Thiazolopyridine derivatives as adenosine receptor antagonists.

Abstract

The present invention relates to thiazolopyridine derivatives falling under the general formula I, and to the use of the compounds of the invention for the treatment and / or prevention of hyperproliferative or infectious diseases and disorders in mammals, particularly humans, and to pharmaceutical compositions containing such compounds.

Description

[Technical field]

[0001] The present invention relates to a compound of general formula I, [ka] and the use of the compounds of the invention for the treatment and / or prevention of hyperproliferative or infectious diseases and disorders in mammals, particularly humans, and pharmaceutical compositions containing such compounds. [Background technology]

[0002] 2. Background of the Invention Adenosine is a ubiquitous modulator with a myriad of physiological activities, specifically within the cardiovascular, nervous, and immune systems. It is structurally and metabolically related to the bioactive nucleotides adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), and cyclic adenosine monophosphate (cAMP), to the biochemical methylating agent S-adenosyl-L-methionine (SAM), and to the coenzymes NAD, FAD, and coenzyme A, as well as to RNA.

[0003] Through cell surface receptors, adenosine modulates a variety of physiological functions, including inducing analgesia, vasodilation, suppressing heart rate and myocardial contractility, inhibiting platelet aggregation, stimulating gluconeogenesis, and inhibiting lipolysis.Research has shown that adenosine can activate adenylate cyclase, open potassium channels, reduce the flow through calcium channels, and inhibit or stimulate phosphoinositide turnover through receptor-mediated mechanisms (Muller CE and Stein B., Current Pharmaceutical Design, 2: 501, 1996;Muller CE, Exp. Opin. Ther. Patents, 7(5): 419, 1997).

[0004] Adenosine receptors belong to the superfamily of G protein-coupled receptors (GPCRs). Four major subtypes of adenosine receptors have been characterized pharmacologically, structurally, and functionally (Fredholm et al., Pharm. Rev., 46: 143-156, 1994): A1, A2, A3, A4, A5, A6, A7, A8, A9, A10, A11, A12, A13, A14, A15, A16, A17, A18, A19, A19, A19, A19, A19, A19, A20, A21, A22, A23, A24, A25, A26, A27, A28, A29, A30, A31, A32, A33, A34, A35, A36, A37, A38, A39, A40 2A , A 2B Although the same adenosine receptor can be coupled to different G proteins, the adenosine A1 and A3 receptors are mostly G i and inhibitory G proteins, termed G0, which inhibit adenylate cyclase and downregulate cellular cAMP levels. In contrast, adenosine A 2A and A 2B The receptor is G S It couples to a stimulatory G protein called α-terminal β-terminal agonist, which activates adenylate cyclase and increases intracellular levels of cAMP (Linden J., Annu. Rev. Pharmacol. Toxicol., 41: 775-87 2001).

[0005] According to the present invention, "adenosine-receptor-selective ligands" are substances that selectively bind to one or more subtypes of adenosine receptors and thus either mimic the action of adenosine (adenosine agonists) or block its action (adenosine antagonists). According to their receptor selectivity, adenosine-receptor-selective ligands can be divided into different categories, for example ligands that selectively bind to A1 or A2 receptors, in the latter case also for example A1 or A2 receptors. 2A Or A 2B The adenosine receptor ligands can be classified as those that selectively bind to the receptor. There can also be adenosine receptor ligands that selectively bind to multiple subtypes of adenosine receptors, such as those that selectively bind to A1 and A2 but not A3 receptors. The above-mentioned receptor selectivity can be determined by the effect of the substance on cell lines that express the receptor subtype in question after stable transfection with the corresponding cDNA (Olah, ME et al., J. Biol. Chem., 267: 10764-10770, 1992). The effect of the substance on such cell lines can be monitored by biochemical measurement of the intracellular messenger cAMP (Klotz, KN et al., Naunyn Schmiedebergs Arch. Pharmacol. 357: 1-9, 1998).

[0006] The A1 receptor system is known to involve activation of phospholipase C and modulation of both potassium and calcium ion channels. The A3 subtype, in addition to its association with adenylate cyclase, also stimulates phospholipase C and thus activates calcium ion channels.

[0007] The A1 receptor (326-328 amino acids) has been cloned from various species (canine, human, rat, dog, chicken, cow, and guinea pig) with 90-95% sequence identity among mammalian species.2A The receptor (amino acids 409-412) has been cloned from dog, rat, human, guinea pig, and mouse. 2B The receptor (332 amino acids) is a 332-amino acid fragment of human and mouse A1 and A2. 2A Human A, which shares 45% homology with the receptor 2B The A3 receptor (317-320 amino acids) was cloned from human, rat, dog, rabbit, and sheep.

[0008] A1 and A 2A It has been proposed that the receptor subtypes play complementary roles in the regulation of adenosine's energy supply. Adenosine, a metabolic product of ATP, diffuses out of the cell and acts locally to activate adenosine receptors, thereby decreasing oxygen demand (A1 and A3) or increasing oxygen supply (A4). 2A ), thus restoring the balance of energy supply / demand in the tissue. The action of both subtypes is to increase the amount of oxygen available to the tissue and to protect the cells against damage caused by short-term oxygen imbalance. One important function of endogenous adenosine is to prevent damage during traumas such as hypoxia, ischemia, hypotension, and seizure activity. It is also known that binding of adenosine receptor agonists to mast cells expressing rat A3 receptors results in increased inositol triphosphate and intracellular calcium concentrations, which enhance antigen-induced secretion of inflammatory mediators. Thus, A3 receptors play a role in mediating asthma attacks and other allergic responses.

[0009] These adenosine receptors are encoded by distinct genes and are classified according to their affinity for adenosine analogues and methylxanthine antagonists (Klinger et al., Cell Signal., 14 (2): 99-108, 2002).

[0010] Regarding the role of adenosine on the nervous system, the first observations were made on the effects of caffeine, the most widely used of all psychotropic drugs. In fact, caffeine is a well-known adenosine receptor antagonist that can enhance the wakefulness and learning capacity of mammals. Adenosine A 2A Receptor pathways are involved in these effects (Fredholm et al., Pharmacol. Rev., 51 (1): 83-133, 1999;Huang et al., Nat Neurosci., 8 (7): 858-9, 2005), with adenosine A 2A The effects of caffeine on receptor signaling pathways are highly specific and potent adenosine A 2A This prompted research into antagonists.

[0011] In mammals, adenosine A 2A The receptor has a restricted distribution in the brain and is found in the striatum, olfactory tubercle, and nucleus accumbens (Dixon et al., Br. J. Pharmacol., 118 (6): 1461-8, 1996). High and intermediate levels of expression can be observed in immune cells, heart, lung, and blood vessels. In the peripheral system, G3 binds adenosine A 2A It appears that the major G protein that binds to the receptor, but in the striatum, the striatal adenosine A 2A The receptors transmit their effects through the G oif It has been shown that this is mediated through the activation of a G protein called G oif is similar to G3 and also couples to adenylate cyclase.

[0012] To date, genetically modified mouse studies and pharmacological analyses have demonstrated that A 2ANot only are the receptors promising therapeutic targets for the treatment of disorders and diseases of the central nervous system (CNS), such as Parkinson's disease, Huntington's disease, attention deficit hyperactivity disorder (ADHD), stroke (ischemic brain injury), and Alzheimer's disease (Fredholm et al., Annu. Rev. Pharmacol. Toxicol., 45: 385-412, 2005;Higgins et al.; Behav. Brain Res. 185: 32-42, 2007;DaIl' Igna et al., Exp. Neurol., 203 (1): 241-5, 2007;Arendash et al., Neuroscience, 142 (4): 941-52, 2006;Trends in Neurosci., 29 (11), 647-654, 2006;Expert Opinion Ther. Patents, 17, 979-991, 2007;Exp. Neurol., 184 (1), 285-284, 2003;Prog. Brain Res, 183, 183-208, 2010;J. Alzheimer Dis., Suppl 1, 1 17-126, 2010;J. Neurosci., 29 (47), 14741-14751, 2009;Neuroscience, 166 (2), 590-603, 2010;J. Pharmacol. Exp. Ther., 330 (1), 294-303, 2009;Frontiers Biosci., 13, 2614-2632, 2008), suggesting that it may also be a promising therapeutic target for the treatment of a variety of psychiatric disorders that are organic in origin (Weiss et al., Neurology, 61 (11 Suppl 6): 88-93, 2003).

[0013] Adenosine A 2A By using receptor knockout mice, adenosine A 2AInactivation of the receptor has been shown to protect against ischemia-induced neuronal death (Chen et al., J. Neurosci., 19 (21): 9192-200, 1999 and Monopoli et al., Neuroreport, 9 (17): 3955-9, 1998) and against the mitochondrial toxin 3-NP (Blum et al., J. Neurosci., 23 (12): 5361-9, 2003). These results support the finding that adenosine A 2A This study provides a rationale for treating ischemia and Huntington's disease with adenosine A antagonists. 2A Blockade of the receptor also has antidepressant effects (El Yacoubi et al., Neuropharmacology, 40 (3): 424-32, 2001). Finally, this blockade prevents memory dysfunction (Cunha et al., Exp. Neurol., 210 (2): 776-81, 2008;Takahashi et al., Front. Biosci., 13: 2614-32, 2008), which may be a promising therapeutic route for the treatment and / or prevention of Alzheimer's disease.

[0014] A 2A For reviews on adenosine receptors, see, e.g., Moreau et al. (Brain Res. Reviews 31: 65-82, 1999) and Svenningsson et al. (Progress in Neurobiology 59: 355-396, 1999).

[0015] To date, several adenosine A 2AReceptor antagonists have shown promise for treating Parkinson's disease. For example, KW-6002 (Istradefylline) has completed phase III clinical trials in the USA after studies that demonstrated its effectiveness in alleviating symptoms of the disease (Bara-Jimenez et al., Neurology, 61 (3): 293-6, 2003 and Hauser et al., Neurology, 61 (3): 297-303, 2003). SCH420814 (Preladenant), now in phase II clinical trials in the USA, produces improvements in motor function in animal models of Parkinson's disease (Neustadt et al., Bioorg. Med. Chem. Lett., 17 (5): 1376-80, 2001) and in human patients (Hunter J. C, poster Boston 2006 - http: / / www.a2apd.org / Speaker abstracts / Hunter.pdf).

[0016] A for treating neurodegenerative diseases 2A In addition to the valuable utility of receptor antagonists, these compounds are being considered for complementary symptomatic indications. 2A Evidence suggests that receptor activation may contribute to the pathophysiology of a wide range of neuropsychiatric and functional disorders, such as depression, excessive daytime sleepiness, restless legs syndrome, attention deficit hyperactivity disorder, and cognitive fatigue (Neurology, 61 (Suppl 6), 82-87, 2003; Behav. Pharmacol., 20 (2), 134-145, 2009; CNS Drug Discov., 2 (1), 1-21, 2007).

[0017] Several authors have suggested that A 2AOther studies have also suggested the use of antagonists in wound healing or atrial fibrillation. 2A This suggests the involvement of adenosine receptors (Am. J. Path., 6, 1774- 1778, 2007;Arthritis & Rheumatism, 54 (8), 2632-2642, 2006).

[0018] A powerful adenosine A previously discovered by pharmaceutical companies 2A Some of the antagonists have progressed into clinical trials and shown positive results, demonstrating that this compound class holds promise for treating not only neurodegenerative disorders such as Parkinson's, Huntington's, or Alzheimer's disease, but also other CNS-related diseases such as depression, restless legs syndrome, sleep disorders, and anxiety disorders (Clin. Neuropharmacol., 33, 55-60, 2010:J. Neurosci., 30 (48), 2010, 16284-16292;Parkinson Relat. Disord., 16 (6), 423-426, 2010;Expert Opinion Ther. Patents, 20(8), 987-1005, 2010;Current Opinion in Drug Discovery & Development, 13 (4), 466-480, 2010 and references cited therein;Mov. Disorders, 25 (2), S305, 2010).

[0019] Known A 2AThe inhibitors are Istradefylline (KW-6002), Preladenant (SCH420814), SCH58261, CGS15943, Tozadenant, Vipadenant (V-2006), V-81444, CPI-444, HTL-1071, PBF-509, Medi-9447, PNQ-370, ZM-241385, ASO-5854, ST-1535, ST-4206, DT1133, and DT-0926, which in most cases were developed for Parkinson's disease.

[0020] Adenosine A 2B Receptors have been cloned from rat hypothalamus (Rivkees and Reppert, 1992), human hippocampus (Pierce et al., 1992), and mouse mast cells (Marquardt et al., 1994) employing standard polymerase chain reaction techniques with degenerate oligonucleotide primers designed to recognize conserved regions of most G protein-coupled receptors. 2B The receptor is rat and mouse A 2B The human A1 and A2 receptors share 86–87% amino acid sequence identity (Rivkees and Reppert, 1992; Pierce et al., 1992; Marquardt et al., 1994). 2A It shares 45% amino acid sequence identity with the rat and mouse A receptors. As expected for closely related species, 2B The receptors share 96% amino acid sequence homology. In comparison, the overall amino acid identity between A1 receptors from various species is 87% (Palmer and Stiles, 1995). 2A The receptors share 90% homology between species (Ongini and Fredholm, 1996), but the most differences are in the second extracellular loop and the long C-terminal domain (Palmer and Stiles, 1995). The lowest identity between species (72%) is found in the A3 receptor sequence (Palmer and Stiles, 1995).

[0021] The adenosine analog NECA remains the most potent A 2B Although it is an agonist (Bruns, 1981; Feoktistov and Biaggioni, 1993, 1997; Brackett and Daly, 1994), it has a concentration that produces a half-maximal effect (EC 50 ) is approximately 2 μM. However, it is nonselective and has a higher affinity to low nanomolar (A1 and A2). 2A ) or EC in the high nanomolar (A3) range 50 , which activates other adenosine receptors. 2B Characterization of a receptor often depends on the lack of efficacy of compounds that are potent and selective agonists of other receptor types. 2B Receptors are characterized by the exclusion method, i.e., by the lack of effectiveness of agonists specific for other receptors. 2A The selective agonist CGS-21680 (Webb et al., 1992) e.g. 2A and A 2B These findings are useful in differentiating between adenosine receptors and those that activate the endothelial cells (Hide et al., 1992; Chern et al., 1993; Feoktistov and Biaggioni, 1995; van der Ploeg et al., 1996).

[0022] Both receptors are positively coupled to adenylyl cyclase and are activated by the nonselective agonist NECA. 2B receptor, but A 2A It is as potent as NECA in activating the receptor, with the EC 50are in the low nanomolar range (Jarvis et al., 1989; Nakane and Chiba, 1990; Webb et al., 1992; Hide et al., 1992; Feoktistov and Biaggioni, 1993; Alexander et al., 1996). 2B The receptor also responds to the A1 selective agonist R-PIA (Feoktistov and Biaggioni, 1993; Brackett and Daly, 1994) and the A3 selective agonist N 6 It also has very low affinity for 3-(3-iodobenzyl)-N-methyl-5'-carbamoyl adenosine (IB-MECA) (Feoktistov and Biaggioni, 1997). The agonist profile NECA>R-PIA=IB-MECA>CGS-21680 was observed in A. 2B The EC ratios of NECA and other agonists for mediated cAMP accumulation were determined in human erythroleukemia (HEL) cells. 50 The difference in A is approximately two orders of magnitude. Thus, the response elicited by NECA at concentrations in the low micromolar range (1-10 μM) is 2B Characteristic for the receptor, no response was elicited by R-PIA, IB-MECA, or CGS-21680.

[0023] A 2B Although receptors generally have a lower affinity for agonists compared to other receptor subtypes, this is not true for antagonists. 2B Although the structure activity relationship of adenosine antagonists at the receptor has not been fully characterized, at least some xanthines have 2BAntagonists of the receptor subtype that are as potent or more potent than antagonists of other subtypes, including DPSPX (1,3-dipropyl-8-sulfophenylxanthine), DPCPX (1,3-dipropyl-8-cyclopentylxanthine), DPX (1,3 diethylphenylxanthine), the antiasthmatic drug enprofylline (3-n-propylxanthine), and the non-xanthine compound 2,4-dioxobenzopteridine (alloxazine), have affinities in the mid-to-high nM range.

[0024] Other known A 2B The inhibitors are ATL801, PSB-605, PSB-1115, ISAM-140, GS6201, MRS1706, and MRS1754. It is disclosed herein that adenosine receptors play a non-redundant role in down-regulating inflammation in vivo by acting as a physiological "STOP" that can limit immune responses, thereby protecting normal tissues from excessive immune damage during the development of various diseases.

