounds of the disclosure by other methods.

REFERENCE EXAMPLE 1

[0342] To explore the effect of DON on cancer, an EL4 mouse lymphoma model was used and showed that DON could markedly inhibit lymphoma growth, suggesting that bone marrow-derived tumors may be exquisitely susceptible to DON. DON (FIG. 1). However, DON had a modest effect in inhibiting the growth of melanoma, which is not a bone marrow-derived tumor (Figure 2).

REFERENCE EXAMPLE 2

[0343] FIG. 3 shows that DON conditioned B16 melanoma to be destroyed by immunotherapy by inhibiting tumor-infiltrating regulatory T cells (Foxp3+).

REFERENCE EXAMPLE 3

Resume

[0344] The glutamine antagonist 6-diazo-5-oxo-L-norleucine (DON, 1) has shown strong anticancer efficacy in preclinical and clinical studies, but its development was halted due to marked systemic toxicities. Here we demonstrate that DON inhibits glutamine metabolism and provides antitumor efficacy in a mouse model of glioblastoma, although toxicity was observed. To improve the therapeutic index of DON, we used a prodrug strategy to increase its cerebral delivery and limit systemic exposure. Although these dual moiety prodrugs exhibited rapid metabolism in mouse plasma, several provided excellent stability in monkey and human plasma. The most stable compound (5c, methyl-POM_DON-isopropyl-ester) was evaluated in monkeys, where it achieved a 10-fold improved brain:plasma ratio versus DON. This strategy may provide a pathway for the utilization of DON in GBM patients.

Introduction

[0345] Glioblastoma multiforme (GBM) is the most common and deadly form of glioma (Ostrom, et al., 2015). Current therapeutic options that increase survival rates in GBM patients after tumor resection are limited to radiation therapy with concomitant administration of the DNA alkylating agent Temodar™ and/or the carmustine-releasing polymer Gliadel™ (Weller, et al. al., 2014). But even with this standard of care, median postoperative survival rates approach 14.6 months (Stupp, et al., 20015) and median 5-year survival is less than 10% (Stupp, et al., 2009 ). Therefore, there is a significant unmet medical need for more effective GBM treatments.

[0346] The ability of GBMs and other cancers to rapidly and persistently proliferate often outpacing vascular supplies requires that they evolve a specialized metabolism (Schulze, et al., 2012; Hensley, et al., 2013). Glutamine metabolism plays an essential role in this specialized metabolism by entering the TCA cycle and then contributing to the biosynthetic pathways (synthesis of nucleotides, proteins and lipids) necessary for uncontrolled cell growth (Hensley, et al., 2013) making make cancer cells dependent on glutamine. This so-called "glutamine addiction" has been well characterized in GBM (Dranoff, et al., 1985; Fogal, et al., 2015; Ru, et al., 2013; Tanaka, et al., 2015) and other types. cancer, including leukemia/lymphomas, lung, triple-negative breast cancer, and pancreatic cancer (Wise, et al., 2010; Hu, et al., 2009). To combat this addiction to glutamine, several groups have explored the utility of selective glutaminase inhibitors (Erickson, et al., 2010; Lee, et al., 2014). Compounds that selectively target glutaminase (GLS1) have recently been developed (McDermott, et al., 2016; Shukla, et al., 2012) and one such compound is in clinical trials (Konopleva, et al., 2015 ). However, the brain penetration of this GLS1 inhibitor is poor (Gross, et al., 2014) and, thus far, the clinical efficacy of this approach has been modest (Harding, et al., 2015).

