RU2790011C2 - Thiazolopyridine derivatives as adenosine receptor antagonists - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to thiazolopyridine derivatives of the general formula I, as well as to methods for their production and the use as A_2A and/or A_2B adenosine receptor inhibitor.

EFFECT: new compounds are obtained, which can be used for the treatment and/or prevention of hyperproliferative or infectious diseases mediated with inhibition of A_2A and/or A_2B adenosine receptor.

14 cl, 7 tbl, 9 ex

Description

The invention relates to thiazolopyridine derivatives of general formula I,

and to the use of the compounds of the present invention for the treatment and/or prevention of hyperproliferative or infectious diseases and disorders in mammals, especially humans, and to pharmaceutical compositions containing such compounds.

Prerequisites for the creation of the invention

Adenosine is a common modulator of numerous physiological actions, especially in the cardiovascular, nervous, and immune systems. Adenosine is associated both structurally and metabolically with bioactive nucleotides: adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP) and cyclic adenosine monophosphate (cAMP), with the biochemical methylation agent S-adenosyl-L-methion (SAM) and structurally with coenzymes NAD, FAD and coenzyme A and with RNA.

Through cell surface receptors, adenosine modulates various physiological functions, including sedation induction, vasodilation, suppression of heart rate and contractility, inhibition of platelet aggregation, stimulation of gluconeogenesis, and inhibition of lipolysis. Studies show that adenosine is able to activate adenylate cyclases, open potassium channels, decrease calcium channel flux, and inhibit or stimulate phosphoinositide metabolism via receptor-mediated mechanisms (Muller CE and Stein B., Current Pharmaceutical Design, 2:501, 1996; Muller CE , Exp Opin Ther Patents 7 (5): 419, 1997).

Adenosine receptors belong to the G-protein coupled receptor (GPCR) superfamily. The four main subtypes of adenosine receptors have been pharmacologically, structurally and functionally characterized (Fredholm et al., Pharm. Rev., 46: 143-156, 1994) and designated as A ₁ , A _2A , A _2B and A ₃ . Although the same adenosine receptor can bind to different G proteins, the A ₁ and A ₃ adenosine receptors typically bind to inhibitory G proteins called G _i and G ₀ , which inhibit adenylate cyclase and reduce cAMP levels in cells. In contrast, adenosine receptors A _2A and A _2B are coupled to stimulatory G proteins called G _S , which activate adenylate cyclase and increase cAMP levels in cells (Linden J., Annu. Rev. Pharmacol. Toxicol., 41: 775- 87 2001).

According to the invention, "adenosine receptor selective ligands" are substances that selectively bind to one or more adenosine receptor subtypes, thereby either mimicking the action of adenosine (adenosine agonists) or blocking its action (adenosine antagonists). According to their receptor selectivity, ligands that are selective for adenosine receptors can be divided into different categories, for example, ligands that selectively bind to A ₁ or A ₂ receptors, and in the case of the latter also, for example, those that selectively bind to A _2A receptors. or _A2B . Also possible are adenosine receptor ligands that selectively bind to multiple adenosine receptor subtypes, for example, ligands that bind selectively to A ₁ and A ₂ but not to A ₃ receptors. The selectivity of the aforementioned receptors can be determined by the effect of substances on cell lines that, after stable transfection of the appropriate cDNA, express the receptor subtypes in question (Olah, ME et al., J. Biol. Chem., 267: 10764-10770, 1992). The effect of substances 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).

The A ₁ receptor system is known to include activation of phospholipase C and modulation of both potassium and calcium ion channels. The A ₃ subtype, in addition to its association with adenylate cyclase, also stimulates phospholipase C and thus activates calcium ion channels.

The A ₁ receptor (326-328 amino acids) has been cloned from various species (canine, human, rat, dog, chicken, cattle, guinea pig) with 90-95% sequence identified among mammalian species. The A _2A receptor (409-412 amino acids) has been cloned from canines, rats, humans, guinea pigs and mice. The A _2B receptor (332 amino acids) was cloned from human and mouse with 45% homology of human A _2B to human A ₁ and A _2A receptors. The A ₃ receptor (317-320 amino acids) has been cloned from human, rat, dog, rabbit and sheep.

The A ₁ and A _2A receptor subtypes are hypothesized to play complementary roles in the regulation of adenosine energy supply. Adenosine, which is a product of ATP metabolism, diffuses out of the cell and acts locally to activate adenosine receptors, decreasing oxygen demand (A ₁ and A ₃ ) or increasing oxygen supply (A _2A ), and thus restoring the balance of energy supply/energy consumption in tissues. The action of both subtypes is to increase the amount of oxygen available to tissues and to protect cells from damage caused by transient oxygen imbalances. One of the important functions of endogenous adenosine is to prevent injury damage such as hypoxia, ischemia, hypotension and seizure activity. In addition, it is known that binding of an adenosine receptor agonist to mast cells expressing the rat A ₃ receptor resulted in an increase in the concentration of inositol triphosphate and intracellular calcium, which increased the antigen-induced secretion of inflammatory mediators. Therefore, the A ₃ receptor plays a role in mediating asthma attacks and other allergic reactions.

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

Regarding the role of adenosine in the nervous system, the first observations were made on the effects of caffeine, which is the most widely used of all psychoactive drugs. In fact, caffeine is a known adenosine receptor antagonist that has been shown to enhance perception and learning in mammals. The adenosine A _2A receptor pathway is responsible for these effects (Fredholm et al., Pharmacol. Rev., 51 (1): 83-133, 1999; Huang et al., Nat Neurosci., 8 (7): 858-9, 2005 ), and the effect of caffeine on the adenosine A _2A receptor signaling pathway, has stimulated research into highly specific and potent adenosine A _2A antagonists.

In mammals, A _2A adenosine receptors have a limited distribution in the brain and are 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, lungs and blood vessels. In the peripheral system, G ₃ appears to be the main G protein associated with the A _2A adenosine receptor, but in the striatum, striatal A _2A adenosine receptors have been shown to mediate their action through activation of a G protein called Goif (KuIl et al. ., MoI Pharmacol., 58 (4): 772-7, 2000), which is similar to G ₃ and also associated with adenylate cyclase.

To date, studies in genetically modified mice and pharmacological analyzes indicate that the A _2A receptor is a promising therapeutic target for the treatment of central nervous system (CNS) disorders and diseases 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 Bi osci., 13, 2614-2632, 2008), as well as for the treatment of various psychoses of organic origin (Weiss et al., Neurology, 61 (11 Suppl 6): 88-93, 2003).

The use of A _2A adenosine receptors in knockout mice has shown that inactivation of the A _2A adenosine receptor protects against ischemia-induced neuronal cell death (Chen et al., J. Neurosci., 19 (21): 9192-200, 1999 and Monopoli et al. , Neuroreport, 9 (17): 3955-9, 1998) and mitochondrial toxin 3-NP (Blum et al., J. Neurosci., 23 (12): 5361-9, 2003). These results provided the basis for the treatment of ischemia and Huntington's disease with adenosine A _2A antagonists. Blockade of A _2A adenosine receptors also has an antidepressant effect (El Yacoubi et al., Neuropharmacology, 40 (3): 424-32, 2001). In conclusion, this blockade prevents memory dysfunction (Cunha et al., Exp. Neurol., 210 (2): 776-81, 2008; Takahashi et al., Front. Biosci., 13: 2614-32, 2008), and this may be a promising therapeutic route for the treatment and/or prevention of Alzheimer's disease.

For a review on A _2A adenosine receptors, see eg Moreau et al. (Brain Res. Reviews 31: 65-82, 1999) and Svenningsson et al. (Progress in Neurobiology 59: 355-396, 1999).

To date, several A _2A adenosine receptor antagonists have shown promising potential for the treatment of Parkinson's disease. As an example, KW-6002 (Istradefillin) completed a phase III clinical trial in the US after studies demonstrated its effectiveness in alleviating disease symptoms (Bara-Himenez et al., Neurology, 61 (3): 293-6, 2003 and Hauser et al., Neurology, 61 (3): 297-303, 2003). SCH420814 (Preladenant) is currently in Phase II clinical trial in the USA and provides improved motor function in animal models of Parkinson's disease (Neustadt et al., Bioorg. Med. Chem. Lett., 17 (5): 1376-80, 2001 ), as well as in human patients (Hunter J. C, poster Boston 2006 - http://www.a2apd.org/Speaker abstracts/Hunter.pdf).

In addition to the utility of A _2A receptor antagonists in the treatment of neurodegenerative diseases, these compounds have been considered for additional symptomatic indications. They are based on evidence that activation of the A _2A receptor may contribute to the pathophysiology of a number of neuropsychiatric disorders and dysfunctions such as depression, excessive daytime sleepiness, restless leg 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).

Some authors propose the use of A _2A antagonists for the treatment of diabetes (WO 1999035147; WO 2001002400). Other studies suggest involvement of A _2A adenosine receptors in wound healing or atrial fibrillation (Am. J. Path., 6, 1774-1778, 2007; Arthritis & Rheumatism, 54(8), 2632-2642, 2006).

Some of the potent adenosine A _2A antagonists discovered in the past by pharmaceutical companies have moved into clinical trials showing positive results and demonstrating the potential of this class of compounds for the treatment of neurodegenerative disorders such as Parkinson's, Huntington's or Alzheimer's, as well as other diseases associated with CNS such as depression, anxiety syndrome, sleep disturbances and anxiety disorder (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 herein; mov. Disorders, 25(2), S305, 2010).

Known A _2A inhibitors are istradefillin (KW-6002), preladenant (SCH420814), SCH58261, CGS15943, tosadenant, vipadenant (V-2006), V-81444 (CPI-444, HTL-1071, PBF-509, Medi-9447, PNQ-370, ZM-241385, ASO-5854, ST-1535, ST-4206, DTI 133 and DT-0926, which are in most cases developed for the treatment of Parkinson's disease.

