[1,2,4]Triazolo[4,3-a]pyrimidin-7(8H)-ones as mitochondrial pyruvate transporter inhibitors for use in the treatment of cancer

Abstract

The present invention relates to compounds, methods, compositions and uses that are capable of inhibiting mitochondrial pyruvate transporter (MPC) activity and that are useful in immunotherapy, in particular in T cell therapy, immune checkpoint inhibitors or anti-cancer vaccines.

Description

[Technical field]

[0001] The present invention relates to the field of immunotherapy, particularly adoptive cell transfer immunotherapy and vaccine therapy. In particular, the present invention relates to mitochondrial pyruvate transporter (MPC) inhibitors and uses thereof. [Background technology]

[0002] Adoptive cell transfer (ACT) immunotherapy is the transfer of T cells into patients to treat diseases such as cancer or viral infections. Chimeric antigen receptor (CAR) T cells are one type of ACT that involves transferred T cells (either the patient's own or a donor's) that are genetically engineered ex vivo to express a chimeric antigen receptor that targets a specific tumor antigen. CAR T therapy has emerged as one of the most promising approaches for relapsed and refractory hematological malignancies. In fact, it has already helped extremely sick patients defeat diffuse large B-cell lymphoma (DLBCL) and acute lymphoblastic leukemia (ALL) where traditional treatments such as chemotherapy have failed (Wang et al., 2017, Journal of Hematology and Oncology, 10(1):53). However, CAR T therapy has shown limited efficacy against solid tumors, which account for the majority of cancer cases (approximately 90% of cancer cases in the United States). In particular, it has been shown that the lack of T cell proliferation and persistence after infusion into patients is largely responsible for the poor efficacy of CAR-T therapy in solid tumors. Also, even in hematological malignancies with high complete remission rates of around 90%, patients remain at risk of relapse due to poor persistence of CAR-T cells in vivo (Gauthier et al., 2017, Curr Res Transl Med, 65(3):93-102). Thus, there is an unmet need to develop CAR T cells that survive, expand and persist in vivo.

[0003] One strategy is to inject T cells with an early memory phenotype. According to their differentiation stage, T cells present a unique phenotype with related functionality and properties. Recently, it has been shown that clinical responses are related to the state of T cell differentiation (Lecuroux et al., 2009, Blood, 113(14):3209-3217). Historically, ACT used terminally differentiated T "effector" cells due to their potent killing capacity. However, T "effector" cells have poor ability to expand and persist in vivo. In mouse tumor models, it has been shown that injecting self-renewing T cells with an early memory phenotype confers stronger and more sustained antitumor responses (Klebanoff et al., 2011, Clin Cancer Res., 17, 5343-5352). Furthermore, it has been shown that inhibition or stimulation of certain metabolic processes alone is sufficient to induce effector to memory differentiation, with corresponding in vivo effects when the cells are reinjected into infected or tumor-bearing mice (Balmer et al., 2016, Immunity, 44, 1312-1324; Phan et al., 2016, Immunity 45, 1024-1037). However, current strategies significantly inhibit T cell proliferation and are therefore not suitable for use as a replenishment during the ex vivo manufacturing process of T cells for ACT immunotherapy. Therefore, there is a need for improved methods to obtain T cells with a memory phenotype.

[0004] Furthermore, there is a need to improve adjuvants for vaccines. Indeed, one of the main goals of prophylactic and therapeutic vaccination in the context of pathogen infection and cancer is the establishment of a strong memory T cell pool. Current adjuvants for vaccination are either aluminum salts and emulsions or more specific pattern recognition receptor agonists, all designed to target and activate antigen-presenting cells (APCs). This does not always result in a highly efficient activation of both humoral and cellular immunity, a necessary requirement for successful virus neutralization and / or elimination (such as SARS-CoV-2) (Pulendran et al., 2021, Nature Reviews Drug Discovery, 20, 454-475).

[0005] Recently, it has been shown that targeting mitochondrial metabolism to boost T cell memory formation and metabolic fitness may be an attractive strategy to improve cancer immunotherapy, including CAR-T therapy (Li et al. 2020, Front Immunol., 11:1834).

[0006] Pyruvate is a metabolite involved in several biological processes and is particularly important in cellular respiration. Pyruvate, the end product of glycolysis in the cytosol, needs to enter mitochondria to fuel the Krebs cycle and boost oxidative phosphorylation and ATP production. To enter mitochondria, pyruvate crosses the outer mitochondrial membrane and reaches the intermembrane space, presumably through a large, relatively nonspecific, voltage-dependent anion channel, which is then transported together with protons across the inner mitochondrial membrane by the mitochondrial pyruvate transporter (MPC) (Papa et al., 1971, FEBS Lett., 12, 285-288). Although the existence of MPC was proposed as a theoretical basis several decades ago, the molecular identification of the MPC complex was only achieved in 2012 (Herzig et al., 2012, Science, 337, 93-96). As the sole point of translocation of pyruvate into the mitochondrial matrix, the MPC plays a key role in coordinating glycolysis and mitochondrial activity, providing a key decision point for modulating cellular energy production and metabolism.

[0007] It has been recently reported that MPCs play pivotal roles in many physiological and pathological processes across the human life span, from embryonic development to age-related neurodegeneration (Buchanan et al., 2020, Biomolecules, 10(8):1162; Zangari et al., 2020, Biomolecules, 10(7):1068). In particular, MPCs are of importance in cancer cell metabolism and tumorigenesis (Ruiz Iglesias et al., 2021, Cancers 2021, 13, 148).

[0008] In particular, genetic inhibition of MPC has been shown to stimulate stem cell proliferation in various tissues, including the gut and skin (Flores et al., 2021, Exp Dermatol., 30(4):448-456; Schell et al., 2017, Nat Cell Biol., 19(9):1027-1036).

[0009] Furthermore, it has been shown that genetic inhibition of MPC is an interesting therapeutic strategy to treat non-alcoholic steatohepatitis (NASH), an increasingly common liver disease in metabolic diseases including type 2 diabetes (Hojlund et al., 2008, Endocrinol Metab Clin North Am., 37(3):713-3), and in cancer (Harisson et al., 2020, J. Hepatol., 72, 613-626), and to enhance existing cancer treatments, including radiation therapy (Corbet et al., 2018, Nat. Commun., 9, 1208).

[0010] Thus, considering the recent development of various strategies in cancer immunotherapy such as cancer vaccines, adoptive cellular immunotherapy, immune checkpoint blockade and oncolytic viruses, despite the limitations encountered in their efficacy, there is an increasing need to develop efficient anticancer therapies for solid tumor cancers, especially for those cancers prone to develop resistance to immunotherapy that would enhance cancer vaccine treatment. Furthermore, there is a need for the development of efficient MPC inhibitors to further understand the role of MPC activity in various disorders. Summary of the Invention

[0011] The present invention is directed to the unexpected finding that pharmacological MPC inhibition by the compounds of the present invention during T cell culture for preparing ACT products can induce an increase in the proportion of activated T cells that are highly involved in acquiring memory phenotype and improved antitumor activity.Therefore, the MPC inhibitors according to the present invention are considered to be of high interest in specific immunotherapy of cancer, in particular in adoptive T cell transfer approach (ACT), in particular for CAR T therapy and / or for cancer vaccine and / or vaccine against infectious diseases.

[0012] The present invention is directed to useful compositions and methods for inhibiting the activity of MPC and therefore, inter alia, for enhancing the efficacy and durability of cancer treatments including ACT, for stimulating stem cell proliferation in various tissues including the gut, skin and brain, for reducing inflammation and fibrosis in several organs including the liver, lung, pancreas, muscle, for reducing tumor incidence, and / or for enhancing existing cancer treatments including immune checkpoint inhibitors and radiation therapy.

[0013] A first aspect of the invention provides a compound of the invention for the prevention and / or treatment of a disease or disorder, wherein said disease or disorder is selected from cancer; an autoimmune disease, such as multiple sclerosis; a metabolic disease, such as type 2 diabetes; a hair loss disorder, such as alopecia; a neurodegenerative disorder, such as Parkinson's disease or Alzheimer's disease; a fibrosis, such as pulmonary fibrosis or non-alcoholic steatohepatitis (NASH); a skin or tissue injury, such as a skin wound or burn; and an acute condition of the brain, such as stroke or brain trauma.

[0014] Another aspect of the present invention provides a compound of the present invention for use in skin or tissue regeneration.

[0015] Another aspect of the present invention provides the use of one or more compounds of the present invention for the preparation of a pharmaceutical composition for the prevention and / or treatment of a disease or disorder, wherein said disease or disorder is selected from cancer; an autoimmune disease, such as multiple sclerosis; a metabolic disease, such as type 2 diabetes; a hair loss disorder, such as alopecia; a neurodegenerative disorder, such as Parkinson's disease or Alzheimer's disease; a fibrosis, such as pulmonary fibrosis or non-alcoholic steatohepatitis (NASH); a skin or tissue injury, such as a skin wound or burn; and an acute condition of the brain, such as stroke or brain trauma.

[0016] Another aspect of the present invention relates to a pharmaceutical composition comprising at least one compound according to the present invention, as well as tautomers, geometric isomers, optically active forms and pharma- ceutically acceptable salts thereof, in combination with at least one anti-cancer immunotherapeutic agent, such as a CAR T cell or an immune checkpoint inhibitor, and at least one pharma- ceutically acceptable carrier, diluent or excipient thereof.

[0017] Another aspect of the present invention relates to a method for treating a subject suffering from a disease or disorder, wherein said disease or disorder is selected from cancer; an autoimmune disease, e.g., multiple sclerosis; a metabolic disease, e.g., type 2 diabetes; a hair loss disorder, e.g., alopecia; a neurodegenerative disorder, e.g., Parkinson's disease or Alzheimer's disease; a fibrosis, e.g., pulmonary fibrosis or non-alcoholic steatohepatitis (NASH); a skin or tissue injury, e.g., a skin wound or burn; and an acute condition of the brain, e.g., a stroke or brain trauma, wherein said method comprises administering to a subject in need thereof an effective amount of one or more compounds of the present invention.

[0018] Another aspect of the invention relates to the described MPC inhibitors, their pharmaceutical formulations and their use as medicaments.

[0019] Another aspect of the present invention relates to an in vitro method for generating and / or maintaining T cells with a memory phenotype.

[0020] Other features and advantages of the present invention will become apparent from the following detailed description. [Brief description of the drawings]

[0021] [Figure 1]Figure 1 represents the treatment protocol (A) and the effect of in vitro treatment of mouse CD8 T cells with a compound that inhibits the mitochondrial pyruvate transporter (MPCi) on memory properties (B-C), as described in Example 3. (A) Schematic representation of in vitro mouse CD8 T cell activation and treatment. (B-C) FACS analysis on day 7 showing the percentage of cells positive for memory markers CD62L (B) and CD127 (C). *p<0.05, **p>0.01, ***p>0.01. Data represent the mean ± sem. [Diagram 2] FIG. 2 depicts the treatment protocol (A) and antitumor activity (B-S) of a compound of the invention (C45) in a mouse melanoma model, as described in Example 4. (B-C) B16-OVA tumor growth (B) and body weight (C) in mice upon transfer of compound 45 or DMSO-treated cells, or in untreated mice. (D-G) Blood analysis 9 days after ACT showing the percentage of transferred cells (D), short-lived effector cells (E), memory progenitor cells (F) and central memory cells (G); (H-K) Analysis of tumor-infiltrating T cells showing the percentage of transferred cells (H), exhausted cells (I), terminally exhausted cells (J) and precursor exhausted cells (K). (L-N) Spleen analysis showing the percentage of transferred cells (L), central memory T cells (M) and T cells expressing TCF1 (N). (O-S) Single cell suspensions of tumors restimulated with Ovalbubin N4 peptide for 4 h. IFNγ (O), TNFα (P), IL2 (Q), Granzyme B (R) and CD107a (S) expression was measured by flow cytometry. ns: not significant, *p<0.05, **p>0.01, ***p>0.001. Data represent mean ± sem. [Diagram 3]FIG. 3 represents the treatment protocol (A) and antitumor activity (B-0) of a compound of the invention (C45) in a mouse melanoma model (transferring 2×10 6 DMSO- or compound 45-treated OT1 T cells instead of only 10 5 ) as described in Example 4. (B-C) B16-OVA tumor growth (B) and body weight (C) in mice upon transfer of compound 45 or DMSO-treated cells or in untreated mice. (D-G) Analysis of tumor-infiltrating T cells showing the number of transferred cells (D), exhausted cells (E), terminally exhausted cells (F) and precursor exhausted cells (G). (H-J) Analysis of spleen showing the number of transferred cells (H), central memory T cells (I) and TCF1-expressing T cells (J). (K-O) Single cell suspensions of tumors restimulated with N4 peptide for 4 hours. IFNγ (K), TNFα (L), IL2 (M), Granzyme B (N) and CD107a (O) expression was measured by flow cytometry. ns: not significant, *p<0.05, **p>0.01, ***p>0.001. Data represent the mean ± sem. [Figure 4]Figures 4 and 5 represent the experimental design (4A) and the effect of the compounds of the invention during the generation of mouse CAR T cells to improve their memory phenotype and antitumor function during adoptive cell transfer therapy (4B-F and 5A-K), as described in Example 5. (4B) Tumor growth curve; (4C) Tumor weight T at dissection; (4D) Number of Her2-CAR T cells per μl of blood; (4E and F) Percentage of short-lived effector cells (4E) and memory progenitor cells (4F) among Her2-CAR T cells. ns: not significant, *p<0.05, **p>0.01. Data represent mean ± sem. (5A) Number of Her2-CAR T cells per tumor-draining lymph node; (5B) percentage of central memory T cells among Her2-CAR T cells in lymph node; (5C) percentage of TCF1 positive cells among Her2-CAR T cells in lymph node; (5D) number of Her2-CAR T cells per spleen; (5E) percentage of central memory T cells among Her2-CAR T cells in spleen; (5F) percentage of TCF1 positive cells among Her2-CAR T cells in spleen; (5G) number of Her2-CAR T cells per mg of tumor. (5H) Percentage of TCF1 positive cells among Her2-CAR T cells in the tumor; (5I and J) Percentage of precursor exhausted (I) and terminal exhausted (5J) T cells among Her2-CAR T cells in the tumor; (5K) Percentage of exhaustion markers (PD1 and TIM3) expressing Her2-CAR T cells in the tumor. ns: not significant, *p<0.05, **p>0.01, ***p<0.001. Data represent mean ± sem. [Diagram 5] Same as above. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022] The expression "solid tumor cancer" includes glioblastoma, lung cancer (small cell and non-small cell), breast cancer, ovarian cancer, cervical cancer, uterine cancer, head and neck cancer, melanoma, hepatocellular carcinoma, colon cancer, rectal cancer, colorectal cancer, kidney cancer, prostate cancer, gastric cancer, bronchial cancer, pancreatic cancer, bladder cancer, liver cancer and brain cancer, in particular glioblastoma.

[0023] The term "liquid tumor cancer" includes lymphomas and leukemias.

[0024] The expression "immunotherapeutics" refers to drugs that support the immune system in fighting diseases such as cancer. There are currently four main categories: T-cell therapies, immune checkpoint inhibitors, monoclonal antibodies and cancer vaccines.

[0025] As used herein, "treatment" and "treating" generally refer to obtaining a desired pharmacological and physiological effect. The term "treatment" as used herein includes any treatment of a disease in a mammal, particularly a human, and includes inhibiting the disease, i.e., arresting its development; or alleviating the disease, i.e., causing a reduction in the disease and / or its symptoms or conditions, such as arresting tumor growth or causing a tumor to shrink.

[0026] The term "subject" as used herein refers to a mammal. For example, mammals contemplated by the present invention include humans, primates, domestic animals such as cows, sheep, pigs, horses, laboratory rodents, dogs, and the like.

[0027] The term "effective amount" as used herein refers to an amount of at least one particle according to the present invention or a pharmaceutical formulation thereof that induces a biological or pharmaceutical response in a tissue, system, animal, or human that is sought. In one embodiment, the effective amount is a "therapeutically effective amount" for the alleviation of symptoms of the disease or condition being treated. Typically, an effective amount can be used to inhibit the growth of cancer cells, i.e., optionally slow the rate of proliferation and / or migration of cancer cells, stop the proliferation and / or migration of cancer cells, or kill cancer cells, so that the rate of growth of cancer cells is reduced compared to the observed or predicted rate of growth of untreated control cancer cells. The term "inhibit growth" can also refer to the reduction or disappearance of the size of cancer cells or tumors, and the reduction of their metastatic potential. Preferably, such inhibition at the cellular level can reduce the size, postpone growth, reduce aggressiveness, or prevent or inhibit metastasis of the cancer in the patient. Those skilled in the art can easily determine whether the growth of cancer cells is inhibited by any of a variety of suitable indications.

[0028] The term "efficacy" of a treatment according to the invention can be measured based on changes in the course of a disease in response to a use or method according to the invention. The efficacy of a treatment of cancer according to the invention can be measured by a reduction in tumor volume and / or an increase in progression-free survival and / or an increase in the health and well-being of a subject (e.g., suppressing cancer). Inhibition of cancer cell growth may be evidenced, for example, by arrest of cancer cells in a particular stage of the cell cycle, for example arrest at the G2 / M phase of the cell cycle. Inhibition of cancer cell growth can also be evidenced using well-known imaging methods, for example magnetic resonance imaging, computed tomography, PET, SPECT, photoacoustic imaging, X-ray and fluorescent imaging / detection. Cancer cell growth can also be determined indirectly, for example, by determining the levels of circulating carcinoembryonic antigen, prostate specific antigen, or other cancer specific antigens that correlate with cancer cell growth.

[0029] In particular, the efficacy of the combination treatment according to the invention can be assessed by the reduction in tumor size or disappearance of any biomarker associated with the tumor or cancer type. In the context of adoptive cell transfer (ACT), efficacy can also be measured by assessing transferred T cell infiltration into the tumor, migration of transferred T cells to lymph nodes, or any change in the T cell "profile" or "differentiation state".

[0030] The term "profile" of a cell according to the invention, in particular a T cell or any peripheral blood mononuclear cell (PBMC), can be measured based on changes in gene expression or cell surface markers in response to a use or method according to the invention, and / or an increase in basal oxygen consumption, maximal respiratory capacity and / or spare respiratory capacity compared to a control.