[0347] Accumulating evidence shows that, unlike selective inhibition of an enzyme using glutamine, the use of broadly antagonizing glutamine is a highly effective means of inhibiting tumor cell growth in vitro and in vivo (Hensley, et al. ., 2013; Ahluwalia, et al., 1990; Dranoff, et al., Cancer Res, 1985). 6-Diazo-5-oxo-L-norleucine (DON), an unnatural amino acid with a structure similar to glutamine, was first isolated from the bacterium Streptomyces in the early 1950s. Due to its diazo group reagent, DON has demonstrated the ability to alkylate several glutamine-utilizing enzymes, such as glutaminase (Thangavelu, et al., 2014) NAD synthase, (Barclay, et al., 1966) and CTP synthase (Hofer, et al., 2001 ) and FGAR aminotransferase (Grayzel, et al., 1960) in the purine and pyrimidine biosynthetic pathways, respectively. In preclinical models, DON strongly inhibited the growth of glutamine-dependent human cancer cells in vitro, reduced tumor size, and improved survival rates in vivo (Ahluwalia, et al., 1990; Cervantes-Madrid, et al. , 2015). Due to robust preclinical data, DON was evaluated in several clinical studies where it demonstrated promising results (Eagan, et al., 1982; Earhart, et al., 1982; Lynch, et al., 1982; Magill, et al., 1957 ; Rahman, et al., 1985; Sklaroff, et al., 1980; Sullivan, et al., 1962). For example, DON administration caused >50% regression or stable disease in late-stage adult patients (Magill, et al. al., 1957) and in children with hematologic malignancies or solid tumors (Sullivan, et al., 1988). Unfortunately, its development was hampered by dose-limiting toxicities, many of which were GI-related (Magill, et al., 1957; Rahman, et al., 1985; Earhart, et al., 1990) since the GI system relies heavily on glutamine utilization.

[0348] One strategy to improve the therapeutic index of DON for GBM therapy would be to increase its cerebral exposure while limiting its systemic exposure and thus its toxicity (Upadhyay, et al., 2014). The prodrug approach is a well-established strategy to alter the pharmacokinetic and tissue distribution of drug molecules, however, synthetically, this approach is a DON exposure.

[0349] Here we describe, for the first time, the profound efficacy of DON in the murine model of GBM, although overt toxicities were observed. In an attempt to increase the therapeutic index of DON, we systematically synthesized various prodrugs of DON. We used three types of amine promoters, including methyl (oxodioxolenyl) carbamate esters (3a-b), dipeptides (4ad), and pivaloyl-oxy-methyl (POM)-based esters (5a-c). Prodrugs containing dual proactives resulted in sufficient chemical stability to allow further evaluation in in vitro metabolic stability assays. While all prodrugs exhibited rapid metabolism in mouse plasma, some provided excellent stability in monkey and human plasma. When evaluated in vivo, the most stable DON prodrug (5c, methyl-POM-DON-isopropyl-ester) achieved a 10-fold improved brain:plasma ratio versus DON in monkeys, providing a potential clinical pathway for use. of DON in GBM patients.

RESULTS AND DISCUSSION

DON showed robust inhibition of glutamine metabolism and antitumor efficacy in a murine GBM model.

[0350] Despite several lines of evidence indicating the potential therapeutic efficacy of targeting glutamine metabolism in GBM, the effect of DON on GBM tumor growth has yet to be determined in vivo. Using the U87 flank xenograft mouse model of GBM, (Eshleman, et al., 2002) we first confirmed that systemic administration of DON (0.8 mg/kg, ip) inhibited glutamine metabolism as reflected in an accumulation of endogenous glutamine in the tumor (Figure 5A; p < 0.05) similar to other model systems. (Willis, et al., 1977; Windmueller, et al., 1974) We next evaluated its antitumor efficacy and found that DON not only arrested tumor growth, but also effectively induced tumor regression. Specifically, vehicle-treated mice showed significant tumor growth over the course of the experiment, while DON (0.8 mg/kg, ip, qd)-treated mice exhibited a >50% reduction in tumor volume (FIG. 5B, main effect of time [F(3,48) = 6.049, p = 0.0014], treatment [F(1,16) = 33.42, p < 0.0001], interaction [F(3, 48) = 21.70, p < 0.0001]). Although DON exhibited excellent antitumor efficacy, all mice receiving DON showed significant signs of toxicity including weight loss (12 ± 4.1%), hunching, ptosis, and lethargy. These results are consistent with other reports of DON efficacy and toxicity both in vitro and in vivo. (Fogal et al., 2015; Cervantes-Madrid, et al., 2015; Potter, et al., 2015)

Simple DON alkyl ester prodrugs were found to be unstable. Mask the carboxylate and amine functionalities of DON necessary to obtain stable prodrugs.