A _2B adenosine 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) using standard polymerase chain reaction techniques with degenerate oligonucleotide primers. designed to recognize conserved regions of most G-protein coupled receptors. The human A _2B receptor shares 86 to 87% amino acid sequence homology with rat and mouse A _2B receptors (Rivkees and Reppert, 1992; Pierce et al., 1992; Marquardt et al., 1994) and 45% amino acid sequence homology with human receptors. A ₁ and A _2A . As expected for closely related species, rat and mouse A _2B receptors share 96% amino acid sequence homology. In comparison, the overall amino acid identity between A ₁ receptors of different species is 87% (Palmer and Stiles, 1995). The A _2A receptors have 90% homology between species (Ongini and Fredholm, 1996), with most differences occurring in the 2nd extracellular loop and the long C-terminal domain (Palmer and Stiles, 1995). The lowest (72%) degree of identity between species is observed for the A ₃ receptor sequences (Palmer and Stiles, 1995).

The adenosine analog NECA remains the most potent A _2B agonist (Bruns, 1981; Feoktistov and Biaggioni, 1993, 1997; Brackett and Daly, 1994), with a concentration that produces a half-maximal effect (EC ₅₀ ) for stimulation of adenyl cyclase of approximately 2 μM. This, however, is non-selective and activates other adenosine receptors with even greater affinity, with EC ₅₀ values in the low nanomolar (A ₁ and A _2A ) or high nanomolar (A ₃ ) range. Therefore, the performance of A _2B receptors often depends on the lack of efficacy of compounds that are potent and selective agonists of other types of receptors. The A _2B receptors have been characterized by exclusion, ie the lack of efficacy of agonists specific to other receptors. The selective A _2A agonist CGS-21680 (Webb et al., 1992), for example, was useful in differentiating between A _2A and A _2B adenosine receptors (Hide et al., 1992; Chern et al., 1993; Feoktistov and Biaggioni, 1995 ; van der Ploeg et al., 1996). Both receptors are positively associated with adenylcyclase and are activated by the non-selective agonist NECA. CGS-21680 is practically ineffective at A _2B receptors, but still as effective as NECA at activating A _2A receptors, with an EC ₅₀ value in the low nanomolar range for both agonists (Jarvis et al., 1989; Nakane and Chiba, 1990; Webb et al., 1992; Hide et al., 1992; Feoktistov and Biaggioni, 1993; Alexander et al., 1996). A _2B receptors also have very low affinity for the A ₁ -selective agonist R-PIA (Feoktistov and Biaggioni, 1993; Brackett and Daly, 1994), as well as for the A ₃ -selective agonist N ⁶ -(3-iodobenzyl)-N- methyl-5'-carbamoyladenosine (IB-MECA) (Feoktistov and Biaggioni, 1997). Agonist profile NECA > R-PIA = IB-MECA > CGS-21680 was determined in human erythroleukemia (HEL) cells for A _2B -mediated cAMP accumulation. The difference between the EC ₅₀ for NECA and the remaining agonists is approximately 2 orders of magnitude. Therefore, responses elicited by NECA at concentrations in the low micromolar range (1-10 μM), but not by R-PIA, IB-MECA or CGS-21680, are characteristic of A _2B receptors. Whereas A _2B receptors generally have a lower affinity for agonists than other receptor subtypes, this does not apply to antagonists. The relation of structural activity of adenosine A _2B receptor antagonists has not been fully characterized, but at least some xanthines are equally or more potent A _2B receptor subtype antagonists than other subtypes. In particular, DPSPX (1,3-dipropyl-8-sulfophenylxanthine), DPCPX (1,3-diproyl-8c-iclopentylxanthine), DPX (1,3-diethylphenylxanthine), the anti-asthma drug enprofylline (3-n-propylxanthine), and nexanthine the compound 2,4-dioxobenzopteridin (alloxazin) have an affinity in the nM range from medium to high.

Other known A _2B inhibitors are ATL801, PSB-605, PSB-1115, ISAM-140, GS6201, MRS1706 and MRS1754.

This document describes that adenosine receptors play a statically indeterminate role in the suppression of inflammation in vivo, acting as a physiological "STOP" (stop mechanism) that can limit the immune response and thereby protect normal tissues from excessive immune damage during the pathogenesis of various diseases.

A _2A receptor antagonists provide long-term enhancement of immune responses by reducing T-cell mediated tolerance to antigenic stimuli, enhancing memory T cell induction, and increasing the efficiency of passive antibody administration for the treatment of cancer and infectious diseases, while A _2A receptor agonists provide long-term reduction of immune responses by increasing T-cell mediated tolerance to antigenic stimuli, in particular to reduce the use of immunosuppressants in certain conditions.

Immunomodulation is a critical aspect of the treatment of a number of diseases and disorders. T cells in particular play a vital role in fighting infections and have the ability to recognize and destroy cancer cells. Enhancement of T-cell mediated responses is a key component for enhancing responses to therapeutic agents. However, it is critical to immunomodulation that any enhancement of the immune response be balanced against the need to prevent autoimmunity as well as chronic inflammation. Chronic inflammation and self-recognition by T cells is the main cause of the pathogenesis of systemic disorders such as rheumatoid arthritis, multiple sclerosis and systemic lupus erythematosus. In addition, long-term immunosuppression is required to prevent rejection of transplanted organs or grafts.

Tumor-induced immunosuppression is a major barrier to the effectiveness of current cancer treatments. Because of their remarkable clinical efficacy against a wider range of cancers, recent advances with immune checkpoint blockade inhibitors such as anti-CTLA-4 and anti-PD-1/PDL1 are revolutionary cancer treatments.

Adenosine is one of the new promising immunosuppressive targets identified in preclinical studies. This metabolite is produced by an ectoenzyme, CD73, expressed on host suppressor cells and tumor cells. Increased CD73 expression correlates with poor prognosis in patients with a number of types of malignancies, including colorectal cancer (Liu et al., J. Surgical Oncol, 2012), gastric cancer (Lu et al., World J. Gastroenterol., 2013), cancer gallbladder (Xiong et al., Cell and Tissue Res., 2014). Preclinical studies have shown that the protumor effects of CD73 may be due (at least in part) to adenosine-mediated immunosuppression. As described above, adenosine binds to the four known receptors A ₁ , A _2A , A _2B , and A ₃ , with activation of the A _2A and A _2B receptors, which are known to suppress the effector functions of many immune cells, i.e., the A _2A receptors. and A _2B induce adenylate cyclase-dependent accumulation of cAMP, leading to immunosuppression. Since A ₁ and A ₃ antagonism will counteract the desired effect, and A ₁ and A ₃ agonists serve as potential cardioprotective agents, selectivity for A ₁ and A ₃ must be achieved (Antonioli et al., Nat. rev. Cancer, 2013, Thiel et al., Microbes and Infection, 2003). In the tumor microenvironment, activation of the A _2A and A _2B receptors has been shown to suppress antitumor immunity and increase the spread of CD73 tumors. In addition, blockade of A _2A or A _2B with small molecule antagonists can reduce tumor metastasis. It has been found that blocking the A _2A receptor can overcome tumor escape mechanisms, including both anergy and regulatory induction of T cells induced by tumor cells, and cause long-term tumor responsiveness to treatment. Ohta et al. demonstrated rejection of approximately 60% of established CL8-1 melanoma tumors in A _2A receptor deficient mice compared with no rejection in normal mice (Ohta, et al.; PNAS 103 (35): 13132-7, 2006). In line with this, investigators also showed improved inhibition of tumor growth, destruction of metastases, and prevention of neovascularization by antitumor T cells after treatment with an A _2A receptor antagonist.

Tumors have been shown to avoid damage by the immune system by preventing T cell activation by inhibiting costimulatory factors in the B7-CD28 and TNF families, and 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. al., Clin Cane Res 13 (13): 4016-4025, 2007). Because A _2A receptor expression is increased in lymphocytes after activation, therapies that elicit effector lymphocyte responses, such as anti-CTLA-4 and anti-PD-1, may also enhance the effects of A _2A -mediated immunosuppression. Immune checkpoint blockade in combination with A _2A antagonists or double A _2A / _2B antagonists increases the magnitude of immune responses to tumors and metastases. Accordingly, the combination of A _2A inhibition with anti-PD-1 therapy enhances IFN-γ production by T cells co-cultured with MC38 tumor cells, improves mouse survival in the 4T1 mammary tumor model, and reduces tumor growth in AT-3ova ^dim CD73 ⁺ tumors. .(Beavis et al., Cancer Immunol. Res., 2015; Mittal et al., Cancer Res., 2014).

In addition, preclinical studies have shown that inhibition of A _2B results in reduced tumor growth and increased survival in mice in Lewis lung carcinoma, MB49 bladder carcinoma, and ortho-4T1 breast carcinoma models (Ryzhov et al., 2009, Cekic et al., 2012), and the combination of A _2B inhibition with anti-PD-1 therapy reduces lung metastasis of B16-F10 melanoma tumors and improves mouse survival in the 4T1 mammary tumor model.

WO 03/050241 describes methods of enhancing the immune response to an antigen by increasing the efficacy of a vaccine or increasing the immune response to a tumor antigen or immune cell-mediated tumor destruction by administering an agent that inhibits extracellular adenosine or inhibits adenosine receptors.

WO 2004/089942, WO 2005/000842 and WO 2006/008041 disclose benzothiazole derivatives, including tosadenant, as A _2A inhibitors for the treatment of Parkinson's disease. WO 2004/092171 and WO 2005/028484 disclose similar thiazolopyridine and pyrazolopyrimidine derivatives also as A _2A inhibitors for the treatment of Parkinson's disease. However, these compounds do not exhibit significant A _2B inhibitory activity and only show good pharmacokinetic properties in rat animal models of Parkinson's disease but not in mouse animal models of cancer. In addition, the compounds do not show that they are able to prevent immunosuppression and, therefore, are able to maintain the inhibition of tumor growth induced by anti-cancer T cells, the reduction or destruction of metastases, and the prevention of neovascularization.