[0031] More specifically, the term "memory phenotype" as used herein is defined as a cell state that at least resembles memory T cells in some aspects. The term "memory-like T cells" is used herein interchangeably with the term "memory phenotype". An important feature associated with memory phenotype is the longevity of cells. Longevity means that cells or precursors survive long enough, for example, without division or with slow division, in a subject that can induce a therapeutic effect. In particular, cells with memory phenotype have stem cell-like properties. Longevity is preferably due to self-renewal, including proliferation. Self-renewal as used herein does not mean a strict meaning, but also includes the ability to maintain a similar, but not necessarily identical, phenotype for a therapeutically relevant period of time. Self-renewal can be maintained for the entire life span or beyond, but as used herein, it is sufficient that it is maintained long enough for therapeutic purposes. A therapeutically relevant period of time means that the transferred cells or their progeny persist in the subject long enough to have a therapeutic effect. While a T cell with a memory phenotype is alive, preferably proliferating, it typically maintains the ability to differentiate into effector cells. Therefore, the ability to generate effector T cells is another important feature associated with the memory phenotype. Therefore, T cells with a memory phenotype can generate a larger number of therapeutically active effector cells than the effector cells themselves, which tend to age and die prematurely. Important functional features associated with the memory phenotype can also be measured relative to other cell populations. For example, longevity, self-renewal and / or the ability to differentiate into effector cells may be compared to effector cells, terminally differentiated cells and / or senescent cells.

[0032] Suitable positively expressed markers for the memory phenotype (memory markers), in particular those of CD8+ T cells, but also at least in part those of CD4+ T cells and / or B cells, are, for example, CCR7, CD62L, CD27, CD28, CD127 and / or TCF1.

[0033] Another important feature associated with the memory phenotype is the ability of cells to respond with increased amplitude of (re)activation upon re-encounter with antigen, as observed, for example, in memory T cells.

[0034] Unless otherwise constrained by the definition of the individual substituents, the term "substituted" ("substituted" or "substituted") refers to a group substituted or substituted with 1 to 5 substituents selected from the group consisting of "C1-C6 alkyl", "C2-C6 alkenyl", "C2-C6 alkynyl", "C3-C8-cycloalkyl", "heterocycloalkyl", "C1-C6 alkylaryl", "C1-C6 alkylheteroaryl", "C1-C6 alkylcycloalkyl", "C1-C6 alkylheterocycloalkyl", "amino", "alkylamino", "arylamino", "heteroarylamino" and "aryloxy", "heteroaryloxy", "urea", "aminosulfonyl", "ammonium", "alkoxy", "acyl", "acylamino", "aminocarbonyl", "aryl", "heteroaryl", "sulfinyl", "sulfonyl", "sulfonamido", "alkoxy", "alkoxycarbonyl", "carbamate", "sulfanyl", "halogen", trihalomethyl, cyano, hydroxy, mercapto, nitro, and the like.

[0035] The term "pharmaceutically acceptable salt or complex" refers to a salt or complex of the compound of the present invention as defined below. Examples of such salts include, but are not limited to, base addition salts formed by reacting the compound of the present invention with an organic or inorganic base, such as hydroxides, carbonates, bicarbonates, etc. of metal cations, such as those selected from the group consisting of alkali metals (sodium, potassium or lithium), alkaline earth metals (e.g., calcium or magnesium), or with organic primary, secondary or tertiary alkylamines. Other examples of such salts include, but are not limited to, acid addition salts formed by reacting the compound of the present invention with an organic or inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, para-toluenesulfonic acid, 2-naphthalenesulfonic acid, camphorsulfonic acid, benzenesulfonic acid, oxalic acid, etc.

[0036] Compounds of the invention also include isotopic isomers of compounds of formula (I), eg, deuterated and C13 analogs.

[0037] "Pharmaceutically active derivative" refers to any compound that, upon administration, is capable of providing to a recipient, either directly or indirectly, an activity disclosed herein.

[0038] Compounds according to the invention In one embodiment, the present invention provides a compound of formula (I) for use in the prevention and / or treatment of a disease or disorder selected from cancer; an autoimmune disease, such as multiple sclerosis; a metabolic disease, such as type 2 diabetes; a hair loss disorder, such as alopecia; a neurodegenerative disorder, such as Parkinson's disease or Alzheimer's disease; a fibrosis, such as pulmonary fibrosis or non-alcoholic steatohepatitis (NASH); a skin or tissue injury, such as a skin wound or burn; and an acute condition of the brain, such as stroke or brain trauma, or for use in skin or tissue regeneration: [ka] wherein R1 is an R5-R6 moiety; R2 is H, optionally substituted C1-C6 alkyl (optionally substituted methyl, optionally substituted ethyl, optionally substituted propyl, such as trifluoropropyl, optionally substituted butyl, optionally substituted t-butyl, etc.), optionally substituted aryl, optionally substituted heteroaryl, optionally substituted aryl C1-C6 alkyl (optionally substituted arylmethyl, such as optionally substituted phenylmethyl), optionally substituted heteroaryl C1-C6 alkyl, optionally substituted C1-C6 alkoxy, such as optionally substituted C1-C6 alkoxy substituted C1-C6 alkyl, such as optionally substituted ethoxy (e.g. optionally substituted ethoxybenzyl, e.g., optionally substituted ethoxyhalogenobenzyl, i.e., 2-chlorobenzyloxyethyl or 3-chlorobenzyloxyethyl, 4-chlorobenzyloxyethyl, or optionally substituted ethoxymethyl), optionally substituted methoxy (e.g., optionally substituted methoxymethyl), optionally substituted C-C heterocycloalkyl, and optionally substituted C-C cycloalkyl, e.g., optionally substituted cyclopropyl; R3 is selected from H and optionally substituted C-C alkyl (e.g., optionally substituted methyl), and at least one of R2 and R3 is not H; R4 is H; R5 is selected from a bond, S, SO2, NR9, and O; R6 is -(CR10R11) n-R7 group (wherein n is an integer from 0 to 2); R7 is an optionally substituted C1-C6 alkyl (e.g., optionally substituted methyl, optionally substituted propyl, optionally substituted t-butyl), optionally substituted C2-C6 alkenyl (e.g., 3-phenylpropylenyl), optionally substituted heterocycle, optionally substituted aryl, for example, optionally substituted phenyl (e.g., 4-nitrophenyl, 2-nitrophenyl). , 3-nitrophenyl, 4-nitrophenyl, 2-fluorophenyl, 3-fluorophenyl, 2,5-difluorophenyl, 2,4-difluorophenyl, 3,5-difluorophenyl, 2,4-difluorophenyl, 2,6-difluorophenyl, 3-(difluoromethyl)phenyl, 2-(difluoromethyl)phenyl, 4-(difluoromethyl)phenyl, 3,4-difluorophenyl, 2-chloro-6-fluorophenyl, 2-chlorophenyl, 2-chloro-5-fluorophenyl, 3-chloro-5-fluorophenyl, 4-chloro-3-fluorophenyl, fluorophenyl, 3-chlorophenyl, 2,4-dichlorophenyl, 3,4-dichlorophenyl, 4-chlorophenyl, 5-fluoro-2-methylphenyl, 4-fluoro-2-benzonitrile, 2,5-dimethylphenyl, 3-methylphenyl, 4-methylphenyl, 3-(trifluoromethyl)phenyl, 4-(trifluoromethyl)phenyl, 5-fluoro-2-methoxyphenyl, 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 4-bromophenyl), optionally substituted naphthalenyl (e.g., optionally substituted optionally substituted naphthalen-1-yl), optionally substituted heteroaryl, for example, optionally substituted furanyl (e.g., optionally substituted furan-3-yl, for example, furanyl, methoxycarbonylfuranyl), optionally substituted pyrazolyl, optionally substituted pyridinyl (e.g., optionally substituted pyridin-4-yl), optionally substituted benzofuranyl (e.g., 1-benzofuranyl), cyano, and C(O)-R8;R8 is selected from optionally substituted amino (e.g., optionally substituted 4-ethoxyphenylamino), optionally substituted alkoxy (e.g., optionally substituted methoxy, optionally substituted ethoxy), and optionally substituted aryl (e.g., optionally substituted phenyl, e.g., phenyl, 2-methoxyphenyl, 2-chloro-6-fluorophenyl); R9 is H or optionally substituted C1-C6 alkyl (e.g., optionally substituted methyl, optionally substituted ethyl); R10 and R11 are independently selected from H and optionally substituted C1-C6 alkyl (e.g., optionally substituted methyl, optionally substituted ethyl); and tautomers, geometric isomers, optically active forms, pharma- ceutically acceptable salts, and pharma- ceutical active derivatives thereof.;

[0039] According to certain embodiments, isotopic isomers of compounds of formula (I), such as deuterated and C13 analogs, are provided.

[0040] According to a particular embodiment, there is provided a compound of formula (I) wherein R1 is an S-R6 moiety.

[0041] According to a particular embodiment, there is provided a compound of formula (I) wherein R1 is a NR9-R6 moiety.

[0042] According to a particular embodiment, there is provided a compound of formula (I) wherein R1 is an NH-R6 moiety.

[0043] According to a particular embodiment, there is provided a compound of formula (I) wherein R1 is an R6 moiety.

[0044] According to a particular embodiment, there is provided a compound of formula (I) wherein R2 is an optionally substituted C1-C6 alkyl.

[0045] In accordance with a particular embodiment, there is provided a compound of formula (I) wherein R2 is optionally substituted propyl.

[0046] According to a particular embodiment, there is provided a compound of formula (I) wherein R3 is an optionally substituted C1-C6 alkyl.

[0047] According to a particular embodiment, there is provided a compound of formula (I) wherein R3 is optionally substituted methyl.

[0048] According to a particular embodiment, there is provided a compound of formula (I) wherein R3 is H.

[0049] According to a particular embodiment, there is provided a compound of formula (I), wherein n is 0.

[0050] According to a particular embodiment, there is provided a compound of formula (I) wherein n is 1.

[0051] According to a particular embodiment, there is provided a compound of formula (I) wherein n is 2.

[0052] According to a particular embodiment, there is provided a compound of formula (I) wherein R7 is an optionally substituted C1-C6 alkyl.

[0053] According to a particular embodiment, there is provided a compound of formula (I) wherein R7 is an optionally substituted C2-C6 alkenyl.

[0054] According to a particular embodiment, there is provided a compound of formula (I) wherein R7 is optionally substituted aryl.

[0055] According to a particular embodiment, there is provided a compound of formula (I) wherein R7 is optionally substituted phenyl.

[0056] According to a particular embodiment, there is provided a compound of formula (I) wherein R7 is an optionally substituted heteroaryl.

[0057] In accordance with a particular embodiment, there is provided a compound of formula (I) wherein R7 is optionally substituted furanyl.

[0058] According to a particular embodiment, there is provided a compound of formula (I) wherein R7 is an optionally substituted pyrazolyl.

[0059] According to a particular embodiment, there is provided a compound of formula (I) wherein R7 is optionally substituted pyridinyl.

[0060] According to a particular embodiment, there is provided a compound of formula (I) wherein R7 is cyano.

[0061] According to a particular embodiment, there is provided a compound of formula (I) wherein R9 is H.

[0062] According to a particular embodiment, there is provided a compound of formula (I) wherein R10 is H.

[0063] According to a particular embodiment, there is provided a compound of formula (I) wherein R10 is an optionally substituted C1-C6 alkyl.

[0064] According to a particular embodiment, there is provided a compound of formula (I), wherein R11 is H.

[0065] According to certain embodiments, there is provided a compound of formula (I) wherein n is an integer selected from 0 to 1, R5 is S, and R7 is an optionally substituted aryl (e.g., an optionally substituted phenyl).

[0066] According to a particular embodiment, there is provided a compound of formula (I), wherein said compound has inhibitory activity against MPC.

[0067] In further particular embodiments, the compounds of the invention are, in particular, those of the following group: 3-[(3,4-difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (1); 3-[(2-chloro-5-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (2); 3-[(3,5-difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (3); 3-{[3-(difluoromethyl)benzyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (4); 3-{[2-(difluoromethyl)benzyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (5); 3-{[4-(difluoromethyl)benzyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (6); 3-{[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile (7); 4-Fluoro-2-{[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile (8); 3-[(5-fluoro-2-methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (9); 3-[(3-chloro-5-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (10); 3-[(4-chloro-3-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (11); 3-[(5-fluoro-2-methylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (12); 3-[(4-fluoro-2-methylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (13); 3-[(2-phenylethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (14); 3-[(furan-3-ylmethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (15); 3-[(2-Methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (16); 3-[(3-methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (17); 4-{[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile (18); 3-[(1-benzofuran-5-ylmethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (19); 3-[(4-Methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (20); 3-[(2,5-difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (21); 3-[(2,4-difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (22); 3-[(2,6-difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (23); 5-Propyl-3-[(1H-pyrazol-4-ylmethyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (24); 3-[(2,6-dimethylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (25); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (26); 3-[(2,5-dimethylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (27); 3-{[2-(2,4-difluorophenyl)ethyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (28); 3-[(2,5-difluorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (29); 5-[2-(benzyloxy)ethyl]-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (30); 3-[(2,5-difluorobenzyl)sulfanyl]-5-(methoxymethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (31); 5-{2-[(4-chlorobenzyl)oxy]ethyl}-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (32); 5-{2-[(2-chlorobenzyl)oxy]ethyl}-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (33); 5-{2-[(3-chlorobenzyl)oxy]ethyl}-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (34); 3-[(2,5-difluorobenzyl)sulfanyl]-5-(2-methoxyethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (35); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (36); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-ethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (37); 5-Butyl-3-[(2-chloro-6-fluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (38); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-cyclopropyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (39); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-(2-methylpropyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (40); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-(cyclopropylmethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (41); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-(methoxymethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (42); 5-[2-(benzyloxy)ethyl]-3-[(2-chloro-6-fluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(1H)-one (43); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (44); 5-Benzyl-3-[(2-chloro-6-fluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (45); 4-{[(5-benzyl-7-oxo-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile (46); 3-(Phenylsulfanyl)-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (47); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-6-methyl-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (48); 3-[(2-chloro-6-fluorobenzyl)amino]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (49); 3-[(2-chloro-6-fluorobenzyl)sulfonyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (50); 3-(2-phenylethyl)-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (51); 3-[(2-chlorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (52); 5-Methyl-3-(methylsulfanyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (53); 5-Methyl-3-[(4-nitrobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (54); 3-{[(2E)-3-phenylprop-2-en-1-yl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (55); N-(4-ethoxyphenyl)-2-[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]acetamide (56); Methyl 5-{[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}furan-2-carboxylate (57); Ethyl 2-[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]propanoate (58); Ethyl 2-[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]butanoate (59); Methyl 2-[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]butanoate (60); Methyl 2-[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]propanoate (61); Benzyl[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]acetate (62); [(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]acetonitrile (63); 3-[(2-oxo-2-phenylethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (64); 3-{[2-(4-methoxyphenyl)-2-oxoethyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (65); N-(5-chloro-2-methoxyphenyl)-2-[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]acetamide (66); 5,6-Dimethyl-3-(propylsulfanyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (67); 5,6-Dimethyl-3-[(3-methylbutyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (68); 5,6-Dimethyl-3-[(4-nitrobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (69); 5,6-Dimethyl-3-[(3-methylbenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (70); 3-[(2,5-dimethylbenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (71); 5,6-Dimethyl-3-[(4-methylbenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (72); 5,6-Dimethyl-3-[(3-nitrobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (73); 3-[(4-chlorobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (74); 3-[(2-chlorobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (75); 5,6-Dimethyl-3-{[3-(trifluoromethyl)benzyl]sulfanyl}[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (76); 5,6-Dimethyl-3-{[4-(trifluoromethyl)benzyl]sulfanyl}[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (77); 3-[(2-fluorobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (78); 3-[(3-chlorobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (79); 3-[(2,4-dichlorobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (80); 3-[(3-chlorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (81); 3-[(3-fluorobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (82); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (83); 3-[(4-bromobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (84); 3-[(4-chlorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (85); 3-(Benzylsulfanyl)-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (86); 3-[(2-methylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (87); 3-[(3-methylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (88); 3-[(4-methylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (89); 3-[(Naphthalen-1-ylmethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (90); 3-[(3-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (91); 3-[(4-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (92); 3-[(2-chlorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (93); 3-[(3,4-dichlorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (94); 3-[(2-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (95); 3-[(2,4-dichlorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (96); 3-[(3-fluorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (97); 3-[(2-fluorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (98); 3-[(2,5-dimethylbenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (99); and 3-[(3-chlorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (100) The compound includes a compound selected from:

[0068] In another further particular embodiment, there is provided a compound for use as a medicament, the compound being selected from compounds (1) to (100) as defined herein.

[0069] In another further particular embodiment, the following group: 3-[(3,4-difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (1); 3-[(2-chloro-5-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (2); 3-[(3,5-difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (3); 3-{[3-(difluoromethyl)benzyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (4); 3-{[2-(difluoromethyl)benzyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (5); 3-{[4-(difluoromethyl)benzyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (6); 3-{[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile (7); 4-Fluoro-2-{[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile (8); 3-[(5-fluoro-2-methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (9); 3-[(3-chloro-5-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (10); 3-[(4-chloro-3-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (11); 3-[(5-fluoro-2-methylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (12); 3-[(4-fluoro-2-methylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (13); 3-[(2-phenylethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (14); 3-[(furan-3-ylmethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (15); 3-[(2-Methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (16); 3-[(3-methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (17); 4-{[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile (18); 3-[(1-benzofuran-5-ylmethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (19); 3-[(4-Methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (20); 3-[(2,5-difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (21); 3-[(2,4-difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (22); 3-[(2,6-difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (23); 5-Propyl-3-[(1H-pyrazol-4-ylmethyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (24); 3-[(2,6-dimethylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (25); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (26); 3-[(2,5-dimethylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (27); 3-{[2-(2,4-difluorophenyl)ethyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (28); 3-[(2,5-difluorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (29); 5-[2-(benzyloxy)ethyl]-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (30); 3-[(2,5-difluorobenzyl)sulfanyl]-5-(methoxymethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (31); 5-{2-[(4-chlorobenzyl)oxy]ethyl}-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (32); 5-{2-[(2-chlorobenzyl)oxy]ethyl}-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (33); 5-{2-[(3-chlorobenzyl)oxy]ethyl}-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (34); 3-[(2,5-difluorobenzyl)sulfanyl]-5-(2-methoxyethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (35); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (36); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-ethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (37); 5-Butyl-3-[(2-chloro-6-fluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (38); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-cyclopropyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (39); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-(2-methylpropyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (40); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-(cyclopropylmethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (41); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-(methoxymethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (42); 5-[2-(benzyloxy)ethyl]-3-[(2-chloro-6-fluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(1H)-one (43); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (44); 5-Benzyl-3-[(2-chloro-6-fluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (45); 4-{[(5-benzyl-7-oxo-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile (46); 3-(Phenylsulfanyl)-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (47); 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-6-methyl-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (48); 3-[(2-chloro-6-fluorobenzyl)amino]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (49); 3-[(2-chloro-6-fluorobenzyl)sulfonyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (50); and 3-(2-Phenylethyl)-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one (51) The compound of the present invention is provided, which is selected from:

[0070] In certain aspects, there is provided a compound of the invention for use in combination with immunotherapy.