[0025] A 2A Receptor antagonists provide long-term enhancement of immune responses by reducing T cell-mediated tolerance to antigenic stimulation, enhancing the induction of memory T cells, and enhancing the effectiveness of passive antibody administration for the treatment of cancer and infectious diseases, while A 2A Receptor agonists provide long-term reduction in immune responses by enhancing T cell-mediated tolerance to antigenic stimulation, inter alia, reducing the use of immunosuppressive drugs in certain conditions.

[0026] Immune modulation is a critical aspect of the treatment of many diseases and disorders. T cells in particular play a crucial role in fighting infection and have the ability to recognize and destroy cancer cells. Boosting T cell-mediated responses is an essential component of boosting responses to therapeutic agents. However, it is crucial in immune modulation that any boost in immune response must be counterbalanced with the need to prevent autoimmunity and chronic inflammation. Chronic inflammation and self-recognition by T cells are the main causes of the development of systemic disorders such as rheumatoid arthritis, multiple sclerosis, and systemic lupus erythematosus. Furthermore, long-term immunosuppression is required to prevent rejection of transplanted organs or grafts.

[0027] Tumor-induced immune suppression is a major hurdle to the efficacy of current cancer therapies. Recent successes with immune checkpoint blockade inhibitors, such as anti-CTLA-4 and anti-PD-1 / PDL1, have revolutionized cancer treatment due to their remarkable clinical efficacy against a broader range of cancers.

[0028] Adenosine is one of the new promising immunosuppressive targets revealed in preclinical studies. This metabolite is produced by the ectoenzyme-CD73 expressed on host suppressor cells and tumor cells. Increased expression of CD73 correlates with poor prognosis in patients with several cancers, including colorectal cancer (Liu et al, J. Surgical Oncol, 2012), gastric cancer (Lu et al., World J. Gastroenterol., 2013), and gallbladder cancer (Xiong et al., Cell and Tissue Res., 2014). Preclinical studies have demonstrated that the protumor effect of CD73 may be driven (at least in part) by adenosine-mediated immunosuppression. As disclosed above, adenosine binds to four known receptors, A1, A2, A3, A4, A5, A6, A7, A8, A9, A10, A11, A12, A13, A14, A15, A16, A17, A18, A19, A20, A21, A22, A23, A24, A25, A26, A27, A28, A29, A30, A31, A32, A33, A34, A35, A36, A37, A38, A39, A39, A40, A41, A42, A43, A44, A45, A46, A47, A48, A49, A49, A51, A52, A53, A54, A55, A56, A57, A58, A59, A61, A62, A63, A64, A65, A66, A67, A68, A69, A71, A72, A73, A74, A75, A75, A76, A77, A78, A79 2A , A 2B A, which is known to bind to A3 and suppress the effector functions of many immune cells. 2A and A 2BThe receptor is activated, i.e., A 2A and A 2B The receptors induce adenylate cyclase-dependent accumulation of cAMP, leading to immunosuppression. Selectivity for A1 and A3 needs to be achieved, as antagonizing A1 and A3 would counter the desired effect, and A1 and A3 agonists serve as potential cardioprotective agents (Antonioli et al., Nat. rev. Cancer, 2013, Thiel et al., Microbes and Infection, 2003).

[0029] In the tumor microenvironment, A 2A and A 2B Activation of both receptors has been demonstrated to suppress antitumor immunity and increase the spread of CD73 tumors. In addition, activation of both receptors with small molecule antagonists has been shown to suppress antitumor immunity and increase the spread of CD73 tumors. 2A Or A 2B Blockade of either of these may reduce tumor metastasis. 2A It was found that blocking the receptor could overcome tumor evasion mechanisms involving both anergy and regulatory T cell induction caused by tumor cells, resulting in long-term sensitivity of tumors to treatment. Ohta et al. found that A1c12+, A2c12+, and A3c12+ were significantly more potent than A1c12+, compared with no rejection in normal mice. 2A demonstrated that approximately 60% of established CL8-1 melanoma tumors were rejected in receptor-deficient mice (Ohta, et al.; PNAS 103 (35): 13132-7, 2006). The researchers also consensually administered A. 2A They also demonstrated improved inhibition of tumor growth, destruction of metastatic cancer, and prevention of neovascularization by antitumor T cells following treatment with the receptor antagonist.

[0030] Tumors have been shown to evade immune destruction by preventing T cell activation through inhibition of B7-CD28 and TNF family costimulatory factors, as well as by recruiting regulatory T cells that inhibit antitumor T cell responses (Wang, Cancer. Semin. Cancer. Biol. 16: 73-79, 2006; Greenwald, et al., Ann. Rev. Immunol. 23: 515-48, 2005; Watts, Ann. Rev. Immunol. 23: 23-68, 2005; Sadum et al., Clin. Cane. Res. 13 (13): 4016-4025, 2007). 2A Where expression of the receptor is increased in lymphocytes and subsequently activated, treatments that generate lymphocyte effector responses, such as anti-CTLA-4 and anti-PD-1, also have been shown to be effective in treating A. 2A The effects of A-mediated immunosuppression may also be increased. 2A Or double A 2A / 2B Immune checkpoint blockade in combination with antagonists increases the magnitude of the immune response against tumors and metastases. 2A Combining blockade with anti-PD-1 treatment enhanced IFN-γ production by T cells in coculture with MC38 tumor cells, improved mouse survival in the 4T1 mammary tumor model, and improved survival in mice treated with AT-3ova dim CD73 + In tumors, it reduces tumor growth (Beavis et al., Cancer Immunol. Res., 2015; Mittal et al., Cancer Res., 2014).

[0031] Furthermore, preclinical studies have shown that A 2B Inhibition of Aβ1 leads to reduced tumor growth and extended mouse survival in Lewis lung carcinoma, MB49 bladder carcinoma, and Ortho4T1 mammary carcinoma models (Ryzhov et al., 2009; Cekic et al., 2012), and 2BIt was demonstrated that combining blockade with anti-PD-1 therapy reduced lung metastasis of B16-F10 melanoma tumors and improved mouse survival in a 4T1 breast cancer model.

[0032] WO 03 / 050241 describes a method for increasing the immune response to an antigen, which method increases the effectiveness of a vaccine or increases the immune response to tumor antigens or immune cell-mediated tumor destruction by administering an agent that inhibits extracellular adenosine or inhibits adenosine receptors.

[0033] WO 2004 / 089942, WO 2005 / 000842, and WO 2006 / 008041 disclose Azotobacter 1,2-dihydropyridine, 1,2-trimethylphenyl ether, and 1,2-dihydropyridine for the treatment of Parkinson's disease. 2A WO 2004 / 092171 and WO 2005 / 028484 disclose similar thiazolopyridine and pyrazolopyrimidine derivatives, as well as A inhibitors for the treatment of Parkinson's disease. 2A However, these compounds do not show significant A 2B It does not show inhibitory activity and shows good pharmacokinetic properties only in rat Parkinson's disease animal model, not in mouse cancer animal model.Furthermore, it does not show that the compound can prevent immunosuppression, and thus support the inhibition of tumor growth induced by antitumor T cells, the reduction or destruction of metastatic cancer, and the prevention of neovascularization.

[0034] Thus, there remains a need for therapies that provide long-term enhancement of immune responses to specific antigens, particularly for the treatment and prevention of hyperproliferative and infectious diseases and disorders, and therefore, it was an object of the present invention to provide a method of treatment that allows for simplified treatment protocols and enhances the immune response to an antigen. It is an object of the present invention to provide improved methods of preventing or treating hyperproliferative and infectious diseases and disorders in a host, and in particular to provide an A antibody that is effective in treating and preventing such diseases. 2AOr double A 2A / 2B It was a particular object of the present invention to provide antagonists. Summary of the Invention

[0035] SUMMARY OF THE PRESENT APPLICATION Surprisingly, the compounds according to the invention have the structure A 2A Adenosine receptor or A 2A and A 2B It has been found that the compounds of the present invention are highly effective inhibitors of both adenosine receptors, while at the same time having high selectivity for the A1 and A3 adenosine receptors, and thus may be used for the treatment of hyperproliferative diseases and disorders, such as cancer and infectious diseases and disorders.

[0036] Specifically, the known adenosine A 2A In contrast to the receptor antagonists tozadenant and similar benzothiazole derivatives, the compounds of the present invention surprisingly have the preferred A activity for the treatment and / or prevention of hyperproliferative and infectious diseases and disorders as disclosed above. 2A / A 2B The compounds of the present invention show dual activity or have at least a high A 2A The inhibitory activity is exhibited together with other surprising advantages disclosed herein that translate into high efficacy in the treatment and / or prevention of hyperproliferative and infectious diseases and disorders.

[0037] In addition, the known adenosine A 2A Compared to the receptor antagonist tozadenant and similar benzothiazole derivatives, the compounds of the present invention surprisingly show better pharmacokinetic properties in mice as a cancer-relevant animal model, said properties being favorable for the treatment and / or prevention of hyperproliferative and infectious diseases and disorders as disclosed above.

[0038] Furthermore, as discussed above, adenosine in the tumor microenvironment is 2AIt can inhibit T cell activity by signaling through the receptor and suppress cytokine secretion by T cells. 2A Specific agonists, like adenosine, inhibit T cell cytokine secretion in vitro and in vivo. 2A Antagonist or A 2A / A 2B Dual antagonists can rescue T cells from this inhibition. 2A In contrast to tozadenant, which is a receptor antagonist, the compounds of the present invention are able to rescue T cells from inhibition and are more potent than adenosine or A receptor antagonists such as CGS-2168, CGS-21680 or NECA. 2A It shows that the compounds of the present invention can prevent the suppression of cytokine secretion induced by specific agonists, which are favorable for the treatment and / or prevention of hyperproliferative and infectious diseases and disorders as disclosed above.Therefore, the compounds of the present invention can surprisingly prevent immunosuppression, and thus support the inhibition of tumor growth, the reduction or destruction of metastatic cancer, and the prevention of neovascularization induced by antitumor T cells.

[0039] The present invention relates to compounds selected from the group consisting of the following, and their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio: [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11] [Table 1-12]

[0040] The compounds of the present invention have the following general formula I: [ka] During the ceremony R 1 teeth, Linear or branched alkyl with 1 to 10 C atoms ,these are , unsubstituted or R 5 and 、1 ~4 C atoms are independently O, S, SO, SO2, NH, NCH3, -OCO-, -NHCONH-, -NHCO-, -NR 6 SO2R 7and / or -CH=CH- groups; and / or in addition, 1 to 10 H atoms may be replaced by F and / or Cl. may be replaced by ,or Monocyclic or bicyclic cyclic alkyl groups having 3 to 7 carbon atoms These are , unsubstituted or R 5 and 、1 ~4 C atoms are independently O, S, SO, SO2, NH, NCH3, -OCO-, -NHCONH-, -NHCO-, -NR 6 SO2R 7 and / or -CH=CH- groups; and / or in addition, 1 to 10 H atoms may be replaced by F and / or Cl. may be replaced by ,or 、 Monocyclic or bicyclic heteroaryl, heterocyclyl, aryl, or cyclic alkylaryl, which are unsubstituted or have the formula: R 5 (mono-, di-, or tri-substituted by and

[0041] R 2 teeth, Linear or branched alkyl groups containing 1 to 10 carbon atoms These are , unsubstituted or R 5 and 、1 ~4 C atoms are independently O, S, SO, SO2, NH, NCH3, -OCO-, -NHCONH-, -NHCO-, -NR 6 SO2R 7and / or -CH=CH- groups; and / or in addition, 1 to 10 H atoms may be replaced by F and / or Cl. may be replaced by ,or Cyclic alkyl groups containing 3 to 7 carbon atoms These are , unsubstituted or R 5 and 、1 ~4 C atoms are independently O, S, SO, SO2, NH, NCH3, -OCO-, -NHCONH-, -NHCO-, -NR 6 SO2R 7 and / or -CH=CH- groups; and / or in addition, 1 to 11 H atoms may be replaced by F and / or Cl. may be replaced by ,or 、 Monocyclic or bicyclic heteroaryl, heterocyclyl, aryl, or cyclic alkylaryl, which are unsubstituted or have the formula: R 5 (mono-, di-, or tri-substituted by and ,

[0042] R 3 is a linear or branched alkyl group having 1 to 6 carbon atoms 、 or 、 O-alkyl, or Cyclic alkyl having 3 to 6 C atoms, these are , unsubstituted or mono-, di- or trisubstituted by H, =S, =NH, =O, OH, cyclic alkyl having 3 to 6 C atoms, COOH, Hal, NH2, SO2CH3, SO2NH2, CN, CONH2, NHCOCH3, NHCONH2 or NO2, R 4is H, D, linear or branched alkyl having 1 to 6 C atoms, CN or Hal,

[0043] R 5 teeth, H, R 6 , =S, =NR 6 , =O, OH, COOH, Hal, NH2, SO2CH3, SO2NH2, CN, CONH2, NHCOCH3, NHCONH2, NO2, or linear or branched alkyl groups having 1 to 10 C atoms. These are , unsubstituted or R 6 and 、1 ~4 C atoms are independently O, S, SO, SO2, NH, NCH3, -OCO-, -NHCONH-, -NHCO-, -NR 6 SO2R 7 and / or -CH=CH- groups; and / or in addition, 1 to 10 H atoms may be replaced by F and / or Cl. may be replaced by ,or Monocyclic or bicyclic cyclic alkyl having 3 to 7 carbon atoms ,these are , unsubstituted or R 6 and 、1 ~4 C atoms are independently O, S, SO, SO2, NH, NCH3, -OCO-, -NHCONH-, -NHCO-, -NR 6 SO2R 7 and / or -CH=CH- groups; and / or in addition, 1 to 10 H atoms may be replaced by F and / or Cl. may be replaced by ,or Monocyclic or bicyclic heteroaryl, heterocyclyl, aryl, or cyclic alkylaryl, which are unsubstituted or have the formula: R 6 (mono-, di-, or tri-substituted by and ,

[0044] R 6 , R 7 are selected independently from the group consisting of H, =S, =NH, =O, OH, COOH, Hal, NH2, SO2CH3, SO2NH2, CN, CONH2, NHCOCH3, NHCONH2, NO2 and linear or branched alkyl having 1 to 10 C atoms, in which 1 to 4 C atoms may be replaced independently from one another by O, S, SO, SO2, NH, NCH3, -OCO-, -NHCONH-, -NHCO-, -COO-, -CONH-, -NCH3CO-, -CONCH3-, -C≡C- and / or -CH=CH- groups and / or in addition 1 to 10 H atoms may be replaced by F and / or Cl, Hal is F, Cl, Br, or I; D is deuterium; and its physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in any proportion.

[0045] Furthermore, the following abbreviations have the following meanings: Boc tert-butoxycarbonyl CBZ Benzyloxycarbonyl DNP 2,4-dinitrophenyl FMOC 9-Fluorenylmethoxycarbonyl imi-DNP 2,4-Dinitrophenyl at the 1-position of the imidazole ring OMe methyl ester POA Phenoxyacetyl DCCI Dicyclohexylcarbodiimide HOBt 1-Hydroxybenzotriazole

[0046] The present invention further relates to pharmaceutical preparations comprising a compound according to the invention and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, as well as mixtures thereof in any ratio.

[0047] The invention also relates to pharmaceutical preparations of this type according to the invention which contain further excipients and / or adjuvants. Additionally, the present invention relates to the above pharmaceutical preparations according to the present invention which comprise at least one further pharma- ceutical active compound.

[0048] Pharmaceutically or physiologically acceptable derivatives are understood to mean, for example, salts of the compounds according to the invention, but also so-called prodrug compounds. Prodrug compounds are understood to mean derivatives of the compounds of the invention that are modified, for example, with alkyl or acyl groups (see also amino-protecting groups and hydroxyl-protecting groups below), sugars, or oligopeptides, and are rapidly cleaved or released in vivo to form effective molecules. They also include biodegradable polymer derivatives of the compounds according to the invention, as described, for example, in Int. J. Pharm. 115 (1995), 61-67.

[0049] The compound of the present invention can be used in its final non-salt form. On the other hand, the present invention also covers the use of pepstatin in the form of its pharmaceutically acceptable salt, which can be derived from various organic and inorganic bases by procedures known in the art. The pharmaceutically acceptable salt form of pepstatin is mostly prepared by conventional methods. When the compound of the present invention contains a carboxyl group, one of its suitable salts can be formed by reacting the compound of the present invention with a suitable base to give the corresponding base addition salt. Such bases are, for example, alkali metal hydroxides including potassium hydroxide, sodium hydroxide, and lithium hydroxide; alkaline earth metal hydroxides such as barium hydroxide and calcium hydroxide; alkali metal alkoxides such as potassium ethoxide and sodium propoxide; and various organic bases such as piperidine, diethanolamine, and N-methylglutamine. The aluminum salt of pepstatin is also included.