Thus, there remains a need for treatments that provide long-term enhancement of immune responses to specific antigens, especially for the treatment and prevention of hyperproliferative and infectious diseases and disorders, and thus the aim of the present invention was to provide treatments that would simplify treatment protocols. and improve immune responses to certain antigens. A specific object of the invention was to provide improved methods for preventing or treating hyperproliferative and infectious diseases and disorders in the host, especially to provide effective A _2A or dual A _2A / _2B antagonists for the treatment and prevention of such diseases.

Brief description of the invention

Surprisingly, it has been found that the thiazolopyridine derivatives according to the invention are highly effective inhibitors of the A _2A adenosine receptor or the A _2A and A _2B adenosine receptors, and at the same time have a high selectivity over the A ₁ and A ₃ adenosine receptors, and thus , the compounds of the present invention can be used to treat hyperproliferative diseases and disorders such as cancer and infectious diseases and disorders.

In particular, in contrast to the known adenosine A _2A receptor antagonist tosadenant and similar benzothiazole derivatives, the compounds of the present invention surprisingly exhibit dual A _2A /A _2B activity which is advantageous for the treatment and/or prevention of hyperproliferative and infectious diseases and disorders as described above. , or the compounds of the present invention exhibit at least high A _2A inhibitory activity along with other unexpected benefits disclosed herein, which result in high efficacy in the treatment and/or prevention of hyperproliferative and infectious diseases and disorders.

In addition, compared with the known adenosine A _2A receptor antagonist tosadenant and similar benzothiazole derivatives, the compounds of the present invention unexpectedly show better pharmacokinetic properties in a mouse as an animal model related to malignancy, which is preferable for the treatment and/or prevention of hyperproliferative and infectious diseases and disorders as described above.

In addition, as described above, adenosine in the tumor microenvironment can inhibit T cell activity by signaling through A _2A receptors and inhibit T cell secretion of cytokines. A _2A -specific agonists such as CGS-21680, like adenosine, inhibit cytokine secretion of T cells in vitro and in vivo. In contrast, potential A _2A antagonists or A _2A /A _2B dual antagonists can rescue T cells from this inhibition. Unlike the known adenosine A _2A receptor antagonist tosadenant, the compounds of the present invention demonstrate that they are able to rescue T cells from inhibition and are able to prevent suppression of cytokine secretion induced by adenosine or A _2A -specific agonists such as CGS-2168, which is preferred. for the treatment and/or prevention of hyperproliferative and infectious diseases and disorders as described above. Thus, the compounds of the present invention are surprisingly able to prevent immunosuppression and therefore are able to maintain the inhibition of tumor growth induced by anti-cancer T cells, the reduction or destruction of metastases, and the prevention of neovascularization.

The invention relates to thiazolopyridine derivatives of general formula I,

Where

R ¹ is straight or branched alkyl which contains 1-10 C atoms, which is unsubstituted or mono-, di- or tri-substituted by R ⁵ , and in which 1-4 C atoms may be replaced, independently of one another, by О, S, SO, SO ₂ , NH, NCH ₃ , -ОСО-, -NHCONH-, -NHCO-, NR ⁶ SO ₂ R ⁷ , COO, CONH-, -NCH ₃ CO-, -CONCH ₃ -, - C≡C- groups and/or -CH=CH- groups, and/or, in addition, 1-10 H atoms can be replaced by F and/or Cl, or mono- or bicyclic alkyl, which contains 3-7 C -atoms, which is unsubstituted or mono-, di- or tri-substituted by R ⁵ , and in which 1-4 C-atoms can be replaced, independently of each other, with O, S, SO, SO ₂ , NH, NCH ₃ , - OCO-, NHCONH, -NHCO-, -NR ⁶ SO ₂ R ⁷ -, -COO-, -CONH-, -NCH ₃ CO-, -CONCH ₃ -, -C≡C- groups and/or -CH=CH - groups and/or, in addition, 1-10 H atoms can be replaced by F and/or Cl, or mono- or bicyclic heteroaryl, heterocyclyl, aryl or cyclic alkylaryl, which contains 3-14 carbon atoms and 0-4 heteroatoms , independently chosen from N, O and S which is unsubstituted or mono-, di- or tri-substituted by R ⁵ ,

R ² is straight or branched alkyl which contains 1-10 C atoms, which is unsubstituted or mono-, di- or tri-substituted by R ⁵ and in which 1-4 C atoms can be replaced, independently of one another, by O , S, SO, SO ₂ , NH, NCH ₃ , -ОСО-, -NHCONH-, -NHCO-, -NR ⁶ SO ₂ R ⁷ -, -COO-, -CONH-, -NCH ₃ CO-, -CONCH _3- , -C≡C- groups and/or -CH=CH- groups, and/or, in addition, 1-10 H atoms can be replaced by F and/or Cl, or cyclic alkyl which contains 3-7 C-atoms, which is unsubstituted or mono-, di- or tri-substituted by R ⁵ and in which 1-4 C-atoms can be replaced, independently of each other, by O, S, SO, SO ₂ , NH, NCH ₃ , - OCO-, -NHCONH-, -NHCO-, R ⁶ SO ₂ R ⁷ -, -COO-, -CONH- -NCH ₃ CO-, -CONCH ₃ -, -C≡C- groups and/or -CH=CH - groups and/or, in addition, 1-11 H atoms can be replaced by F and/or Cl, or mono- or bicyclic heteroaryl, heterocyclyl, aryl or cyclic alkylaryl, which contains 3-14 carbon atoms and 0-4 heteroatoms , independently selected from N, O and S, to which is unsubstituted or mono-, di- or trisubstituted R ⁵ ,

R ³ is linear or branched alkyl or O-alkyl which contains 1-6 C atoms, or cyclic alkyl which contains 3-6 C atoms which is unsubstituted or mono-, di- or tri-substituted by H, =S, \u003d NH, \u003d O, OH, cyclic alkyl, which contains 3-6 C atoms, COOH, Hal, NH ₂ , SO ₂ CH ₃ , SO ₂ NH ₂ , CN, CONH ₂ , NHCOCH ₃ , NHCONH ₂ or NO ₂ ,

R ⁴ is H, D, linear or branched alkyl which contains 1-6 C atoms or Hal,

R ⁵ is H, R ⁶ , =S, =NR ⁶ , =O, OH, COOH, Hal, NH ₂ , SO ₂ CH ₃ , SO ₂ NH ₂ , CN, CONH ₂ , NHCOCH ₃ , NHCONH ₂ , NO ₂ , or linear or branched alkyl which contains 1-10 C atoms, which is unsubstituted or mono-, di- or tri-substituted by R ⁶ , and in which 1-4 C atoms can be replaced, independently of one another, by O , S, SO, SO ₂ , NH, NCH ₃ , -ОСО-, -NHCONH-, -NHCO-, -NR ⁶ SO ₂ R ⁷ -, -COO-, -CONH-, -NCH ₃ CO-, -CONCH ₃ -, -C≡C- groups and/or -CH=CH- groups, and/or, in addition, 1-10 H atoms can be replaced by F and/or Cl, or mono- or bicyclic alkyl, which contains 3-7 C atoms which is unsubstituted or mono-, di- or tri-substituted by R ⁶ , and in which 1-4 C atoms can be replaced, independently of one another, with O, S, SO, SO ₂ , NH, NCH ₃ , -ОСО-, -NHCONH-, -NHCO-, -NR ⁶ SO ₂ R ⁷ -, -COO-, -CONH-, -NCH ₃ CO-, -CONCH ₃ -, -C≡C- groups and /or -CH=CH- groups and/or, in addition, 1-10 H atoms can be replaced by F and/or Cl, or mono- or bicyclic heteroaryl, het erocyclyl, aryl or cyclic alkylaryl which contains 3-14 carbon atoms and 0-4 heteroatoms independently selected from N, O and S, which is unsubstituted or mono-, di- or tri-substituted by R ⁶ ,

R ⁶ , R ⁷ are independently selected from the group consisting of H, ═S, ═NH, ═O, OH, COOH, Hal, NH ₂ , SO ₂ CH ₃ , SO ₂ NH ₂ , CN, CONH ₂ , NHCOCH ₃ , NHCONH ₂ , NO ₂ and linear or branched alkyl, which contains 1-10 C-atoms, in which 1-4 C-atoms can be replaced, independently of each other, with O, S, SO, SO ₂ , NH, NCH ₃ , -ОСО-, -NHCONH-, -NHCO-, -COO-, -CONH-, -NCH ₃ CO-, -CONCH ₃ -, -C=C- groups and/or -CH=CH - groups, and/or, in addition, 1-10 H atoms can be replaced by F and/or Cl,

Hal is F, Cl, Br, or I,

D is deuterium,

and their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all proportions.

Preferably the invention relates to a compound of formula I, where

R ¹ is straight or branched alkyl which contains 1-10 C atoms, which is unsubstituted or mono-, di- or tri-substituted by R ⁴ , and in which 1-4 C atoms may be replaced, independently of one another, by О, S, SO, SO ₂ , NH, NCH ₃ , -ОСО-, -NHCONH-, -NHCO- -NR ⁵ SO ₂ R ⁶ -, -COO-, -CONH-, -NCH ₃ CO-, -CONCH _3- , -C≡C- groups and/or -CH=CH- groups, and/or, in addition, 1-10 H atoms can be replaced by F and/or Cl, or one of the following structures:

which is unsubstituted or mono-, di- or tri-substituted by R ⁵

and where R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above.

Particularly preferably, the invention relates to a compound of formula I, wherein

R ¹ is one of the following structures:

and where R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above

Preferably the invention relates to a compound of formula I, where

R ¹ is phenyl, methylpyrazole or dihydropyran,

and R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above.

Particularly preferably, the invention relates to a compound of formula I, where R ² represents one of the following structures:

which is unsubstituted or mono-, di- or tri-substituted by R ⁶

and where R ¹ , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above.

Particularly preferably, the invention relates to a compound of formula I, wherein

R ² is one of the following structures:

and where R ¹ , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above.