[0351] A prodrug strategy is often employed to improve tissue penetration and change the pharmacokinetic parameters of effective drugs. In fact, prodrug strategies are common in drug development, with 5-7% of approved drugs worldwide being prodrugs. (Rautio, et al., 2008) Our initial prodrug strategy for DON involved masking the carboxylic acid with simple alkyl esters such as ethyl 2a and isopropyl 2b. The synthesis of these two derivatives was straightforward providing compounds 2a and 2b in good yield. We are surprised that these simple DON alkyl esters have not been previously reported in the chemical literature, given that the chemistry and utility of DON have been described by numerous groups for over 60 years. (Magill, et al., 1957; Dion, et al., 1956; Magill, et al., 1956; Coffey, et al., 1956) One possible reason is that we found 2a and 2b to be unstable, slowing down the cycling to form unique diazoimines 9a and 9b. These two unique derivatives were found to be chemically stable even at acidic pH, precluding their use as DON prodrugs.

[0352] Given the instability of single ester prodrugs, we next masked both the primary amine and carboxylate of DON with prodrug moieties. This dual promotion strategy was streamlined to eliminate cyclization potential and potentially further improve lipophilicity. We used three amine promoters including (oxodioxolenyl)methyl carbamate esters (FIG. 4, 3a-b), dipeptides (4a-d) and pivaloyloxymethyl (POM)-based esters (5a-c). These promotions 59 because they target different metabolic enzymes including paraoxonase, aminopeptidases, and carboxylesterases, respectively. To impart a higher metabolic stability of the POM derivative (Table 1, 5a), we prepared the corresponding methyl-POM analogs (5b, 5c). All prodrugs containing dual proactives exhibited sufficient chemical stability to allow further evaluation.

All DON prodrugs were rapidly metabolized in mouse plasma, however 5b and 5c were found to be stable in human and monkey plasma.

[0353] Table 2 describes the plasma stability of DON prodrugs 3a-b, 4a-d and 5a-c. All prodrugs were completely metabolized in mouse plasma within the 60 min incubation time. However, in human and monkey plasma, prodrugs 5b and 5c, with methyl-POM on the amine and ethyl or isopropyl ester on the carboxylate, respectively, demonstrated moderate/high stability with 60-75% of the prodrug remaining in the plasma. monkey plasma, and the remaining 80-90% in human plasma within the 60 min incubation time. Since 5c had the best stability profile in human plasma, it was selected for further evaluation in pharmacokinetic studies and compared to DON for its ability to penetrate the brain and release DON.

Table 2. Plasma stability of DON prodrugs after 60 min incubation in mouse, monkey, and human plasma.

Major prodrug 5c enhanced brain delivery of DON in monkeys, but not in mice.

[0354] As expected for a DON prodrug that is completely metabolized in mouse plasma, we found that oral administration of DON(1) (0.8 mg/kg) and 5c (0.8 mg/kg equivalent) showed DON concentration profiles in plasma (FIG. 6) and brain (FIG. 6) similar when dosed in mice. The AUCü-t of DON after administration of DON and 5c in plasma was 1.25 nmol*h/mL and 1.22 nmol*h/mL respectively, suggesting rapid and complete release of DON from 5c in alive. Similarly, in mouse brain, the AUCü-t of DON after DON or 5c administration was 0.57 nmol*h/g and 0.69 nmol*h/g, respectively, with brain/plasma approximately 0.46 for DON vs. 0.56 for prodrug 5c. These pharmacokinetic results corroborated in vitro metabolism studies suggesting that 5c was completely converted to DON in mouse plasma.

[0355] Following the mouse studies, we evaluated the pharmacokinetics of DON and 5c (destereoisomer 1) in monkeys, as monkeys better mimicked the stability profile of human plasma. In pig-tailed macaques, i.v. DON and 5c (1.6 mg/kg DON equivalent) demonstrated significantly different plasma DON profiles (Figure 7). DON administration provided elevated plasma exposures with AUC0-t of 42.7 nmol*h/mL. In contrast, 5c administration produced a ~7-fold lower plasma DON exposure with an AUC0-t of 5.71 nmol*h/mL. The opposite observation was seen in the CSF where enhanced DON levels were seen after 5c administration. In the CSF at 30 min post-dose, DON administration resulted in 0.33 nmol/g DON while 5c delivered 1.43 nmol/g DON. When the ratio of plasma to CSF was compared at 30 min, 5c demonstrated a 10-fold improvement of DON CSF delivery versus DON (FlG. 7).