[0050] Additionally, base salts of the compounds of the present invention include, but are not intended to be limiting, aluminum, ammonium, calcium, copper, iron(III), iron(II), lithium, magnesium, manganese(III), manganese(II), potassium, sodium, and zinc salts.

[0051] Preferred among the above salts are ammonium; the alkali metal salts sodium and potassium, and the alkaline earth metal salts calcium and magnesium. Salts of the compounds of the present invention derived from pharma- ceutically acceptable organic non-toxic bases are suitable for use with primary, secondary, and tertiary amines, as well as substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, chloroprocaine, choline, N,N'-dibenzylethylenediamine (benzathine), dicyclohexylamine, diethanolamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmonoethanol, N-methyl ... The term "amino acids" includes, but is not intended to represent a limitation, salts of: morphine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lidocaine, lysine, meglumine, N-methyl-D-glucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethanolamine, triethylamine, trimethylamine, tripropylamine, and tris(hydroxymethyl)methylamine (tromethamine).

[0052] As mentioned, pharma- ceutically acceptable base addition salts of pepstatin are formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Preferred metals are sodium, potassium, magnesium, and calcium. Preferred organic amines are N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methyl-D-glucamine, and procaine.

[0053] The base addition salts of the compounds of the present invention are prepared by contacting the free acid form with a sufficient amount of the desired base to cause the formation of the salt in a conventional manner. The free acid can be regenerated by contacting the salt form with an acid in a conventional manner and isolating the free acid. The free acid forms differ in some respects from their corresponding salt forms with respect to certain physical properties, such as solubility in polar solvents; however, for purposes of the present invention, the salts otherwise correspond to their respective free acid forms.

[0054] In view of the above, it is clear that the term "pharmaceutical acceptable salt" in this context is to be understood as meaning the active compound comprising the compound of the present invention in one of its salt forms, especially when this salt form gives the active compound improved pharmacokinetic properties compared to the free form of the active compound or any other salt form of the active compound used before.The pharmaceutical acceptable salt form of the active compound can also give the active compound desirable pharmacokinetic properties that the active compound did not have before for the first time, and can even have a positive effect on the pharmacodynamics of the active compound with regard to its therapeutic effectiveness in the body.

[0055] Solvates of the compounds of the invention are understood to mean adducts of the inert solvent molecule pepstatin, said adducts being formed due to their mutual attractive force. Solvates are, for example, hydrates, such as mono- or dihydrates, or alcoholates, i.e. addition compounds with alcohols, such as, for example, methanol or ethanol.

[0056] All physiologically acceptable salts, derivatives, solvates and stereoisomers of these compounds, as well as mixtures thereof in any proportion, also fall within the scope of the invention. The compounds of the present invention may contain one or more chiral centers, as all stereoisomers, enantiomers, diastereomers, etc. of the compounds of the present invention are also claimed in the present invention.

[0057] The invention also relates to the optically active forms (stereoisomers), the enantiomers, the racemates, the diastereomers and the hydrates and solvates of these compounds.

[0058] The compounds of the invention according to the present invention may be chiral due to their molecular structure and may therefore exist in various enantiomeric forms.They may therefore be racemic or optically active.The pharmaceutical effectiveness of the racemic or stereoisomers of the compounds according to the present invention may differ, and it may be desirable to use enantiomers.In these cases, not only the final products but even intermediates may be separated into enantiomeric compounds by chemical or physical means known to those skilled in the art or already employed in the synthesis itself.

[0059] Pharmaceutically or physiologically acceptable derivatives are understood to mean, for example, salts of the compounds according to the invention, but also so-called prodrug compounds.Prodrug compounds are understood to mean the compounds of the invention that are modified, for example, with alkyl or acyl groups (see also amino-protecting groups and hydroxyl-protecting groups below), sugars, or oligopeptides, and are rapidly cleaved or released in vivo to form the effective compounds according to the invention.They also include biodegradable polymer derivatives of the compounds according to the invention, as described, for example, in Int. J. Pharm. 115 (1995), 61-67.

[0060] Suitable acid addition salts are all physiologically or pharmacologically acceptable inorganic or organic salts of acids, such as halides, especially the hydrochlorides or hydrobromides, lactates, sulfates, citrates, tartrates, maleates, fumarates, oxalates, acetates, phosphates, methylsulfonates or p-toluenesulfonates.

[0061] Very particular preference is given to the hydrochlorides, trifluoroacetates or bistrifluoroacetates of the compounds according to the invention.

[0062] Solvates of the compounds of the invention are taken to mean adducts of inert solvent molecules onto the compounds of the invention, said adducts forming due to their mutual attractive force. Solvates are, for example, hydrates, such as mono- or dihydrates, or alcoholates, i.e. addition compounds with alcohols, such as, for example, methanol or ethanol.

[0063] It is further intended that the compounds of the present invention include their isotopically labeled forms. Isotopically labeled forms of the compounds of the present invention are identical to the compounds except for the fact that one or more atoms of the compounds are replaced by an atom or atoms having an atomic mass or mass number different from the atomic mass or mass number of the normally occurring atom. Examples of isotopes that are readily commercially available and can be incorporated into the compounds of the present invention by well-known methods include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, for example, respectively: 2 H, 3 H, 13 C. 14 C. 15 N, 18 O. 17 O. 31 P, 32 P, 35 S, 18 F and 36 Compounds of the invention, prodrugs thereof, or any pharma- ceutically acceptable salts thereof that contain one or more of the aforementioned isotopes and / or other isotopes of other atoms are intended to be part of this invention.

[0064] Isotopically labeled compounds of the invention can be used in a number of beneficial ways. For example, isotopically labeled compounds of the invention can be used in 3 H or 14 Radioactive isotopes such as C have been incorporated therein and are suitable for drug and / or substrate tissue distribution assays. 3 H) and carbon-14 ( 14C) is particularly preferred due to its easy preparation and excellent detectability. Heavier isotopes, such as deuterium ( 2 Incorporation of H) into the compounds of the present invention has therapeutic advantages due to the higher metabolic stability of the isotopically labeled compounds. Higher metabolic stability translates directly into increased half-life in vivo or smaller dosages, which under most circumstances represent preferred aspects of the present invention. Isotopically labeled compounds of the present invention can be generally prepared by carrying out the procedures disclosed in the synthetic schemes and related descriptions in the Examples section and Preparation section of the text, substituting readily available isotopically labeled reactants for non-isotopically labeled reactants.

[0065] deuterium( 2 H) may also be incorporated into the compounds of the invention for the purpose of manipulating the oxidative metabolism of the compounds through the primary kinetic isotope effect. The primary kinetic isotope effect is the change in the rate of a chemical reaction due to the exchange of an isotopic nuclei, which in turn is caused by the change in the ground energy required for covalent bond formation after this isotope exchange. The exchange of a heavier isotope usually results in a lowering of the ground energy of the chemical bond, thus causing a slowing down of the rate-limiting bond cleavage. If the bond cleavage occurs along the coordinate of a multi-product reaction in or near the saddle-point region, the product distribution ratio can be substantially altered. To illustrate: if deuterium is attached to a carbon atom at a non-exchangeable position, k M / k D A rate difference of 2-7 is typical. If this rate difference is successfully applied to a compound of the invention that is susceptible to oxidation, the in vivo profile of the compound may thereby be dramatically modified, resulting in improved pharmacokinetic properties.

[0066] When discovering and developing therapeutic agents, those skilled in the art attempt to optimize pharmacokinetic parameters while retaining the desired in vitro properties. It is reasonable to assume that many compounds with poor pharmacokinetic profiles are prone to oxidative metabolism. Currently available in vitro liver microsomal assays provide valuable information about the course of this type of oxidative metabolism, which in turn allows for the rational design of deuterated compounds of the invention with improved stability through resistance to such oxidative metabolism. Significant improvements in the pharmacokinetic profile of the compounds of the invention are thereby obtained, including improvements in in vivo half-life (T / 2), concentration at maximum therapeutic effect (C max ), area under the dose-response curve (AUC), and F; and quantitatively in terms of reduced clearance, dose, and material cost.

[0067] The following is intended to illustrate the above: Compounds of the invention having multiple potential sites of oxidative metabolic attack (e.g., benzylic hydrogen atoms and hydrogen atoms attached to nitrogen atoms) are prepared as a series of analogs in which various combinations of hydrogen atoms are replaced by deuterium atoms (such that some, most or all of these hydrogen atoms are replaced by deuterium atoms). Determination of the half-life allows for a good and correct determination of the extent to which resistance to oxidative metabolism has been improved. In this way, it is determined that the half-life of the parent compound can be extended by up to 100% as a result of this type of deuterium-hydrogen exchange.

[0068] The replacement of hydrogen with deuterium in the compounds of the invention can also be used to achieve favorable modification of the metabolic spectrum of the starting compound to reduce or eliminate undesired toxic metabolites. For example, if a toxic metabolite arises through oxidative carbon-hydrogen (CH) bond cleavage, it can be reasonably assumed that a deuterated analog would greatly reduce or eliminate the production of the undesired metabolite, even if the specific oxidation is not the rate-limiting step. Further information on the state of the art regarding deuterium-hydrogen exchange is given, for example, in Hanzlik et al., J. Org. Chem. 55, 3992-3997, 1990; Reider et al., J. Org. Chem. 52, 3326-3334, 1987; Foster, Adv. Drug Res. 14, 1-40, 1985; Gillette et al., Biochemistry 33(10) 2927-2937, 1994; and Jarman et al., Carcinogenesis 16(4), 683-688, 1993.

[0069] The present invention also relates to mixtures of the compounds of the invention according to the invention, for example mixtures of two diastereomers, for example in a ratio of 1:1, 1:2, 1:3, 1:4, 1:5, 1:10, 1:100 or 1:1000. These are particularly preferred mixtures of two stereoisomeric compounds. However, mixtures of two or more compounds of the invention are also preferred.

[0070] In addition, the present invention provides a) a compound of formula II is reduced to give a compound of formula III, a compound of formula III is reacted with a compound of formula IV at elevated temperature to give a compound of formula V, a compound of formula V is converted to a compound of formula VI employing the use of a catalyst and a base, a compound of formula VI is converted to a compound of formula VII by bromination, a compound of formula VII is converted to a compound of formula VIII under essentially basic conditions, and a compound of formula VIII is reacted with a compound of formula IX under standard amidation or carbamide forming conditions to give a compound of formula I, [ka]

[0071] b) a compound of formula V is reacted with a compound of formula X under Suzuki-type reaction conditions to give a compound of formula VI, which is converted to a compound of formula VII by bromination, which is converted to a compound of formula VIII under essentially basic conditions, and which is reacted with a compound of formula IX under standard amidation or carbamide forming conditions to give a compound of formula I; [ka]

[0072] c) a compound of formula XII is iodized to give a compound of formula XIII, a compound of formula XIII is converted to a compound of formula XIV by treatment with a base and an electrophile, a compound of formula XIV is converted to a compound of formula XV by reduction, a compound of formula XV is reacted with a compound of formula IV at elevated temperature to give a compound of formula XVI, a compound of formula XVI is converted under catalytic conditions to a compound of formula XVII, a compound of formula XVII is converted to a compound of formula VIII under basic conditions, and a compound of formula VIII is reacted with a compound of formula IX under standard amidation or carbamide forming conditions to give a compound of formula I; [ka]

[0073] d) a base of a compound of the invention is converted into one of its salts by treatment with an acid, or e) An acid of a compound of the invention may be converted to one of its salts by treatment with a base. The present invention relates to a process for the preparation of the compounds of the present invention, characterized in that

[0074] It is also possible to carry out the reactions in each case stepwise and to adapt protecting group concepts to modify the sequence of linking of the building blocks. The starting materials or compounds are generally known. If they are novel, they can be prepared in a manner known per se.

[0075] If desired, the starting materials can also be formed in situ by not isolating them from the reaction mixture, but instead immediately converting them further into the compounds of the invention.

[0076] The compounds of the present invention are preferably obtained by liberating them from their functional derivatives by solvolysis, especially by hydrolysis, or by hydrogenolysis.Preferred starting materials for solvolysis or hydrogenolysis are those that contain the corresponding protected amino, carboxyl, and / or hydroxyl groups instead of one or more free amino, carboxyl, and / or hydroxyl groups, preferably those that carry an amino protecting group instead of the H atom connected to the N atom.Furthermore preferred are starting materials that carry a hydroxyl protecting group instead of the H atom of the hydroxyl group.Preferred are also starting materials that carry a protected carboxyl group instead of the free carboxyl group.It is also possible that a multiplicity of identical or different protected amino, carboxyl, and / or hydroxyl groups are present in the molecule of the starting material.When the protecting groups present are different from each other, they can in many cases be selectively cleaved off.

[0077] The term "amino protecting group" is generally known and relates to a group suitable for protecting (blocking) an amino group from chemical reaction, but which is easily removed after the desired chemical reaction has been carried out elsewhere in the molecule. Typical of such groups are, inter alia, unsubstituted or substituted acyl groups, also unsubstituted or substituted aryl groups (e.g. 2,4-dinitrophenyl), or aralkyl groups (e.g. benzyl, 4-nitrobenzyl, triphenylmethyl). Since the amino protecting groups are removed after the desired reaction or reaction sequence, their type and size become even less critical, but preferred are those having 1 to 20, especially 1 to 8 C atoms. The term "acyl group" should be understood in the broadest sense in the context of the present process. It covers acyl groups derived from aliphatic, araliphatic, aromatic or heterocyclic carboxylic or sulfonic acids, especially alkoxycarbonyl groups, aryloxycarbonyl groups, and especially aralkoxycarbonyl groups. Examples of such acyl groups are alkanoyl such as acetyl, propionyl, butyryl, aralkanoyl such as phenylacetyl, aroyl such as benzoyl or toluyl, aryloxyalkanoyl such as phenoxyacetyl, methoxycarbonyl, ethoxycarbonyl, alkoxycarbonyl such as 2,2,2-trichloroethoxycarbonyl, BOC, 2-iodoethoxycarbonyl, aralkoxycarbonyl such as CBZ, 4-methoxybenzyloxycarbonyl or FMOC. Preferred acyl groups are CBZ, FMOC, benzyl, and acetyl.

[0078] The terms "acid protecting group" or "carboxyl protecting group" are likewise commonly known and relate to groups suitable for protecting the -COOH group from chemical reaction, but which can be easily removed after the desired chemical reaction has been carried out elsewhere in the molecule. Instead of the free acid, it is typical to use esters, for example substituted and unsubstituted alkyl esters (such as methyl, ethyl, tert-butyl, and their substituted derivatives), substituted and unsubstituted benzyl esters or silyl esters. The type and size of the acid protecting group are not critical, but preferred are those having 1 to 20, especially 1 to 10, C atoms.

[0079] The term "hydroxyl protecting group" is likewise generally known and relates to a group suitable for protecting a hydroxyl group from chemical reaction, but which can be easily removed after the desired chemical reaction has been carried out elsewhere in the molecule. Typical of such groups are the abovementioned unsubstituted or substituted aryl, aralkyl or acyl groups, but also alkyl groups. The type and size of the hydroxyl protecting group is not critical, but preferred are those having 1-20, especially 1-10, C atoms. Examples of hydroxyl protecting groups are, among others, benzyl, p-nitrobenzoyl, p-toluenesulfonyl and acetyl, where benzyl and acetyl are preferred.

[0080] Further representative examples of amino-, acid-, and hydroxyl-protecting groups can be found, for example, in “Greene's Protective Groups in Organic Synthesis”, fourth edition, Wiley-Interscience, 2007. Functional derivatives of the compounds of the invention to be used as starting materials may be prepared by known methods of amino acid and peptide synthesis, for example as described in the standard works and patent applications.

[0081] The compounds of the invention are liberated from their functional derivatives with the aid of, for example, a strong acid, depending on the protecting group used, advantageously using trifluoroacetic acid or perchloric acid, but also other strong inorganic acids such as hydrochloric acid or sulfuric acid, strong organic acids such as p-trichloroacetic acid, or sulfonic acids such as benzoyl- or p-toluenesulfonic acid. The presence of additional inert solvents and / or catalysts is possible, but is not always necessary.

[0082] Depending on the respective synthetic route, the starting materials may optionally be reacted in the presence of an inert solvent.

[0083] Suitable inert solvents are, for example, heptane, hexane, petroleum ether, DMSO, benzene, toluene, xylene, trichloroethylene, 1,2-dichloroethane, carbon tetrachloride, chloroform or dichloromethane; alcohols, such as methanol, ethanol, isopropanol, n-propanol, n-butanol or tert-butanol; ethers, such as diethyl ether, diisopropyl ether (preferably for substitution on the indole nitrogen), tetrahydrofuran (THF) or dioxane; glycol ethers, such as ethylene glycol monomethyl or monoethyl ether, ethylene glycol diethyl ether (diglyme); ketones, such as acetone or butanone; amides, such as acetamide, dimethylacetamide, N-methylpyrrolidone (NMP) or dimethylformamide (DMF); nitriles, such as acetonitrile; esters, such as ethyl acetate, carboxylic acids or acid anhydrides, for example acetic acid or acetic anhydride, nitro compounds, such as nitromethane or nitrobenzene, optionally also mixtures of said solvents with one another or with water.