Preferably the invention relates to a compound of formula I, where

R ³ is one of the following structures:

and R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above.

Preferably the invention relates to a compound of formula I, where

R ³ is O-alkyl which contains 1-6 C atoms which is unsubstituted or mono-, di- or tri-substituted by F,

and R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above.

Preferably the invention relates to a compound of formula I, where R ³ represents OMe,

and R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above.

Particularly preferably, the invention relates to a compound of formula I, wherein

R ¹ is phenyl, methylpyrazole or dihydropyran,

R ³ is OMe,

and R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined above.

Preferably the invention relates to a compound of formula I, where

R ⁴ is H, D, methyl, ethyl, F, Br or Cl,

and where R ¹ , R ² , R ³ , R ⁵ , R ⁶ and R ⁷ are as defined above.

Preferably the invention relates to a compound of formula I, where

R ⁴ is H, D, methyl, F, Br or Cl,

and where R ¹ , R ² , R ³ , R ⁵ , R ⁶ and R ⁷ are as defined above.

Preferably the invention relates to a compound of formula I, where

R ⁴ is H,

and where R ¹ , R ² , R ³ , R ⁵ , R ⁶ and R ⁷ are as defined above.

Particularly preferably, the invention relates to a compound selected from the group consisting of the following:

and their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all ratios.

All of the above preferred, particularly preferred and most preferred meanings of the above radicals of compounds of formula I are to be understood in such a way that these preferred, particularly preferred and most preferred meanings or embodiments can be combined with each other in any possible combination to give compounds of formula I, and that preferred, particularly preferred and most preferred compounds of formula I of this type are thus explicitly disclosed in this document.

Hal means fluorine, chlorine, bromine or iodine, in particular fluorine, bromine or chlorine.

D or ² H means deuterium.

или атом кислорода, присоединенный к атому углерода двойной связью.-(C=O)- or =O means the presence of carbonyl oxygen and means

or an oxygen atom attached to a carbon atom by a double bond.

Alkyl means a saturated straight chain or branched hydrocarbon chain and contains 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. Alkyl preferably means methyl, in addition ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or mpem-butyl, in addition also pentyl, 1-,2- or 3-methylbutyl, 1,1-, 1,2- or 2,2-dimethylpropyl, 1-ethylpropyl, hexyl, 1-, 2-, 3- or 4-methylpentyl, 1,1-, 1,2-, 1,3-, 2,2-, 2,3- or 3,3-dimethylbutyl, 1- or 2-ethylbutyl, 1-ethyl-1-methylpropyl, 1-ethyl-2-methylpropyl, 1,1,2- or 1,2,2-trimethylpropyl, linear or branched heptyl, octyl , nonyl or decyl, even more preferably, for example, trifluoromethyl.

Cyclic alkyl or cycloalkyl means a saturated cyclic hydrocarbon chain and contains 3-10, preferably 3-7 carbon atoms and preferably means cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl. Cycloalkyl also means a partially unsaturated cyclic alkyl such as, for example, cyclohexenyl or cyclohexynyl.

Alkenyl means an unsaturated straight (linear) or branched hydrocarbon chain and contains 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms.

O-alkyl or OA means a linear or branched alkoxy containing 1-6 carbon atoms, and preferably means methoxy, in addition also, for example, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy .

Alkyloxycarbonyl refers to straight or branched chain carboxylic acid esters derived from the present invention, i.e. methyloxycarbonyl (MeOCO-), ethyloxycarbonyl or butyloxycarbonyl.

Alkylcarbonyl refers to a straight or branched chain alkyl and carboxylic acid group.

Aryl, Ar or aromatic ring means a mono- or polycyclic aromatic or fully unsaturated cyclic hydrocarbon chain, for example, unsubstituted phenyl, naphthyl or biphenyl, in addition, preferably phenyl, naphthyl or biphenyl, each of which is mono-, di- or tri-substituted, e.g. via A, fluorine, chlorine, bromine, iodine, hydroxyl, methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, nitro, cyano, formyl, acetyl, propionyl, trifluoromethyl, amino, methylamino, ethylamino, dimethylamino, diethylamino, benzyloxy , sulfonamido, methylsulfonamido, ethylsulfonamido, propylsulfonamido, butylsulfonamido, dimethylsulfonamido, phenylsulfonamido, carboxyl, methoxycarbonyl, ethoxycarbonyl, aminocarbonyl.

Heterocycle and heterocyclyl refer to saturated or unsaturated non-aromatic rings or ring systems containing at least one heteroatom selected from O, S and N, including, in addition, oxidized forms of sulfur, namely SO and SO ₂ . Examples of heterocycles include tetrahydrofuran (THF), dihydrofuran, 1,4-dioxane, morpholine, 1,4-dithiane, piperazine, piperidine, 1,3-dioxolane, imidazolidine, imidazoline, pyrroline, pyrrolidine, tetrahydropyran, dihydropyran, oxathiolane, dithiolane, 1,3-dioxane, 1,3-dithian, oxatian, thiomorpholine, and the like.

Heteroaryl means an aromatic or partially aromatic heterocycle which contains at least one ring heteroatom selected from O, S and N. Heteroaryls thus include heteroaryls fused to other types of rings such as aryls, cycloalkyls and heterocycles that are not aromatic. Examples of heteroaryl groups include: pyrrolyl, isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, oxadiazolyl, thiadiazolyl, thiazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, triazinyl, thienyl, pyrimidyl, benzisoxazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, indolinebenzofuranyl, , indazolyl, isoxazolyl, isoindolyl, dihydrobenzothienyl, indolizinyl, cinnolinyl, phthalazinyl, quinazolinyl, naphthyridinyl, carbazolyl, benzodioxynyl, benzodioxolyl, quinoxalinyl, purinyl, furazanyl, thiophenyl, isobenzylfuranyl, benzimidazolyl, benzimidazolyl, benzofuranyl, benzothienyl, indollyquinyl, and thiophenyl .P. In the case of heterocyclyl and heteroaryl groups, rings and ring systems containing from 3 to 15 atoms are included which form 1-3 rings.

Mono- or bicyclic saturated, unsaturated or aromatic heterocycle preferably means unsubstituted or mono-, di- or tri-substituted 2- or 3-furyl, 2- or 3-thienyl, 1-, 2- or 3-pyrrolyl, 1-, 2- , 4-or 5-imidazolyl, 1-, 3-, 4- or 5-pyrazolyl, 2-, 4- or 5-oxazolyl, 3-, 4- or 5-isoxazolyl, 2-, 4- or 5-thiazolyl, 3-, 4- or 5-isothiazolyl, 2-, 3- or 4-pyridyl, 2-, 4-, 5- or 6-pyrimidinyl, more preferably 1,2,3-triazole-1-, -4 - or -5-yl, 1,2,4-triazol-1-, -3- or 5-yl, 1- or 5-tetrazolyl, 1,2,3-oxadiazol-4- or -5-yl, 1 ,2,4-oxadiazol-3- or -5-yl, 1,3,4-thiadiazol-2- or -5-yl, 1,2,4-thiadiazol-3- or -5-yl, 1,2 ,3-thiadiazol-4- or -5-yl, 3- or 4-pyridazinyl, pyrazinyl, 1-, 2-, 3-, 4-, 5-, 6- or 7-indolyl, 4- or 5-isoindolyl , 1-, 2-, 4- or 5-benzimidazolyl, 1-, 3-, 4-, 5-, 6- or 7-benzopyrazolyl, 2-, 4-, 5-, 6- or 7-benzoxazolyl, 3 -, 4-, 5-, 6- or 7-benzisoxazolyl, 2-, 4-, 5-, 6- or 7-benzothiazolyl, 2-, 4-, 5-, 6- or 7-benzisothiazolyl, 4-, 5-, 6- or 7-benz-2,1,3-oxadiazolyl, 2-, 3-, 4-, 5-, 6-, 7- or 8-quinolyl, 1-, 3-, 4-, 5-, 6 -, 7- or 8-isoquinolyl, 3-, 4-, 5-, 6-, 7- or 8-cinnolinyl, 2-, 4-, 5-, 6-, 7- or 8-quinazolinyl, 5- or 6-quinoxalinyl, 2-, 3-, 5-, 6-, 7- or 8-2H-benzo-1,4-oxazinyl, even more preferably 1,3-benzodioxol-5-yl, 1,4-benzodioxan- 6-yl, 2,1,3-benzothiadiazol-4- or -5-yl or 2,1,3-benzoxadiazol-5-yl.

Heterocyclic radicals can also be partially or completely hydrogenated and mean, for example, 2,3-dihydro-2-, -3-, -4- or -5-furyl, 2,5-dihydro-2-, -3-, - 4- or 5-furyl, tetrahydro-2- or -3-furyl, 1,3-dioxolan-4-yl, tetrahydro-2- or -3-thienyl, 2,3-dihydro-1-, -2-, -3-, -4- or -5-pyrrolyl, 2,5-dihydro-1-, -2-, -3-, -4- or -5-pyrrolyl, 1-, 2- or 3-pyrrolidinyl, tetrahydro -1-, -2- or -4-imidazolyl, 2,3-dihydro-1-, -2-, -3-, -4- or -5-pyrazolyl, tetrahydro-1-, -3- or -4 -pyrazolyl, 1,4-dihydro-1-, -2-, -3- or -4-pyridyl, 1,2,3,4-tetrahydro-1-, -2-, -3-, -4-, -5- or -6-pyridyl, 1-, 2-, 3- or 4-piperidinyl, 2-, 3- or 4-morpholinyl, tetrahydro-2-, -3- or -4-pyranyl, 1,4- dioxanyl, 1,3-dioxan-2-, -4- or -5-yl, hexahydro-1-, -3- or -4-pyridazinyl, hexahydro-1-, -2-, -4- or -5- pyrimidinyl, 1-, 2- or 3-piperazinyl, 1,2,3,4-tetrahydro-1-, -2-, -3-, -4-, -5-, -6-, -7- or - 8-quinolyl, 1,2,3,4-tetrahydro-1-, -2-, -3-, -4-, -5-, -6-, -7- or -8-isoquinolyl, 2-, 3 -, 5-, 6-, 7- or 8-3,4-dihydro-2H-benzo-1,4-oxazinyl, even more preferably 2,3-methylenedioxyphenyl, 3,4-methylenedioxyphenyl, 2,3-ethylenedioxyphenyl, 3,4-ethylenedioxyphenyl, 3, 4-(difluoromethylenedioxy)phenyl, 2,3-dihydrobenzofuran-5- or 6-yl, 2,3-(2-oxomethylenedioxy)phenyl or also 3,4-dihydro-2H-1,5-benzodioxepin-6- or - 7-yl, further preferably 2,3-dihydrobenzofuranyl or 2,3-dihydro-2-oxofuranyl.