[0071] According to a particular embodiment, there is provided a compound of the invention for combination with T cell transfer therapy, e.g., chimeric antigen receptor (CAR) T cell, tumor infiltrating lymphocyte (TIL) or T cell receptor (TCR) therapy.

[0072] According to further particular embodiments, there is provided a compound of the invention for combination with a cancer vaccine or at least one immune checkpoint inhibitor.

[0073] In certain aspects, there is provided a compound of the invention for combination with an immune checkpoint inhibitor.

[0074] According to a particular embodiment, the immune checkpoint inhibitor is selected from a PD1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a VISTA inhibitor, a CD155 / TIGIT inhibitor, a TIM-3 inhibitor.

[0075] According to a particular embodiment, the immune checkpoint inhibitor according to the invention is a PD-1 inhibitor.

[0076] According to certain embodiments, there is provided a compound of the invention for combination with an immunotherapeutic agent, said immunotherapeutic agent being at least one monoclonal antibody, such as rituximab and blinatumomab.

[0077] According to certain embodiments, there are provided compounds of the invention for combination with an immunotherapeutic agent, wherein said immunotherapeutic agent is at least one cytokine, such as interferon and aldesleukin.

[0078] According to another more particular aspect, there is provided a compound of the invention for use in combination with an anti-cancer vaccine.

[0079] According to a particular embodiment, the anti-cancer vaccine is selected from DNA, RNA, peptide and oncolytic viral vaccines.

[0080] In certain embodiments, there is provided a compound of the invention for combination with a cancer vaccine, such as an oncolytic or anti-herpes simplex vaccine.

[0081] According to another further particular embodiment, there is provided a compound of the invention for use in activating both humoral and cellular immunity, a prerequisite for successful virus neutralization and / or elimination (such as SARS-CoV-2).

[0082] According to a further aspect, there is provided a compound of the invention for use as a vaccine adjuvant component, complementary to classical adjuvants, to improve the longevity of both cellular and humoral immune responses upon vaccination.

[0083] According to another further particular embodiment, depending on the cancer type, the compounds of the present invention are provided for combination with chemotherapeutic agents, such as alkylating agents, nitrosoureas, antimetabolites, plant alkaloids and natural products, antitumor antibiotics, hormonal agents, biological response modifiers.

[0084] According to another further particular embodiment, there is provided a compound of the invention for use in combination with radiation therapy, including intensity modulated radiation therapy (IMRT), volumetric arc therapy (VMAT) and image-guided radiation therapy (IGRT).

[0085] Synthesis of compounds according to the invention The compounds of the present invention can be prepared from readily available starting materials using the following general methods and procedures. Where typical or preferred experimental conditions (i.e., reaction temperatures, times, moles of reagents, solvents, etc.) are given, it will be recognized that other experimental conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvents used, but such conditions can be determined by one skilled in the art using routine optimization procedures.

[0086] General synthetic approaches to obtain compounds of formula (I) are depicted in Schemes 1 and 2 below. [ka]

[0087] R2 and R3 functionalized β-ketoacetates (iv) are accessible from readily available starting materials via a wide range of transformations, including but not limited to, base-catalyzed alkylation or electrophilic substitution of simple acetylacetates (i) or alpha-unsubstituted β-ketoacetates (ii), or via acetylation of acetates (iii). Base-catalyzed condensation with S-methylisothiourea provides intermediate 2-methylsulfanyl-1H-pyrimidin-4-ones (v), which undergo nucleophilic aromatic substitution with hydrazine to give 2-hydrazino-1H-pyrimidin-4-ones (vi). Intermediate (vi) can undergo cyclization with carbon disulfide to 3-thioxo-2,8-dihydro-[1,2,4]triazolo[4,3-a]pyrimidin-7-one (vii), which can then undergo various transformations, including but not limited to base-mediated S-alkylation or metal-mediated (e.g., via copper-iodine) S-arylation, to give the final compound of formula (I), in particular formula (Ia). In the case of the preparation of isotopically isomers, isotopically labeled building blocks, such as C13 or C14 carbon disulfide, N15 hydrazine, and the like, can be used in Scheme 1.

[0088] Intermediate (vi) can undergo reaction with an isothiocyanate to give aminothiourea (viii), which is then cyclized in the presence of DCC or a similar peptide coupling agent to give the final compound of formula (I), particularly formula (Ib). [ka]

[0089] 2,4-Dichloro-6-methoxypyrimidines (ix) can undergo regioselective alkylation with metal alkyls, such as Grignard reagents, to give functionalized 2-chloro-6-methoxypyrimidines (x), which can then undergo nucleophilic aromatic substitution with hydrazine to give 2-hydrazino-6-methoxy-pyrimidine intermediates (xi). Cyclization of (xi) with carbon disulfide gives 7-methoxy-2H-[1,2,4]triazolo[4,3-a]pyrimidine-3-thiones (xii), which can undergo further functionalization on the thio group via alkylation or metal-mediated arylation to give 3-substituted 7-methoxy-3-sulfanyl-[1,2,4]triazolo[4,3-a]pyrimidines (xiii). Intermediate (xiii) may undergo acid mediated demethylation at the 7-position, for example via hydrobromic acid, to give the final compound of formula (I), in particular of formula (Ia), or compound (xiii) may undergo further functionalization, for example meta-chloroperbenzoic acid (m-CPBA) mediated oxidation to 7-methoxy-3-sulfonyl-[1,2,4]triazolo[4,3-a]pyrimidine (xiv), which may then undergo acid mediated demethylation at the 7-position to give the final compound of formula (I), in particular of formula (Ic).

[0090] Intermediate (xi) can be reacted with a carboxylic acid under amide coupling conditions (e.g., EDC / HOBt) to form an acetohydride intermediate (xv), which can then be cyclized, for example, by dehydration mediated via Burgess reagent to a 7-methoxy-3-alkyl-[1,2,4]triazolo[4,3-a]pyrimidine or 7-methoxy-3-aryl-[1,2,4]triazolo[4,3-a]pyrimidine intermediate (xvi), which can then undergo acid mediated demethylation at the 7-position to give the final compound of formula (I), in particular formula (Id).

[0091] Further functionalization or functional group manipulation of side chains on the final compounds or intermediates can be achieved by one skilled in the art to expand the scope of final compounds of formula (I) that can be obtained via Scheme 1 and Scheme 2.

[0092] composition The present invention provides pharmaceutical or therapeutic agents as compositions and methods for treating patients, preferably mammalian patients, most preferably human patients, suffering from solid tumor cancers that are resistant or susceptible to resistance to immunotherapy.

[0093] The pharmaceutical compositions of the present invention can contain one or more compounds in any form as described herein. The compositions of the present invention may further include one or more additional pharma- ceutically acceptable ingredients, such as alum, solubilizers, stabilizers, antibacterial agents, buffers, colorants, flavoring agents, adjuvants, and the like.

[0094] The compounds of the present invention may be put into the form of pharmaceutical compositions and unit dosages thereof together with conventionally used adjuvants, carriers, diluents or excipients, and in such form may be used as solids, such as powders in sachets, tablets or filled capsules, or liquids, such as solutions, suspensions, emulsions, elixirs, nasal sprays or capsules filled therewith, all for oral use, or in the form of sterile injectable solutions for parenteral (including subcutaneous) use. Such pharmaceutical compositions and unit dosage forms thereof may contain the ingredients in conventional proportions, with or without additional active compounds or substances, and such unit dosage forms may contain any suitable effective amount of the active ingredient equivalent to the intended daily dosage range used. The compositions according to the present invention are preferably oral, sublingual, nasal and subcutaneous.

[0095] The compositions of the present invention may also be liquid formulations, including, but not limited to, aqueous or oily suspensions, solutions, emulsions, syrups, sprays, and elixirs. Liquid forms suitable for oral administration may contain a suitable aqueous or non-aqueous vehicle, together with buffers, suspending and dispersing agents, colorants, flavorings, and the like. The compositions may also be formulated as dry products for reconstitution with water or other suitable vehicles before use. Such liquid preparations may contain auxiliary agents, including, but not limited to, suspending agents, emulsifying agents, non-aqueous vehicles, and preservatives. Suspending agents include, but are not limited to, sorbitol syrup, methylcellulose, glucose / sugar syrup, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel, and hydrogenated edible fats. Emulsifying agents include, but are not limited to, lecithin, sorbitan monooleate, and acacia. Non-aqueous vehicles include, but are not limited to, edible oils, almond oil, fractionated coconut oil, oily esters, propylene glycol, and ethyl alcohol. Preservatives include, but are not limited to, methyl or propyl p-hydroxybenzoate, and sorbic acid. Additional materials and processing techniques are provided in Remington: The Science & Practice of Pharmacy, 23rd Edition, 2020, edited by Adeboye Adejare, Academic Press, which is incorporated herein by reference.

[0096] The solid composition of the present invention may be in the form of powder in a sachet, tablet, or lozenge, formulated in a conventional manner. For example, sachets, tablets, and capsules for oral or sublingual administration may contain conventional excipients, including but not limited to binders, fillers, lubricants, disintegrants, and wetting agents. Binders include but are not limited to syrup, acacia, gelatin, sorbitol, tragacanth, mucilage of starch, and polyvinylpyrrolidone. Fillers include but are not limited to lactose, sugar, microcrystalline cellulose, corn starch, calcium phosphate, and sorbitol. Lubricants include but are not limited to magnesium stearate, stearic acid, talc, polyethylene glycol, and silica. Disintegrants include but are not limited to potato starch, and sodium starch glycolate. Wetting agents include but are not limited to sodium lauryl sulfate. Tablets may be coated according to methods well known in the art.

[0097] Injectable compositions are typically based on injectable sterile saline or phosphate-buffered saline or other injectable carriers known in the art.

[0098] The compositions of the present invention may also be formulated for parenteral administration, including, but not limited to, by injection or continuous infusion.The formulations for injection may be in the form of a suspension, solution, or emulsion in an oily or aqueous medium, and may contain formulating agents, including, but not limited to, suspending agents, stabilizing agents, and dispersing agents.The compositions may also be provided in powder form for reconstitution with a suitable medium, including, but not limited to, sterile pyrogen-free water.

[0099] The composition of the present invention may also be formulated as a depot preparation, which may be administered by implantation or by intramuscular injection.The composition may be formulated with suitable polymeric or hydrophobic materials (e.g., as an emulsion in an acceptable oil), with ion exchange resins, or as a sparingly soluble derivative (e.g., as a sparingly soluble salt).

[0100] The compounds of the invention can also be administered in sustained release forms or from sustained release drug delivery systems. A description of representative sustained release materials can be found in incorporated materials in Remington's Pharmaceutical Sciences.

[0101] Mode of Administration The compositions of the present invention may be administered in any manner, including but not limited to, topically, orally, parenterally, sublingually, buccally, intranasally, intralesionally, intracerebroventricularly, or combinations thereof.Parenteral administration includes but is not limited to subcutaneously and intramuscularly.The compositions of the present invention may also be administered in the form of an implant, which allows delayed release of the composition, as well as slow, controlled iv infusion.In certain embodiments, one or more compounds of the present invention are administered orally.

[0102] The dosage administered to an individual, either as a single or multiple doses, will vary depending on a variety of factors, including pharmacokinetic properties, the condition and characteristics of the patient (age, weight, health, body size), the severity of symptoms, frequency of treatment and the desired effect.

[0103] combination According to one embodiment of the present invention, the compounds according to the invention and their pharmaceutical formulations are administered or used in combination with an anti-cancer immunotherapeutic agent, in particular an anti-cancer vaccine, or at least one immune checkpoint inhibitor, such as at least one PD-1, PD-L1 or CTLA4 inhibitor.

[0104] The present invention encompasses the administration of a compound of the invention or a pharmaceutical formulation thereof, wherein said compound of the invention or a pharmaceutical formulation thereof is administered to an individual prior to or simultaneously with an anti-cancer immunotherapeutic agent, e.g., simultaneously in the same formulation, or separately in different formulations, in particular via different formulation routes.

[0105] According to a particular embodiment of the invention, the compounds according to the invention and pharmaceutical formulations thereof are administered chronically (e.g., daily or weekly) for the duration of the treatment, prior to administration of an anti-cancer immunotherapeutic agent or an anti-angiogenic treatment.

[0106] According to another particular aspect of the invention, the compounds according to the invention and their pharmaceutical formulations are administered simultaneously with anti-cancer immunotherapeutic agents.

[0107] According to another particular embodiment of the present invention, the anti-cancer immunotherapeutic agent can be administered in combination with a therapeutically effective amount of other therapeutic regimens or co-agents (e.g., multi-drug regimens) that are useful for the treatment of cancer, for example, in combination with substances that are useful for treating, stabilizing, preventing, and / or delaying cancer, for example, substances used in conventional chemotherapy for solid tumors, for controlling the establishment of metastasis, or any other molecule that acts by causing programmed cell death.In particular, according to another particular embodiment of the present invention, the anti-cancer immunotherapeutic agent can be administered in combination with a therapeutically effective amount of other therapeutic regimens or co-agents (e.g., multi-drug regimens) that are useful for the treatment of cancer.

[0108] Compounds of the invention or pharmaceutical formulations thereof that are administered simultaneously with said anti-cancer immunotherapeutic agents can be administered in or within the same or different compositions and by the same or different routes of administration.

[0109] patient In one embodiment, the subject according to the invention is a subject suffering from a solid tumor cancer, in particular a poorly responsive solid tumor cancer that has demonstrated or is likely to demonstrate resistance to immunotherapy.

[0110] In certain embodiments, the subject according to the invention is a subject suffering from a solid tumor cancer selected from lung cancer (small cell and non-small cell), breast cancer, ovarian cancer, cervical cancer, uterine cancer, head and neck cancer, melanoma, hepatocellular carcinoma, colon cancer, rectal cancer, colorectal cancer, renal cancer, prostate cancer, gastric cancer, bronchial cancer, pancreatic cancer, bladder cancer, liver cancer and brain cancer, in particular glioblastoma.

[0111] In another specific embodiment, a subject according to the invention is a subject suffering from a head and neck tumor.

[0112] In another specific embodiment, a subject according to the present invention is a subject suffering from melanoma.

[0113] In another specific embodiment, a subject according to the invention is a subject suffering from colon cancer.

[0114] In another specific embodiment, a subject according to the present invention is a subject suffering from lung cancer.

[0115] In another specific embodiment, a subject according to the invention is a subject suffering from breast cancer.

[0116] In another specific embodiment, a subject according to the invention is a subject suffering from hepatocellular carcinoma or liver cancer.

[0117] In another specific embodiment, a subject according to the invention is a subject suffering from rectal or colorectal cancer.

[0118] In another specific embodiment, a subject according to the present invention is a subject suffering from renal cancer.

[0119] In another specific embodiment, a subject according to the present invention is a subject suffering from pancreatic cancer.

[0120] In another particular embodiment, the subject according to the invention is a subject suffering from brain cancer, in particular glioblastoma.

[0121] In another specific embodiment, the subject according to the invention is a subject with a solid tumor cancer who is at risk of developing resistance or partial resistance to anti-cancer immunotherapy due to another concomitant treatment or genetic predisposition.

[0122] In another specific embodiment, a subject according to the invention is a subject suffering from a metabolic disease, eg, type 2 diabetes.

[0123] In another specific embodiment, a subject according to the invention is a subject suffering from fibrosis, eg, pulmonary fibrosis or non-alcoholic steatohepatitis (NASH).

[0124] In another specific embodiment, a subject according to the invention is a subject suffering from an autoimmune disease, for example multiple sclerosis.

[0125] In another specific embodiment, a subject according to the present invention is a subject suffering from a hair loss disorder, e.g., alopecia.

[0126] In another specific embodiment, a subject according to the invention is a subject suffering from a neurodegenerative disorder, such as Parkinson's disease or Alzheimer's disease.

[0127] In another specific embodiment, a subject according to the invention is a subject suffering from a skin or tissue injury, for example a skin wound or a burn.

[0128] In another specific embodiment, a subject according to the invention is a subject suffering from an acute condition of the brain, such as a stroke or brain trauma.

[0129] Use according to the invention In certain embodiments, the present invention provides compounds, methods, uses and compositions useful for the prevention and / or treatment of hair loss disorders.

[0130] In another specific embodiment, the present invention provides compounds, methods, uses and compositions useful for the prevention and / or treatment of skin or tissue injury, such as skin wounds or burns.Typically, the compounds of the present invention are applied topically to the skin or tissue.

[0131] Typically, according to certain embodiments, the compounds of the present invention are applied topically to the skin.

[0132] In another specific embodiment, the present invention provides compounds, methods, uses and compositions useful for the prevention and / or treatment of autoimmune diseases, such as multiple sclerosis.

[0133] In another particular embodiment, the present invention provides compounds, methods, uses and compositions useful for the prevention and / or treatment of neurodegenerative disorders, such as Parkinson's disease or Alzheimer's disease.

[0134] In another particular embodiment, the present invention provides compounds, methods, uses and compositions useful for the prevention and / or treatment of fibrosis, such as pulmonary fibrosis or non-alcoholic steatohepatitis (NASH).

[0135] In another particular embodiment, the present invention provides compounds, methods, uses and compositions useful for the prevention and / or treatment of acute pathologies of the brain, such as stroke or traumatic brain injury.

[0136] In another particular embodiment, the present invention provides compounds, methods, uses and compositions useful for the prevention and / or treatment of metabolic diseases, such as type 2 diabetes.