CONCLUSION

[0356] We demonstrated a profound efficacy of DON in a murine model of GBM, although overt toxicity was observed. To improve the therapeutic index of DON, we used a prodrug strategy to increase its cerebral delivery and limit its systemic exposure. While our dual-promoting prodrugs exhibited rapid metabolism in mice, our lead prodrug 5c provided excellent stability in human and monkey plasma, and achieved a 10-fold improved brain:plasma ratio versus DON in monkeys. This strategy may provide a pathway for the utilization of DON in GBM patients.

Efficacy studies in mice

[0357] All efficacy studies in mice were performed in accordance with protocol #MO13M69 approved by the Johns Hopkins University Animal Care and Use Committee. Nude female mice (HR Foxnlnu mice) between 25 and 30 g were obtained (Harlan Sprague Dawley Inc, Indianapolis, Indiana), and maintained on a 12-h light-dark cycle with ad libitum access to food and water. U87 human glioma cells were injected sc (5 x 10 6 cells in 100 ml PBS) at four separate sites on the flanks of each mouse. When the tumors grew to a mean volume of about 200 mm3, the mice were randomized into vehicle (buffered saline with HEPES, ip) or DON (1; 0.8 mg/kg, ip). In one cohort, mice were given a single dose of the appropriate solution two hours after which tumor glutamine levels were quantitated as described previously.49 Briefly, tumors were harvested, snap-frozen, and they were homogenized in liquid N2 and then subjected to metabolite extraction using methanol and DI water. Quantitation was performed using the Agilent 6520 Quadrupole-Time-of-Flight (Q-TOF) mass spectrometer with Agilent 1290 HPLC and the Agilent Mass Hunter and Agilent Qualitative and Quantitative Analysis software packages. Glutamine content was averaged by group for each individual tumor (n=3-4/group), represented as relative intensity, and analyzed by one-tailed t-test. In a second cohort, efficacy experiments were performed. Mice were injected once a day for six days; Tumor volumes were measured using digital calipers and calculated according to the Formula: [volume = (largest tumor dimension) x (smallest tumor dimension) 2 x 0.52] at 2, 4 and 6 days later from the start of treatment. Each individual tumor (n=8-10/group) was normalized to its pre-treatment volume, averaged, and analyzed by repeated measures two-way analysis of variance (ANOVA). If significant, a Bonferroni post hoc test was subsequently applied. Significance was defined as p < 0.05.

In vitro metabolic stability studies

[0358] For metabolic stability, plasma from rodent (mouse) and non-rodent (human and monkey) species was used. For stability, prodrugs (10 pm) were added into each plasma matrix and incubated on an orbital shaker at 37°C. At predetermined times (0 and 60 min), 100 mL aliquots of the mixture were removed in duplicate and the reaction stopped by the addition of three times the volume of ice-cold acetonitrile spiked with the internal standard (losartan 5 pm). Samples were shaken for 30 s and centrifuged at 12,000 g for 10 min. 50 ml of the supernatant was diluted with 50 ml of water and transferred to a 250 ml polypropylene vial sealed with a teflon cap. The disappearance of the prodrug was monitored over time using a liquid chromatography-tandem mass spectrometry (LC/MS/MS) method as described below.

Pharmacokinetic studies in mice

[0359] All pharmacokinetic studies in mice were performed in accordance with the protocol (#MO13M113) approved by the Johns Hopkins University Animal Care and Use Committee. C57BL/6 mice were obtained between 25 and 30 g from Harlan and maintained on a 12 hour light-dark cycle with ad libitum access to food and water. To assess the brain and plasma pharmacokinetics of DON and its 5c prodrug, C57BL/6 was administered from 8 to 12 weeks of age (1; 0.8 mg/kg, po in phosphate-buffered saline) and its 5c prodrug (a 0.8mg/kg). DON equivalent (1), po in phosphate buffered saline with 5% EtOH and 5% Tween-80. Mice were sacrificed by pentobarbital injection at 10, 30, and 90 minutes after drug administration, and blood was collected by cardiac puncture and placed in ice-cold EDTA-coated BD microvessels. Blood samples were centrifuged at 2,000 g for 15 minutes, and plasma was extracted and stored at -80 °C. Brain tissues were collected after blood collection and immediately frozen in liquid nitrogen and stored at -80 °C until LC/MS analysis.