[0084] The amount of solvent is not critical; per gram of the compound of the invention to be reacted, 10 g to 500 g of solvent may preferably be added.

[0085] It may be advantageous to add an acid-binding agent, such as an alkali or alkaline earth metal hydroxide, carbonate, or bicarbonate, or an alkali or alkaline earth metal salt of another weak acid, preferably the potassium, sodium, or calcium salt, or an organic base, such as, for example, triethylamine, diethylamine, pyrimidine, or quinoline, or an excess of an amine component.

[0086] The resulting compounds according to the invention can be separated from the corresponding solutions in which they were prepared (for example by centrifugation and washing) and stored in other compositions after separation, or they can remain directly in the solutions in which they were prepared. The resulting compounds according to the invention can also be absorbed in a desired solvent for specific use.

[0087] The reaction duration depends on the reaction conditions selected. In general, the reaction time is between 0.5 hours and 10 days, preferably between 1 and 24 hours. When using microwaves, the reaction time can be reduced to values ​​between 1 and 60 minutes.

[0088] The compounds of the present invention, as well as the starting materials for their preparation, are prepared by known methods, for example, under known reaction conditions suitable for the reaction, as described in the literature (for example, in standard works such as Houben-Weyl, Methoden der organischen Chemie [Methods of Organic Chemistry], Georg-Thieme-Verlag, Stuttgart). The use of per se known variants, which are not described in more detail here, may also be made here.

[0089] Conventional work-up steps, such as the addition of water to the reaction mixture and extracts, make it possible to obtain the compound after removal of the solvent. For further purification of the product, it may be advantageous to follow this with distillation or crystallization, or to carry out chromatographic purification.

[0090] The acids of the present invention can be converted into the relevant addition salts using a base, for example, by reaction of an equal amount of acid and base in an inert solvent such as ethanol, including evaporation. Suitable bases for this reaction are those which give particularly physiologically acceptable salts. Thus, the acids of the present invention can be converted into the corresponding metal salts, particularly alkali metal salts or alkaline earth metal salts, or into the corresponding ammonium salts using a base, such as, for example, sodium hydroxide, potassium hydroxide, sodium carbonate, or potassium carbonate. Organic bases which give physiologically acceptable salts, such as, for example, ethanolamine, are also suitable for this reaction.

[0091] On the other hand, the bases of the present invention can be converted into the relevant acid addition salts using an acid, for example by reaction of equal amounts of base and acid in an inert solvent such as ethanol, followed by evaporation. Suitable acids for this reaction are, inter alia, those which give physiologically acceptable salts. Thus, it is possible to use inorganic acids, such as sulfuric acid, nitric acid, hydrohalic acids (such as hydrochloric acid or hydrobromic acid), phosphoric acid (such as orthophosphoric acid), sulfamic acid, and also organic acids, in particular aliphatic, cycloaliphatic, araliphatic, aromatic or heterocyclic, monobasic or polybasic carboxylic acids, sulfonic acids or sulfuric acids, such as formic acid, acetic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, lactic acid, tartaric acid, malic acid, citric acid, gluconic acid, ascorbic acid, nicotinic acid, isonicotinic acid, methanesulfonic acid or ethanesulfonic acid, ethanedisulfonic acid, 2-hydroxysulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenemonosulfonic acid and disulfonic acid, or laurylsulfuric acid. Salts with physiologically unacceptable acids, such as picrates, can be used for the isolation and / or purification of the compounds of the invention.

[0092] The compounds of the invention have been found to be well tolerated and to have valuable pharmacological properties. A 2A and A 2BAdenosine receptors such as have been shown to downregulate immune responses during inflammation and protect tissues from immune damage, and inhibition of signaling through adenosine receptors can be used to enhance and prolong immune responses.

[0093] A method of enhancing immune response is provided herein. In one example, the method enhances the damage to desired and targeted tissue (such as tumor, cancer damage). Disclosed herein is a method of inhibiting one or more processes that lead to the production of extracellular adenosine and adenosine-induced signaling through adenosine receptors. For example, the enhancement of immune response, local tissue inflammation, and targeted tissue destruction is achieved by: inhibiting or reducing the hypoxic condition of the local tissue that produces adenosine; by degrading (or inactivating) accumulated extracellular adenosine; by preventing or reducing the expression of adenosine receptors on immune cells; and / or inhibiting / antagonizing the signaling by adenosine ligands through adenosine receptors. The results disclosed herein demonstrate that in vivo administration of agents that interfere with the "hypoxia -> adenosine accumulation -> immunosuppressive adenosine receptor signaling to immune cells" pathway in subjects suffering from various diseases (e.g., cancer and sepsis) can result in in vivo treatment of tumors or improved immunity.

[0094] In one example, the method comprises administering one or more inhibitors of extracellular adenosine and / or adenosine receptor inhibitors, such as adenosine receptor antagonists.To increase the effectiveness of vaccines, one or more adenosine receptor inhibitors and / or inhibitors of extracellular adenosine can be administered in conjunction with vaccines.In one example, one or more adenosine receptor inhibitors or inhibitors of extracellular adenosine are administered to increase immune response / inflammation.In another example, the method is provided to achieve targeted tissue damage, such as for tumor destruction.

[0095] Thus, the present invention also provides adenosine or other A 2A and / or A 2B The use of a compound according to the invention for the preparation of a medicament for the treatment and / or prophylaxis of diseases which are caused, promoted and / or propagated by receptor agonists.

[0096] The present invention therefore also relates to a medicament comprising at least one compound according to the invention and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, as well as mixtures thereof in any ratio, for use in the treatment and / or prophylaxis of physiological and / or pathophysiological conditions.

[0097] Particularly preferred is adenosine A 2A and / or A 2B Physiological and / or pathophysiological conditions associated with the receptor.

[0098] Physiological and / or pathophysiological conditions are understood to mean medically relevant physiological and / or pathophysiological conditions, especially diseases, such as, for example, diseases or illnesses, and medical disorders, complaints, symptoms, or complications.

[0099] The present invention further relates to a medicament comprising at least one compound according to the invention, and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio, for use in a method for the treatment and / or prophylaxis of physiological and / or pathophysiological conditions selected from the group consisting of hyperproliferative and infectious diseases and disorders.

[0100] The present invention further relates to a medicament comprising at least one compound according to the invention, and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio, for use in a method for the treatment and / or prophylaxis of a physiological and / or pathophysiological condition selected from the group consisting of hyperproliferative and infectious diseases and disorders, where the hyperproliferative disease or disorder is cancer.

[0101] The present invention therefore particularly and preferably relates to a medicament comprising at least one compound according to the invention and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, as well as mixtures thereof in any ratio, in which cancer is selected from the group consisting of acute and chronic lymphocytic leukemia, acute granulocytic leukemia, adrenal cortical carcinoma, bladder cancer, brain cancer, breast cancer, cervical cancer, cervical hyperplasia, cervical cancer, choriocarcinoma, chronic granulocytic leukemia, chronic lymphocytic leukemia, colon cancer, endometrial cancer, esophageal cancer, essential thrombocytosis, genitourinary cancer, glioma, glioblastoma, hairy cell leukemia, head and neck cancer, Hodgkin's disease, Kaposi's sarcoma, lung cancer, lymphoma, malignant carcinoid cancer, malignant hypercalcemia, malignant melanoma, malignant pancreatic insulinoma, medullary thyroid carcinoma, black blood cell carcinoma, thyroid cancer ... The cancer is selected from the group consisting of chromoma, multiple myeloma, mycosis fungoides, myeloid and lymphocytic leukemia, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, osteosarcoma, ovarian cancer, pancreatic cancer, polycythemia vera, primary brain tumors, primary macroglobulinemia, prostate cancer, renal cell carcinoma, rhabdomyosarcoma, skin cancer, small cell lung cancer, sarcoma of soft tissue, squamous cell carcinoma, gastric cancer, testicular cancer, thyroid cancer, and Wilms' tumor.

[0102] The present invention further preferably relates to a medicament comprising at least one compound according to the invention, and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio, for use in a method for the treatment and / or prophylaxis of physiological and / or pathophysiological conditions selected from the group consisting of hyperproliferative and infectious diseases and disorders, wherein the hyperproliferative disease or disorder is selected from the group consisting of age-related macular degeneration, Crohn's disease, cirrhosis of the liver, chronic inflammatory-related disorders, proliferative diabetic retinopathy, proliferative vitreoretinopathy, retinopathy of prematurity, granulomatous diseases, immune hyperproliferation associated with organ or tissue transplantation, and immune proliferative diseases or disorders selected from the group consisting of inflammatory bowel disease, psoriasis, rheumatoid arthritis, systemic lupus erythematosus (SLE), vascular hyperproliferation secondary to retinal hypoxia, and vasculitis.

[0103] The present invention further preferably relates to a medicament comprising at least one compound according to the invention, and / or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, and mixtures thereof in any ratio, for use in a method for the treatment and / or prophylaxis of a physiological and / or pathophysiological condition selected from the group consisting of hyperproliferative and infectious diseases and disorders, wherein the infectious disease or disorder is selected from the group consisting of:

[0104] a) virus-induced infectious diseases caused by retroviruses, hepadnaviruses, herpesviruses, flaviviridae, and / or adenoviruses, where the retrovirus is selected from lentiviruses or oncoretroviruses, where the lentivirus is selected from the group consisting of HIV-1, HIV-2, FIV, BIV, SIVs, SHIV, CAEV, VMV, and EIAV, and the oncoretrovirus is selected from the group consisting of HTLV-I, HTLV-II, and BLV, the hepadnavirus is selected from the group consisting of HBV, GSHV, and WHV, the herpesvirus is selected from the group consisting of HSV I, HSV II, EBV, VZV, HCMV, or HHV8, and the flaviviridae is selected from the group consisting of HCV, West Nile, and Yellow Fever;

[0105] b) bacterial infectious diseases caused by gram-positive bacteria, where the gram-positive bacteria include methicillin-susceptible and methicillin-resistant staphylococci (including Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus saprophyticus, and coagulase-negative staphylococci), glycopeptides-intermediate susceptible Staphylococcus aureus (GISA), penicillin-susceptible and penicillin-resistant streptococci (Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus avium, Streptococcus bovis, Streptococcus lactis, Streptococcus sanguis, and group C streptococci (GCS), group G streptococci (GGS), and viridans streptococci), enterococci (vancomycin-susceptible and vancomycin-resistant strains such as Enterococcus faecalis and Enterococcus faecium), Clostridium difficile, Iisteria monocytogenes, Corynebacterium jeikeium, Chlamydia spp (including C. pneumoniae), and Mycobacterium tuberculosis;

[0106] c) bacterial infections caused by gram-negative bacteria, where the gram-negative bacteria are selected from the group consisting of Enterobacteriaceae including Escherichia spp. (including Escherichia coli), Klebsiella spp., Enterobacter spp., Citrobacter spp., Serratia spp., Proteus spp., Providencia spp., Salmonella spp., Shigella spp., Pseudomonas (including P. aeruginosa), Moraxella spp. (including M. catarrhalis), Haemophilus spp., and Neisseria spp.;

[0107] d) Infectious diseases induced by intracellular active parasites selected from the group consisting of the phylum Apicomplexa, or Sarcomastigophora (including Trypanosoma, Plasmodia, Leishmania, Babesia, or Theileria), Cryptosporidia, Sacrocystida, Amoebia, Coccidia, and Trichomonadia.

[0108] The medicaments disclosed above are intended to encompass the corresponding uses of the compounds according to the invention for the preparation of medicaments for the treatment and / or prophylaxis of physiological and / or pathophysiological conditions.

[0109] In addition, the above disclosed medicaments are intended to encompass corresponding methods for the treatment and / or prophylaxis of the above physiological and / or pathophysiological conditions, in which at least one compound according to the invention is administered to a patient in need of such treatment.

[0110] The compounds according to the invention preferably exhibit advantageous biological activity, which can be easily demonstrated in enzyme assays and animal experiments, as described in the examples. In such enzyme-based assays, the compounds according to the invention preferably have IC in the suitable range, preferably in the micromolar range, more preferably in the nanomolar range. 50 These exert and cause inhibitory effects that are mostly evidenced by the values.

[0111] The compounds according to the invention can be administered to humans or animals, especially mammals, such as monkeys, dogs, cats, rats or mice, and can be used in the therapeutic treatment of the human or animal body and against the above-mentioned diseases. They can also be used as diagnostic agents or as reagents.

[0112] Furthermore, the compounds according to the invention are 2A and / or A 2B They can be used to isolate receptors and to investigate their activity or expression. 2A and / or A 2B The present invention is therefore particularly suitable for use in diagnostic methods for diseases related to disturbed activity of the adenosine A receptor. 2A and / or A 2B For the isolation of receptors and for the investigation of their activity or expression, or for the adenosine A 2A and / or A 2B The use of the compounds according to the invention as binders as well as inhibitors of receptors.

[0113] For diagnostic purposes, the compounds according to the invention can be, for example, radioactively labeled. Examples of radioactive labels are: 3 H, 14 C. 231 I, and 125I. A preferred labeling method is the iodogen method (Fraker et al., 1978). In addition, the compounds according to the present invention can be labeled with enzymes, fluorophores, and chemophores. Examples of enzymes are alkaline phosphatase, β-galactosidase, and glucose oxidase, examples of fluorophores are fluorescein, and examples of chemophores are luminol, and automated detection systems for, e.g., fluorescence color development are described in, e.g., US 4,125,828 and US 4,207,554.

[0114] The present invention further relates to pharmaceutical compositions containing the compounds of the present invention, as well as to adenosine A 2A and / or A 2B It also relates to their use for the treatment and / or prophylaxis of diseases and disorders in which partial or total inactivation of the receptor may be beneficial.

[0115] The compounds of the present invention can be used for the preparation of pharmaceutical preparations, especially by non-chemical methods. In this case, they are combined with at least one solid, liquid and / or semi-liquid excipient or adjuvant, optionally with one or more additional active compound(s), to form a suitable dosage form.

[0116] The present invention therefore also relates to pharmaceutical preparations comprising at least one compound of the invention and / or its physiologically acceptable salts, derivatives, solvates and stereoisomers, as well as mixtures thereof in any ratio. In particular, the present invention also relates to pharmaceutical preparations comprising further excipients and / or adjuvants, as well as to pharmaceutical preparations comprising at least one further pharma- ceutical active compound.

[0117] In particular, the present invention also relates to a process for the preparation of a pharmaceutical preparation, characterized in that the compound of the invention, and / or one of its physiologically acceptable salts, derivatives, solvates and stereoisomers, as well as mixtures thereof in any ratio, together with solid, liquid or semi-liquid excipients or adjuvants, and optionally further active compounds, are brought into a suitable dosage form.

[0118] The pharmaceutical preparations according to the invention can be used as human medicine or veterinary medicine. The patient or host can belong to any mammalian species, such as primate species, specifically humans; rodents, including mice, rats, and hamsters; rabbits; horses, cows, dogs, cats, etc. Animal models are intended for experimental investigations, and they provide models for the treatment of human diseases.

[0119] Suitable carrier substances are organic or inorganic substances which are suitable for enteral (e.g. oral), parenteral or topical administration and which do not react with the new compounds, such as water, vegetable oils (such as sunflower oil or cod liver oil), benzyl alcohol, polyethylene glycol, gelatin, carbohydrates (such as lactose or starch), magnesium stearate, talc, lanolin or petrolatum. Those skilled in the art are familiar with the advantages of their expertise which adjuvants are suitable for the desired pharmaceutical preparation. Besides solvents, such as water, physiological saline solutions, or alcohols (such as ethanol, propanol or glycerol), sugar solutions (glucose or mannitol solutions) or mixtures of said solvents, gel-forming agents, tableting aids and other active ingredient carriers, it is also possible to use, for example, lubricants, stabilizers and / or wetting agents, emulsifiers, salts for influencing the osmotic pressure, antioxidants, dispersants, antifoaming agents, buffer substances, flavors and / or aromas or flavor correctors, preservatives, solubilizers or dyes. If desired, the preparations or medicaments according to the invention may also contain one or more further active compounds, such as one or more vitamins.

[0120] If desired, the preparations or medicaments according to the invention may also comprise one or more further active compounds and / or one or more action enhancers (adjuvants). The terms "pharmaceutical formulation" and "pharmaceutical preparation" are used synonymously for the purposes of this invention.