[0137] Typically, and according to certain embodiments, the compounds of the present invention are administered orally.

[0138] According to a particular embodiment, there is provided a method for inducing or increasing an immune response to immunotherapy, in particular anti-cancer immunotherapy, or vaccine therapy, said method comprising administering to a subject in need thereof an effective amount of one or more compounds of the invention or a pharmaceutical formulation thereof in combination with an immunotherapeutic agent.

[0139] In certain embodiments, the present invention provides compounds, methods, uses and compositions useful for the treatment of solid tumor cancer in a combination form, where at least one compound of the present invention is administered in combination with a vaccine, in particular an anti-cancer vaccine, such as an oncolytic vaccine or an anti-herpes simplex virus vaccine.

[0140] According to certain aspects, methods are provided for treating a subject suffering from cancer, the method comprising administering to a subject in need thereof an effective amount of one or more compounds of the present invention in combination with an anti-cancer immunotherapeutic agent.

[0141] In certain embodiments, the present invention provides compounds, methods, uses and compositions useful for the treatment of solid tumor cancer in a combination form, where at least one compound of the present invention is administered in combination with at least one anti-cancer immunotherapeutic agent.

[0142] According to certain aspects, methods are provided for treating a subject suffering from cancer, the method comprising administering to a subject in need thereof an effective amount of one or more compounds of the present invention in combination with an anti-cancer immunotherapeutic agent.

[0143] According to a particular embodiment, there is provided an in vitro method for obtaining and / or maintaining T cells with a memory phenotype, said method comprising the steps of: - providing at least one T cell, e.g., a CD8+ or CD4+ T cell, having the capacity to differentiate into a memory phenotype; - contacting said at least one T cell with at least one compound according to the invention or a mixture thereof; - culturing the cells in a T cell culture medium; - isolating the resulting T cells A method is provided, comprising:

[0144] According to a particular embodiment, said at least one compound from the present invention is included in the culture medium used to culture the T cells, and the T cells are then cultured in the presence of said at least one compound from the present invention.

[0145] In another embodiment, the at least one compound of the present invention is added to the culture medium after the T cell culture has been initiated, e.g. after or while the cells are seeded or incubated, but ideally shortly after initiation of the culture.

[0146] According to a particular embodiment, the T cells are contacted with a compound of the invention at least during activation, preferably for example for the first 3 or 4 days from the start of culture and / or activation.

[0147] According to another particular embodiment, the T cells may be contacted with the compounds of the invention throughout the culture period.

[0148] According to another particular embodiment, the compound of the invention is washed out after the initial activation step (priming step) and the culture is continued in the absence of the compound of the invention, e.g., with a medium containing IL-2 and IL-7.

[0149] The compound of the present invention is preferably present in the culture medium from the beginning of the culture. Also, preferably, T cells are at least contacted with the compound of the present invention, but it is not strictly necessary that the compound of the present invention is present during the entire activation phase. Therefore, the compound of the present invention is used to culture T cells, in particular to generate and / or maintain T cells with memory phenotype, as described herein.

[0150] The activated T cells according to the invention may then be used to improve commonly performed anti-cancer T cell immunotherapy.

[0151] All references cited herein are incorporated by reference in their entirety. The invention having been described, the following examples are given by way of illustration and not by way of limitation. EXAMPLES

[0152] Several methods for preparing the compounds of the present invention are described in the following examples. Unless otherwise specified, all starting materials were obtained from commercial suppliers and used without further purification. Specifically, the following abbreviations may be used in the examples and throughout the specification:

[0153] The following abbreviations have the following definitions: AMU (atomic mass unit); CDI (carbonyldiimidazole); DCC (dicyclohexylcarbodiimide); DCM (dichloromethane); DIPEA (diisopropylethylamine); DMF (dimethylformamide); DMSO (dimethylsulfoxide); EDC (1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide methiodide); HOBt (1-hydroxybenzotriazole); IPA (isopropyl alcohol); HMBC (heteronuclear multiple bond correlation); m-CPBA (meta-chloroperbenzoic acid); TEA (triethylamine).

[0154] Example 1: Synthesis of compounds of the present invention The compounds of the present invention were synthesized as described herein.

[0155] a) Compounds of formula (I) wherein R7 is optionally substituted phenyl Compounds 1-28 were synthesized according to Scheme 1 herein. [ka]

[0156] In particular, according to Scheme 3 below, where R is selected from one or more halogen (fluoro, chloro), cyano, halogenoC1-C6 alkyl, phenyl or benzyl groups substituted with C1-C6 alkyl, or from heteroaromatic groups such as benzofuran, furan, pyrazole. [ka]

[0157] Scheme 3, Step 1: Preparation of intermediate (1c) (2-(methylthio)-6-propylpyrimidin-4(1H)-one) To a stirred solution of ethyl butyryl acetate (1a) (2.5 g, 15.803 mmol) and 2-methyl-2-thiopseudourea hemisulfate (1b) (2.42 g, 17.38 mmol) in water (20 mL) was added Na2CO3 (2.68 g, 25.28 mmol) and stirred at room temperature for 48 h. A white precipitate formed which was filtered and triturated with ether to give the desired intermediate (1c) in 48.08% yield, 1.4 g, as a white solid. 1 H NMR (400MHz, DMSO) δ: 0.88 (t, 3H), 1.52-1.65 (m, 2H), 2.37 (t, 2H), 2.46 (s, 3H), 5.91 (s, 1H), 12.47 (brs, 1H); LCMS (method-1): R t = 2.47 min; [M+H] = 185 (5 min run time).

[0158] Scheme 3, Step 2: Preparation of intermediate (1d) To a stirred solution of 1c (1.4 g, 7.59 mmole) and hydrazine hydrate (3.79 g, 75.98 mmole) in ethanol (10 ml) was added and refluxed for 6 h. The reaction mixture was evaporated in vacuo and triturated with EtOH to give intermediate 1d as a white solid in 62.6% yield, 800 mg. 1 H NMR (400MHz, DMSO) δ: 0.86 (t, 3H), 1.50-1.59 (m, 2H), 2.22 (t, 2H), 4.48 (brs, 2H), 5.35 (s, 1H), 8.45 (brs, 1H), 9.91 (brs, 1H); LCMS (method-1): R t= 1.24 min; [M+H] = 169 (5 min run time).

[0159] Scheme 3, Step 3: Preparation of intermediate (1e) To a stirred solution of 1d (2 g, 11.90 mmol) in pyridine (55 mL) was added CS2 (11 mL) and refluxed for 6 h. The reaction mixture was evaporated in vacuo, azeotroped with toluene, and finally triturated with EtOH to give intermediate 1e in 31.96% yield, 800 mg, as an off-white solid. 1 H NMR (400MHz, DMSO) δ: 0.95 (t, 3H), 1.56-1.73 (m, 2H), 3.36 (t, 2H), 5.82 (s, 1H), 12.54 (brs, 1H), 13.74 (brs, 1H); LCMS (method-1): R t = 1.77 min; [M+H] = 211 (5 min run time).

[0160] Preparation of compounds (1) to (28) of the present invention according to Scheme 3, Step 4: To a stirred solution of intermediate (1e) (1 equiv.) and TEA (2.5 equiv.) in EtOH (20 mL / mmol), substituted benzyl bromide (1 equiv.) was added and stirred at room temperature for 18 h. The reaction mixture was evaporated in vacuo and partitioned between 10% MeOH-DCM and water. The organic phase was dried, concentrated and purified first by Combi-flash (0.1-0.2% MeOH-DCM) and finally by preparative TLC to give the corresponding compound of the invention.

[0161] The compounds were characterized using the methods detailed below:

[0162] Compound (1) : 42 mg, white solid (26% yield). 1 H NMR (400MHz, DMSO) δ:0.94(t, 3H), 1.56(m, 2H), 2.85(t, 2H), 4.45(s, 2H), 5.88(s, 1H), 7.21(m, 1H), 7.38(m, 1H), 7.45(m, 1H); 12.68(br s, 1H), LCMS (method-2):R t = 2.48 min; [M+H] = 353 (3 min run time) HPLC = 99.60%.

[0163] Compound (2) :80mg, white solid (yield 32%). 1 H NMR (400 MHz, DMSO) δ: 0.93 (t, 3H), 1.52-1.57 (t, 2H), 2.82 (t, 2H), 4.45 (s, 2H), 5.88 (s, 1H), 7.18-7.22 (t, 1H), 7.27 (d, 1H), 7.50-7.53 (m, 1H); 12.73 (s, 1H). LCMS (Method-1): R t =2.86 points; [M+H]=337 (5 minutes of travel time). HPLC=99.64%.

[0164] Compound (3) :70 mg, white solid (yield 29%). 1 H NMR (400 MHz, DMSO) δ: 0.95 (t, 3H), 1.54-1.60 (t, 2H), 2.86 (t, 2H), 4.45 (s, 2H), 5.87 (s, 1H), 7.09-7.16 (t, 3H), 7.27 (d, 1H), 12.69 (s, 1H). LCMS (Method-1): R t =2.37 points; [M+H]=337. HPLC=98.97%.

[0165] Compound (4) :105 mg, white solid (yield 42%). 1 H NMR (400MHz, DMSO) δ: 0.94 (t, 3H), 1.50-1.55 (t, 2H), 2.80 (t, 2H), 4.50 (s, 2H), 5.83 (s, 1H), 6.84(s, 1H), 6.98(s, 1H), 7.12(s, 1H), 7.45(brs, 2H), 7.50(brs, 2H), 12.68(s, 1H). LCMS(Method-1):R t =1.54 minutes; [M+H]=351. HPLC=98.18%.

[0166] Compound (5) :105 mg, white solid (yield 42%). 1H NMR (400MHz, DMSO) δ: 0.90 (t, 3H), 1.49-1.54 (t, 2H), 2.77 (t, 2H), 4.58 (s, 2H), 5.85 (s, 1H), 7.17-7.39 (m, 2H), 7.45 (brs, 2H), 7.60 (d, 1H), 7.45 (brs, 2H), 12.69 (s, 1H). LCMS(Method-1):R t =1.56 min; [M+H]=351. HPLC=97.96%.

[0167] Compound (6) :105 mg, white solid (yield 42%). 1 H NMR (400 MHz, DMSO) δ: 0.92 (t, 3H), 1.49-1.57 (t, 2H), 2.82 (t, 2H), 4.50 (s, 2H), 5.85 (s, 1H), 6.99 (m, 2H), 7.49 (brs, 4H), 12.67 (s, 1H). LCMS (Method-1): R t =1.55 min; [M+H]=351. HPLC=99.65%.

[0168] Compound (7) : 48 mg, Efufuto solid (yield 31%). 1 H NMR (400 MHz, DMSO) δ: 0.94 (t, 3H), 1.56 (m, 2H), 2.84 (t, 2H), 4.48 (s, 2H), 5.85 (s, 1H), 7.52 (t, 1H), 7.71 (m, 2H), 7.82 (s, 1H), 12.61 (br s, 1H). LCMS (Method-5): R t =2.21 minutes;[M+H]=326.2 (5 minutes travel time). HPLC=99.26%.

[0169] Compound (8) :30mg, white solid (yield 18%). 1 H NMR (400 MHz, DMSO) δ: 0.96 (t, 3H), 1.56-1.62 (t, 2H), 2.90 (t, 2H), 4.51 (s, 2H), 5.90 (s, 1H), 7.34-7.47 (m, 2H), 7.93 (m, 1H), 12.74 (s, 1H). LCMS (Method-1): R t =1.50 min; [M+H]=344. HPLC=99.36%.

[0170] Compound (9) :60mg, white solid (yield 24%). 1 H NMR (400 MHz, DMSO) δ: 0.90 (t, 3H), 1.49-1.56 (t, 2H), 2.80 (t, 2H), 3.71 (s, 3H) 4.29 (s, 2H), 5.86 (s, 1H), 6.96-7.11 (m, 3H), 12.74 (s, 1H). LCMS (Method-1): R t =1.55 min; [M+H]=349. HPLC=99.64%.

[0171] Compound (10) :50mg, white solid (yield 20%). 1 H NMR (400 MHz, DMSO) δ: 0.93 (t, 3H), 1.54-1.59 (t, 2H), 2.85 (t, 2H), 4.44 (s, 2H), 5.87 (s, 1H), 7.21-7.34 (m, 3H), 12.69 (s, 1H). LCMS (Method-1): R t =2.48 minutes; [M+H]=353. HPLC=99.10%.

[0172] Compound (11) :50mg, white solid (yield 20%). 1 H NMR (400 MHz, DMSO) δ: 0.92 (t, 3H), 1.53-1.59 (t, 2H), 2.85 (t, 2H), 4.45 (s, 2H), 5.86 (s, 1H), 7.21-7.54 (m, 3H), 12.68 (s, 1H). LCMS (Method-1): R = 2.47 minutes; [M+H] = 353. HPLC = 99.17%.

[0173] Compound (12) :36 mg, white solid (yield 15%). 1 H NMR (400 MHz, DMSO) δ: 0.92 (t, 3H), 1.51-1.56 (t, 2H), 2.30 (s, 3H) 2.80 (t, 2H), 4.42 (s, 2H), 5.85 (s, 1H), 7.00-7.24 (m, 3H), 12.71 (s, 1H). LCMS (Method-1): R t =1.58 minutes; [M+H]=333. HPLC=99.24%.

[0174] Compound (13) :60mg, white solid (yield 25%). 1 H NMR (400 MHz, DMSO) δ: 0.91 (t, 3H), 1.49-1.55 (t, 2H), 2.35 (s, 3H) 2.79 (t, 2H), 4.39 (s, 2H), 5.79 (s, 1H), 6.89-7.19 (m, 3H), 12.69 (s, 1H). LCMS (Method-1): R t =1.60 minutes; [M+H]=333. HPLC=97.64%.

[0175] Compound (14) :80mg, white solid (yield 27%). 1 H NMR (400 MHz, DMSO) δ: 0.93 (t, 3H), 1.56-1.61 (m, 2H), 2.86 (t, 2H), 3.01 (t, 2H), 3.48 (t, 2H), 5.86 (s, 1H), 7.18-7.30 (m, 5H), 12.65 (brs, 1H); LCMS (method-1): R t =1.56 minutes; [M+H]=315 (3 minutes of travel time). HPLC=99.51%.

[0176] Compound (15) :27 mg, white solid (yield 20%). 1 H NMR (400 MHz, DMSO) δ: 0.95 (t, 3H), 1.56-1.62 (m, 2H), 2.88 (t, 2H), 4.30 (s, 2H), 5.87 (s, 1H), 6.44 (s, 1H), 7.59 (d, 2H), 12.66 (brs, 1H); LCMS (method-1): R t =1.48 minutes; [M+H]=291 (3 minutes of travel time). HPLC=97.01%.

[0177] Compound (16) :40mg, white solid (yield 25%). 1 H NMR (400 MHz, DMSO) δ: 0.88 (t, 3H), 1.47 1.52 (m, 2H), 2.76 (t, 2H), 3.73 (s, 3H), 4.31 (s, 2H), 5.82 (s, 1H), 6.82 (d, 1H), 6.97 (t, 1H), 7.14 (d, 1H), 7.27 (t, 1H), 12.69 (brs, 1H); LCMS (method-2): Rt = 1.60 minutes; [M+H] = 331 (execution time of 3 minutes). HPLC = 96.61%.

[0178] Compound (17) : 80 mg, white solid (yield 25%). 1 H NMR (400 MHz, DMSO) δ: 0.85 (t, 3H), 1.48 - 1.56 (m, 2H), 2.80 (t, 2H), 3.69 (s, 3H), 4.39 (s, 2H), 5.83 (s, 1H), 6.82 - 6.88 (m, 3H), 7.21 (t, 1H), 12.68 (brs, 1H); LCMS (method - 1): R t = 1.52 minutes; [M+H] = 331 (execution time of 3 minutes). HPLC = 99.38%.

[0179] Compound (18) : 36 mg, white solid (yield 23%). 1 H NMR (400 MHz, DMSO) δ: 0.94 (t, 3H), 1.51 - 1.58 (m, 2H), 2.84 (t, 2H), 4.53 (s, 2H), 5.87 (s, 1H), 7.55 (d, 2H), 7.77 (d, 2H), 12.67 (brs, 1H); LCMS (method - 2): R t = 1.53 minutes; [M+H] = 326 (execution time of 3 minutes). HPLC = 99.56%.

[0180] Compound (19) : 60 mg, white solid (yield 25%). 1 H NMR (400 MHz, DMSO) δ: 0.90 (t, 3H), 1.49 - 1.55 (m, 2H), 2.81 (t, 2H), 4.55 (s, 2H), 5.82 (s, 1H), 6.92 (s, 1H), 7.27 (d, 1H), 7.52 (d, 1H), 7.61 (s, 1H), 7.98 (d, 1H), 12.65 (brs, 1H); LCMS (method - 3): R t = 2.87 minutes; [M+H] = 341 (execution time of 5 minutes). HPLC = 97.65%.

[0181] Compound(20) : 40 mg, white solid (yield 25%) 11H NMR (400 MHz, DMSO) δ: 0.92 (t, 3H), 1.50 - 1.59 (m, 2H), 2.82 (t, 2H), 3.71 (s, 3H), 4.39 (s, 2H), 5.84 (s, 1H), 6.85 (d, 2H), 7.24 (d, 2H), 12.66 (brs, 1H). LCMS (Method - 3): R t = 2.81 min; [M + H] = 331 (retention time of 5 min). HPLC = 98.85%.

[0182] Compound (21) : 30 mg, white solid (yield 19%). 1 1H NMR (400 MHz, DMSO) δ: 0.94 (t, 3H), 1.53 - 1.59 (m, 2H), 2.86 (t, 2H), 4.42 (s, 2H), 5.89 (s, 1H), 7.16 - 7.28 (m, 3H), 12.73 (brs, 1H); LCMS (Method - 1): R t = 1.52 min; [M + H] = 337 (retention time of 3 min). HPLC = 99.75%.

[0183] Compound (22) : 30 mg, white solid (yield 19%). 1 1H NMR (400 MHz, DMSO) δ: 0.92 (t, 3H), 1.51 - 1.57 (m, 2H), 2.83 (t, 2H), 4.39 (s, 2H), 5.83 (s, 1H), 7.00 - 7.04 (m, 1H), 7.21 - 7.27 (m, 1H), 7.37 - 7.43 (m, 1H), 12.73 (brs, 1H); LCMS (Method - 1): R t = 2.37 min; [M + H] = 337 (retention time of 3 min). HPLC = 99.31%.