Pharmacokinetic studies in non-human primates

[0360] All monkey studies were performed in accordance with the protocol (#PR15M298) approved by the Johns Hopkins University Animal Care and Use Committee. Two female pig-tailed macaques (approximately 3.5 kg, drug naïve) were housed adjacent to each other in stainless steel cages in a social interaction rack (contains 4 cages, each 32.5" in diameter). width x 28" depth x 32" height) maintaining a temperature of 64-84°C. °F, humidity of 30-70% with alternating light/dark cycles of 14-10 hours per the Animal Welfare Law of the USDA (9 CFR Parts 1,2 and 3) Food was provided daily in amounts appropriate for the size and age of the animals and reverse osmosis purified water provided ad libitum through a lixit valve in the cage. enriched feed Monday through Friday.Prior to drug administration, macaques were sedated with ketamine administered as an intramuscular injection prior to administration of the test article.Sedation was maintained through collection of blood and cerebrospinal fluid samples. (CSF) with ketamine at an initial rate of 15 mg/kg with additional doses of 20-30 mg during the first hour. At later times, ketamine was administered at 10-15 mg/kg. DON (50 pm HEPES-buffered saline) and 5c (Diastereoisomer 1), (50 mM HEPES-buffered saline containing 5% ethanol and 5% Tween) (equivalent to 1.6 mg/kg) to animals at a dosage volume of 1 ml/kg intravenously. CSF sample (50 mL target) was obtained by percutaneous puncture of the cisterna magna 30 min post-dose. Blood samples (1 mL) were collected at 15, 30, 1, 2, 4, and 6 h post-dose by percutaneous puncture of a peripheral vein. The samples were processed for plasma (centrifuged at a temperature of 4 °C, at 3000xg, for 10 minutes). All samples were kept refrigerated on ice throughout the processing. Samples were collected in microcentrifuge tubes, snap frozen, and placed in a freezer to maintain -80 °C until LC/MS analysis.

DON bioassay

[0361] We have previously published a highly sensitive method for the analysis of DON in biological matrices (Alt, et al., 2015). However, due to the chemical lability of DON and its prodrugs, a test method was developed and validated. milder derivatization employing dabsyl chloride. Briefly, DON was extracted from the samples (50 mg) with 250 |j| of methanol containing glutamate-d5 (10 µM ISTD) by vortexing in low retention tubes. The samples were centrifuged at 16,000 X g for 5 minutes to precipitate the proteins. The supernatants (200 jl) were transferred to a new tube and dried at 45 °C under vacuum for 1 hour. To each tube was added 50 µL of 0.2 M sodium bicarbonate buffer (pH 9.0) and 100 µL of 10 mM dabsyl chloride in acetone. After vortexing, samples were incubated at 60 °C for 15 min to derivatize. Samples (2 µl) were injected and separated on an Agilent 1290 equipped with an Agilent Eclipse plus C18 RRHD 2.1 X 100mm column in a 2.5 minute gradient of 20-95% acetonitrile 0.1% formic acid and they were quantified in an Agilent 6520 QTOF mass spectrometer. Calibration curves in the range of 0.005-17.1 µg/mL in plasma and CSF for DON were constructed from the ratio of the peak area of the analyte to the internal standard by linear regression with a weighting factor of 1/( nominal concentration). A correlation coefficient greater than 0.99 was obtained in all analytical runs. The predicted mean relative standard deviation for the back-calculated concentrations of the standards and quality controls for all analytes was within the range of 85 to 115%, except for the lowest concentration which was within the range of 80 to 120% with a overall accuracy and precision of 6.7% and 6.6% respectively.

pharmacokinetic analysis

[0362] Median concentration-time data was used for pharmacokinetic analysis. The non-compartmental analysis module in WinNonlin® (version 5.3) was used to evaluate the pharmacokinetic parameters. Peak plasma concentrations (Cmax) and time to Cmax (Tmax) were the observed values. The area under the curve (AUC) was calculated using the log-linear trapezoidal rule to the end of sample collection (AUClast).

REFERENCE EXAMPLE 4

[0363] In head-to-head comparisons, 25 was found to be markedly more effective than a clinical stage selective glutaminase inhibitor CB-839.

FIG. 8 illustrates that 25 (5 day dosing beginning on day 7) is superior to CB-839 (30 day dosing beginning on day 1) in a CT26 tumor model.

[0364] FIG. 9 illustrates that 25 (4 days from day 6) is superior to CB-839 (continuous dosing twice daily from day 1 prior to engraftment) in a CT26 tumor model. Mice received 25 (1.9 mg/kg) daily on days 6-9 compared to BID glutaminase inhibitor on days 1-15.