[0121] As used herein, "pharmacologically tolerated" refers to drugs, precipitating agents, excipients, adjuvants, stabilizers, solvents, and other agents that facilitate the administration to a mammal of a pharmaceutical preparation obtained therefrom without undesirable physiological side effects such as nausea, dizziness, digestive problems, or the like.

[0122] In pharmaceutical preparations for parenteral administration, the formulation, the adjuvants employed, and the primary packaging require isotonicity, euhydration and tolerability, as well as safety (low toxicity).Surprisingly, the compounds according to the present invention preferably have the advantage that they can be used directly, and therefore no further purification steps are required before the use of the compounds according to the present invention in pharmaceutical preparations, for example for the removal of toxicologically unacceptable agents such as high concentrations of organic solvents or other toxicologically unacceptable adjuvants.

[0123] The present invention also particularly preferably relates to pharmaceutical preparations comprising at least one compound according to the invention in precipitated amorphous form, precipitated crystalline form or in dissolved or suspended form, and optionally excipients and / or adjuvants and / or further pharma- ceutical active compounds.

[0124] The compounds according to the invention preferably allow the preparation of highly concentrated formulations without the occurrence of undesirable and undesirable aggregation of the compounds according to the invention. Thus, ready-to-use solutions with high contents of active ingredient can be prepared using the compounds according to the invention with or in aqueous media.

[0125] The compounds and / or their physiologically acceptable salts and solvates may also be lyophilized and the resulting lyophilizates may be used, for example, for the preparation of injectable preparations.

[0126] Aqueous preparations can be prepared by dissolving or suspending the compound according to the invention in an aqueous solution and optionally adding an adjuvant.To this end, a defined volume of stock solution containing the further adjuvant at a defined concentration is advantageously added to a solution or suspension having a compound according to the invention at a defined concentration, and the mixture is optionally diluted with water to a precalculated concentration.Alternatively, the adjuvant can be added in solid form.The amount of stock solution and / or water required in each case can then be added to the resulting aqueous solution or suspension.The compound according to the invention can also advantageously be dissolved or suspended directly in a solution containing all further adjuvants.

[0127] Solutions or suspensions containing the compounds according to the invention and having a pH of 4-10, preferably a pH of 5-9, and an osmolality of 250-350 mOsmol / kg can be advantageously prepared. Thus, the pharmaceutical preparations can be administered directly, intravenously, intraarterially, intraarticularly, subcutaneously or transdermally, virtually painlessly. In addition, the preparations can also be added to infusion fluids, such as glucose solutions, isotonic saline solutions or Ringer's solutions, which can also contain further active compounds, thus allowing relatively large amounts of active compounds to be administered.

[0128] Pharmaceutical preparations according to the invention may also contain mixtures of multiple compounds according to the invention.

[0129] The preparations according to the invention are physiologically well tolerated, easy to prepare, can be accurately dispensed, and are preferably stable with respect to assays, degradation products, and aggregates during storage and transportation, as well as during multiple freezing and thawing processes. They can be stored in a stable manner, preferably at refrigerator temperature (2-8° C.) and at room temperature (23-27° C.) and 60% relative atmospheric humidity (RH), for a period of at least 3 months to 2 years.

[0130] For example, the compounds according to the present invention can be stored in a stable manner by drying and, when necessary, can be converted into ready-to-use pharmaceutical preparations by dissolving or suspending.Possible drying methods are, for example, but not limited to these examples, nitrogen gas drying, vacuum oven drying, freeze drying, washing with organic solvent followed by air drying, liquid-bed drying, fluidised-bed drying, spray drying, roller drying, layer drying, air drying at room temperature, and further methods.

[0131] The term "effective amount" refers to an amount of a drug or pharma- ceutical active compound that elicits the biological or medical response desired or desired by, for example, a researcher or physician, in a tissue, system, animal, or human.

[0132] In addition, the term "therapeutically effective amount" refers to an amount that provides the following causal effects: improved treatment, cure, prevention, or elimination of a disease, syndrome, condition, complaint, disorder, or prevention of side effects, or also reduction in the progression of a disease, complaint, or disorder, compared to a corresponding subject not receiving this amount. The term "therapeutically effective amount" also encompasses an amount effective to increase normal physiological function.

[0133] When using the preparations or medicaments according to the invention, the compounds according to the invention and / or their physiologically acceptable salts and solvates are generally used similarly to known commercial preparations or preparations, preferably in dosages between 0.1 mg and 500 mg, in particular between 5 mg and 300 mg per use unit. The daily dose is preferably between 0.001 and 250 mg / kg body weight, in particular between 0.01 and 100 mg / kg body weight. The preparations can be administered one or more times per day, for example two, three or four times per day. However, the individual dose for a patient depends on a number of individual factors, such as the effectiveness of the specific compound used, age, body weight, general health, sex, nutritional absorption, time and method of administration, excretion rate, combination with other medicaments, and the severity and duration of the specific disease.

[0134] The measure of the uptake of a pharmacokinetically active compound into the body is its bioavailability. When a pharmacokinetically active compound is delivered to the body in the form of an injection solution intravenously, its absolute bioavailability, i.e. the proportion of the drug that reaches the systemic blood, i.e. the major circulation, in unchanged form, is 100%. In the case of oral administration of a therapeutically active compound, the active compound is generally in solid form in the formulation. It must therefore first be dissolved so that it can overcome the barriers of entry, such as the gastrointestinal tract, the oral mucosa, the nasal membrane or the skin, especially the stratum corneum, or so that it can be absorbed by the body. Data on pharmacokinetics, i.e. bioavailability, can be obtained similarly to the method of J. Shaffer et al., J. Pharm. Sciences, 88 (1999), 313-318.

[0135] Furthermore, a medicament of this type can be prepared by one of the processes generally known in the art of pharmacy.

[0136] The medicaments may be adapted for administration via any suitable route desired, for example, oral (including buccal or sublingual), rectal, pulmonary, intranasal, topical (including buccal, sublingual, or transdermal), vaginal, or parenteral (including subcutaneous, intramuscular, intravenous, intradermal, especially intraarticular) routes. Medicaments of this type may be prepared using any process known in the pharmaceutical art, for example, by combining the active compound with the excipient(s) or adjuvant(s).

[0137] Parenteral administration is preferably suitable for the administration of the medicaments according to the invention. In the case of parenteral administration, intra-articular administration is particularly preferred.

[0138] The present invention therefore also relates to the use of a pharmaceutical preparation according to the invention for intra-articular administration in a method for the treatment and / or prophylaxis of physiological and / or pathophysiological conditions, preferably selected from the group consisting of osteoarthritis, traumatic cartilage damage, arthritis, pain, allodynia or hyperalgesia.

[0139] Intra-articular administration has the advantage that the compounds according to the invention can be administered directly into the synovial fluid near the articular cartilage and can also diffuse from there into the cartilage tissue. Thus, the pharmaceutical preparations according to the invention can also be injected directly into the joint space and thus exert their action directly at the intended site of action. The compounds according to the invention are also suitable for the preparation of medicaments to be administered parenterally, with slow, sustained and / or controlled release of the active compound. Thus, they are also suitable for the preparation of delayed release formulations, which are advantageous for patients, since administration can be carried out at relatively long time intervals.

[0140] Medicaments adapted for parenteral administration include aqueous and non-aqueous sterile injection solutions containing antioxidants, buffers, bacteriostats, and solutes with which the formulation is made isotonic with the blood or synovial fluid of the recipient to be treated; and aqueous and non-aqueous sterile suspensions which may contain suspending media and thickening agents. The formulations may be delivered in single-dose or multi-dose containers, such as sealed ampoules and vials. The formulations may also be stored in a freeze-dried (lyophilised) state, requiring only the addition of a sterile carrier liquid, such as water for injection, immediately prior to use. Injection solutions and suspensions prepared according to formulations may be prepared from sterile powders, granules, and tablets.

[0141] The compounds according to the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine, or phosphatidylcholines.

[0142] The compounds according to the present invention can also be coupled to soluble polymers as excipients for targeted drugs. Such polymers can cover polyvinylpyrrolidone, pyran copolymers, polyhydroxypropylmethacrylamidephenol, polyhydroxyethylaspartamidephenol, or polyethylene oxide polylysine substituted with palmitoyl radicals. The compounds according to the present invention can also be coupled to classes of biodegradable polymers suitable for achieving sustained release of drugs, such as polylactic acid, poly-epsilon-caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydroxypyrans, polycyanoacrylates, polylactic acid-glycolic acid copolymers, such as dextran and methacrylate conjugates, polyphosphoesters, various polysaccharides, and polyamines, and poly-ε-caprolactone, albumin, chitosan, collagen, or modified gelatin, as well as crosslinked or amphiphilic hydrogel block copolymers.

[0143] Suitable for enteral administration (oral or rectal) are, inter alia, tablets, dragees, capsules, syrups, juices, drops or suppositories, while suitable for topical use are ointments, creams, pastes, lotions, gels, sprays, foams, aerosols, solutions (for example in alcohols such as ethanol or isopropanol, acetonitrile, DMF, dimethylacetamide, 1,2-propanediol, or in mixtures thereof with each other and / or with water) or powders. Also particularly suitable for topical use are liposomal preparations.

[0144] In the case of a formulation giving an ointment, the active compound may be employed with either a paraffinic or a water-miscible cream base. Alternatively, the active compound may be formulated into a cream with an oil-in-water cream base or a water-in-oil base.

[0145] Medicaments adapted for transdermal administration may be delivered as separate patches for widespread and intimate contact with the recipient's epidermis, thus, for example, the active compound may be delivered from the patch by iontophoresis, as generally described in Pharmaceutical Research, 3(6), 318 (1986).

[0146] It goes without saying that in addition to the components specifically mentioned above, the medicaments according to the invention may also contain other agents conventional in the art for the particular type of pharmaceutical formulation.

[0147] The present invention also provides a) an effective amount of a compound of the present invention, and / or its physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in any ratio; b) an effective amount of a further pharma- ceutical active compound; This relates to a set (kit) consisting of individual packs of

[0148] The sets include suitable containers, such as boxes or cartons, individual bottles, bags, or ampoules. The sets may, for example, include individual ampoules, each of which contains an effective amount of a compound of the invention, and / or its pharma- ceutically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in any ratio, and an effective amount of a further pharma- ceutically active compound, in dissolved or lyophilized form.

[0149] Furthermore, medicaments according to the present invention may be used to provide an additive or synergistic effect in certain known treatments and / or to restore the effectiveness of certain existing treatments.

[0150] Besides the compounds according to the invention, the pharmaceutical preparations according to the invention may also contain further pharma- ceutical active compounds, for example said further pharma- ceutical active compounds for use in the treatment of cancer, other antitumor medicaments. For the treatment of the other diseases mentioned, the pharmaceutical preparations according to the invention may also contain, besides the compounds according to the invention, further pharma- ceutical active compounds known to the skilled person in their treatment.

[0151] In one main embodiment, a method is provided for enhancing immune response in a host in need thereof. The immune response can be enhanced in the host by reducing T cell tolerance, including increasing IFN-γ release, decreasing regulatory T cell production or activation, or increasing antigen-specific memory T cell production. In one embodiment, the method comprises administering a compound of the present invention to the host in combination with or instead of an antibody. In specific subembodiments, the antibody is a therapeutic antibody. In a specific embodiment, a method is provided for enhancing the effectiveness of passive antibody therapy, comprising administering a compound of the present invention in combination with or instead of one or more passive antibodies. This method can enhance the effectiveness of antibody therapy for the treatment of abnormal cell proliferation disorders, such as cancer, or enhance the effectiveness of therapy in the treatment or prevention of infectious diseases. The compound of the present invention can be administered in combination with or instead of an antibody, such as, for example, Rituximab, Herceptin, or Erbitux.

[0152] In another main aspect, there is provided a method of treating or preventing abnormal cell proliferation, said method comprising administering a compound of the present invention in the substantial absence of another anti-cancer agent in a host in need thereof.

[0153] In another main aspect, there is provided a method of treating or preventing abnormal cell proliferation in a host in need thereof, said method comprising administering to the host a first compound of the present invention substantially in combination with a first anticancer agent, and subsequently administering to the host a first compound of the present invention in combination with a first anticancer agent, 2A and / or A2B and administering a second antagonist of the receptor. In one sub-embodiment, the second antagonist is administered in the substantial absence of another anti-cancer agent. In another main embodiment, a method of treating or preventing abnormal cell proliferation in a host in need thereof is provided, the method comprising administering a compound of the present invention to the host substantially in combination with a first anti-cancer agent, and subsequently administering a second anti-cancer agent in the absence of an antagonist.

[0154] Thus, the cancer treatment disclosed herein may be carried out as a treatment with the compounds of the invention or said treatment in combination with surgery, radiation therapy or chemotherapy. This type of chemotherapy may include the use of one or more active compounds from the following categories of antitumor active compounds: (i) alkylating active compounds (e.g., cisplatin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulfan, and nitrosoureas); antimetabolites (e.g., antifolates such as fluoropyrimidines such as 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside, hydroxyurea, and gemcitabine); antitumor antibiotics (e.g., anthracyclines such as adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin C, dactinomycin, and mitracyclines); mycin); antimitotic active compounds (e.g., vinca alkaloids such as vincristine, vinblastine, vindesine, and vinorelbine, and taxoids such as taxol and taxotere); topoisomerase inhibitors (e.g., epipodophyllotoxins such as etoposide and teniposide, amsacrine, topotecan, irinotecan, and camptothecin), and cell differentiation active compounds (e.g., all-trans retinoic acid, 13-cis-retinoic acid, and fenretinide), and antiproliferative / antineoplastic / DNA damage active compounds and combinations thereof as used in medical oncology;

[0155] (ii) cytostatically active compounds, such as antiestrogens (e.g. tamoxifen, toremifene, raloxifene, droloxifene, and iodoxyfene), estrogen receptor modulators (e.g. fulvestrant), antiandrogens (e.g. bicalutamide, flutamide, nilutamide, and cyproterone acetate), LHRH antagonists or agonists (e.g. goserelin, leuprorelin, and buserelin), progesterones (e.g. megestrol acetate), aromatase inhibitors (e.g. anastrozole, letrozole, vorazole, and exemestane), and inhibitors of 5α-reductase, such as finasteride;

[0156] (iii) active compounds that inhibit cancer invasion, including, for example, metalloproteinase inhibitors such as marimastat, and inhibitors of urokinase-type plasminogen activator receptor function;

[0157] (iv) inhibitors of growth factor function, such as growth factor antibodies, growth factor receptor antibodies, such as the anti-erbb2 antibody trastuzumab [Herceptin™] and the anti-erbb1 antibody cetuximab [C225]), farnesyltransferase inhibitors, tyrosine kinase inhibitors, and serine / threonine kinase inhibitors, such as inhibitors of the epidermal growth factor family (e.g., N-(3-chlorophenyl)-2-(4-methylphenyl)-2-propanediol (DMPO) and N-(3-chlorophenyl)-2-propanediol (DMPO) EGFR family tyrosine kinase inhibitors, such as N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine (gefitinib, AZD1839), N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine (erlotinib, OSI-774), and 6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)quinazolin-4-amine (CI 1033), e.g. inhibitors of the platelet-derived growth factor family, as well as inhibitors of the hepatocyte growth factor family, e.g.

[0158] (v) bevacizumab, angiostatin, endostatin, linomide, batimastat, captopril, cartilage-derived inhibitor, genistein, interleukin 12, lavendustin, medroxyprogesterone acetate, recombinant human platelet factor 4, tecogalan, thrombospondin, TNP-470, anti-VEGF monoclonal antibodies, soluble VEGF receptor chimeric proteins, anti-VEGF receptor antibodies, anti-PDGF receptors, inhibitors of integrins, tyrosine kinase inhibitors, serine / threonine kinase inhibitors, antisense oligonucleotides, antisense oligodeoxynucleotides, siRNA, anti-VEGF aptamers, pigment epithelium-derived factor, and the like, as well as the compounds described in International Patent Applications WO 97 / 22596, WO 97 / 30035, WO 97 / 32856, and WO antiangiogenic compounds, such as those disclosed in 98 / 13354;

[0159] (vi) vessel-destroying agents, such as combretastatin A4 and the compounds published in International Patent Applications WO 99 / 02166, WO 00 / 40529, WO 00 / 41669, WO 01 / 92224, WO 02 / 04434, and WO 02 / 08213; (vii) antisense therapy, e.g., ISIS 2503, anti-Ras antisense, etc., directed to the targets mentioned above;

[0160] (viii) gene therapy approaches, including, for example, approaches for the replacement of abnormal modifier genes, such as abnormal p53 or abnormal BRCA1 or BRCA2, GDEPT approaches (gene-directed enzyme prodrug therapy), such as approaches using cytosine deaminase, thymidine kinase, or bacterial nitroreductase enzymes, and approaches for increasing a patient's tolerance to chemotherapy or radiation therapy, such as multidrug resistance gene therapy; and

[0161] (ix) immunotherapeutic approaches, including ex-vivo and in-vivo approaches to increase the immunogenicity of a patient's tumor cells, such as transfection with cytokines such as interleukin 2, interleukin 4, or granulocyte-macrophage colony-stimulating factor, approaches to reduce T-cell anergy, approaches using transfected immune cells such as cytokine-transfected dendritic cells, approaches for the use of cytokine-transfected tumor cell lines, and approaches for the use of anti-idiotypic antibodies;

[0162] (x) For example, abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, anastrozole, arsenic trioxide, asparaginase, live BCG bacteria, bevacizumab, bexarotene, bleomycin, bortezomib, busulfan, calsterone, camptothecin, capecitabine, carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cinacalcet, cisplatin, cladribine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, darbepoetin alfa, daunorubicin, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone, epirubicin, epoetin Chemotherapeutic agents, including alpha, estramustine, etoposide, exemestane, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, and gemcitabine.