[0184] Compound (23) : 35 mg, white solid (yield 22%). 1 1H NMR (400 MHz, DMSO) δ: 0.92 (t, 3H), 1.50 - 1.57 (m, 2H), 2.86 (t, 2H), 4.36 (s, 2H), 5.89 (s, 1H), 7.09 (t, 2H) 7.37 - 7.43 (m, 1H), 12.72 (brs, 1H); LCMS (Method - 1): R t = 2.32 min; [M + H] = 337 (retention time of 5 min). HPLC = 98.74%.

[0185] Compound (24) :70 mg, white solid (yield 17%). 1 H NMR (400 MHz, DMSO) δ: 0.94 (t, 3H), 1.54-1.60 (m, 2H), 2.85 (t, 2H), 4.35 (s, 2H), 5.85 (s, 1H), 7.55 (s, 2H), 12.71 (brs, 1H) LCMS (Method-1): R t =1.829 minutes; [M+H]=291 (5 minutes travel time). HPLC=98.45%.

[0186] Compound (25) :30mg, white solid (yield 19%). 1 H NMR (400 MHz, DMSO) δ: 0.91 (t, 3H), 1.52-1.62 (m, 2H), 2.33 (s, 6H), 2.82 (t, 2H), 4.49 (s, 2H), 5.87 (s, 1H), 7.04-7.13 (m, 3H), 12.69 (brs, 1H); LCMS (method-2): R t =1.69 minutes; [M+H]=329 (3 minutes travel time). HPLC=99.6%.

[0187] Compound (26) 230mg, white solid (yield 27.38%). 1 H NMR (400 MHz, DMSO) δ: 0.91 (t, 3H), 1.50-1.55 (m, 2H), 2.82 (t, 2H), 4.41 (s, 2H), 5.88 (s, 1H), 7.20-7.24 (m, 1H), 7.33-7.40 (m, 2H), 12.77 (brs, 1H); LCMS (method-1): R t =1.58 minutes; [M+H]=353 (3 minutes of travel time). HPLC=99.82%.

[0188] Compound (27) 234mg, white solid (yield 29.9%) 1H NMR (400MHz, DMSO) δ:0.88(t, 3H), 1.49-1.54(m, 2H), 2.15(s, 3H), 2.29(s, 3H), 2.74(t, 2H), 4.36(s, 2H), 5.81(s, 1H), 6.86(s, 1H), 6.98 6.99(m, 1H), 7.06-7.08(m, 2H), 12.77(brs, 1H);LCMS(Method-1):R t = 1.63 min; [M+H] = 329 (3 min run time). HPLC = 99.79%.

[0189] Compound (28) 40 mg, white solid (12% yield). 1 H NMR (400MHz, DMSO) δ: 0.93(t, 3H), 1.55-1.61(t, 2H), 2.83(t, 2H) 3.03(t, 2H), 3.47(t, 2H), 5.87(s, 1H), 7.00-7.42(m, 3H), 12.69(s, 1H). LCMS (Method-1):R t =1.67 minutes; [M+H]=351. HPLC=98.34%.

[0190] b) Compounds of formula (I) wherein R2 is optionally substituted alkyl. Compounds 29-46 were synthesized according to Scheme 1 herein. [ka]

[0191] In particular, according to Scheme 4 below, wherein R2 is selected from cycloalkyl, benzyl, optionally substituted alkyl, such as 3,3,3-trifluoropropyl, -CH2OCH3 or -CH2CH2OCH2A (wherein A is selected from methyl, or optionally substituted CH2Ph). [ka]

[0192] Preparation of compound (29) Scheme 4, Step 1: Representative preparation of intermediate (2c) with R2 = Me To a stirred solution of ethyl acetoacetate (2a, R2=Me) [3 g, 1.0 equiv] in water [24 mL, 8 vol] was added 2-methyl-2-thiourea (2b) [7.0 g, 1.1 equiv] followed by sodium carbonate [3.9 g, 1.6 equiv]. The reaction was stirred at room temperature for 48 hours. A white solid precipitated, which was filtered, washed with diethyl ether (2 vol) and dried under vacuum to give intermediate (2c R2=Me) as a white solid (1.8 gm, 50% yield). 1 H NMR (400MHz, DMSO-d6): δ 2.11 (s, 3H), 2.41 (s, 3H), 5.84 (s, 1H). LCMS (Method-2): Product: RT=1.57 min, m / z=157(M+H).

[0193] Scheme 4, Step 2: Representative preparation of intermediate (2d) with R2 = Me To a stirred solution of intermediate (2c, R2=Me) [800 mg, 1.0 equiv] in ethanol [8 mL, 10 vol] was added 100% hydrazine hydrate [4.9 mL, 30 equiv] at room temperature. The reaction mass was heated at 80° C. for 24 h. The reaction mass was monitored by TLC / LCMS. After complete consumption of starting material, the reaction mixture was distilled under reduced pressure and co-distilled with toluene. The solid was recrystallized using ethanol to precipitate a white solid. The solid was filtered, washed with ethanol and dried under reduced pressure to give intermediate (2d, R2=Me) as a white solid (420 mg, 58% yield). LCMS (Method-2): Product: RT=0.8 min, m / z=141 (M+H).

[0194] Scheme 4, Step 3: Representative preparation of intermediate (2e) with R2 = Me To a stirred solution of intermediate (2d, R2=Me) [400 mg, 1.0 equiv] in pyridine [11 mL, 27 vol] was added carbon disulfide [2.2 mL, 5.5 vol] at room temperature. The reaction mass was heated at 100° C. for 24 h. The reaction mass was monitored by TLC / LCMS. After completion of starting material, the reaction mixture was distilled under reduced pressure and co-distilled with toluene. The solid was recrystallized using ethanol to precipitate a white solid. The solid was filtered, the solid was washed with ethanol and dried under reduced pressure to give intermediate (2e, R2=Me) as a white solid (150 mg, 29% yield).1 H NMR (400MHz, DMSO-d6): δ 2.11 (s, 3H), 5.28 (s, 1H), 5.83 (s, 1H). LCMS (Method-2): Product: RT=0.52 min, m / =183.2(M+H).

[0195] Preparation of compound (29) according to Scheme 4, step 4: To a stirred solution of intermediate (2e, R2=Me) [350 mg, 1.0 equiv] in ethanol [15 mL, 50 vol], triethylamine [0.3 mL, 1.1 equiv] was added followed by 2,5-difluorobenzyl bromide [397 mg, 1.0 equiv] at room temperature. The reaction mass was stirred at RT for 24 h. The reaction mass was monitored by TLC / LCMS. After completion of starting material, the reaction mixture was distilled under reduced pressure. Then water was added and extracted with 3×10 vol of 20% MeOH / DCM. The organic layer was washed with brine solution, dried over Na2SO4 and concentrated under reduced pressure. The crude was purified by column chromatography on 100-200 mesh silica gel eluting with ethyl acetate / hexane. The compound was again purified by preparative TLC to give compound (29) as a solid (35 mg, 6% yield) (R=CH3): 1H NMR (400 MHz, DMSO-d6): δ 2.66 (s, 3H), 4.36 (s, 2H), 5.94 (s, 1H), 7.18-7.25 (m, 3H). LCMS (Method-2): Product: RT=1.44 min, m / z=309.3 (M+H).

[0196] Compounds 30-35 were prepared similarly to compound 29 via Scheme 4, steps 1-4, starting from variously substituted intermediates 2a.

[0197] Compound(30) Starting from intermediate (2a, (R2 = CH2CH2OCH2Ph): 13% yield. 1H NMR (400 MHz, DMSO-d6): δ 3.24-3.27 (t, 2H), 3.71-3.73 (t, 2H), 4.37 (s, 2H), 4.48 (s, 2H), 5.96 (s, 1H), 7.17-7.31 (m, 8H), 12.75 (bs, 1H). LCMS (Method-5): Product: RT = 2.53 minutes, m / z = 429.1 (M+H). HPLC: 99.18%.

[0198] Compound (31) The intermediate (2a, R2 = CH2OCH3) was also generated: yield 12%. 1H NMR (400MHz, DMSO-d6): δ 3.31 (s, 3H), 4.27 (s, 2H), 4.62 (s, 2H), 6.06 (s, 1H), 7.12-7.19 (m, 3H). LCMS (Method-5): Product: RT = 2.14 minutes, m / z = 339.1 (M+H).

[0199] Compound (32) The intermediate (2a, (R2=CH2CH2OCH2(4-Cl-Ph)) is produced: the yield is 5%. 1 H NMR (400 MHz, DMSO-d6): δ 3.24-3.27 (t, 2H), 3.70-3.73 (t, 2H), 4.37 (s, 2H), 4.47 (d, 2H), 5.94 (s, 1H), 7.17-7.24 (m, 3H), 7.28-7.30 (d, 2H), 7.36-7.38 (d, 2H), 12.75 (s, 1H). LCMS (Method-5): Product: RT = 2.66 minutes, m / z = 463.1 (M+H). HPLC: 96.47%.

[0200] Compound (33) , the intermediate (2a, (R2=CH2CH2OCH2(2-Cl-Ph))) is produced: the yield is 25%. 1 H NMR (400 MHz, DMSO-d6): δ 3.27-3.31 (t, 2H), 3.78-3.81 (t, 2H), 4.38 (s, 2H), 4.55 (s, 2H), 5.98 (s, 1H), 7.14-7.23 (m, 3H), 7.24-7.31 (d, 2H), 7.40-7.42 (t, 2H), 12.75 (bs, 1H). LCMS (Method-5): Product: RT = 2.63 minutes, m / z = 463.1 (M+H). HPLC: 99.64%.

[0201] Compound (34) Starting from intermediate (2a, (R2 = CH2CH2OCH2(3-Cl-Ph)): 15% yield. 1 H NMR (400MHz, DMSO-d6): δ 3.26-3.29(t, 2H), 3.72-3.75(t, 2H), 4.38(s, 2H), 4.49(s, 2H), 5.97(s, 1H), 7.15-7.37(m, 7H), 12.76(bs, 1H). LCMS (Method-5): Product: RT=2.65 min, m / z=463(M+H). HPLC:99.11%.

[0202] Compound (35) Starting from intermediate (2a, (R2 = CH2CH2OCH3): 13% yield. 1 H NMR (400MHz, DMSO-d6): δ 3.24-3.27(t, 2H), 3.71-3.73(t, 2H), 4.37(s, 2H), 4.48(s, 2H), 5.96(s, 1H), 7.17-7.31(m, 8H), 12.75(bs, 1H). LCMS (Method-5): Product: RT=2.53 min, m / z=429.1 (M+H). HPLC:99.18%.

[0203] Preparation of compound (36) Scheme 4, Step 1: Representative preparation of intermediate (2c) with R2 = Me To a stirred solution of ethyl acetoacetate (2a, R2=Me) (5 g, 38.462 mmol) and 2-methyl-2-thiopseudourea sulfate (12.83 g, 46.15 mmol) in water (10 mL) was added Na2CO3 (10.19 g, 96.15 mmol) and stirred at room temperature for 48 h. A white precipitate formed which was filtered and triturated with diethyl ether to give the desired intermediate (2c, R2=Me) in 86% yield, 5.2 g, as a white solid. 1 H NMR (400MHz, DMSO-d6)δ 12.50(br s, 1H), 5.95(s, 1H), 2.46(s, 3H), 2.32(s, 3H)

[0204] Scheme 4, Step 2: Representative preparation of intermediate (2c) with R2 = Me Intermediate (2c, R2=Me) (1.3 g, 8.33 mmol) was taken in a sealed tube to which hydrazine 1 (M) in THF (18 ml, 16.66 mmol) was added and the reaction mixture was heated at 100° C. for 16 hours. TLC showed the formation of a polar spot. Crude LCMS data confirmed the formation of the product. The reaction mixture was evaporated in vacuum and triturated with diethyl ether to give intermediate (2d, R=Me) as a pink solid in 56% yield, 650 mg. 1 H NMR (400MHz, DMSO-d6) δ 8.32 (br s, 2H), 5.36 (s, 1H), 4.54 (br, 2H), 1.99 (s, 3H)

[0205] Scheme 4, Step 3: Representative preparation of intermediate (2c) with R2 = Me To a stirred solution of intermediate (2d, R=Me) (650 mg, 4.64 mmol) in pyridine (11 mL) was added CS2 (7 mL, 1 ml / mmol) and refluxed for 16 h. The reaction mixture was evaporated in vacuo, azeotroped with toluene, and finally triturated with ethanol and filtered to give intermediate (2e, R=Me) in 53% yield, 450 mg, as a yellow solid. 1 H NMR (400MHz, DMSO-d6)δ 13.69(s, 1H), 12.49(s, 1H), 5.83(s, 1H), 2.85(s, 3H)

[0206] Preparation of compound (36) according to scheme 4, step 4: To a stirred solution of intermediate (2e, R=Me) (230 mg, 1.26 mmol) and TEA (0.46 mL, 3.16 mmol) in EtOH (12 mL), 2-chloro-6-fluorobenzyl bromide (225 mg, 1.011 mmol) was added at 0° C. and stirred at room temperature for 1 h. TLC showed the formation of a non-polar spot (desired disubstituted compound). The reaction mixture was evaporated in vacuum, then diluted with dichloromethane, extracted twice with water, and the organic layer was passed through anhydrous sodium sulfate. The organic layer was dried, concentrated, and purified by Combi-flash chromatography (4% MeOH-DCM) to give compound 36 as a white solid in 22% yield, 91 mg. NBHMBC confirmed the formation of the right region isomer.1 H NMR (400MHz, DMSO-d6) δ 12.75 (brs, 1H), 7.33-7.40 (m, 2H), 7.20-7.29 (t, 1H), 5.98 (s, 1H), 4.4 (s, 2H), 2.54 (S, 3H). LCMS (Method-4):R t = 2.16 min; [M+H] = 325 (5 min run time). HPLC = 99.82%.

[0207] Compounds 37-46 were prepared similarly to compound 36 via Scheme 4, steps 1-4, starting from variously substituted intermediates 2a.

[0208] Compound(37) Starting from intermediate 2a (R2 = CH2CH3) 1 H NMR (400MHz, DMSO-d6) δ 12.6 (brs, 1H), 7.34-7.41 (m, 2H), 7.21 -7.25 (t, 1H), 5.88 (s, 1H), 4.43 (s, 2H), 2.93-2.98 (q, 2H), 1.12-1.16 (t, 3H). LCMS (Method-1):R t = 1.64 min; [M+H] = 339 (3 min run time). HPLC = 95.18%.

[0209] Compound (38) Starting from intermediate (2a, (R2 = CH2CH2CH2CH3) 1 H NMR (400MHz, DMSO-d6)δ 12.76(brs, 1H), 7.37(m, 2H), 7.22(t, 1H), 5.91(s, 1H), 4.42(s, 2H), 2.84(t, 2H), 1.48(m, 2H), 1.32(m, 2H), 0.87(3, 3H). LCMS (Method-1):R t = 1.60 min; [M+H] = 367.3 (3 min run time). HPLC = 95.68%.

[0210] Compound (39) Starting from intermediate 2a (R2 = cyclopropyl) 11H NMR (400 MHz, DMSO-d6) δ 12.6 (brs, 1H), 7.35 - 7.42 (m, 2H), 7.23 - 7.27 (t, 1H), 5.81 (s, 1H), 4.51 (s, 2H), 2.50 (s, 1H), 1.02 - 1.04 (d, 2H), 0.96 (brs, 1H). LCMS (Method-4): R t = 2.32 min; [M+H] = 351 (retention time of 3 min). HPLC = 95.76%.

[0211] Compound(40) Starting from intermediate (2a, (R2 = CH2CH(CH3)2)) 1 1H NMR (400 MHz, DMSO-d6) δ 12.81 (brs, 1H), 7.34 - 7.41 (m, 2H), 7.20 - 7.25 (t, 1H), 4.43 (s, 2H), 2.66 - 2.68 (d, 2H), 1.79 - 1.84 (m, 1H), 0.86 - 0.88 (d, 6H). LCMS (Method-5): R t = 1.62 min; [M+H] = 367 (retention time of 3 min). HPLC = 97.66%.

[0212] Compound (41) Starting from intermediate (2a, (R2 = CH2-cyclopropyl)) 1 1H NMR (400 MHz, DMSO-d6) δ 12.82 (brs, 1H), 7.34 - 7.41 (m, 2H), 7.21 - 7.25 (t, 1H), 6.08 (s, 1H), 4.39 (s, 2H), 2.80 - 2.82 (d, 2H), 0.95 (brs, 1H), 0.55 - 0.57 (d, 2H), 0.13 - 0.14 (d, 2H). LCMS (Method-4): R t = 2.42 min; [M+H] = 365 (retention time of 5 min). HPLC = 97.46%.

[0213] Compound (42) Starting from intermediate (2a, (R2 = CH2OCH3)) 1 1H NMR (400 MHz, DMSO-d6) δ 12.9 (brs, 1H), 7.34 - 7.42 (m, 2H), 7.20 - 7.24 (t, 1H), 6.1 (s, 1H), 4.67 (s, 2H), 4.37 (s, 2H), 3.32 (s, 3H). LCMS (Method-4): Rt = 2.19 min; [M+H] = 355 (5 min run time). HPLC = 96.03%.

[0214] Compound (43) Starting from intermediate 2a (R2 = CH2CH2OCH2Ph) 1 H NMR (400MHz, DMSO-d6) δ 12.82 (brs, 1H), 7.34 (m, 8H), 5.98 (s, 1H), 4.47 (s, 2H), 4.39 (s, 2H), 3.71 (t, 2H), 3.29 (t, 2H). LCMS (Method-6):R t = 1.82 min; [M+H] = 445.3 (3 min run time). HPLC = 98.23%.

[0215] Compound (44) Starting from intermediate 2a (R2 = CH2CH2CF3) 1 H NMR (400MHz, DMSO-d6) δ 7.33-7.40(m, 2H), 7.20-7.24(t, 1H), 6.04(s, 1H), 4.39(s, 2H), 3.16-3.19(t, 2H), 2.6-2.67(m, 2H). LCMS (Method-4):R t = 2.43 min; [M+H] = 407 (5 min run time). HPLC = 99.06%.