The heterocycle also means, for example, 2-oxopiperidin-1-yl, 2-oxopyrrolidin-1-yl, 2-oxo-1H-pyridin-1-yl, 3-oxomorpholin-4-yl, 4-oxo-1H -pyridin-1-yl, 2,6-dioxopiperidin-1-yl, 2-oxo-piperazin-1-yl, 2,6-dioxopiperazin-1-yl, 2,5-dioxopyrrolidin-1-yl, 2-oxo-1,3 -oxazolidin-3-yl, 3-oxo-2H-pyridazin-2-yl, 2-caprolactam-1-yl (=2-oxoazepan-1-yl), 2-hydroxy-6-oxopiperazin-1-yl, 2 -methoxy-6-oxopiperazin-1-yl or 2-azabicyclo[2.2.2]octan-3-on-2-yl.

Heterocycloalkyl in this case means a fully hydrogenated or saturated heterocycle, heterocycloalkenyl (one or more double bonds) or heterocycloalkynyl (one or more triple bonds) means a partially or incompletely hydrogenated or unsaturated heterocycle, heteroaryl means an aromatic or fully unsaturated heterocycle.

A cyclic alkylaryl group in connection with the present invention means that one or two Ar aromatic rings are fused on an unsubstituted or mono- or disubstituted cyclic alkyl in which one or two CH ₂ groups and/or, in addition, 1-11 hydrogen atoms may be replaced, as, for example, takes place in the radicals shown below:

и

And

In addition, the abbreviations below 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

ROA - phenoxyacetyl

DCCI - Dicyclohexylcarbodiimide

HOBt - 1-hydroxybenzotriazole

Thus, the invention relates to a pharmaceutical preparation containing a compound according to the present invention and/or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all ratios.

The invention also relates to a pharmaceutical preparation according to an invention of this type additionally containing excipients and/or excipients.

In addition, the invention relates to the aforementioned pharmaceutical preparation according to the invention, containing at least one additional active drug compound.

By pharmaceutically or physiologically acceptable derivatives are meant, for example, salts of the compounds of the present invention, as well as so-called prodrugs. By prodrugs is meant derivatives of a compound of the present invention which have been modified with, for example, alkyl or acyl groups (see also amino and hydroxyl protecting below), sugars or oligopeptides and which are rapidly degraded or released in the body to form effective molecules. They also include biodegradable polymeric derivatives of the compounds of the present invention, as described, for example, in Int. J Pharm. 115 (1995), 61-67.

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 salts, which can be obtained from various organic and inorganic bases using methods well known in the art. Pharmaceutically acceptable salt forms of pepstatin are obtained mainly by using traditional methods. In case the compound of the present invention contains a carboxyl group, one of its suitable salts may 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 alcoholates such as potassium ethoxide and sodium propoxide; and various organic bases such as piperidine, diethanolamine and N-methylglutamine. Also included are aluminum salts with pepstatin.

In addition, base salts of the compound of the present invention include, but are not limited to, aluminum, ammonium, calcium, copper, iron(III), iron(II), lithium, magnesium, manganese(III), manganese(II), potassium salts. , sodium and zinc.

Of the above salts, preference is given to ammonium salts; alkali metal salts - sodium and potassium, and alkaline earth metal salts - calcium and magnesium. Salts of the compounds of the present invention which are derived from pharmaceutically acceptable organic non-toxic bases include, but are not limited to, salts of primary, secondary and tertiary amines, substituted amines, also including natural substituted amines, cyclic amines, and basic ion exchange resins, for example, arginine, betaine, caffeine, chlorprocaine, choline, N,N'-dibenzylethylenediamine (benzatine), dicyclohexylamine, diethanolamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lidocaine, lysine, meglumine, N-methyl-D-glucamine, morpholine, piperazine, piperidine, polyamine gums, procaine, purines, theobromine, triethanolamine, triethylamine, trimethylamine, tripropylamine and tris(hydroxymethyl)methylamine (tromethamine ).

As already mentioned, pharmaceutically acceptable pepstatin base addition salts are formed with metals or amines such as alkali metals and alkaline earth metals or organic amines. Preferred metals are sodium, potassium, magnesium and calcium. Preferred organic amines are N,N'-dibenzylethylenediamine, chlorprocaine, choline, diethanolamine, ethylenediamine, N-methyl-D-glucamine and procaine.

Base addition salts to the compounds of the present invention are prepared by contacting the free acid form with a sufficient amount of the desired base to form the salt in the usual manner. The free acid can be recovered by contacting the salt form with the acid and isolating the free acid in the usual manner. The free acid forms differ somewhat from their corresponding salt forms in terms of certain physical properties, such as solubility in polar solvents; for the purposes of the present invention, however, the salts otherwise correspond to their respective free acid forms.

In the light of the above, it can be seen that the expression "pharmaceutically acceptable salt" in the context of this application means an active compound, which is a compound of the present invention in the form of one of its salts, in particular if such a salt form gives this active compound improved pharmacokinetic properties compared to the free form of this active compound or any other salt form of this active compound previously used. A pharmaceutically acceptable salt form of the active compound may also provide the active compound for the first time with a desired pharmacokinetic property that it did not previously possess, and may even positively influence the pharmacodynamics of the active compound in relation to its therapeutic efficacy in the body.

Solvates of the compound of the present invention imply adductions of inert solvent and pepstatin molecules, which are formed due to their mutual attraction. Solvates are, for example, hydrates, such as monohydrates or dihydrates, or alcoholates, i.e. addition products of alcohols, such as, for example, methanol or ethanol, to compounds.

All physiologically acceptable salts, derivatives, solvates and stereoisomers of these compounds, including mixtures thereof in all proportions, are also in accordance with the present invention.

The compounds of general formula I may contain one or more centers of chirality, whereby all stereoisomers, enantiomers, diastereomers, etc., of the compounds of general formula I are also covered by the present invention.

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

The compounds of formula I according to the invention may be chiral due to their molecular structure and, accordingly, may occur in various enantiomeric forms. Thus, they may be in racemic or optically active form. Since the pharmaceutical efficacy of the racemates or stereoisomers of the compounds according to the invention may differ, it may be desirable to use enantiomers. In these cases, both the end product and even the intermediates can be separated into enantiomeric compounds by chemical or physical methods known to the person skilled in the art, or already used as such in the synthesis.

Pharmaceutically or physiologically acceptable derivatives include, for example, salts of the compounds according to the invention, as well as so-called prodrugs. Prodrugs are compounds of formula I which have been modified, for example, with alkyl or acyl groups (see also amino and hydroxyl protecting groups below), sugars or oligopeptides, and which are rapidly degraded or released in the body to form effective compounds according to the invention. These also include biodegradable polymeric derivatives of the compounds according to the invention, as described, for example, in Int. H. Pharm. 115 (1995), 61-67.

Suitable acid addition salts are inorganic and organic salts of all physiologically or pharmacologically acceptable acids, for example halides, in particular hydrochlorides or hydrobromides, lactates, sulfates, citrates, tartrates, maleates, fumarates, oxalates, acetates, phosphates, methylsulfonates or p-toluenesulfonates. .

Most preference is given to the hydrochlorides, trifluoroacetates or bistrifluoroacetates of the compounds according to the invention.

Solvates of the compounds of formula I mean the adductions of inert solvent molecules on the compounds of formula I, which are formed due to their force of mutual attraction. Solvates are, for example, hydrates, such as monohydrates or dihydrates, or alcoholates, i.e. addition products of alcohols, such as, for example, methanol or ethanol, to compounds.

In addition, the compound of formula I is contemplated to include its isotopically labeled forms. The isotopically labeled form of a compound of formula I is identical to said compound except for the fact that one or more atoms of said compound have been replaced by an atom or atoms which have an atomic mass or mass number different from the atomic mass or mass number of said atom which usually found in nature. Examples of isotopes which are readily available commercially and which can be introduced into a compound of formula I using well known methods include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, for example ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ O, ¹⁷ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, and ³⁶ CI, respectively. A compound of formula I, a prodrug or a pharmaceutically acceptable salt thereof, which contains one or more of the above isotopes and/or other isotopes of other atoms, is also within the scope of the present invention. The isotopically labeled compound of formula I can be used in a number of useful ways. For example, an isotopically labeled compound of Formula I, for example, into which a radioactive isotope such as ³ H or ¹⁴ C has been introduced, will be useful in drug and/or substrate tissue distribution assays. Such radioactive isotopes, i.e. tritium ( ³ H) and carbon-14 ( ¹⁴ C) are particularly preferred due to their ease of preparation and high detectability. Incorporation of heavier isotopes, eg deuterium ( ² H), into a compound of formula I will provide therapeutic advantages based on the greater metabolic stability of said isotopically labeled compound. The greater metabolic stability manifests itself directly as an increase in in vivo half-life or a reduction in the required dose, which under most conditions would represent a preferred embodiment of said invention. An isotopically labeled compound of formula I can generally be prepared by carrying out the procedures disclosed in the synthetic schemes and in the description relating to them in the sections relating to examples and methods of preparation described in this application, by replacing the non-isotopically labeled reagent with its corresponding readily available labeled reagent. isotope.