[0163] Medicaments from Table 1 may be preferably, but not exclusively, combined with the compounds of the present invention. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5]

[0164] Without further embodiments, it is believed that a person skilled in the art can utilize the above description in its broadest scope, and the preferred embodiments should therefore be regarded merely as a descriptive disclosure that is in no way limiting in any absolute sense.

[0165] Thus, the following examples are intended to illustrate the invention without limiting it. Unless otherwise indicated, percentage data refer to percentages by weight. All temperatures are given in degrees Celsius. "Conventional work-up": water is added if necessary, the pH is adjusted if necessary to a value between 2 and 10, the mixture is extracted with ethyl acetate or dichloromethane depending on the composition of the final product, the phases are separated, the organic phase is dried over sodium sulfate, filtered and evaporated, and the product is purified by chromatography on silica gel and / or by crystallization.

[0166] Rf value on silica gel; Mass spectrometry: EI (electron impact ionization): M + , FAB (Fast Atom Collision): (M+H) + , THF (tetrahydrofuran), NMP (N-methylpyrrolidone), DMSO (dimethyl sulfoxide), EA (ethyl acetate), MeOH (methanol), TLC (thin layer chromatography)

[0167] List of abbreviations AUC Area under the plasma drug concentration-time curve C max maximum plasma concentration CL Clearance CV Coefficient of variation CYP Cytochrome P450 DMSO Dimethyl sulfoxide F. Bioavailability f a Absorption rate iv LC-MS / MS Liquid Chromatography Tandem Mass Spectrometry LLOQ Lower limit of quantification NC Not calculated ND Not Determined PEG Polyethylene glycol Pgp Permeability glycoprotein PK Pharmacokinetic (or pharmacokinetic) po per os (oral) t 1 / 2 Half-life t max Time to reach maximum plasma concentration of drug UPLC Ultra-Performance Liquid Chromatography V ss Volume of distribution (at steady state) v / v volume to volume

[0168] Example 1: Examples of compounds of the present invention The invention particularly relates to the compounds of Table 2, and to their physiologically acceptable salts, derivatives, solvates, prodrugs, and stereoisomers, and mixtures thereof in any ratio.

[0169] Table 2 - Examples of compounds of the present invention [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5]

Table 3-6

Table 3-7

Table 3-8

Table 3-9

Table 3-10

Table 3-11

Table 3-12

Table 3-13

Table 3-14

Table 3-15

Table 3-16

Table 3-17

Table 3-18

Table 3-19

Table 3-20

Table 3-21

Table 3-22

Table 3-23

Table 3-24

Table 3-25

Table 3-26

Table 3-27

Table 3-28

Table 3-29

Table 3-30

Table 3-31

Table 3-32

[0170] Table 3 - NMR profiles of compounds of the present invention The numbers listed here correspond to the numbers given to the compounds disclosed in Table 2. [Table 4-1] [Table 4-2] [Table 4-3] [Table 4-4] [Table 4-5] [Table 4-6] [Table 4-7] [Table 4-8] [Table 4-9] [Table 4-10] [Table 4-11] [Table 4-12] [Table 4-13] [Table 4-14] [Table 4-15]

[0171] Example 2: Preparation and Analytical Methods of Compounds of the Invention All solvents used were commercially available and were used without further purification. Reactions were typically carried out using anhydrous solvents under an inert atmosphere of nitrogen. Flash column chromatography was generally performed using Silica gel 60 (0.035-0.070 mm particle size).

[0172] All NMR experiments were recorded either on a Bruker Mercury Plus 400 NMR Spectrometer equipped with a Bruker 400 BBFO probe at 400 MHz for proton NMR, or on a Bruker Mercury Plus 300 NMR Spectrometer equipped with a Bruker 300 BBFO probe at 300 MHz for proton NMR. All deuterated solvents typically contained 0.03%-0.05% v / v tetramethylsilane, which was used as the reference signal ( 1 H and 13C was set to δ = 0.00).

[0173] LC-MS analysis was performed on a SHIMADZU LC-MS instrument consisting of a UFLC 20-AD system and an LCMS 2020 MS detector. The column used was Shim-pack XR-ODS, 2.2 μm, 3.0 × 50 mm. A linear gradient was applied, starting with 95% A (A: 0.05% TFA in water) and ending with 100% B (B: 0.05% TFA in acetonitrile) over 2.2 min. The total run time was 3.6 min. The column temperature was 40 °C with a flow rate of 1.0 mL / min. The Diode Array detector scanned from 200 to 400 nm. The mass spectrometer was equipped with an electrospray ion source (ES) operated in positive or negative mode. The mass spectrometer scanned between m / z 90 and 900 with a scan time of 0.6 s. Unless otherwise stated.

[0174] 1. (5R)-N-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-thiazolo[4,5-c]pyridin-2-yl]-7-oxa-2-azaspiro[4.5]decane-2-carboxamide 24 and (5S)-N-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-thiazolo[4,5-c]pyridin-2-yl]-7-oxa-2-azaspiro[4.5]decane-2-carboxamide 25 [ka]

[0175] a. 4-Chloro-5-iodo-3-nitropyridin-2-ol In a 250 mL round bottom flask was placed 4-chloro-3-nitropyridin-2-ol (10.0 g, 54.4 mmol, 95%), N-iodosuccinimide (NIS, 14.2 g, 59.9 mmol, 95%) in acetonitrile (115 mL). The solution was stirred overnight at 80° C. in an oil bath for 1 h. The mixture was concentrated and the precipitate formed was collected by filtration. The residue was washed twice with petroleum ether (500 mL) which was dried under vacuum at 60° C. overnight. This gave 4-chloro-5-iodo-3-nitropyridin-2-ol (16.5 g, 97.9%, 97% purity) as a yellow solid. MS: m / z=300.9 [M+H] + .

[0176] b. 4-Chloro-5-iodo-2-methoxy-3-nitropyridine In a 500 mL round bottom flask were placed 4-chloro-5-iodo-3-nitropyridin-2-ol (16.5 g, 53.3 mmol, 97%), Ag2CO3 (15.5 g, 53.3 mmol, 95%) in toluene (310 mL). To this suspension was added CH3I (15.9 g, 107 mmol, 95%) at 50 °C and the mixture was stirred at 80 °C for 4 h. The precipitate was collected by filtration and discarded. The filtrate was evaporated to dryness under vacuum and the residue was purified by silica gel chromatography with ethyl acetate / petroleum ether (15:85). This gave 4-chloro-5-iodo-2-methoxy-3-nitropyridine (9.90 g, 52.6%, 89% purity) as a light yellow solid. MS: m / z = 315.5 [M+H] + .

[0177] c. 4-Chloro-5-iodo-2-methoxypyridin-3-amine In a 250 mL three-necked round bottom flask were placed 4-chloro-5-iodo-2-methoxy-3-nitropyridine (9.90 g, 28.0 mmol, 89%), iron (16.5 g, 281 mmol, 95%) and NH4Cl (9.40 g, 174 mmol, 99%) in ethanol (152 mL) and water (30 mL). The mixture was stirred in an oil bath at 80° C. for 2 h. The reaction mixture was filtered over Celite, washed with ethanol and the mother liquor was concentrated to dryness. The residue was stirred with 100 ml of water for 30 min and dried in vacuum at 60° C. This gave 4-chloro-5-iodo-2-methoxypyridin-3-amine (7.20 g, 75%, 83% purity) as an off-white solid, which was used in the next step without further purification. MS: m / z=285.9 [M+H] + .

[0178] dN-[7-iodo-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide In a 500 mL round bottom flask, 4-chloro-5-iodo-2-methoxypyridin-3-amine (7.20 g, 21.0 mmol, 83%) was placed in acetone (150 mL) and benzoyl isothiocyanate (5.21 g, 31.5 mmol, 99%) was added dropwise at room temperature. The solution was stirred in an oil bath at 50° C. for 1 h. The solid was collected by filtration, washed with acetone, and dried in vacuum to give N-[7-iodo-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (8.73 g, 91%, 90% purity) as a white solid. MS: m / z=412.2 [M+H] + .

[0179] eN-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide To a solution of N-[7-iodo-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (6.00 g, 13.1 mmol, 90%) and 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (6.13 g, 27.7 mmol, 95%) in dioxane (200 mL) and water (40.00 mL) was added NaOH (2.90 g, 68.9 mmol, 95%) and Pd(dppf)Cl2*dichloromethane (1.20 g, 1.40 mmol, 95%). After stirring for 1 h at 100 °C under nitrogen atmosphere, the mixture was concentrated to dryness under vacuum. The residue was purified using silica gel chromatography with ethyl acetate / hexane (95:5). This gave 3.32 g (62%, 90% purity) of N-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide as a colorless solid. MS: m / z=368.1 [M+H] + .

[0180] f.7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-amine To a stirred mixture of N-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (3.27 g, 8.00 mmol, 90%) in water / methanol (1:1, 300 mL) was added NaOH (3.36 g, 80.0 mmol, 95%) at room temperature under nitrogen atmosphere. The mixture was stirred overnight at 90° C. under nitrogen atmosphere and evaporated to dryness. The residue was taken up in water and extracted three times with dichloromethane (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and evaporated to dryness. The residue was purified by silica gel column chromatography, eluting with petroleum ether / ethyl acetate (1:1) to provide 7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-amine (1.50 g, 68%, 96% purity) as a light brownish solid. MS: m / z=264.1 [M+H] + .

[0181] g. Phenyl N-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5c]pyridin-2-yl]-N-(phenoxycarbonyl)carbamate To a stirred solution of 7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-amine (600 mg, 2.19 mmol, 96%) and phenyl chloroformate (1.81 g, 11.0 mmol, 95%) in THF (50 mL) was added K2CO3 (1.59 g, 11.0 mmol, 95%) and pyridine (913 mg, 11.0 mmol, 95%) at room temperature under nitrogen atmosphere. The mixture was stirred at 50 °C for 6 h and then quenched by addition of water (300 mL) after recooling to room temperature. The mixture was extracted three times with dichloromethane (200 mL) and the combined organic layers were washed once with brine (200 mL), dried over anhydrous Na2SO4, filtered, and evaporated to dryness under reduced pressure. This gave phenyl N-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]-N-(phenoxycarbonyl)carbamate (1.00 g, 69%, 76% purity) as a pale yellow solid. The crude product was used directly in the next step without further purification. MS: m / z=504.1 [M+H] + .

[0182] hN-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]-7-oxa-2-azaspiro[4.5]decane-2-carboxamide To a mixture of phenyl N-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]-N-(phenoxycarbonyl)carbamate (1.00 g, 1.52 mmol, 76.0) and bis(7-oxa-2-azaspiro[4.5]decane), oxalic acid (1.19 g, 3.03 mmol, 95%) in THF (50 mL) was added diisopropylethylamine (1.24 g, 9.09 mmol, 95%) at room temperature under nitrogen atmosphere. The mixture was stirred at 60° C. for 1 h. After recooling to room temperature, the mixture was extracted twice with dichloromethane (100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and evaporated to dryness. The residue was purified using silica gel column chromatography, eluting with petroleum ether / ethyl acetate (1:1) to provide N-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]-7-oxa-2-azaspiro[4.5]decane-2-carboxamide (600 mg, 92%) as a white solid. HPLC: 99.9% purity, RT=1.17 min. MS: m / z=431.1 [M+H]+. 1H NMR (300 MHz, DMSO-d6) δ 11.37 (s, 1H), 7.95 (s, 1H), 6.25 (s, 1H), 4.30-4.29 (m, 2H), 3.99 (s, 3H), 3.89 (t, J=5.4Hz, 2H), 3.61-3.29 (m, 8H), 2.55-2.51 (m, 2H), 1.82-1.54 (m, 6H).

[0183] i. (5R)-N-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-thiazolo[4,5-c]pyridin-2-yl]-7-oxa-2-azaspiro[4.5]decane-2-carboxamide 24 and (5S)-N-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-thiazolo[4,5-c]pyridin-2-yl]-7-oxa-2-azaspiro[4.5]decane-2-carboxamide 25 N-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]-7-oxa-2-azaspiro[4.5]decane-2-carboxamide (450 mg, 1.044 mmol, 1 equiv, 99.9%) was purified by chiral preparative HPLC (Preparative HPLC-032, column: ChiralPak IA, 2*25 cm, 5 μm; mobile phase, dichloromethane:ethanol (20:80); detector, UV). This gave (5R)-N-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]-7-oxa-2-azaspiro[4.5]decane-2-carboxamide (178 mg, 39%) as a white solid. HPLC: 99.7% purity, RT (chiral) = 3.86 min, 100% ee. MS: m / z = 431.2 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 11.36 (s,1H), 7.94 (s,1H), 6.24 (s,1H), 4.29-4.27 (m, 2H), 3.97 (s,3H), 3.88 (t, J=5.2 Hz, 2H), 3.51-3.19 (m, 8H), 2.55-2.50 (m, 2H), 1.83-1.53 ​​(m, 6H) and (5S)-N-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]-7-oxa-2-azaspiro[4.5]decane-2-carboxamide (171 mg, 38%) as a white solid. HPLC: 99.8% purity, RT (chiral) = 5.23 min, 99.9% ee. MS: m / z = 431.2 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 11.35 (s,1H), 7.94 (s,1H), 6.24 (s,1H), 4.29-4.28 (m, 2H), 3.99 (s, 3H), 3.88-3.85 (m, 2H), 3.61-3.29 (m, 8H), 2.55-2.50 (m,2H), 1.83-1.53 ​​(m,6H).

[0184] 2. N-[7-(3-aminophenyl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]-8-oxa-2-azaspiro[4.5]decane-2-carboxamide 39 [ka]

[0185] jN-[3-(2-benzamido-4-methoxy-1,3-benzothiazol-7-yl)phenyl]carbamate To a solution of N-(7-iodo-4-methoxy-1,3-benzothiazol-2-yl)benzamide (400 mg, 0.878 mmol, 90%) and (3-[[(tert-butoxy)carbonyl]amino]phenyl)boronic acid (328 mg, 1.32 mmol, 95%) in 1,4-dioxane and water was added NaOH (369 mg, 8.78 mmol, 95%) and Pd(dppf)Cl2*dichloromethane (113 mg, 0.132 mmol, 95%) under an inert atmosphere of nitrogen. After stirring at 100 °C under nitrogen overnight, the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether / ethyl acetate (0-100%, 40 min) to provide tert-butyl N-[3-(2-benzamido-4-methoxy-1,3-benzothiazol-7-yl)phenyl]carbamate (390 mg, 86%, 92% purity) as a yellow solid. MS: m / z=477.0 [M+H] + .

[0186] K. tert-Butyl N-[3-(2-amino-4-methoxy-thiazolo[4,5-c]pyridin-7-yl)phenyl]carbamate To a stirred solution of tert-butyl N-(3-[2-benzamido-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-7-yl]phenyl)carbamate (390 mg, 0.753 mmol, 92%) in MeOH was added dropwise NaOH (318 mg, 7.55 mmol, 95%) dissolved in water (10 mL) at room temperature. The mixture was stirred at 90° C. overnight, concentrated under vacuum, and the aqueous layer was extracted three times with dichloromethane (30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated to dryness under reduced pressure to yield tert-butyl N-[3-(2-amino-4-methoxy-thiazolo[4,5-c]pyridin-7-yl)phenyl]carbamate (220 mg, 54%, 69% purity) as a yellow solid. The crude product was used directly in the next step without further purification. MS: m / z=372.9 [M+H]+.