[0216] Compound (45) Starting from intermediate 2a (R2 = CH2Ph) 1 H NMR (400MHz, DMSO-d6) δ 12.90(brs, 1H), 7.34-7.43(m, 5H), 7.19-7.31(m, 3H), 5.44(s, 1H), 4.34(s, 2H), 4.26(s, 2H). LCMS (Method-5):R t = 1.60 min; [M+H] = 401 (3 min run time). HPLC = 99.46%.

[0217] Compound(46) Starting from intermediate (2a, (R2 = CH2Ph) and replacing 2-chloro-6-fluorobenzyl bromide with 4-cyanobenzyl bromide in step 4: 60 mg, white solid (25% yield). 1H NMR (400MHz, DMSO) δ: 4.34(s, 2H), 4.41(s, 1H), 5.39(s, 1H), 7.22-7.37(m, 5H), 7.47(d, 2H), 7.75(d, 2H), 12.69(s, 1H). LCMS (Method-1):R t =1.56 minutes; [M+H]=374. HPLC=98.93%.

[0218] When not commercially available, the individual intermediates 2a for compounds 30-46 were supplied as follows.

[0219] Intermediate 2a for compounds 30, 32-34 and 43 was synthesized according to Scheme 5. [ka]

[0220] Scheme 5, Step 1 Preparation of intermediate (3b) 4-Chlorobenzyl alcohol intermediate (3a, R=4-Cl) [10 g, 1.0 equiv.] in dry hexane [100 ml, 10 vol.] was stirred at room temperature for 10 min, then paraformaldehyde [2.7 g, 1.0 equiv.] was added and stirred at RT for 10 min. The reaction mass was cooled to 0° C. and purged with dry HCl gas for 1 h. The reaction mixture was then continued at 5° C. for 24 h. The reaction mass was monitored by 1H NNMR. The reaction mixture was filtered through a sintered funnel, dried over Na2SO4, and concentrated under reduced pressure to give intermediate (3b, R=4-Cl) as a liquid (10.9 g, 81% yield). 1 H NMR (400MHz, DMSO-d6): δ 4.69 (s, 2H), 5.50 (s, 2H), 7.27-7.34 (m, 4H).

[0221] 3b where R is H: purchased from a commercial supplier.

[0222] For 3b where R is 2-chloro, the 2-chlorobenzyl alcohol intermediate (3a, R=2-Cl) was used, leading to the corresponding intermediate (3b R=2-Cl): 82% yield. 1H NMR (400MHz, DMSO-d6): δ 4.84(s, 2H), 5.57(s, 2H), 7.24-7.28(m, 2H), 7.36-7.39(t, 1H), 7.43-7.45(t, 1H).

[0223] For 3b where R is 3-chloro, the 3-chlorobenzyl alcohol intermediate (3a, R=3-Cl) was used, leading to the corresponding intermediate compound (3b R=3-Cl): 85% yield. 1 H NMR (400MHz, DMSO-d6): δ 4.70(s, 2H), 5.51(s, 2H), 7.21-7.29(m, 1H), 7.29-7.30(d, 2H), 7.35(s, 1H).

[0224] Scheme 5, Step 1 Intermediate (2a, R2=CH 2 CH 2 OCH 2 Preparation of Ar NaH [1.6 g, 1.05 equiv] was taken under nitrogen atmosphere and dry THF [50 mL, 10 vol] was added slowly. The reaction mixture was cooled to 0°C and then ethyl acetoacetate [5 g, 1.0 equiv] in dry THF was added dropwise. After stirring for 10 min, 2.5 M nBuLi [17 ml, 1.1 equiv] was added dropwise at 0°C. The reaction mixture was stirred for 15 min and then 3a (R=4-Cl) [6.5 g, 0.9 equiv] was added dropwise. The reaction mixture was brought to room temperature and stirred for 1-1.5 h and then quenched at 0°C with ice-cold water followed by 6N HCl, pH=2-3. The reaction mixture was extracted with ethyl acetate (3×10 vol). The combined organic layers were washed with brine solution, dried over Na2SO4 and concentrated under reduced pressure. The crude material was purified by column chromatography on 100-200 mesh silica gel eluted with hexane. 20% EA / Hex. Pure fractions were collected and distilled under reduced pressure to give pure intermediate 2a (R2 = CH2CHOCH2-(4-Cl-Ph)) as a liquid (3.5 g, 32% yield). 1H NMR (400 MHz, DMSO-d6): δ 1.21-1.27 (t, 3H), 1.71-1.74 (t, 1H), 2.80-2.83 (t, 2H), 3.46 (s, 1H), 3.71-3.74 (t, 2H), 4.14-4.20 (q, 2H), 4.45 (s, 2H), 4.65-4.66 (d, 2H), 7.22-7.33 (m, 4H).

[0225] The following intermediate 2a was synthesized using the same approach from 3b as described above.

[0226] Yield 39% for intermediate 2a (R2=CH2CH2OCH2-(2-Cl-Ph)). 1H NMR (400MHz, DMSO-d6):δ 1.21-1.29(t, 3H), 1.90-93-1.74(t, 1H), 2.26(s, 1H), 3.43(s, 1H), 3.71-3.74(t, 2H), 4.1 5-4.22(q, 1H), 4.77-4.79(d, 2H), 7.23-7.29(m, 2H), 7.34-7.36(d, 1H), 7.46-7.48(d, 1H).

[0227] Yield 30% for intermediate 2a (R2=CH2CH2OCH2Ph). 1H NMR (400MHz, DMSO-d6): δ 1.21-1.24(t, 3H), 2.02(s, 2H), 4.11-4.20(m, 2H), 4.68-4.69(d, 1H), 4.73(s, 1H), 5.21(s, 2H), 7.24-7.36(m, 5H).

[0228] Yield 21% for intermediate 2a (R2=CH2CH2OCH2-(3-Cl-Ph)). 1H NMR (400MHz, DMSO-d6):δ 1.21-1.27(t, 3H), 2.24(s, 2H), 3.49-3.54(t, 2H), 4.11-4.20(q, 2H), 4.58(s, 1H), 4.77( s, 1H), 5.28(s, 2H), 7.19-7.22(m, 1H), 7.24(s, 1H), 7.32-7.36(t, 1H), 7.41-7.48dd, 1H).

[0229] Intermediate 2a (R2 = CH2CH2OMe) for generating compound 35 was prepared as follows: To a stirred solution of potassium ethyl malonate [12.5 g, 1.0 equiv] in DCM [10 vol], TEA [12.0 ml, 1.1 equiv] was added. The reaction mixture was cooled to 10°C under N2 atmosphere and stirred for 30 min. Magnesium chloride [1.2 equiv] was added and stirred at RT for 2.5 h. The reaction mixture was cooled to 0°C and 3-methoxypropyl chloride (0.5 equiv) was added slowly and stirred at RT for 18 h. The reaction mixture was distilled under reduced pressure, then 15% HCl was added and extracted with DCM (3 x 10 vol). The organic layer was washed with brine solution, dried over Na2SO4 and concentrated under reduced pressure. The crude compound was purified by column chromatography on 100-200 mesh silica gel eluted with EA / Hex. Pure fractions were collected from 10-20% EA / hexane and distilled under reduced pressure to give intermediate 2a (R = CH2CH2OMe) (2.9 g, 23% yield). 1H NMR (400MHz, DMSO-d6): δ 1.16-1.20(t, 3H), 2.72-2.75(t, 2H), 3.22(s, 3H), 3.51-3.54(t, 2H), 3.59(s, 2H), 4.06-4.11(q, 2H).

[0230] Intermediate 2a (R2=CH2CH2CF3) for producing compound 44 was prepared as follows: Meldrum's acid (0.986g, 6.94mmol) was added to a stirred solution of 4,4,4-trifluorobutyric acid (1.0g, 6.94mmol) in 15ml DCM and stirred at room temperature for 10 minutes. To it, DCC and DMAP were added and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was filtered through a celite bed and extracted with 1(N) HCl. The combined organic portion was passed through anhydrous sodium sulfate and evaporated under vacuum, and the crude was sent to the next step without further purification. The crude was then dissolved in methanol, catalytic p-TSA was added and it was refluxed for 1.5 hours. The reaction mixture was evaporated to dryness and the crude material was used directly without further purification.

[0231] c) Compounds of formula (I) where R5 is S and n=0; R2 and R3 are both alkyl; and R5 is NH. Compounds 47-49 were synthesized according to Scheme 1 herein. [ka]

[0232] Compound 47 was prepared according to Scheme 6 below: [ka]

[0233] Scheme 6 for the preparation of compound (47) To a stirred solution of 5-propyl-3-thioxo-2,8-dihydro-[1,2,4]triazolo[4,3-a]pyrimidin-7(3H)-one (intermediate 1e from Scheme 3, 500 mg, 2.38 mmol) and iodobenzene (728 mg, 3.57 mmol) in DMSO (5 mL) was added K3PO4 (1.51 g, 7.14 mmol), and the reaction mixture was then degassed for 10 min under argon atmosphere in a sealed tube. CuI (45 mg, 0.23 mmol) and 1,10-phenanthroline (41 mg, 0.23 mmol) were then added, and the reaction mixture was heated at 120° C. for 72 h. The reaction mixture was evaporated in gene-vacuo and purified by Combi-flash (0.5% MeOH-DCM) to give compound 47 as a white solid (200 mg, 29% yield). 1 H NMR (400MHz, DMSO) δ: 0.93(t, 3H), 1.54-1.61(t, 2H), 2.96(t, 2H), 5.98(s, 1H), 7.23-7.38(m, 5H), 12.90(s, 1H). LCMS (Method-1):R t =2.16 minutes; [M+H]=287. HPLC=99.75%.

[0234] Compound 48 was prepared according to Scheme 7 below: [ka]

[0235] Scheme 7, Step 1: Intermediate (4a) To a stirred solution of ethyl butyrate (5 g, 31.62 mmol) in DMF (100 mL) at 0° C. was added K2CO3 (10.91 g, 79.06 mmol) and MeI (1.97 mL, 31.62 mmol) and stirred at rt for 48 h. The reaction mixture was partitioned between ethyl acetate and aqueous NH4Cl. The organic portion was dried, concentrated and purified by Combi-flash (eluted with 2% EA-Hex) to give the desired intermediate 4a as a colorless liquid in 22.03% yield, 1.2 g. 1H NMR (400MHz, DMSO) δ: 0.82(t, 3H), 1.17(d, 3H), 1.43-1.52(m, 2H), 2.45(s, 3H), 3.66-3.71(m, 1H), 4.07-4.12(m, 2H);

[0236] Scheme 7, Step 2: Intermediate (4b) To a stirred solution of intermediate 4a (1.2 g, 6.96 mmol) and 2-methyl-2-thiopseudourea hemisulfate (1.06 g, 7.66 mmol) in water (20 mL) was added Na2CO3 (1.18 g, 11.15 mmol) and stirred at rt for 48 h. A white precipitate was slowly removed and attempted filtration was unsuccessful as white material stuck to the walls of the sinter funnel. Therefore, the reaction mixture was partitioned between 10% IPA-MeOH and water. The organic portion was dried, concentrated and triturated with ether to give the desired intermediate 4b as a white solid in 57.9% yield, 800 mg. 1 H NMR (400MHz, DMSO) δ: 0.90(t, 3H), 1.54-1.63(m, 2H), 1.87(s, 3H), 2.26(s, 3H), 2.42(t, 2H), 8.71(brs, 1H);

[0237] Scheme 7, Step 3: Intermediate (4c) To a stirred solution of intermediate 4b (800 mg, 4.03 mmol) in ethanol (10 mL) was added NH2NH2.H2O (2.01 mL, 40.35 mmol) and refluxed for 24 h. The reaction mixture was evaporated to dryness. Neat NH2NH2.H2O (5 mL) was added and refluxed for 16 h. LCMS showed the formation of the major product. The reaction mixture was evaporated to complete dryness and triturated with EtOH and ether to give the desired intermediate 4c as an off-white solid (crude). 1 H NMR (400MHz, DMSO) δ: 0.85 (t, 3H), 1.49-1.57 (m, 2H), 1.79 (s, 3H), 2.34 (t, 2H), 3.91 (brs, 2H), 7.09 (brs, 1H); LCMS (method-2): R t = 1.32 min; [M+H] = 183 (3 min run time, NHOAc+MeCN).

[0238] Scheme 7, Step 4: Intermediate (4d) To a stirred solution of crude 2-hydrazinyl-5-methyl-6-propylpyrimidin-4(1H)-one (400 mg, 2.19 mmol) in pyridine (10 mL) was added CS2 (2.2 mL) and refluxed for 6 h. The reaction mixture was evaporated in vacuum, azeotroped with toluene, and finally triturated with EtOH to give the crude target compound as an off-white solid. This crude material (Intermediate 4d, 180 mg) was used for the next step without further purification.

[0239] Scheme 7, step 5 for preparing compound (48) To a stirred solution of crude intermediate 4d (180 mg, 0.80 mmol) and TEA (167 μL, 1.20 mmol) in EtOH (10 mL) was added 2-chloro,6-fluorobenzyl bromide (99 μL, 0.72 mmol) and stirred at rt for 18 h. The reaction mixture was evaporated to dryness and partitioned between 10% IPA-DCM and water. The organic portion was dried over anhydrous Na2SO4 and purified first by Combi-flash (1% MeOH-DCM) and finally by preparative TLC (mobile phase 5% MeOH-DCM) to give the desired compound (48) as an off-white solid (20 mg) in 6.79% yield. 1 H NMR (400MHz, DMSO) δ: 0.85 (t, 3H), 1.46-1.48 (m, 2H), 1.93 (s, 3H), 2.94 (t, 2H), 4.38 (s, 2H), 7.22-7.38 (m, 3H), 12.87 (brs, 1H); LCMS (method-1): R t = 1.58 min; [M+H] = 367 (3 min run time, HCOOH+MeCN). HPLC = 98.60%.

[0240] Compound 49 was prepared according to Scheme 8 below: [ka]

[0241] Scheme 8, Step 1: Intermediate (5b) To a stirred solution of intermediate 1d (Scheme 3, 700 mg, 4.16 mmol) in EtOH (25 mL) was added 1-chloro-3-fluoro-2-(isothiocyanatomethyl)benzene (5a, 1.25 g, 6.25 mmol) and refluxed for 18 h. The reaction mixture was evaporated to dryness and washed with EtOH. The white solid material was filtered and concentrated to give crude intermediate 5b as an off-white solid (600 mg). LCMS (Method-3): R t = 2.82 min [M+H] = 370 (5 min run time, NHOAc + MeCN).

[0242] Scheme 8, step 2 for preparing compound (49) To a stirred solution of intermediate 5b (600 mg, 1.62 mmol) in dioxane (25 mL) was added DCC (402 mg, 1.95 mmol) and refluxed for 18 h. TLC showed complete consumption of starting material and the formation of two polar spots. The reaction mixture was evaporated to dryness and partitioned between 10% IPA-DCM and saturated NaHCO3 solution. The organic portion was dried over anhydrous Na2SO4 and purified first by Combi-flash (1% MeOH-DCM) to give (49) as an off-white solid (60 mg) in 11% yield. 1 H NMR (400MHz, DMSO) δ:0.79(t, 3H), 1.53-1.58(m, 2H), 2.79(t, 2H), 4.49(s, 2H) ), 5.70(s, 1H), 6.12(brs, 1H), 7.23-7.40(m, 3H), 12.22(s, 1H);LCMS(Method-1):R t = 1.46 min [M+H] = 336 (3 min run time, HCOOH+MeCN). HPLC = 99.18%.

[0243] d) R5 is SO 2 or a bond. Compounds 50-51 were synthesized according to Scheme 2 herein. [ka]

[0244] Compound 50 was prepared according to Scheme 9 below. [ka]

[0245] Scheme 9, Step 1: Intermediate (6b) To a stirred solution of 2,4-dichloro-6-methoxypyrimidine (6a, 500 mg, 2.81 mmol) and Fe(acac)3 (99.22 mg, 0.28 mmol) in THF (25 mL) at 0° C., PrMgBr (0.41 ml, 2.81 mmol) was added and stirred at rt for 48 h. The reaction mixture was partitioned between ethyl acetate and aqueous NH4Cl. The organic portion was dried, concentrated and purified by Combi-flash (eluted with 2% EA-Hex) to give the desired intermediate 6b in 55% yield, 300 mg, as a colorless viscous liquid. 1 H NMR (400 MHz, DMSO) δ: 0.88 (t, 3H), 1.61-1.66 (m, 2H), 2.60 (t, 2H), 3.91 (s, 3H), 6.84 (s, 1H); positive nOe was observed between the aromatic proton (6.84 ppm) and the adjacent CH2 (2.60 ppm) proton. LCMS (Method-3): R t = 3.41 min [M+H] = 187 (5 min run time, NHOAc+MeCN).

[0246] Scheme 9, Step 2: Intermediate (6c) To a stirred solution of intermediate 6b (300 mg, 1.61 mmol) in dioxane (20 mL) was added NH2NH2 (4.03 mL, 40.30 mmol, 1 M in THF) and refluxed for 24 h. The reaction mixture was evaporated to dryness and triturated with ether to give the desired intermediate 6c as a white solid (150 mg, crude). 1 H NMR (400MHz, DMSO) δ: 0.88 (t, 3H), 1.57-1.66 (m, 2H), 2.40 (t, 2H), 3.81 (s, 3H), 4.10 (brs, 2H), 5.90 (s, 1H), 7.92 (s, 1H).

[0247] Scheme 9, Step 3: Intermediate (6d) To a stirred solution of crude intermediate 6c (1 g, 5.49 mmol) in pyridine (25 mL) was added CS2 (5.5 mL) and refluxed for 6 h. The reaction mixture was evaporated in vacuum, azeotroped with toluene, and finally triturated with EtOH to give crude target intermediate 6d as a light yellow solid (600 mg). This crude material (600 mg) was used for the next step without further purification. 1 H NMR (400MHz, DMSO) δ: 0.95 (t, 3H), 1.70-1.76 (m, 2H), 3.55 (t, 2H), 3.92 (s, 3H), 6.35 (s, 1H), 14.09 (s, 1H).