[0365] FIG. 10 illustrates that 25 (daily on days 7-22) is superior to CB-839 (continuous dosing twice daily on days 1-29) in a 4T1 breast cancer model. Mice received 25 (1.0 mg.kg/d) daily for days 7-22 compared to BID glutaminase inhibitor for days 1-29.

REFERENCE EXAMPLE 5

[0366] DON prodrugs demonstrate efficacy in multiple tumor types, including efficacy in B and T cell lymphomas, colon cancers, breast cancer, and melanoma. Daily dosing provides effective monotherapy. Dosing every other day leads to minimal resistance. FIG. 11 illustrates that dosing of 1 mg/kg followed by 0.3 mg/kg leads to a complete and long-lasting response in the MC38 tumor.

[0367] FIG. 12 illustrates that 25 provides a robust response and improves overall survival in multiple tumor models, including CT26 Colon Cancer.

[0368] FIG. 13 illustrates that 25 provides a robust response and improves overall survival in multiple tumor models, including 4T1 Breast Cancer.

[0369] FIG. 14 illustrates that 25-cured mice only immunologically reject tumors upon rechallenge, demonstrating that monotherapy with certain DON prodrugs, such as 25 monotherapy, is immunotherapy. FIG. 15 further illustrates that 25 is immunotherapy.

EXAMPLE 6

[0370] DON/DON prodrugs condition tumors for immunotherapy and significantly enhance response to checkpoint inhibitors, adoptive cell transfer, and A2aR inhibition

[0371] Conclusions: DON/DON prodrugs robustly enhance the immune-mediated response to anti-PD1 therapy, the response to adoptive cell transfer, and the response to adenosine A2a receptor blockade. Immunotherapy fails in patients with rapidly progressive disease due to delayed response. 30-40% of patients treated with anti-PDI therapy progress rapidly in the first few months, chemotherapy showed an early survival advantage in NSCLC probably due to its rapid effect on rapidly growing disease compared with immunotherapy, and the Conditioning therapies and adjuvants to immunotherapy must be able to control tumor growth to be effective in these patients.

[0372] FIG. 16 illustrates that inhibition of glutamine ( eg, using DON) reduces oxygen consumption and lactate production of tumor cells. FIG. 17 illustrates that glutamine inhibition ( eg, using DON) also improved the CD8/Treg ratio in the tumor and reduces hypoxia in TILs.

[0373] FIG. 18 illustrates that 25 conditions tumors to be eliminated by anti-PDI therapy in Model MC38, and that 25 rescues anti-PDI failures. Furthermore, FIG. 19 illustrates that even in the most difficult CT26 model, 25 the response to anti-PDl improves. Similarly, FIG. 20 illustrates that inhibition of glutamine metabolism potentiates the antitumor response to A2aR inhibition. Finally, the figure. 21 illustrates that inhibition of glutamine metabolism improves the efficacy of adoptive cell therapy (ACT) in a B16-OVA model.

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[0375] All publications, patent applications, patents and other references mentioned in the specification are indicative of the level of those skilled in the art to which the subject matter of the present disclosure pertains. It will be understood that, although reference is made herein to a number of patent applications, patents and other references, such reference does not constitute an admission that any of these documents form part of the common general knowledge in the art. In the event of a conflict between the specification and any of the incorporated references, the specification shall control. The standard meanings of terms accepted in the art are used herein unless otherwise indicated. In this document, standard abbreviations are used for various terms.

Claims (12)

1. A compound having Formula (I):

or a pharmaceutically acceptable salt thereof, wherein: X is -(CH2)n-; n is 1; R1 is selected from the group consisting of C1-6 alkyl and substituted C1-6 alkyl; R2 is selected from the group consisting of an amino acid, an N-substituted amino acid, -C(=O)-Y-(CR3R4)m-NR5R6, and -C(=O)-O-(CR3R4)m-O-C(=O) -R10; Y is -O- or a link; R2' is selected from the group consisting of H, C1-C6 alkyl, and substituted C1-C6 alkyl; each R3 and R4 are independently H, C1-C6 alkyl, substituted C1-C6 alkyl, aryl, substituted aryl, -(CR3R4)m-NR5R6, or