[0187] l. Phenyl N-[7-(3-[[(tert-butoxy)carbonyl]amino]phenyl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]-N-(phenoxycarbonyl)carbamate To a stirred mixture of tert-butyl N-(3-[2-amino-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-7-yl]phenyl)carbamate (220 mg, 0.405 mmol, 69%) and K2CO3 (294 mg, 2.02 mmol, 95%) in THF (15 mL) was added dropwise phenyl carbonochloridate (333 mg, 2.02 mmol, 95%) and pyridine (168 mg, 2.02 mmol, 95%) at room temperature under nitrogen atmosphere. The mixture was stirred at 50 °C for an additional 4 h. The reaction mixture was concentrated in vacuo, diluted with water (30 mL), and extracted three times with dichloromethane (30 mL). The combined organic layers were dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to yield phenyl N-[7-(3-[[(tert-butoxy)carbonyl]amino]phenyl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]-N-(phenoxycarbonyl)carbamate (350 mg, 74%, 52% purity) as a yellow solid. The crude product was used directly in the next step without further purification. MS: m / z=613.3 [M+H]+.

[0188] mN-[4-methoxy-7-(1-methyl-1H-pyrazol-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-4-(methoxymethyl)benzamide, 14 To a stirred mixture of phenyl N-[7-(3-[[(tert-butoxy)carbonyl]amino]phenyl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]-N-(phenoxycarbonyl)carbamate (350 mg, 0.297 mmol, 52%) and 8-oxa-2-azaspiro[4.5]decane hydrochloride (111 mg, 0.594 mmol, 95%) in THF was added diisopropylethylamine (242 mg, 1.78 mmol, 95%) dropwise at room temperature under a nitrogen atmosphere. The mixture was stirred at 60° C. overnight and then concentrated in vacuo. The residue was purified by silica gel column chromatography, eluting with petroleum ether / ethyl acetate (1:1) to provide tert-butyl N-[3-[4-methoxy-2-([8-oxa-2-azaspiro[4.5]decane-2-carbonyl]amino)-[1,3]thiazolo[4,5-c]pyridin-7-yl]phenyl]carbamate (176 mg, 89%, 81% purity) as a yellow solid. MS: m / z=540.3 [M+H]+.

[0189] N-[7-(3-aminophenyl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]-8-oxa-2-azaspiro[4.5]decane-2-carboxamide 39 To a stirred mixture of tert-butyl N-[3-[4-methoxy-2-([8-oxa-2-azaspiro[4.5]decane-2-carbonyl]amino)-[1,3]thiazolo[4,5-c]pyridin-7-yl]phenyl]carbamate (176 mg, 0.264 mmol, 81%) in MeOH was added dropwise a 4N solution of HCl in 1,4-dioxane (2.00 mL, 8.00 mmol, 95%) at 0° C. The mixture was stirred at room temperature for an additional 2 h, concentrated to dryness, and purified by preparative HPLC (2#SHIMADZU (HPLC-01), Column: XBridge Prep OBD C18 Column, 30x150mm 5μm; Mobile phase: Water / ACN (phase B 30% to 60%, 8min); Detector, 254UV) to produce N-[7-(3-aminophenyl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]-8-oxa-2-azaspiro[4.5]decane-2-carboxamide (60.0mg, 80%, 99% purity) as a white solid. HPLC: 99.0% purity, RT=3.19min. MS: m / z=440.1 [M+H]+. 1H NMR(400 MHz, DMSO-d6, ppm) : 11.31 (s, 1H), 7.93 (s, 1H), 7.16 (t, J = 7.8 Hz, 1H), 6.84 (t, J = 2.0 Hz, 1H), 6.78 - 6.73 (m, 1H), 6.66 - 6.59 (m, 1H), 5.30 (s, 2H), 4.02 (s, 3H), 3.69 - 3.44 (m, 4H), 1.81 (s, 2H), 1.50 (t, J = 5.4 Hz, 4H).

[0190] 3. N-(6-fluoro-4-methoxy-7-tetrahydropyran-4-yl-thiazolo[4,5-c]pyridin-2-yl)-8-oxa-2-azaspiro[4.5]decane-2-carboxamide [ka]

[0191] oN-[4-Methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide In a 500 mL round bottom flask, Pd / C (10%, 19.6 g) was added to a solution of N-[7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxy-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (4.10 g, 9.90 mmol, 89%) in 100 mL MeOH under nitrogen atmosphere. The mixture was stirred at 50° C. under hydrogen atmosphere for 3 days, filtered through a celite pad, and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography, eluted with petroleum ether / ethyl acetate (1:1) to provide N-[4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (2.60 g, 54%, 76% purity) as a yellow solid. MS: m / z=370.1 [M+H]+.

[0192] pN-[6-bromo-4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide In a 50 mL round bottom flask was placed N-[4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (2.60 g, 5.34 mmol, 76%), N-bromosuccinimide (1.20 g, 6.40 mmol, 95%) in DMF (30 mL). The resulting solution was stirred at room temperature for 18 h. The reaction was then quenched by the addition of water and concentrated to dryness. The residue was applied onto a silica gel column with ethyl acetate / petroleum ether (0-50%) to give N-[6-bromo-4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (2.50 g, 92%, 88% pure) as a yellow solid. MS: m / z=449.1 [M+H]+.

[0193] qN-[4-Methoxy-7-(oxan-4-yl)-6-(trimethylstannyl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide In a 50 mL pressure tank reactor purged and maintained with an inert atmosphere of nitrogen, N-[6-bromo-4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (2.50 g, 4.92 mmol, 88%), Pd(PPh3)4 (1.44 g, 1.18 mmol, 95%) and hexamethyldistannane (1.70 g, 4.92 mmol, 95%) in dioxane (35 mL) were placed. The resulting mixture was stirred at 110 °C for 1 h. The reaction was then quenched by the addition of water and concentrated to dryness. The residue was applied onto a silica gel column with ethyl acetate / petroleum ether (0-50%). This yielded 2.30 g (70%, 79% purity) of N-[4-methoxy-7-(oxan-4-yl)-6-(trimethylstannyl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide as a white solid. MS: m / z=449.1 [M+H]+.

[0194] rN-[6-fluoro-4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide In a 100 mL round bottom flask was placed N-[4-methoxy-7-(oxan-4-yl)-6-(trimethylstannyl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (2.30 g, 3.43 mmol, 79%), Selectfluor® (2.56 g, 6.85 mmol, 95%) in acetonitrile (50 mL). The resulting solution was stirred at room temperature for 18 h. The reaction was then quenched by the addition of water and concentrated to dryness. The residue was applied onto a silica gel column with ethyl acetate / hexane (0-50%). This yielded 1.50 g (91%, 81% purity) of N-[6-fluoro-4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide as a white solid. MS: m / z=388.1 [M+H]+.

[0195] s.6-Fluoro-4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-amine N-[6-fluoro-4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (1.50 g, 3.12 mmol, 81%) in MeOH (20 mL) was placed in a 20 mL sealed tube purged and maintained with an inert atmosphere of nitrogen. NaOH (1.27 g, 31.2 mmol, 98%) dissolved in water (20 mL) was added at rt and the resulting solution was stirred at 100° C. for 16 h. The resulting mixture was concentrated and the aqueous solution was extracted three times with 100 mL of dichloromethane. After filtration, the filtrate was evaporated to dryness and used without further purification to yield 6-fluoro-4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-amine (320 mg, 32%, 89% purity) as an off-white solid. MS: m / z=284.1 [M+H]+.

[0196] t. Phenyl N-[6-fluoro-4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-N-(phenoxycarbonyl)carbamate To a mixture of 6-fluoro-4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-amine (25 mg, 0.078 mmol, 89%) and K2CO3 (56.9 mg, 0.391 mmol, 95%) in THF (3.00 mL) was added dropwise phenyl chloroformate (64.5 mg, 0.391 mmol, 95%) and pyridine (32.6 mg, 0.391 mmol, 95%) at room temperature. The resulting mixture was stirred at 50 °C under nitrogen atmosphere for 6 h. The reaction was quenched by the addition of water (10 mL) and the resulting mixture was extracted twice with dichloromethane (10 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and evaporated to dryness under reduced pressure to give N-[6-fluoro-4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-N-(phenoxycarbonyl)carbamate (60.0 mg, 73%, 50% purity). The crude product was used directly in the next step without further purification. MS: m / z=524.1 [M+H]+.

[0197] N-[6-fluoro-4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-8-oxa-2-azaspiro[4.5]decane-2-carboxamide To a mixture of phenyl N-[6-fluoro-4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-N-(phenoxycarbonyl)carbamate (50 mg, 0.047 mmol, 50%) and 8-oxa-2-azaspiro[4.5]decane hydrochloride (26.5 mg, 0.142 mmol, 95%) in THF (3.00 mL) was added diisopropylethylamine (38.6 mg, 0.284 mmol, 95%) at room temperature. The resulting mixture was stirred at 60° C. under nitrogen atmosphere for 16 h. The mixture was diluted with water (10 mL) and extracted three times with dichloromethane (10 mL). The combined organic layer was dried over anhydrous Na2SO4, filtered, and evaporated to dryness under reduced pressure. The crude product was purified by preparative HPLC (2#SHIMADZU (HPLC-01): Column: XBridge Prep OBD C18 30x150mm, 5μm; Mobile phase: Water / Acetonitrile, Detector: UV). This resulted in N-[6-Fluoro-4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-8-oxa-2-azaspiro[4.5]decane-2-carboxamide (12.4mg, 57%) as a light yellow solid. HPLC: 97.9% purity, RT=5.93min. MS: m / z=451.2 [M+H]+. 1H NMR (400 MHz, DMSO-d6, ppm): 11.31 (s, 1H), 3.99-3.94 (m, 5H), 3.61-3.45 (m, 10H), 3.12 (t, J=12.4 Hz, 1H), 2.03 (q, J=12.5 Hz, 2H), 1.82 (s, 2H), 1.65-1.62 (m,2H),1.50 (s, 4H).

[0198] 4. N-[5-(difluoromethyl)-4-oxo-7-phenyl-5 lambda 4-[1,3]thiazolo[4,5-c]pyridin-2-yl]-8-oxa-2-azaspiro[4.5]decane-2-carboxamide [ka]

[0199] v N-[4-hydroxy-7-phenyl-[1,3]thiazolo[4,5-c]pyridin-2-yl]-8-oxa-2-azaspiro[4.5]decane-2-carboxamide In a 25 mL round-bottom flask was placed N-[4-methoxy-7-phenyl-[1,3]thiazolo[4,5-c]pyridin-2-yl]-8-oxa-2-azaspiro[4.5]decane-2-carboxamide (200 mg, 0.458 mmol, 97%) in dichloromethane (8 mL). BBr3 (0.138 mL, 0.138 mmol, 1 M in dichloromethane) was added dropwise at 0° C., and the resulting solution was stirred at 0° C. for an additional 2 h. The reaction was then quenched by the addition of ice water. The mixture was extracted three times with dichloromethane (10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and evaporated to dryness under reduced pressure. The residue was applied onto a silica gel column with dichloromethane / methanol (0-30%) to yield 150 mg (78%, 98% purity) of N-[4-hydroxy-7-phenyl-[1,3]thiazolo[4,5-c]pyridin-2-yl]-8-oxa-2-azaspiro[4.5]decane-2-carboxamide as a white solid. MS: m / z=411.2 [M+H]+.

[0200] w N-[5-(difluoromethyl)-4-oxo-7-phenyl-5 lambda 4-[1,3]thiazolo[4,5-c]pyridin-2-yl]-8-oxa-2-azaspiro[4.5]decane-2-carboxamide In a 50 mL sealed tube purged and maintained with an inert atmosphere of nitrogen, N-[4-hydroxy-7-phenyl-[1,3]thiazolo[4,5-c]pyridin-2-yl]-8-oxa-2-azaspiro[4.5]decane-2-carboxamide (100 mg, 0.238 mmol, 98%), anhydrous Na2SO4 (3.37 mg, 0.024 mmol, 98%) in acetonitrile (5 mL) was placed. To this mixture, 2,2-difluoro-2-(fluorosulfonyl)acetic acid (50.8 mg, 0.285 mmol, 98%) was added at room temperature, and the resulting solution was stirred at room temperature for an additional 3 h. The reaction was then quenched by the addition of water (20 mL). The mixture was extracted three times with dichloromethane (10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and evaporated to dryness under reduced pressure. The crude product was purified by preparative HPLC (2#SHIMADZU (HPLC-01): Column; Xselect CSH OBD Column 30*150mm, 5μm, Mobile phase: water / acetonitrile; Detector: UV). This gave N-[5-(difluoromethyl)-4-oxo-7-phenyl-5lambda 4-[1,3]thiazolo[4,5-c]pyridin-2-yl]-8-oxa-2-azaspiro[4.5]decane-2-carboxamide (20.0mg,17%) as a white solid. Mp=225~226℃. HPLC: 95.1% purity, RT=6.27min. MS: m / z=461.2 [M+H]+. 1H NMR (400 MHz, DMSO-d6, ppm) 11.57 (s, 1H), 8.08 (d, J = 8.6 Hz, 1H), 7.89 (s, 1H), 7.72 (d, J = 7.9 Hz, 2H), 7.54 (dt, J = 32.6, 7.5 Hz, 3H), 3.52 (s, 7H), 3.30 (s, 1H), 1.81 (d, J = 38.2 Hz, 2H), 1.50 (s, 4H).

[0201] Example 3: Compounds of the invention are tested for inhibitory activity against human adenosine receptors in recombinant cells. Human A 2A , A 2BFunctional activity of A, A1, and A3 receptors was determined by quantification of cAMP, the second messenger of adenosine receptors. For this purpose, human A 2A Or A 2B Recombinant HEK293 cells expressing either of the receptors (both Gs-coupled) were seeded into 394-well microtiter plates and test compounds and agonists (NECA) were added. After 15 min of incubation, HTRF reagent (cAMP dynamic 2, Cis Bio) was added and cellular cAMP levels were determined using an ENVISION (Perkin Elmer) plate reader.

[0202] For human A1 and A3 receptors, recombinant CHO cells expressing either the A1 or A3 receptor were used. As both receptors are coupled to Gi proteins, the following assay protocol was adapted: Cells were seeded into 384-well plates and forskolin, test compounds, and agonists (CPA for A1 receptors and IB-MECA for A3 receptors) were added. After 30 min of incubation, HTRF reagent (cAMP dynamic 2, Cis Bio) was added and cellular cAMP levels were determined using an ENVISION (Perkin Elmer) plate reader. The resulting raw data was normalized to inhibitor and neutral controls (DMSO) and the normalized data was fitted using GeneData software.

[0203] The compounds of the present invention bind to adenosine A1 and A3 receptors. 2A and A 2B They show high selectivity for the receptor (see, for example, the data for some of the examples of compounds of the invention in Table 4).

[0204] Specifically, the known adenosine A 2A In contrast to the receptor antagonists tozadenant and similar benzothiazole derivatives (see Table 5), the compounds of the present invention surprisingly exhibited the following properties: 2A / A2B The compounds of the present invention exhibit dual activity (see Table 4), which is preferred for the treatment and / or prevention of hyperproliferative and infectious diseases and disorders as disclosed above. Alternatively, the compounds of the present invention exhibit at least high A activity, together with other surprising advantages disclosed herein. 2A They exhibit inhibitory activity, which translates into high efficacy in the treatment and / or prevention of hyperproliferative and infectious diseases and disorders.

[0205] Table 4 - Compounds of the invention [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4]

[0206] Table 5 - Prior art compounds specifically disclosed in WO 2005 / 028484 and WO 2005 / 000842 [Table 6-1] [Table 6-2] [Table 6-3] [Table 6-4]

[0207] Example 4: Endogenous human A 2A Testing the effect of the compounds of the present invention on the receptor Endogenous Gs-coupled human A 2A The functional activity of the receptor was measured in T cells where this receptor is highly expressed. Receptor activity was determined by quantification of cAMP, the second messenger of the adenosine receptor.

[0208] Briefly, human pan T cells were isolated from human PBMCs derived from fresh whole blood (MACS Pan T Cell Isolation Kit, Miltenyi Biotec). T cells were seeded in 384-well microtiter plates and treated with test compounds. After 10 min of incubation at room temperature, A 2A The adenosine receptor agonist CGS-21680 was added and the plates were incubated for another 45 min. Finally, HTRF reagent (cAMP Femto Kit, CisBio) was added to the wells and cellular cAMP levels were determined 1 h later using an ENVISION (Perkin Elmer) plate reader.

[0209] The resulting raw data was normalized to inhibitor and neutral controls (DMSO) and the normalized data was fitted using Genedata Screener software.

[0210] The compounds of the present invention also include 2A A expressed on human T cells incubated with the adenosine receptor agonist CGS-21680 2A It has been shown that the compounds of the present invention also inhibit the receptor (as measured by quantifying cAMP), which is favorable for the treatment and / or prevention of hyperproliferative and infectious diseases and disorders as disclosed above.Therefore, the compounds of the present invention can surprisingly prevent immunosuppression, and thus support the inhibition of tumor growth induced by antitumor T cells, the reduction or destruction of metastatic cancer, and the prevention of neovascularization.