In order to change the oxidative metabolism of the compound through the primary kinetic isotope effect, deuterium ( ² H) can also be introduced into the compound of formula I. The primary kinetic isotope effect is a change in the rate of a chemical reaction that occurs due to the exchange of isotopic nuclei, which in turn is caused by a change in ground state energies, which is necessary for the formation of a covalent bond after said isotopic exchange. An exchange for a heavier isotope will typically result in a decrease in the ground state energy for the chemical bond, thus causing a decrease in the rate of the rate-limiting bond breaking step. When a bond break occurs at or near the saddle configuration along the multi-product formation reaction coordinate, the distribution coefficient of the products can change significantly. For example, if deuterium binds to a carbon atom in a position in which the exchange does not occur, the rate differences k _M /k _D =2-7 are typical. If such a difference in rate is successfully applied to a compound of formula I which is susceptible to oxidation, the in vivo profile of that compound can thus be significantly altered, resulting in improved pharmacokinetic properties.

In the process of discovering and improving therapeutic agents, one of ordinary skill in the art is looking for ways to optimize pharmacokinetic parameters until the desired in vitro properties are obtained. It is reasonable to assume that many compounds with weak pharmacokinetic profiles are susceptible to oxidative metabolism. In vitro assays with liver microsomes, which are now available, provide valuable information on the process of this type of oxidative metabolism, which in turn provides a rational model of deuterium labeled compounds of formula I with improved stability up to resistance to such oxidative metabolism. Thus, a significant improvement in the pharmacokinetic profiles of the compounds of formula I is obtained, which can be quantified in terms of increases in in vivo half-life (T / 2), in concentration at maximum therapeutic effect (C _max ), area under the dose response curve (AUC ) and F; in terms of reduction in clearance, dose and material costs.

The following is intended to illustrate the above: a compound of formula I which has numerous potential sites for oxidative metabolism, e.g., benzyl hydrogens and hydrogens bonded to a nitrogen atom, is prepared as a series of analogues in which various combinations of hydrogens are substituted for deuterium in such a way that as a result some, most or all of these hydrogen atoms are replaced by deuterium atoms. Determining the half-life provides a suitable and accurate measure of the degree of improvement in resistance to oxidative metabolism. Thus, it is determined that the half-life of the parent compound can be extended up to 100% as a result of this replacement of hydrogen with deuterium.

The substitution of hydrogen for deuterium in a compound of formula I can also be used to achieve a favorable change in the metabolite spectrum of the parent compound in order to reduce or eliminate unwanted toxic metabolites. For example, if a toxic metabolite results from oxidative cleavage of a carbon-hydrogen C-H bond, then it can be rationally assumed that the deuterium-labeled analog will significantly reduce or eliminate the production of the undesired metabolite, even if the oxidation alone is not a rate-limiting step. Additional information relating to the prior art in relation to the exchange of hydrogen for deuterium is given, for example, in Hanzlik and others, J. Org. Chem. 55, 3992-3997, 1990; Reider et al., J. Org. Chem. 52, 3326-3334, 1987; Foster, Adv. DrugRes. 14, 1-40, 1985; Gillette et al., Biochemistry 33 (10) 2927-2937, 1994; and Jarman et al. Carcinogenesis 16(4), 683-688, 1993.

The invention also relates to mixtures of compounds of formula I 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 mixtures are particularly preferably mixtures of two stereoisomeric compounds. However, preference is also given to mixtures of two or more compounds of formula I.

In addition, the invention relates to a process for the preparation of compounds of formula I, which is characterized in that

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 using 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 urea formation conditions to give a compound of formula I,

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, 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 the compound formula VIII is reacted with a compound of formula IX under standard amidation or urea formation conditions to give a compound of formula I,

c) a compound of formula XII is iodinated 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 obtain 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 urea formation conditions to obtain compounds of formula I,

d) the base of the compound of formula I is converted to one of its salts by treatment with an acid, or

e) the acid of the compound of formula I is converted into one of its salts by treatment with a base.

In addition, in each case, the reactions can be carried out in stages, and it is also possible to modify the reaction sequence of the combination of structural elements with the adaptation of the concept of using protective groups.

The starting materials or starting compounds are well known. If they are new, they can be obtained using well-known methods.

If necessary, the starting materials can also be formed in situ without isolating them from the reaction mixture, but instead immediately converting them further into the compounds of formula I.

The compounds of formula I are preferably obtained by releasing them from functional derivatives by solvolysis, in particular hydrolysis, or by hydrogenolysis. Preferred starting materials for solvolysis or hydrogenolysis are substances which contain appropriate protected amino, carboxyl and/or hydroxyl groups in place of one or more free amino, carboxyl and/or hydroxyl groups, preferably substances which carry an amino protecting group in place of the H atom that is attached to the N atom. In addition, preference is given to starting materials which carry a hydroxyl protecting group instead of the H atom of the hydroxyl group. Preference is also given to starting materials which carry a protected carboxyl group instead of a free carboxyl group.

Also, a plurality of the same or different protected amino, carboxyl and/or hydroxyl groups may be present in the parent compound molecule. If the protecting groups present differ from one another, they can in many cases be cleaved off selectively.

The term "amino protecting group" is generally known and refers to groups that are suitable for protecting (blocking) an amino group from chemical reactions, but which can be easily removed after the desired chemical reaction has been carried out in another part of the molecule. Typical of such groups are, in particular, unsubstituted or substituted acyl groups, in addition unsubstituted or substituted aryl (eg 2,4-dinitrophenyl) or aralkyl groups (eg benzyl, 4-nitrobenzyl, triphenylmethyl). In addition, since amino protecting groups are removed after the desired reaction or sequence of reactions, their type and size are not critical, but preference is given to groups containing 1-20, in particular 1-8 carbon atoms. The term "acyl group" should be understood in the broadest sense in connection with the present method. It embraces acyl groups derived from aliphatic, araliphatic, aromatic or heterocyclic carboxylic acids or sulfonic acids and in particular alkoxycarbonyl, aryloxycarbonyl and in particular 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, alkoxycarbonyl such as methoxycarbonyl, ethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl , BOC, 2-iodoethoxycarbonyl, aralkoxycarbonyl such as CBZ, 4-methoxybenzyloxycarbonyl or FMOC. Preferred acyl groups are CBZ, FMOC, benzyl and acetyl.

The term "acid protecting group" or "carboxyl protecting group" is also commonly known and refers to groups that are suitable for protecting the -COOH group from chemical reactions, but which can be easily removed after the desired chemical reaction has been carried out in another part of the molecule. The use of esters in place of free acids, for example substituted and unsubstituted alkyl esters (such as methyl, ethyl, t-butyl and their substituted derivatives), substituted and unsubstituted benzyl esters or silyl esters, is typical. The type and size of the acid protecting group is not critical, but preference is given to groups containing 1-20, in particular 1-10 carbon atoms.

The term "hydroxyl protecting group" is also commonly known and refers to groups that are suitable for protecting the hydroxyl group from chemical reactions, but which can be easily removed after the desired chemical reaction has been carried out in another part of the molecule. Typical of such groups are the aforementioned unsubstituted or substituted aryl, aralkyl or acyl groups, in addition also alkyl groups. The type and size of the hydroxyl protecting groups are not critical, but preference is given to groups containing 1-20, in particular 1-10 carbon atoms. Examples of hydroxyl protecting groups are, inter alia, benzyl, p-nitrobenzoyl, p-toluenesulfonyl and acetyl, with benzyl and acetyl being preferred.

Additional typical 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 formula I for use as starting materials can be obtained by known methods for the synthesis of amino acids and peptides, as described, for example, in these standard works and patent applications.

The compounds of formula I are released from their functional derivatives, depending on the protecting group used, for example with strong acids, preferably with trifluoroacetic acid or perchloric acid, but also with other strong inorganic acids such as hydrochloric acid or sulfuric acid, strong organic acids such as trichloroacetic acid, or sulfonic acids such as benzoyl or p-toluenesulfonic acid. The presence of an additional inert solvent and/or catalyst is possible, but not always necessary.

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

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 t-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 ether or monoethyl ether, diethylene glycol dimethyl 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 such as, for example, acetic acid or acetic anhydride, nitro compounds such as nitromethane or nitrobenzene, optionally also mixtures of said solvents with one another or mixtures with water.

The amount of solvent is not critical; preferably, 10 g to 500 g of solvent may be added per g of compound of formula I to be reacted.

It may be preferable to add an acid-binding agent, for example an alkali metal or alkaline earth metal hydroxide, carbonate or bicarbonate, or other alkali or alkaline earth metal salts of weak acids, preferably potassium, sodium or calcium salts, or to add an organic base such as, for example, triethylamine, dimethylamine, pyridine or quinoline, or an excess of the amine component.

The compounds obtained according to the invention can be separated from the corresponding solution in which they were obtained (for example, by centrifugation and washing) and can be stored in another composition after separation, or they can be left directly in the solution in which they were received. The compounds obtained according to the invention can also be incorporated into the desired solvents for a particular application.

The duration of the reactions depends on the chosen reaction conditions. In general, the duration of the reactions is from 0.5 hour to 10 days, preferably from 1 to 24 hours. When using a microwave oven, the reaction time can be reduced to values from 1 to 60 minutes.

The compounds of formula I, as well as the starting materials for their preparation, are also prepared by known methods 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), for example under reaction conditions which are known and suitable for said reactions. In this case, it is also possible to use well-known variants, which are not described here in more detail.

Conventional work-up steps, such as, for example, adding water to the reaction mixture and extracting, make it possible to obtain compounds after removal of the solvent. It may be preferable if further purification of the product is followed by distillation or crystallization, or by chromatographic purification.