m is an integer selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, and 8; R5 and R6 are independently H or alkyl; Y Rio is selected from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, monosaccharide, acylated monosaccharide, aryl, substituted aryl, heteroaryl, and substituted heteroaryl, for use in the treatment of cancer, wherein the compound having Formula (I) is to be administered with immune checkpoint blockade therapy, adoptive cell therapy, or an adenosine A2aR inhibitor. 2. The compound for use according to claim 1, wherein R1 is selected from the group consisting of methyl, ethyl, and isopropyl. The compound for use according to claims 1 or 2, wherein: R2 is selected from the group consisting of -C(=O)-Y-(CR3R4)m-NR5R6 and -C(=O)-O-(CR3R4)m-O-C(=O)-R10; Y is a link; m is an integer selected from the group consisting of 0, 1,2, 3, 4, 5, 6, 7, and 8; each R3 and R4 is independently H, C1-C6 alkyl or substituted C1-C6 alkyl, aryl or substituted aryl; and R10 is alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, or substituted heteroaryl. 4. The compound for use according to claim 1, wherein R2 is an amino acid. The compound for use according to claim 1, wherein the compound having Formula (I) is selected from the group consisting of:





; Y

The compound for use according to any of claims 1-5, wherein the cancer is a newly diagnosed, recurrent and/or refractory cancer selected from the group consisting of cellnasopharyngeal cancer, synovial cancer, hepatocellular cancer, renal cancer, connective tissue cancer, melanoma, lung cancer, bowel cancer, colon cancer, rectal cancer, colorectal cancer, brain cancer, throat cancer, oral cancer, liver cancer, bone cancer, pancreatic cancer, choriocarcinoma , gastrinoma, pheochromocytoma, prolactinoma, T-cell leukemia/lymphoma, neuroma, von Hippel-Lindau disease, Zollinger-Ellison syndrome, adrenal cancer, anal cancer, bile duct cancer, bladder cancer, ureteral cancer, cancer brain, oligodendroglioma, neuroblastoma, meningioma, spinal cord tumor, bone cancer, osteochondroma, chondrosarcoma, Ewing's sarcoma, cancer of unknown primary site, carcinoid, g-tract carcinoid astrointestinal, fibrosarcoma, breast cancer, Paget's disease, cervical cancer, colorectal cancer, rectal cancer, esophageal cancer, gallbladder cancer, head cancer, eye cancer, neck cancer, kidney cancer, tumor of Wilms, liver cancer, Kaposi's sarcoma, prostate cancer, lung cancer, testicular cancer, Hodgkin's disease, non-Hodgkin's lymphoma, oral cancer, skin cancer, mesothelioma, multiple myeloma, ovarian cancer, endocrine pancreatic cancer, glucagonoma, pancreatic cancer, parathyroid cancer, penile cancer, pituitary cancer, soft tissue sarcoma, retinoblastoma, small bowel cancer, stomach cancer, thymus cancer, thyroid cancer, trophoblastic cancer, hydatidiform mole, cancer of the uterus, endometrial cancer, vaginal cancer, vulvar cancer, acoustic neuroma, mycosis fungoides, insulinoma, carcinoid syndrome, somatostatinoma, gum cancer, heart cancer, lip cancer, meningeal cancer, mouth cancer, nerve cancer, palate cancer, parotid gland cancer, peritoneal cancer, pharyngeal cancer, pleural cancer, salivary gland cancer, tongue cancer, and tonsil cancer. The compound for use of any one of claims 1-6, wherein the compound is to be administered with checkpoint blockade therapy. The compound for use of claim 7, wherein the checkpoint blockade therapy is selected from the group consisting of PD-1 antagonists, PD-L1 antagonists, CTLA-4 antagonists, Lag-3 antagonists , CD137 antagonists, KIR antagonists, Tim3 antagonists, Ox40 agonists, and B7-H3 antagonists. The compound for use of claim 7, wherein the compound is to be administered with nivolumab, pembrolizumab, or pidilizumab. The compound for use of any one of claims 1-6, wherein the compound is to be administered with adoptive cell therapy. The compound for use of claim 10, wherein the compound is to be administered with T cells, CD8+ cells, CD4+ cells, NK cells, delta-gamma T cells, marrow infiltrating lymphocytes (MIL), regulatory T cells, peripheral t. blood mononuclear cells or tumor-infiltrating lymphocytes (TILs). The compound for use of any one of claims 1-6, wherein the compound is to be administered with an adenosine A2aR inhibitor.