[0248] Scheme 9, Step 4: Intermediate (6e) To a stirred solution of crude intermediate 6d (100 mg, 0.44 mmol) and TEA (93 μL, 0.67 mmol) in EtOH (10 mL) was added 2-chloro,6-fluorobenzyl bromide (71 μL, 0.44 mmol) and stirred at rt for 18 h. The reaction mixture was evaporated to dryness and partitioned between 10% IPA-DCM and water. The organic portion was dried over anhydrous Na2SO4 and purified first by Combi-flash (1% MeOH-DCM) and finally by preparative TLC (mobile phase 5% MeOH-DCM) to give intermediate 6e as an off-white solid (80 mg) in 42.76% yield. 1 H NMR (400 MHz, DMSO) δ: 0.91 (t, 3H), 1.56-1.58 (m, 2H), 3.02 (t, 3H), 3.97 (s, 3H), 4.41 (s, 2H), 6.51 (s, 1H), 7.15-7.38 (m, 3H); HMBC confirmed the formation of the desired isomer. LCMS (Method-2): t = 1.76 min [M+H] = 367 (3 min run time, NHOAc + MeCN). HPLC = 99.83%

[0249] Scheme 9, Step 5: Intermediate (6f) To a stirred solution of 3-((2-chloro-6-fluorobenzyl)thio)-7-methoxy-5-propyl-[1,2,4]triazolo[4,3-a]pyrimidine intermediate 6e (200 mg, 0.54 mmol) in DCM (25 mL) was added m-CPBA (312 mg, 1.09 mmol, 60%) and stirred at rt for 18 h. The reaction mixture was partitioned between saturated NaHCO3 solution and DCM. The organic layer was dried and concentrated to give 80 mg of crude intermediate 6f. LCMS (Method-1): R t =2.18 minutes; [M+H]=399.

[0250] Scheme 9, step 6 for preparing compound (50) HBr-AcOH (10 mL) was added to intermediate 6f (80 mg, 0.20 mmol) and heated at 80° C. for 18 h. TLC showed complete consumption of starting material and formation of a polar spot. The reaction mixture was evaporated to dryness and partitioned between 10% IPA-DCM and saturated NaHCO3 solution. The organic portion was dried over anhydrous Na2SO4 and purified first by Combi-flash (1% MeOH-DCM) to give compound (50) as a white solid (10 mg) in 13% yield. 1 H NMR (400MHz, DMSO) δ: 0.92 (t, 3H), 1.62-1.66 (m, 2H), 2.90 (t, 2H), 5.41 (s, 2H), 6.06 (s, 1H), 7.37-7.57 (m, 3H). LCMS (Method-1):R t =3.72 minutes; [M+H]=385. HPLC=94.78%.

[0251] Compound 51 was prepared according to Scheme 10 below. [ka]

[0252] Scheme 10, Step 1: Intermediate (7a) To a stirred solution of intermediate 6c (from Scheme 9, 700 mg, 3.84 mmol) and 3-phenylpropanoic acid (577 mg, 3.84 mmol) in DMF (25 mL) was added EDC.HCl (1.1 g, 5.76 mmol), HOBt (796 mg, 5.76 mmol) and DIPEA (1.48 g, 11.53 mmol) and stirred for 18 h. The reaction mixture was evaporated to dryness and partitioned between DCM and saturated NaHCO3 solution. The organic portion was dried over anhydrous Na2SO4 and purified first by Combi-flash (70% EA-Hex) to give the desired intermediate 7a as an off-white solid (500 mg) in 42% yield. LCMS (Method-2): R t = 1.77 min [M+H] = 315 (3 min run time, NHOAc + MeCN).

[0253] Scheme 10, Step 1: Intermediate (7b) To a stirred solution of intermediate 7a (500 mg, 1.59 mmol) in THF (25 ml) was added Burgess reagent (758 mg, 3.18 mmol) and stirred for 48 h. LCMS showed the formation of the desired product and some unreacted SM. The reaction mixture was evaporated to dryness and partitioned between DCM and saturated NaHCO3 solution. The organic portion was dried over anhydrous Na2SO4 and purified first by Combi-flash (60% EA-Hex) to give the desired intermediate 7b as an off-white solid (200 mg) in 42% yield. LCMS (Method-2): R t = 1.75 min [M+H] = 297 (3 min run time, NHOAc+MeCN).

[0254] Scheme 10, step 3 for preparing compound (51) HBr-AcOH (10 mL) was added to intermediate 7b (200 mg, 0.67 mmol) and heated at 80° C. for 18 h. TLC showed complete consumption of starting material and formation of a polar spot. The reaction mixture was evaporated to dryness and partitioned between 10% IPA-DCM and saturated NaHCO3 solution. The organic portion was dried over anhydrous Na2SO4 and purified first by Combi-flash (1% MeOH-DCM) to give the desired compound (51) as an off-white solid (50 mg) in 26% yield. 1 H NMR (400MHz, DMSO) δ: 0.93 (t, 3H), 1.58-1.64 (m, 2H), 2.86 (t, 2H), 3.12 (t, 2H), 5.83 (s, 1H), 7.20-7.31 (m, 5H), 12.64 (s, 1H); LCMS (method-2): R t = 1.58 min [M+H] = 283 (3 min run time, NHOAc + MeCN). HPLC = 94.22%.

[0255] Compounds 52-100 (Table 1) can be readily obtained by one skilled in the art from Schemes 1 and 2, particularly via alkylation of intermediate 1e with a suitable electrophile, such as benzyl bromide, as in step 4 of Scheme 3. Compounds were obtained for testing via commercial sources.

[0256] [Table 1-1]

[0257] [Table 1-2]

[0258] [Table 1-3]

[0259] [Table 1-4]

[0260] [Table 1-5]

[0261] [Table 1-6]

[0262] [Table 1-7]

[0263] [Table 1-8]

[0264] [Table 1-9]

[0265] [Table 1-10]

[0266] LCMS method LC-MS method 1: Column - YMC-Triat C18 (33 x 2.1 mm, 3 mm), (Mobile phase: 98% [HCOOH in 0.05% water] and 2% [CH3CN] for 0.75 min, then 90% [HCOOH in 0.05% water] and 10% [CH3CN] for 1.0 min, then 2% [HCOOH in 0.05% water] and 98% [CH3CN] for 2.0 min, this mobile phase composition was held for a maximum of 2.5 min, and finally returned to the initial condition for 3.0 min). Flow = 1.0 ml / min.

[0267] LC-MS method 2: LCMS / MS-API 2000 / Q trap

[0268] Monitoring Methodology API 2000 mass spectrometer (single quadrupole mass spectrometer) manufactured by Applied Biosystems Ionization method: electrospray; Polarity: positive ion. Capillary (kV) 5.5, DP (V) 50.00, Inlet voltage (V) 10, Focus voltage (V) 400, Source temperature 200°C, Ion source gas 1 (Psi) 40, Ion source gas 2 (Psi) 50, Curtain gas (Psi) 40 Mass range: 100-800 AMU; UV wavelength range: 220-260 nm; Method (Shimadzu Prominance system) using the following HPLC gradient conditions: Solvent A: NH4OAc in 10 Mm water and Solvent B: Acetonitrile. Flow rate: 1.2 ml / min.

[0269] [Table 2]

[0270] Column Type : Zorbax Extend C18; Column length: 50 mm; Column inner diameter: 4.6 mm; Particle size: 5 mm

[0271] Mass condition :Ionization technique: ESI (electrospray ionization) using API (atmospheric pressure ionization) source; Cluster separation potential: 10-70V depending on compound ionization; Mass range: 100-800amu; Scan type: Q1; Polarity: +ve; Ion source: turbospray; Ion spray voltage: +5500; Mass source temperature: 200°C.

[0272] LC-MS method 3: LCMS / MS-API 2000 / Q trap-monitoring method API 2000 mass spectrometer (single quadrupole mass spectrometer) manufactured by Applied Biosystems Ionization method: electrospray; Polarity: positive ion. Capillary (kV) 5.5, DP (V) 50.00, Inlet voltage (V) 10, Focus voltage (V) 400, Source temperature 200 °C, Ion source gas 1 (Psi) 40, Ion source gas (Psi) 50, Curtain gas (Psi) 40; Mass range: 100-800 amu; UV wavelength range: 220-260 nm; Method Shimadzu Prominence using the following HPLC gradient conditions (Solvent A: NHOAc in 10 Mm water and Solvent B: Acetonitrile). Flow rate: 1.2 ml / min.

[0273] [Table 3]

[0274] Column Type : Xbridge C18; column length: 50 mm; column inner diameter: 4.6 mm; particle size: 5 microns.

[0275] Mass condition :Ionization technique: ESI (electrospray ionization) using API (atmospheric pressure ionization) source; Cluster separation potential: 10-70V depending on compound ionization; Mass range: 100-800amu; Scan type: Q1; Polarity: +ve; Ion source: turbospray; Ion spray voltage: +5500; Mass source temperature: 200°C.

[0276] LC-MS method 4: UPLC-MS method: column : XBRIDGE C18; column length: 50mm; column inner diameter: 4.6mm; particle size: 5mm.

[0277] Gradient conditions Column temperature: 50° C.; injection volume: 0.5 μL; Waters ACQUITY UPLC with the following HPLC gradient conditions (Solvent A: 5 mM NH4OAc in water and Solvent B: 5 mM NH4OAc in acetonitrile:water--90:10).

[0278] [Table 4]

[0279] Mass conditions: ACQUITY SQD mass spectrometer manufactured by Waters (single quadrupole mass spectrometer, Waters ACQUITY SQD2); ionization method: electrospray; polarity: positive ion; capillary (kV) 3.50, cone (V) 40.00, source temperature 150 °C, desolvation temperature 450 °C, cone gas flow (L / hr) 50, desolvation gas flow (L / hr) 750; mass range: 100-800 Da; DAD (diode array detection); wavelength range (nm): 210-400 nm; HPLC: Waters AQUITY UPLC.

[0280] LC-MS Method 5: Agilent UPLC-MS: Column - YMC Triart C18 (2.1 x 33 mm, 3 mm)

[0281] Gradient conditions : Flow rate: 1.2 ml / min; Column temperature: 50 °C; Injection volume: 0.4 μL Solvent A: 0.01% HCOOH in water and Solvent B: 0.01% HCOOH in CH3CN, (Mobile phase: 95% [0.01% HCOOH in water] and 5% [0.01% HCOOH in CH3CN] for 0.50 min, then 1% [0.01% HCOOH in water] and 99% [0.01% HCOOH in CH3CN] held for 3.0 min, this composition held for up to 4.00 min, and finally returned to initial conditions at 4.10 min and held for 4.50 min).

[0282] [Table 5]

[0283] Mass condition: Agilent SQD mass spectrometer (single quadrupole mass spectrometer); ionization method: electrospray; polarity: negative ion capillary (kV) 4.00, fragmentor (V) 150.00, threshold -200, drying gas temperature 350 °C, drying gas flow (L / min) 12, nebulizer pressure (Psi) 50.

[0284] Example 2: Inhibitory activity against MPC The inhibition of the mitochondrial pyruvate transporter (MPC) by the compounds of the present invention was tested as follows, and their inhibitory activity was measured using IC 50 expressed as a value.

[0285] reagent Seahorse Xfe24 Flux Analyser[Agilent] Assay medium: PBS / CaCl2 / MgCl2 containing 1 mM pyruvate (Eurobio CS1PBS00-01).

[0286] The following working reagents were made up in assay medium at 10× final concentration: - Oligomycin A stock: 10 mM in DMSO. Working solution: 20μM (4μl / 2ml) -fCCP (carbonyl cyanide-(trifluoromethoxy)phenylhydrazone) Stock: 10 mM in DMSO Working solution: 10μM (2μl / 2ml) -Rotenone 10mM in DMSO Working solution: 10μM -Antimycin Stock: 10 mM in EtOH Working solution: 10uM (Pool the rotenone + antimycin reagents: (2 μl rotenone + 2 μl antimycin in 2 ml assay medium)

[0287] The day before Cells were plated in dedicated Seahorse plates. For HeLa cells, the seeding was 4 × 10 in 500 ml of DMEM, 10% FBS, 1% pen / strep / glutamine. 4 10 cells / well. 3 wells were left blank (medium alone) for internal calibration. Assay cartridges were hydrated overnight at 37° C. in 1 ml / well of XF Calibrant solution.

[0288] Assay Date Reagent Plate Reagents / dilutions prepared in assay medium. 50 μl of a compound of the invention is injected into 450 μl of washed cells, so that the compound is made up in assay medium at 10× the final desired concentration. For dose response analysis, compounds are tested at final concentrations of 30, 5, 0.83, 0.14, 0.023 μM (1:6 serial dilutions from 30 mM). Positive control: 30 mM (parent hit compound that gave rise to the actual series) Negative control: assay medium alone.

[0289] The seahorse reagent plate is prepared as follows and transferred to the Xfe24 analyser for the calibration step. Port A Compound (50 μl) Port B Oligomycin A (55 μl) Port C fCCP [(carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone]] (62 μl) Port D rotenone + antimycin (68 μl).

[0290] Cell Plates The medium is almost completely removed from the wells without allowing the cells to dry.

[0291] Cells are washed twice with 300 μl of assay medium. 450 μl of fresh assay medium is added to each well. Plates are transferred to an Xfe24 analyser for measurement of pyruvate-dependent OCR. All values ​​are normalized to the initial basal respiration rate in each well, measured before compound addition and designated as 100%. IC 50 Determinations are based on the average of triplicate measurements performed in the presence of fCCP, with 30 μM of the parent hit compound representing maximal inhibition and PBS / Ca / Mg representing zero inhibition.

[0292] Table 2 below shows the IC of the compounds of the present invention. 50 Represents.

[0293] [Table 6]

[0294] Example 3: Effect of MPC inhibition during in vitro priming of CD8 T cells on memory marker expression OT1 splenocytes were cultured for 3 days at a concentration of 106 cells per mL in RPMI medium (Gibco 61870-01) supplemented with 10% FBS (Gibco 10270-106), 1% penicillin / streptomycin (Gibco 15070-063), 50 μM β-mercaptoethanol, 1% HEPES (Gibco 15630-080), 1× non-essential amino acids (Gibco 11140-035), 1% L-glutamine (Gibco 25030-081), and 1 mM sodium pyruvate (Gibco 11360-039). The cells were additionally supplemented with 100U / ml hIL-2 (Glaxo-IMB), 1 μg / ml ovalbumin N4 peptide (257-264) (a model peptide antigen derived from ovalbumin), and different concentrations of either compound 45 or 47, or their solvent DMSO as a control. On day 3, splenocytes were harvested, washed, split, and the cells were cultured for an additional 4 days with 100U / ml hIL-2 and hIL-7 (Peprotech 200-07) supplemented with either compound 45 or 47, or DMSO. On day 7, flow cytometry analysis was performed for surface marker expression.

[0295] To evaluate the effect of MPC inhibition on T cell differentiation, OT1 splenocytes were cultured for 3 days in the presence of 1 μg / ml ovalbumin-derived N4 peptide, 100 IU / ml recombinant human IL-2 (rhIL-2) and MPC inhibitors Compound 45 or 47, or control DMSO. Cells were cultured for an additional 4 days in the presence of 100 IU / ml IL-2 and 10 ng / ml rhIL-7 (Figure 1A), as well as inhibitors or DMSO. When analyzing the surface expression of the central memory marker CD62L (Figure 1B) and the pro-survival receptor CD127 (Figure 1C) by flow cytometry on day 7, an increase was observed in cells treated with the highest dose of the compound of the invention compared to DMSO.

[0296] Thus, these data demonstrate that treatment of CD8 T cells with the compounds of the invention results in enhanced memory characteristics.

[0297] Example 4: Adoptive cell transfer therapy of CD8 T cells treated with compounds of the invention The efficacy of the compounds of the invention is tested in an adoptive cell transfer therapy model using CD8 T cells treated as follows, and in a melanoma tumor model.

[0298] Adoptive cell transfer Activated CD45.1+ OT-1 splenocytes were cultured in vitro for 7 days, harvested, and purified on a Ficoll gradient to separate dead and live splenocytes as described above. Live splenocytes were counted using 0.4% trypan blue staining. 100,000 or 2,000,000 live splenocytes were transferred into CD45.2+ host mice by tail vein injection.

[0299] To assess the functional capacity of the generated memory T cells, compound 45-treated cells were further tested in a mouse model of melanoma.

[0300] Melanoma Tumor Models B16-OVA cells were cultured in DMEM (GIBCO) with 10% FBS and 1% P / S before their subcutaneous injection into the mouse flank. Each mouse received 100,000 cells in a volume of 200 μl PBS. Six days after B16-OVA cell injection, tumors were measured and mice were randomized and lymphodepleted by irradiation (5Gray). Seven days after B16-Ova injection, mice were adoptively transferred using the ACT protocol previously described. After ACT, mice were vaccinated with CpG (50 μg / mouse) and N4 Ova peptide (10 μg / mouse) diluted in PBS to obtain a total volume of 100 μl / mouse that was injected subcutaneously in the tail. Tumors were measured every 2 days and tumor volume was calculated according to the formula: V=π×[d2×D] / 6, where d is the tumor minor axis and D is the tumor major axis. On day 26 after tumor engraftment, tumors and spleens were dissected and then stained for flow cytometry analysis.

[0301] Briefly, 105 ovalbumin-expressing B16 melanoma cells were subcutaneously injected into 6-week-old mice. Six days after engraftment, when palpable tumors were present, mice were irradiated with 5 Gy. The next day, 105 compound 45 or DMSO-treated OT1 cells were intravenously injected, followed by subcutaneous vaccination with 50 μg CpG and 10 μg N4 Ova peptide (Figure 2A). It has previously been shown that memory CD8+ T cells are more potent in controlling tumor growth, and indeed, B16 tumor growth was reduced in mice receiving OT1 CD8+ T cells pretreated with compound 45 compared to DMSO (Figure 2B). However, tumor weight was not significantly reduced due to high variability (Figure 2C). When blood was analyzed 9 days after adoptive cell transfer, an increased number of transferred cells could be observed when treated with compound 45 in vitro compared to DMSO (Figure 2D). The phenotype of compound 45-treated cells was biased toward memory, as there was a reduction in the percentage of short-lived effector T cells (Figure 2E), as well as an increase in memory precursor effector T cells and cells with a central memory phenotype (Figures 2F and 2G). Upon analysis of the tumors at dissection, more compound 45-treated T cells infiltrated the tumor (Figure 2H). However, tumor-infiltrating compound 45-treated T cells were not significantly more or less exhausted compared to DMSO-treated T cells (Figure 2I). There was no difference in terminal exhausted or precursor exhausted T cells (Figures 2J and 2K). In the spleen, the number of compound 45-treated T cells was also increased (Figure 2L), and they showed an increase in the central memory phenotype (Figure 2M), but TCF1 expression was not significantly altered (Figure 2N). When tumor single cell populations were restimulated with OVA peptide in vitro, OT1 T cells treated with compound 45 did not show a significant increase in IFNy, TNF, IL2 or granzyme B expression (Figure 2O-R), but showed increased expression of CD107a (LAMP1) on the cell surface membrane and increased degranulation, which correlated with improved cytotoxicity (Figure 2S).