An acid of formula I can be converted to the corresponding addition salt using a base, for example by reacting equivalent amounts of acid and base in an inert solvent such as ethanol, followed by evaporation. Suitable bases for this reaction are in particular those which give physiologically acceptable salts. Thus, an acid of formula I can be converted to the corresponding metal salt, in particular an alkali or alkaline earth metal salt, using a base (eg sodium hydroxide, potassium hydroxide, sodium carbonate or potassium carbonate) or to the corresponding ammonium salt. Organic bases which give physiologically acceptable salts, such as, for example, ethanolamine, are also suitable for this reaction.

Alternatively, a base of formula I can be converted to the corresponding acid addition salt using an acid, for example by reacting equivalent amounts of base and acid in an inert solvent such as ethanol, followed by evaporation. Suitable acids for this reaction are in particular those which give physiologically acceptable salts. Thus, it is possible to use inorganic acids, for example sulfuric acid, nitric acid, hydrohalic acids such as hydrochloric acid or hydrobromic acid, phosphoric acid such as phosphoric acid, sulfamic acid and, in addition, organic acids, in particular aliphatic, alicyclic, araliphatic, aromatic or heterocyclic, mono- or polybasic carboxylic, sulfonic 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, methane or ethanesulfonic acid, ethanedisulfonic acid, 2-hydroxysulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalene mono- and disulfonic acids or lauryl sulfuric acid. Salts with physiologically unacceptable acids, such as picrates, can be used to isolate and/or purify compounds of formula I.

The compounds of formula I have been found to be well tolerated and have valuable pharmacological properties.

Since adenosine receptors such as A _2A and A _2B have been shown to down-regulate the immune response during inflammation and protect tissues from immune damage, inhibition of adenosine receptor signaling can be used to enhance and prolong the immune response.

Provided herein are methods for enhancing an immune response. In one example, the method increases the desired and targeted tissue damage, such as tumor damage, such as cancer. Disclosed herein are methods of inhibiting one or more processes that promote the production of extracellular adenosine and adenosine-driven signaling through adenosine receptors. For example, enhancement of the immune response, local tissue inflammation, and targeted tissue destruction is achieved: by inhibiting or reducing local hypoxia of adenosine-producing tissue; by decomposing (or rendering inactive) accumulated extracellular adenosine; by preventing or reducing the expression of adenosine receptors on immune cells; and/or by inhibiting/antagonizing adenosine ligand signaling through adenosine receptors. The results disclosed herein demonstrate that in vivo administration of agents that disrupt the "hypoxia -> adenosine accumulation -> immunosuppressive adenosine receptor signaling to immune cells" pathway in subjects suffering from various diseases (e.g., cancer and sepsis) can in vivo lead to the provision of tumor treatment or improved immunization.

In one example, the method includes administering one or more extracellular adenosine and/or adenosine receptor inhibitors, such as an adenosine receptor antagonist. To improve the effectiveness of the vaccine, one or more inhibitors of adenosine receptors and/or inhibitors of extracellular adenosine can be administered in combination with the vaccine. In one example, one or more adenosine receptor inhibitors or extracellular adenosine inhibitors are administered to enhance the immune response/inflammation. In another example, the method achieves targeted tissue damage, such as tumor destruction.

Therefore, the invention also relates to the use of the compounds according to the invention for the preparation of a medicament for the treatment and/or prevention of diseases caused, provoked and/or enhanced by adenosine or other A _2A and/or A _2B receptor agonists.

Thus, the invention also relates in particular to a medicament containing at least one compound according to the invention and/or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all proportions, for applications for the treatment and/or prevention of physiological and/or pathophysiological conditions.

Particular preference is given in particular to physiological and/or pathophysiological conditions that are associated with adenosine receptors A _2A and/or A _2B .

Physiological and/or pathophysiological conditions mean physiological and/or pathophysiological conditions that are of medical importance, such as, for example, diseases or disorders and medical disorders, complaints, symptoms or complications, and the like, in particular diseases.

The invention further relates to a medicament containing at least one compound according to the invention and/or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all proportions, for use in the treatment and/or prevention of physiological and/or pathophysiological conditions selected from the group consisting of hyperproliferative and infectious diseases and disorders.

The invention further relates to a medicinal product containing at least one compound according to the invention and/or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all proportions, for use in treatment and/or prophylaxis physiological and/or pathophysiological conditions selected from the group consisting of hyperproliferative and infectious diseases and disorders, where the hyperproliferative disease or disorder is a malignant neoplasm.

Thus, the invention particularly preferably relates to a medicinal product containing at least one compound according to the invention and/or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all ratios, as as defined above, wherein the malignancy is selected from the group consisting of acute and chronic lymphocytic leukemia, acute granulocytic leukemia, adrenal cortex cancer, 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, urogenital carcinoma, glioma, glioblastoma, hairy cell leukemia, head and neck carcinoma, Hodgkin's disease, Kaposi's sarcoma, lung carcinoma, lymphoma, malignant carcinoid tumor and, malignant hypercalcemia, malignant melanoma, malignant pancreatic insulinoma, medullary thyroid carcinoma, melanoma, multiple myeloma, mycosis fungoides, myeloid and lymphocytic leukemia, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, osteogenic sarcoma, ovarian carcinoma, pancreatic carcinoma, polycythemia vera, primary brain carcinoma, primary macroglobulinemia, prostate cancer, renal cell carcinoma, rhabdomyosarcoma, skin cancer, small cell lung cancer, soft tissue sarcoma, squamous cell carcinoma, gastric cancer, testicular cancer, thyroid cancer, and Wilms' tumor.

The invention even more preferably relates to a drug containing at least one compound according to the invention and/or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all proportions, for use in treatment and/ or prevention of physiological and/or pathophysiological conditions selected from the group consisting of hyperproliferative and infectious diseases and disorders, where the hyperproliferative disease or disorder is selected from the group consisting of age-related macular degeneration, Crohn's disease, cirrhosis, chronic inflammation-related disorders, proliferative diabetic retinopathy, proliferative vitreoretinopathy, retrolental fibroplasia, granulomatosis, immune hyperproliferation associated with organ or tissue transplantation, and an immunoproliferative disease or disorder selected from the group consisting of inflammatory bowel disease, soriasis, rheumatoid arthritis, systemic lupus erythematosus (SLE), vascular hyperproliferation against the background of retinal hypoxia and vasculitis.

The invention even more preferably relates to a drug containing at least one compound according to the invention and/or one of its physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all proportions, for use in treatment and/ or prevention of physiological and/or pathophysiological conditions selected from the group consisting of hyperproliferative and infectious diseases and disorders, where the infectious disease or disorder is selected from the group consisting of

a) virus-induced infectious diseases that are caused by retroviruses, hepadnaviruses, herpesviruses, flaviviruses and/or adenoviruses, where retroviruses are selected from lentiviruses or oncoretroviruses, where lentiviruses are selected from the group consisting of HIV-1, HIV-2, FIV, BIV, SIV viruses , SHIV, GAEV, VMV and EIAV, and oncoretroviruses are selected from the group consisting of HTLV-I, HTLV-II and BLV, hepadnaviruses are selected from the group consisting of HBV, GSHV and WHV, herpesviruses are selected from the group consisting of HSV I, HSV II, EBV, VZV, HCMV or HHV 8, and flaviviruses are selected from the group consisting of HCV, West Nile virus and yellow fever virus,

b) bacterial infectious diseases that are caused by gram-positive bacteria, where gram-positive bacteria are selected from the group consisting of methicillin-sensitive and methicillin-resistant staphylococci resistant staphylococci (including Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus saprophytics and coagulococcus-negative staphylococci) , Staphylococcus aureus with intermediate sensitivity to glycopeptides (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 (including vancomycin-susceptible and vancomycin-resistant strains such as Enterococcus faecalis and Enterococcus faecium), Clostridium difficile, l isteria monocytogenes, Corynebacterium jeikeium, Chlamydia spp (including C.pneumoniae) and Mycobacterium tuberculosis,

c) bacterial infectious diseases that are caused by gram-negative bacteria, where the gram-negative bacteria are selected from the group consisting of the genus Enterobacteriacae, including Escherichia spp. (including Escherichia coli), Klebsiella spp., Enterobacter spp., Citrobacter spp., Serratia spp., Proteus spp., Providencia spp., Salmonella spp., Shigella spp., Pseudomonas genus (including P. aeruginosa), Moraxella spp. (including M. catarrhalis), Haemophilus spp. and Neisseria spp.,

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.

The medicaments disclosed above are intended to include the respective use of the compounds according to the invention for the preparation of a medicament for the treatment and/or prevention of the above physiological and/or pathophysiological conditions.

It is further contemplated that the medicaments disclosed above comprise an appropriate method for the treatment and/or prevention of the above physiological and/or pathophysiological conditions, wherein at least one compound according to the invention is administered to a patient in need of such treatment.

Compounds according to the invention preferably exhibit beneficial biological activity, which can be readily demonstrated in enzyme assays and animal experiments as described in the examples. In such enzyme activity assays, the compounds according to the invention preferably exhibit and elicit an inhibitory effect, which is usually confirmed by IC ₅₀ values in the suitable range, preferably in the micromolar range and more preferably in the nanomolar range.

The compounds according to the invention can be administered to humans or animals, in particular to mammals such as monkeys, dogs, cats, rats or mice, and can be used for therapeutic treatment in humans or animals and for combating the aforementioned diseases. In addition, they can be used as diagnostic tools or as reagents.

In addition, the compounds according to the invention can be used to isolate and study the activity or expression of A _2A and/or A _2B adenosine receptors. In addition, they are particularly suitable for use in methods for diagnosing diseases against the background of impaired activity of the A _2A and/or A _2B adenosine receptors. Therefore, the invention also relates to the use of the compounds according to the invention for isolating and assaying the activity or expression of A _2A and/or A _2B adenosine receptors, or as binders and inhibitors of A _2A and/or A _2B adenosine receptors ^.