[0211] Example 5: Testing the pharmacokinetic properties of compounds of the invention in rats and mice The aim of this study was to obtain information regarding the pharmacokinetic properties of compounds of the invention in female Wistar rats / mice after single intravenous and oral administration.

[0212] material and method: Animal experiments (in-life phase) Female Wistar rats / mice (n=6) received either a single intravenous (bolus) injection or oral administration (by gavage) of the test compounds. Doses of 0.2 and 10 mg / kg (per compound) were given intravenously and orally (per os), respectively, as a solution in DMSO (0.2%) / PEG 200 (40%) / water for iv administration and as a suspension in Methocel (0.5%) / Tween 20 (0.25%) in water for oral dosing. Serial blood samples were taken sublingually from three animals per dosing route after 0.1 (iv only), 0.25 (po only), 0.5, 1, 2, 4, 6, and 24 h under isoflurane inhalation and further processed to obtain plasma. Urine and fecal samples from three rats per dosing route were also collected over the time interval 0-24 h and pooled for analysis.

[0213] Bioanalytics: Compound concentrations in plasma and feces were quantified using a UPLC method with tandem mass spectrometry detection (LC-MS / MS) previously developed at the "Institute of Drug Metabolism and Pharmacokinetics". The LC-MS / MS system consisted of a Waters Acquity UPLC interfaced with an AB Sciex mass spectrometer API 5500 Q-trap. UPLC separation was performed on a reversed-phase column (HSS T3, 1.8 μM, 2.1×50 mm) using a gradient of the mobile phase with 0.1% formic acid and acetonitrile as eluents. Compound detection was performed using multiple reaction monitoring in positive ionization mode. Plasma samples were spiked with an internal standard (20 μl) and the analytes were extracted from the matrix using tertiary butyl methyl ether (tBME). The organic phase was evaporated to dryness under a stream of nitrogen. The residue was dissolved in acetonitrile / 0.1% formic acid for LC-MS / MS analysis. Fecal samples were homogenized in four volumes of an ethanol / water mixture (4:1, v / v). Aliquots of the aqueous ethanol extracts were spiked with an internal standard, diluted with acetonitrile / water (1:1, v / v) and directly injected into the LC-MS / MS system.

[0214] Pharmacokinetic evaluation: Pharmacokinetic parameter C max and t max were obtained from the data obtained. Area under the curve (AUC), clearance (CL), volume (V), half-life (t 1 / 2 ), F, and all dose-normalized values ​​were calculated using the custom-written software "DDS-TOX". "DDS-TOX" values ​​were evaluated for several compounds and shown to be comparable to those given by the validated software WinNonLin. AUC values ​​were calculated by non-compartmental analysis using the linear up / log down method. Numerical data for mean plasma concentrations and derived pharmacokinetic parameters were rounded to three significant figures for presentation. Oral bioavailability and excretion data - expressed as % of dose - are presented using two significant figures.

[0215] Adenosine A, also known as 2A In contrast to the receptor antagonist tozadenant and similar benzothiazole derivatives, the compounds of the present invention surprisingly show better pharmacokinetic properties in mice as a cancer-relevant animal model (see Table 6), which is favorable for the treatment and / or prevention of hyperproliferative and infectious diseases and disorders as disclosed above.

[0216] Table 6 [Table 7]

[0217] Example 6: Testing the effect of compounds of the invention on mouse T cells background: Adenosine (Ado) in the tumor microenvironment is 2A Signaling through the receptor can inhibit T cell activity and suppress cytokine secretion by T cells. 2A Specific agonists act similarly, inhibiting T cell cytokine secretion in vitro and in vivo. 2A Antagonist or A 2A / A 2B Dual antagonists can rescue T cells from this inhibition. Herein, we use pan-T cells from mouse spleens to investigate the potency of latent Aβ. 2A Antagonist or A 2A / A 2B We describe an in vitro system that we established to screen dual antagonists for their activity. The method described uses potential Aβ antagonists to assess their potentiation of T cell cytokine production. 2A Or A 2A / A 2B A with dual antagonists 2AIt involves the use of CD3 / CD28 pre-coated beads to stimulate pan T cells purified from mouse splenocytes in combination with the addition of agonist.

[0218] mT Cell Assay, cAMP, Assay Description: Pan-T cells are isolated and purified from six black mice by manual dissociation using Miltenyi's Pan-T cell isolation kit. The isolated T cells are activated in 96-well plates for 48 hours using anti-CD3 / CD28 beads in T cell proliferation medium. After 48 hours, T cells are pooled, counted, and suspended in serum-free medium containing 0.1% BSA. Cells are plated at 50,000 cells per well in 10ul of medium and incubated at 37°C for 2 hours. Test compounds are dispensed into the wells in a 10-step dose response starting at a concentration of 10uM. Following compound addition, the agonist NECA is added to all wells at a concentration of 1uM. Plates are incubated at 37°C for 30 minutes and assayed for cAMP levels using Cisbio cAMP Dynamic2 reagent kit by adding 10ul of kit reagent to each well. Plates are incubated at room temperature for 1 hour and HTRF signals are read on an Envision plate reader. Raw data is analyzed in Genedata Screener and the resulting data is loaded into a database.

[0219] mT cell assay, IL-2, Assay description: Briefly, mouse pan-T cells are purified from the spleens of BALB / c mice using the pan-T cell isolation kit Mouse II (MACS Miltenyi biotech Cat# Order no. 130-095-130) according to the manufacturer's protocol. Purified T cells are seeded in Nunc™ 96-well polystyrene round-bottom microwell plates in RPMI medium with 10% heat-inactivated fetal bovine serum. Cells are rested for 1 h at 37°C before activation with CD3 / CD28 pre-coated beads (Dynabeads™ Mouse T-Activator CD3 / CD28; Cat# 11456D). After 30 min, cells are treated with varying doses of the test antagonist(s). Cells are incubated for 1 h at 37°C ... 2A An additional 30 min incubation is performed at 37°C before treatment with the agonist NECA (1 μM) or neutral control (DMSO). After 24 h incubation, IL-2 levels in the supernatants are measured by ELISA (R&D systems Cat# DY402 (IL-2); DY485 (IFN-γ)) according to the manufacturer's protocol. Once concentrations are calculated, the difference in cytokine concentrations between the DMSO control and the agonist alone control is calculated and the percentage of rescue by each concentration of antagonist is calculated by using Microsoft Excel. These percentages of antagonist dose-dependent cytokine rescue are plotted using GraphPad Prism software to obtain IC 50 Calculate.

[0220] Adenosine A, also known as 2A In contrast to tozadenant and similar compounds that are receptor antagonists (see Table 8), the compounds of the present invention are capable of rescuing T cells from inhibition and are also capable of inhibiting A receptors such as adenosine or NECA. 2AIt is also possible to prevent the suppression of cytokine secretion as induced by specific agonists (see Table 7), which is preferable for the treatment and / or prevention of hyperproliferative and infectious diseases and disorders as disclosed above. Thus, the compounds of the present invention can surprisingly prevent immunosuppression, and thus support the inhibition of tumor growth induced by antitumor T cells, the reduction or destruction of metastatic cancer, and the prevention of neovascularization.

[0221] Table 7 - Compounds of the invention [Table 8-1] [Table 8-2] [Table 8-3] [Table 8-4] [Table 8-5]

[0222] Table 8 - Prior art compounds specifically disclosed in WO 2005 / 028484 and WO 2005 / 000842 [Table 9-1] [Table 9-2] [Table 9-3]

[0223] Example 7: Testing the effect of compounds of the invention on human T cells Endogenous Gs-coupled human A 2A The functional activity of the receptor was measured in T cells where this receptor is highly expressed. Receptor activity was determined by quantification of cAMP, the second messenger of the adenosine receptor.

[0224] Assay description Briefly, human pan T cells were isolated from human PBMCs derived from fresh whole blood (MACS Pan T Cell Isolation Kit, Miltenyi Biotec). T cells were seeded in 384-well microtiter plates and treated with test compounds. After 10 min of incubation at room temperature, A 2A The adenosine receptor agonist NECA was added and the plates were incubated for an additional 45 min. Finally, HTRF reagent (cAMP Femto Kit, CisBio) was added to the wells and cellular cAMP levels were determined after 1 h using an ENVISION (Perkin Elmer) plate reader.

[0225] The resulting raw data was normalized to inhibitor and neutral controls (DMSO) and the normalized data was fitted using Genedata Screener software.

[0226] The compounds of the present invention are 2A A expressed on human T cells incubated with the adenosine receptor agonist NECA 2A It shows that the compound of the present invention can inhibit the receptor (as measured by quantifying cAMP), which is favorable for the treatment and / or prevention of hyperproliferative and infectious diseases and disorders as disclosed above.Therefore, the compound of the present invention can surprisingly prevent immunosuppression, and thus support the inhibition of tumor growth induced by antitumor T cells, the reduction or destruction of metastatic cancer, and the prevention of neovascularization.

[0227] Table 9 - Compounds of the invention [Table 10-1] [Table 10-2] [Table 10-3] [Table 10-4] [Table 10-5] [Table 10-6] [Table 10-7]

[0228] Table 10 - Prior art compounds specifically disclosed in WO 2005 / 028484 and WO 2005 / 000842 [Table 11-1] [Table 11-2] [Table 11-3]

[0229] Example 8: Injection vial A solution of 100 g of the compound of the invention and 5 g of disodium hydrogen phosphate in 3 liters of bidistilled water is adjusted to pH 6.5 using 2N hydrochloric acid, filtered under sterile conditions, transferred into injection vials, lyophilized under sterile conditions and sealed under sterile conditions, each injection vial containing 5 mg of the compound of the invention.

[0230] Example 9: Solution A solution is prepared from 1 g of the compound of the invention, 9.38 g of NaH2PO4·2H2O, 28.48 g of Na2HPO4·12H2O, and 0.1 g of benzalkonium chloride in 940 ml of double-distilled water. The pH is adjusted to 6.8, the solution made up to 1 liter, and sterilized by radiation.

[0231] Example 10: Ampoule A solution of 1 kg of the compound of the invention in 60 L of double-distilled water is filtered under sterile conditions, transferred into ampoules, lyophilized under sterile conditions and sealed under sterile conditions, each ampoule containing 10 mg of the compound of the invention.

Claims

1. below, 【Table 1-1】 【Table 1-2】 【Table 1-3】 【Table 1-4】 【Table 1-5】 【Table 1-6】 【Table 1-7】 【Table 1-8】 【Table 1-9】 【Table 1-10】 or a physiologically acceptable salt, solvate or stereoisomer thereof, or mixtures thereof in any ratio.

2. Adenosine A ２Ａ And / or A ２Ｂ 2. A compound according to claim 1 or a physiologically acceptable salt, solvate or stereoisomer thereof or a mixture thereof in any ratio as a receptor inhibitor.

3. 10. A pharmaceutical preparation comprising at least one compound according to claim 1, or a physiologically acceptable salt, solvate or stereoisomer thereof, or a mixture thereof in any ratio.

4. The pharmaceutical preparation according to claim 3 , further comprising an excipient and / or an adjuvant.

5. 13. A pharmaceutical preparation comprising at least one compound according to claim 1, or a physiologically acceptable salt, solvate, or stereoisomer thereof, or mixtures thereof in any ratio, and at least one further pharma- ceutical active compound.

6. A process for the preparation of a pharmaceutical preparation, characterized in that the compound according to claim 1, or one of its physiologically acceptable salts, solvates or stereoisomers, or mixtures thereof in any ratio, is brought into a suitable dosage form together with solid, liquid or semi-liquid excipients or adjuvants.

7. A medicament comprising at least one compound according to claim 1 or one of its physiologically acceptable salts, solvates or stereoisomers, or mixtures thereof in any ratio, for use in the treatment and / or prophylaxis of physiological and / or pathophysiological conditions.

8. A medicament comprising at least one compound according to claim 1 or one of its physiologically acceptable salts, solvates or stereoisomers, or mixtures thereof in any ratio, for use in the treatment and / or prophylaxis of physiological and / or pathophysiological conditions selected from the group consisting of hyperproliferative and infectious diseases and disorders.

9. The method of claim 8, wherein the hyperproliferative disease or disorder is cancer.

10. Cancers include acute and chronic lymphocytic leukemia, acute granulocytic leukemia, adrenal cortical carcinoma, bladder cancer, brain cancer, breast cancer, cervical cancer, cervical hyperplasia, cervical cancer, choriocarcinoma, chronic granulocytic leukemia, chronic lymphocytic leukemia, colorectal cancer, endometrial cancer, esophageal cancer, essential thrombocytosis, genitourinary cancer, glioma, glioblastoma, hairy cell leukemia, head and neck cancer, Hodgkin's disease, Kaposi's sarcoma, lung cancer, lymphoma, malignant carcinoid cancer, malignant hypercalcemia, malignant melanoma, malignant pancreatic insulinoma, and pulmonary hypercalcemia.

10. The pharmaceutical composition of claim 9, wherein the cancer is selected from the group consisting of linoma, medullary thyroid carcinoma, melanoma, multiple myeloma, mycosis fungoides, myeloid and lymphocytic leukemia, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, osteosarcoma, ovarian cancer, pancreatic cancer, polycythemia vera, primary brain tumor, primary macroglobulinemia, prostate cancer, renal cell carcinoma, rhabdomyosarcoma, skin cancer, small cell lung cancer, sarcoma of soft tissue, squamous cell carcinoma, gastric cancer, testicular cancer, thyroid cancer, and Wilms' tumor.

11. The pharmaceutical composition of claim 8, wherein the hyperproliferative disease or disorder is selected from the group consisting of age-related macular degeneration, Crohn's disease, cirrhosis of the liver, disorders related to chronic inflammation, proliferative diabetic retinopathy, proliferative vitreoretinopathy, retinopathy of prematurity, granulomatous disease, immune hyperproliferation associated with organ or tissue transplantation, and immune proliferative diseases or disorders (selected from the group consisting of inflammatory bowel disease, psoriasis, rheumatoid arthritis, systemic lupus erythematosus (SLE), vascular hyperproliferation secondary to retinal hypoxia, and vasculitis).

12. An infectious disease or disorder i. virus-induced infectious diseases caused by retroviruses, hepadnaviruses, herpesviruses, flaviviridae, and / or adenoviruses, wherein the retrovirus is selected from lentiviruses or oncoretroviruses, wherein the lentiviruses are selected from the group consisting of HIV-1, HIV-2, FIV, BIV, SIVs, SHIV, CAEV, VMV, and EIAV, and the oncoretroviruses are selected from the group consisting of HTLV-I, HTLV-II, and BLV, the hepadnaviruses are selected from the group consisting of HBV, GSHV, and WHV, the herpesviruses are selected from the group consisting of HSV I, HSV II, EBV, VZV, HCMV, or HHV8, and the flaviviridae are selected from the group consisting of HCV, West Nile, and yellow fever; ii. Bacterial infections caused by gram-positive bacteria, where the gram-positive bacteria are methicillin-susceptible and methicillin-resistant staphylococci (including Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus saprophyticus, and coagulase-negative staphylococci), glycopeptide-intermediate-susceptible Staphylococcus aureus (GISA), penicillin-susceptible and penicillin-resistant streptococci (including Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus avium, Streptococcus bovis, Streptococcus lactis, Streptococcus sanguis, and group C streptococci (GCS), group G streptococci (GGS), and viridans streptococci), enterococci (Enterococcus faecalis and Enterococcus faecium), Clostridium difficile, Iisteria monocytogenes, Corynebacterium jeikeium, Chlamydia spp (including C. pneumoniae), and Mycobacterium tuberculosis; iii. Bacterial infections caused by gram-negative bacteria, wherein the gram-negative bacteria are selected from the group consisting of Enterobacteriaceae including Escherichia spp. (including Escherichia coli), Klebsiella spp., Enterobacter spp., Citrobacter spp., Serratia spp., Proteus spp., Providencia spp., Salmonella spp., Shigella spp., Pseudomonas including P. aeruginosa, Moraxella spp. (including M. catarrhalis), Haemophilus spp., and Neisseria spp.; iv. Infectious diseases induced by intracellular active parasites selected from the group consisting of the phylum Apicomplexa, or Sarcomastigophora (including Trypanosoma, Plasmodia, Leishmania, Babesia, or Theileria), Cryptosporidia, Sacrocystida, Amoebia, Coccidia, and Trichomonadia. The pharmaceutical composition according to claim 8, selected from the group consisting of:

13. a) an effective amount of a compound according to claim 1 or 2, or a physiologically acceptable salt, solvate, or stereoisomer thereof, or mixtures thereof in any ratio, and b) an effective amount of a further pharma- ceutical active compound; A set (kit) consisting of individual packs.

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