[0302] These data demonstrate that CD8 T cells treated with the compounds of the invention exhibit enhanced anti-tumor activity.

[0303] To observe a stronger antitumor response of the transferred OT1 T cells, we performed the previous experiment at 10 5 (Figure 3A) instead of 2 x 10 6 The study was repeated by transferring 100 DMSO- or Compound 45-treated OT1 T cells. Under these conditions, B16 tumor growth and weight were significantly greater in OT1 CD8 T cells pretreated with Compound 45 compared to DMSO. + In mice that received T cells, the T cell count was strongly reduced (Figures 3B and 3C). When tumors were analyzed at the time of dissection, no differences in T cell infiltration were observed (Figure 3D). Surprisingly, tumor-infiltrating compound 45-treated T cells were more exhausted compared to DMSO-treated T cells (Figure 3E). However, there were no differences in terminally exhausted or precursor exhausted T cells (Figures 3F and 3G). In the spleen, the number of compound 45-treated T cells was reduced (Figure 3H), but they showed an increase in the central memory phenotype (Figure 3I), and a trend towards increased TCF1 expression (Figure 3J). When single cell populations from tumors were restimulated in vitro with OVA peptide, compound 45-treated OT1 T cells showed no differences in IFNy, TNF, or IL2 expression (Figures 3K-3M), but increased granzyme B expression (Figure 3N). There were no differences in CD107a expression (Figure 3O).

[0304] These data support that adoptive cell transfer therapy of CD8 T cells treated with the compounds of the invention can better control tumor growth.

[0305] Example 5: Effect of MPC inhibition during the generation of mouse CAR T cells during adoptive cell transfer therapy The above data were obtained in mice using CD8 T cells isolated from transgenic OT1 mice that have been engineered to express one unique T cell receptor that recognizes a peptide sequence of chicken ovalbumin protein (N4 peptide) when presented on an MHC class I molecule.

[0306] The usefulness of the compounds of the invention when applied during the generation of mouse CAR T cells was further explored by using a CAR construct that recognizes human HER2, an oncogene frequently implicated in human breast cancer, containing a 4-1BB costimulatory domain.

[0307] Retroviral preparation (protocol modified from Tschumi et al., 2018, J Immunother Cancer.;6(1):71) For each retroviral preparation, 8 × 10 6 Phoenix ECO cells (ATCC, CRL-3214) were plated in RPMI medium supplemented with 10% FCS, 10 mM HEPES and 50 U / ml penicillin-streptomycin in a T150 tissue culture flask. The next day, cells were transfected with 21 μg of retroviral construct using Turbofect transfection reagent (Thermo Fischer Scientific) according to the manufacturer's protocol. Medium was changed daily and collected 48 and 72 hours after transfection. The 48 and 72 hour viral supernatants were pooled and precipitated at 22,000 rcf for 2 hours at 4°C. Finally, the retroviral pellet was resuspended in 2 ml of whole RPMI medium and divided into eight aliquots of 250 μl each, which were flash frozen on dry ice and stored at -80°C.

[0308] T cell transduction (protocol modified from Tschumi et al., 2018, above) Spleens from wild-type CD45.1.2 mice were disrupted on a 70 μm cell strainer. CD8 T cells were purified using the EasySep™ Mouse CD8+ T Cell Isolation Kit (StemCell) according to the manufacturer's protocol. 0.5×106 CD8 T cells were plated in 0.5 ml of complete RPMI 1640 medium supplemented with 10% FCS, antibiotics and 50 IU / ml recombinant human IL-2 in a 48-well plate and exposed to either DMSO or 20 μM compound 45. Mouse T cells were activated with activators CD3 / CD28 Dynabeads (Gibco) at a ratio of 2 beads per cell. Retroviral infection was performed for 24 hours at 37°C. Untreated 48-well plates were coated with 20 μg / ml recombinant human fibronectin (Takara Clontech) for 24 h at 4°C, followed by PBS 2% BSA for 30 min at RT and finally washing with PBS. One aliquot of concentrated retrovirus was plated onto each fibronectin-coated 48-well plate and centrifuged at 2,000 rcf for 90 min at 32°C. 0.5 × 10 6 24 hour activated CD8 T cells were added on top of the virus and spun at 400rcf for 10 minutes at 32°C. On day 3, the medium was replaced with 10IU / ml recombinant human IL-2, 10ng / ml recombinant human IL-7 and 10ng / ml recombinant human IL-15 containing either DMSO or 20M compound 45. Cells were then split every 2 days.

[0309] Adoptive cell transfer (protocol modified from Tschumi et al., 2018, above) CD45.2 C57BL / 6 mice were injected with 4 × 10 5 B16F10 tumors were implanted subcutaneously. Six days later, mice were lymphodepleted by ip injection of 100 mg / kg cyclophosphamide (Sigma Aldrich, C7397) and homogenous groups were established with respect to tumor volume. T cells (5×10 6) were adoptively transferred iv the following day. Tumor volumes were measured three times a week with calipers and calculated using the formula: V = π x [d2 x D] / 6, where d is the tumor minor axis and D is the tumor major axis. Tumors were harvested and separated from the skin. Single cell suspensions were obtained using a Mouse Tumor Dissociation Kit (Miltenyi, 130-096-730) according to the manufacturer's protocol. Spleens and draining lymph nodes were disrupted on 70 μm cell strainers. Single cell suspensions were stained with antibodies before flow cytometry analysis.

[0310] Polyclonal CD8 T cells from wild-type mice were activated in the presence of DMSO or 20 μM compound 45 and then retrovirally transduced with the HER2-CAR construct (Figure 4A). When adoptively transferring T cells (ACT) in mice bearing HER2-expressing B16 melanoma tumors, only ACT of compound 45-treated HER2CAR T cells was able to significantly suppress tumor growth (Figures 4B and 4C). When blood was analyzed 12 days after ACT, no difference in the number of CAR T cells was observed (Figure 4D), however, the percentage of compound 45-HER2CAR T cells forming short-lived effector T cells was significantly reduced (Figure 4E). No difference in the percentage of memory precursor effector T cells (Figure 4F), TCF1 expressing T cells (Figure 4G) could be detected. A trend towards increased engraftment of compound 45-HER2CAR T cells was observed in the tumor-draining lymph nodes, but this increase was significant in the spleen (Figures 5A and 5D). No differences in central memory T cell differentiation (Figures 5B and 5E) or HER2CAR T cells expressing the memory-specific transcription factor TCF1 (Figures 5C and 5F) could be observed in either the draining lymph nodes or the spleen. Tumors in mice treated with compound 45-HER2CAR contained significantly more CAR T cells (Figure 5G). More tumor-infiltrating compound 45-treated CAR T cells expressed TCF1 (Figure 5H). Interestingly, looking at the co-expression of TCF1 and PD1, compound 45-treated CAR T cells formed more precursor exhausted T cells (TCF1 positive) and fewer terminally differentiated exhausted T cells (TCF1 negative) (Figures 5I and 5J), indicating an increase in a stem cell-like phenotype that may benefit from combination therapy with checkpoint blockade immunotherapy. Finally, some compound 45-treated CAR T cells expressed the inhibitory molecules PD1 and TIM3, indicative of a less exhausted phenotype (Figure 5K).

[0311] Taken together, these data support that the compounds of the invention are useful during the generation of CAR T cells by improving their memory phenotype and antitumor function during adoptive cell transfer therapy.

Claims

1. A compound of the following formula (I): 【Chemical 1】 [Wherein, R1 is the R5-R6 moiety; R2 is H, C which may optionally be substituted １ -C ６ alkyl, aryl which may optionally be substituted, heteroaryl which may optionally be substituted, arylC １ -C ６ alkyl, heteroarylC １ -C ６ alkyl, C which may optionally be substituted １ -C ６ alkoxy, for example, C which may optionally be substituted １ -C ６ alkoxy-substituted C １ -C ６ alkyl, C which may optionally be substituted ３ -C ８ heterocycloalkyl, and C which may optionally be substituted ３ -C ８ cycloalkyl, for example, selected from cyclopropyl which may optionally be substituted; R3 is H and C which may optionally be substituted １ -C ６ alkyl, and at least one of R2 and R3 is not H; R4 is H; R5 is a bond, S, SO ２ , NR9 and O; R6 is -(CR10R11) ｎ -R7 group (wherein n is an integer from 0 to 2); R7 is C which may optionally be substituted １ -C ６ alkyl, C which may optionally be substituted ２ -C ６ Selected from alkenyl, an optionally substituted heterocycle, an optionally substituted aryl, for example, optionally substituted phenyl, optionally substituted heteroaryl, cyano, and -C(O)-R8; R8 is selected from optionally substituted amino, optionally substituted alkoxy, and optionally substituted aryl; R9 is H or optionally substituted C １ -C ６ ; R10 and R11 are independently selected from H and optionally substituted C １ -C ６ alkyl], or a tautomer, geometric isomer, optically active form, pharmaceutically acceptable salt or pharmaceutically active derivative thereof, for the prevention and / or treatment of cancer; autoimmune diseases, such as multiple sclerosis; metabolic diseases, such as type 2 diabetes; hair loss disorders, such as alopecia; neurodegenerative disorders, such as Parkinson's disease or Alzheimer's disease; fibrosis, such as pulmonary fibrosis or non-alcoholic steatohepatitis (NASH); skin or tissue injury, such as skin wounds or burns; and acute brain conditions, such as stroke or traumatic brain injury, or for skin or tissue regeneration, a pharmaceutical composition.

2. The pharmaceutical composition according to Claim 1, wherein R1 is an S-R6 moiety.

3. The pharmaceutical composition according to Claim 1, wherein R1 is an NR9-R6 moiety.

4. The pharmaceutical composition according to Claim 1, wherein R1 is an R6 moiety.

5. R2 is optionally substituted C １ -C ６ alkyl, and the pharmaceutical composition according to claim 1.

6. R3 is C which may optionally be substituted １ -C ６ The pharmaceutical composition according to claim 1, wherein the R3 is alkyl

7. The pharmaceutical composition according to Claim 1, wherein R3 is H.

8. The pharmaceutical composition according to Claim 1, wherein R4 is H.

9. R7 is optionally substituted C １ -C ６ alkyl, and the pharmaceutical composition according to claim 1.

10. R7 is optionally substituted C ２ -C ６ The pharmaceutical composition according to claim 1, wherein it is alkenyl.

11. The pharmaceutical composition according to Claim 1, wherein R7 is an aryl which may be optionally substituted.

12. The pharmaceutical composition according to Claim 1, wherein n is an integer selected from 0 and 1, R5 is S, and R7 is an aryl which may be optionally substituted.

13. The pharmaceutical composition according to Claim 1, wherein R7 is a heteroaryl which may be optionally substituted.

14. The compound is from the following group: 3-[(3,4-Difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chloro-5-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(3,5-Difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-{[3-(Difluoromethyl)benzyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-{[2-(Difluoromethyl)benzyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-{[4-(Difluoromethyl)benzyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-{[(7-Oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile; 4-Fluoro-2-{[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile; 3-[(5-Fluoro-2-methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(3-Chloro-5-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(4-Chloro-3-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(5-Fluoro-2-methylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one); 3-[(4-Fluoro-2-methylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Phenylethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(Furan-3-ylmethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(3-Methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 4-{[(7-Oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile; 3-[(1-Benzofuran-5-ylmethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(4-Methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,5-Difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,4-Difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,6-Difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-Propyl-3-[(1H-pyrazol-4-ylmethyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,6-Dimethylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chloro-6-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,5-Dimethylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-{[2-(2,4-Difluorophenyl)ethyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,5-Difluorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-[2-(Benzyloxy)ethyl]-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,5-Difluorobenzyl)sulfanyl]-5-(methoxymethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-{2-[(4-Chlorobenzyl)oxy]ethyl}-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-{2-[(2-Chlorobenzyl)oxy]ethyl}-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-{2-[(3-Chlorobenzyl)oxy]ethyl}-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,5-Difluorobenzyl)sulfanyl]-5-(2-methoxyethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chloro-6-fluorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chloro-6-fluorobenzyl)sulfanyl]-5-ethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-Butyl-3-[(2-chloro-6-fluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chloro-6-fluorobenzyl)sulfanyl]-5-cyclopropyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chloro-6-fluorobenzyl)sulfanyl]-5-(2-methylpropyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chloro-6-fluorobenzyl)sulfanyl]-5-(cyclopropylmethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chloro-6-fluorobenzyl)sulfanyl]-5-(methoxymethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-[2-(Benzyloxy)ethyl]-3-[(2-chloro-6-fluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(1H)-one; 3-[(2-Chloro-6-fluorobenzyl)sulfanyl]-5-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-Benzyl-3-[(2-chloro-6-fluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 4-{[(5-Benzyl-7-oxo-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile; 3-(Phenylsulfanyl)-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chloro-6-fluorobenzyl)sulfanyl]-6-methyl-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chloro-6-fluorobenzyl)amino]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chloro-6-fluorobenzyl)sulfonyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-(2-Phenylethyl)-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chlorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-Methyl-3-(methylsulfanyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-Methyl-3-[(4-nitrobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-{[(2E)-3-Phenylprop-2-en-1-yl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; N-(4-Ethoxyphenyl)-2-[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]acetamide; Methyl 5-{[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}furan-2-carboxylate; Ethyl 2-[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]propanoate; Ethyl 2-[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]butanoate; Methyl 2-[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]butanoate; Methyl 2-[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]propanoate; Benzyl [(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]acetate; [(7-Oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]acetonitrile; 3-[(2-Oxo-2-phenylethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-{[2-(4-Methoxyphenyl)-2-oxoethyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; N-(5-Chloro-2-methoxyphenyl)-2-[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]acetamide; 5,6-Dimethyl-3-(propylsulfanyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5,6-Dimethyl-3-[(3-methylbutyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5,6-Dimethyl-3-[(4-nitrobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5,6-Dimethyl-3-[(3-methylbenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,5-Dimethylbenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5,6-Dimethyl-3-[(4-methylbenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5,6-Dimethyl-3-[(3-nitrobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(4-Chlorobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chlorobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5,6-Dimethyl-3-{[3-(trifluoromethyl)benzyl]sulfanyl}[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5,6-Dimethyl-3-{[4-(trifluoromethyl)benzyl]sulfanyl}[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Fluorobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(3-Chlorobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,4-Dichlorobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(3-Chlorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(3-Fluorobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chloro-6-fluorobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(4-Bromobenzyl)sulfanyl]-5,6-dimethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(4-Chlorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-(Benzylsulfanyl)-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Methylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(3-Methylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(4-Methylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(Naphthalen-1-ylmethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(3-Fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(4-Fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chlorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(3,4-Dichlorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,4-Dichlorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(3-Fluorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Fluorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,5-Dimethylbenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; and 3-[(3-Chlorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one The pharmaceutical composition according to any one of claims 1 to 13, selected from

15. The following group: 3-[(3,4-Difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chloro-5-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(3,5-Difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-{[3-(Difluoromethyl)benzyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-{[2-(Difluoromethyl)benzyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-{[4-(Difluoromethyl)benzyl]sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-{[(7-Oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile; 4-Fluoro-2-{[(7-oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile; 3-[(5-Fluoro-2-methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(3-Chloro-5-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(4-Chloro-3-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(5-Fluoro-2-methylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(4-Fluoro-2-methylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Phenylethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(Furan-3-ylmethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(3-Methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 4-{[(7-Oxo-5-propyl-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile; 3-[(1-Benzofuran-5-ylmethyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(4-Methoxybenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,5-Difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,4-Difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,6-Difluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-Propyl-3-[(1H-pyrazol-4-ylmethyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,6-Dimethylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-Chloro-6-fluorobenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,5-Dimethylbenzyl)sulfanyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-{(2-(2,4-Difluorophenyl)ethyl)sulfanyl}-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,5-Difluorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-[2-(Benzyloxy)ethyl]-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,5-Difluorobenzyl)sulfanyl]-5-(methoxymethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-{2-[(4-Chlorobenzyl)oxy]ethyl}-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-{2-[(2-Chlorobenzyl)oxy]ethyl}-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-{2-[(3-chlorobenzyl)oxy]ethyl}-3-[(2,5-difluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2,5-difluorobenzyl)sulfanyl]-5-(2-methoxyethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-methyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-ethyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-butyl-3-[(2-chloro-6-fluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-cyclopropyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-(2-methylpropyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-(cyclopropylmethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-(methoxymethyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-[2-(benzyloxy)ethyl]-3-[(2-chloro-6-fluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(1H)-one; 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-5-(3,3,3-trifluoropropyl)[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 5-benzyl-3-[(2-chloro-6-fluorobenzyl)sulfanyl][1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 4-{[(5-benzyl-7-oxo-7,8-dihydro[1,2,4]triazolo[4,3-a]pyrimidin-3-yl)sulfanyl]methyl}benzonitrile; 3-(phenylsulfanyl)-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-chloro-6-fluorobenzyl)sulfanyl]-6-methyl-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-chloro-6-fluorobenzyl)amino]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; 3-[(2-chloro-6-fluorobenzyl)sulfonyl]-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one; and 3-(2-phenylethyl)-5-propyl[1,2,4]triazolo[4,3-a]pyrimidin-7(8H)-one A compound selected from.

16. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition further comprises at least one anti-cancer immunotherapeutic agent, such as a CAR T cell or an immune checkpoint inhibitor, and at least one of its pharmaceutically acceptable carriers, diluents or excipients. A pharmaceutical composition.

17. An in vitro method for obtaining and / or maintaining T cells having a memory phenotype, the method comprising the following steps: - Providing at least one T cell capable of differentiating into a memory phenotype, such as a CD8+ or CD4+ T cell; - Contacting the at least one T cell with a compound according to any one of claims 1 to 13 or a mixture thereof; - Culturing the cells in a T cell culture medium; - Isolating the obtained T cells A method comprising.

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