For diagnostic purposes, the compounds according to the invention, for example, may be radiolabeled. Examples of radioactive labels are ³ H, ¹⁴ C, ²³¹ I and ¹²⁵ I. The preferred labeling method is the iodogen method (Fraker et al., 1978). In addition, the compounds according to the invention can be labeled with enzymes, fluorophores and chemophores. Examples of enzymes are alkaline phosphatase, β-galactosidase and glucose oxidase, an example of a fluorophore is fluorescein, an example of a chemophore is luminol, and automated detection systems such as fluorescent staining are described in, for example, US 4,125,828 and US 4,207,554.

Moreover, the present invention relates to pharmaceutical compositions containing the compounds of the present invention and their use in the treatment and/or prevention of diseases and disorders in which partial or complete inactivation of A _2A and/or A _2B adenosine receptors may be beneficial.

The compounds of formula I can be used for the preparation of pharmaceutical preparations, in particular by non-chemical methods. In this case, they are administered in a suitable dosage form together with at least one solid, liquid and/or semi-liquid excipient or excipient and, optionally, in combination with one or more additional active compound(s). yami).

Therefore, the invention also relates to pharmaceutical preparations containing at least one compound of formula I and/or one of its physiologically acceptable salts, derivatives, solvates and stereoisomers, including mixtures thereof in all proportions. In particular, the invention also relates to pharmaceutical preparations which additionally contain excipients and/or excipients, as well as to pharmaceutical preparations which contain at least one additional active drug compound.

In particular, the invention also relates to a process for preparing a pharmaceutical preparation, characterized in that a compound of formula I and/or one of its physiologically acceptable salts, derivatives, solvates and stereoisomers, including mixtures thereof in all proportions, is administered in a suitable dosage form together with a solid , a liquid or semi-liquid excipient or excipient, and optionally with an additional active drug compound.

Pharmaceutical preparations according to the invention can be used as medicines in human and veterinary medicine. The patient or host may be any mammalian species such as, for example, primates, especially humans; rodents, including mice, rats and hamsters; rabbits; horses, cattle, dogs, cats, etc. Animal models are of interest for experimental studies where they provide a model for the treatment of human disease.

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 novel compounds, e.g. water, vegetable oils (such as sunflower oil or fish oil), benzyl alcohols, polyethylene glycols, gelatin, carbohydrates such as lactose or starch, magnesium stearate, talc, lanolin or petroleum jelly. Through his expert knowledge, the person skilled in the art is aware of which excipients are suitable for the desired drug formulation. In addition to solvents, such as water, physiological saline or alcohols, such as, for example, ethanol, propanol or glycerol, sugar solutions, such as glucose or mannitol solutions, or a mixture of these solvents, gelling agents, tablet auxiliaries and others can also be used. carriers of the active ingredient, for example, lubricants, stabilizers and/or wetting agents, emulsifiers, salts to influence the osmotic pressure, antioxidants, dispersants, antifoams, buffering agents, flavoring and/or aromatic substances or taste correctors, preservatives, solubilizers or coloring agents. If necessary, preparations or drugs in accordance with the invention may contain one or more additional active compounds, for example, one or more vitamins.

If necessary, preparations or medicinal products in accordance with the invention may contain one or more additional active compounds and/or one or more action enhancers (auxiliaries).

The terms "pharmaceutical composition" and "pharmaceutical preparation" for the purposes of the present invention are used as synonyms.

As used herein, "pharmaceutically acceptable" refers to drugs, precipitating agents, excipients, excipients, stabilizers, diluents, and other agents that facilitate the administration of pharmaceutical preparations derived therefrom to a mammal without undesirable physiological side effects, such as, for example, nausea, dizziness, digestive problems, or the like.

In the case of pharmaceutical preparations for parenteral administration, there is a need for isotonicity, normal hydration and tolerability and safety of the formulation (low toxicity), the excipients used and the primary packaging. Surprisingly, the compounds according to the invention preferably have the advantage that direct application is possible and additional purification steps to remove toxicologically unacceptable substances, such as, for example, high concentrations of organic solvents or other toxicologically unacceptable excipients, are thus not necessary before the use of compounds according to the invention in pharmaceutical formulations.

The invention particularly preferably also relates to pharmaceutical preparations containing at least one compound according to the invention in precipitated non-crystalline, precipitated crystalline or dissolved or suspended form, and optionally excipients and/or excipients and/or additional pharmaceutically active compounds.

Preferably, the compounds according to the invention allow the preparation of highly concentrated formulations without harmful, undesirable aggregation of the compounds according to the invention. Thus, with the compounds according to the invention it is possible to obtain ready-to-use solutions having a high content of active ingredients with aqueous solvents or in an aqueous medium.

The compounds and/or their physiologically acceptable salts and solvates can also be lyophilized and the resulting lyophilisates used, for example, for the preparation of injections.

Aqueous preparations can be obtained by dissolving or suspending the compounds according to the invention in an aqueous solution and optionally adding excipients. To this end, certain volumes of stock solutions containing said additional excipients in a certain concentration are preferably added to a solution or suspension having a certain concentration of the compounds according to the invention, and the mixture is optionally diluted with water to a pre-calculated concentration. Alternatively, excipients may be added in solid form. The amounts of stock solutions and/or water required in each case may then be added to the resulting aqueous solution or suspension. The compounds according to the invention can preferably also be dissolved or suspended directly in a solution including all additional excipients.

Preferably, solutions or suspensions can be prepared which include compounds according to the invention and have a pH in the range of 4 to 10, preferably 5 to 9, and an osmolality in the range of 250 to 350 mOsmol/kg. The pharmaceutical preparation can thus be administered directly, essentially without pain, intravenously, intra-arterially, intra-articularly, subcutaneously or transdermally. In addition, the drug can also be added to infusion solutions, such as, for example, glucose solution, isotonic saline or Ringer's solution, which may also contain additional active compounds, which also makes it possible to administer relatively large amounts of the active compound.

Pharmaceutical preparations according to the invention may also contain mixtures of several compounds according to the invention.

The preparations according to the invention are well tolerated physiologically, they are easy to prepare, they can be well distributed and preferably they are stable with respect to assay conditions, degradation products and aggregates during storage and transport, and under repeated freezing and thawing procedures. They can preferably be stored in a stable condition for at least three months to two years at refrigerator temperature (2-8°C) and at room temperature (23-27°C) and relative air humidity (R.H. ) 60%.

For example, compounds according to the invention can be stored in a stable state by drying and, if necessary, converted into a ready-to-use pharmaceutical preparation by dissolution or suspension. Possible drying methods are, for example, but not limited to, nitrogen gas drying, vacuum oven drying, lyophilization, washing with organic solvents and subsequent air drying, liquid bed drying, fluidized bed drying, spray drying, roller drying, layer drying, air drying at room temperature and further methods.

The term "effective amount" means the amount of a drug or pharmaceutical active ingredient that causes a biological or medical response in a tissue, system, animal or human that is sought or desired, for example, by a researcher or physician.

In addition, the term "therapeutically effective amount" means an amount which, compared with a corresponding subject who has not received this amount, has the following effects: improving treatment, curing, preventing or eliminating a disease, syndrome, disease state, complaints, disturbance or prevention side effects or also slowing down the progression of the disease, easing complaints or disorders. The term "therapeutically effective amount" also encompasses amounts that are effective to enhance normal physiological function.

When using preparations or medicaments according to the invention, the compounds according to the invention and/or their physiologically acceptable salts and solvates are usually used in analogy to known, commercially available preparations or formulations, preferably in dosages between 0.1 and 500 mg, in particular , 5 and 300 mg, per unit used. The daily dose is preferably between 0.001 and 250 mg/kg, in particular 0.01 and 100 mg/kg, body weight. The drug can be administered one or more times a day, for example two, three or four times a day. However, the individual dose for a patient depends on a large number of individual factors such as, for example, the efficacy of the particular compound used, age, body weight, general health, sex, nutrition, time and route of administration, rate of excretion, combination with other drugs, and the severity and duration of a particular disease.

A measure of the uptake of an active compound of a drug in the body is its bioavailability. If the active compound of the drug is delivered to the body intravenously as an injection solution, its absolute bioavailability, i.e. the proportion of the pharmaceutical that reaches the systemic blood, i.e. the main circulatory system, in unchanged form, is 100%. In the case of oral administration of a therapeutically active compound, the active compound is usually in the form of a solid in the formulation and therefore must first be dissolved in order to be able to cross barriers, for example, the gastrointestinal tract, oral mucosa, nasal mucosa or skin. , in particular, the stratum corneum of the epidermis, or be absorbed by the body. Pharmacokinetic data, i.e. bioavailability data can be obtained similarly to the method described in . J. Shaffer et al., J. Pharm. Sciences, 88 (1999), 313-318.

In addition, drugs of this type can be obtained using one of the methods that are well known in the pharmaceutical prior art.

Drugs may be adapted for administration by any suitable route desired, for example, oral (including buccal or sublingual), rectal, pulmonary, nasal, topical (including buccal, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous , intradermal and, in particular, intra-articular) routes of administration. Drugs of this type can be obtained using any method known in the pharmaceutical field, for example, by combining the active compound with the excipient(s) or auxiliary(s) substance(s).

Preferably, parenteral administration is suitable for administering the medicaments according to the invention. In the case of parenteral administration, intra-articular administration is particularly preferred.

Thus, the invention preferably also relates to the use of a pharmaceutical preparation according to the invention for intra-articular administration for the treatment and/or prevention of physiological and/or pathophysiological conditions selected from the group consisting of osteoarthritis, traumatic cartilage injuries, arthritis, pain, allodynia or hyperalgesia. .

The advantage of intra-articular administration is that the compound according to the invention can be injected directly into the synovial fluid in close proximity to the articular cartilage and is also able to spread from there into the cartilage tissue. The pharmaceutical preparations according to the invention can thus also be administered directly to the joint space and thus act directly at the site of action, as intended. The compounds according to the invention are also suitable for the preparation of parenteral formulations having a slow, sustained and/or controlled release of the active compound. Thus, they are also suitable for the preparation of sustained release formulations, which are preferred by the patient since the administration only needs to be repeated at relatively long intervals.

Medicinal products adapted for parenteral administration include aqueous and non-aqueous sterile injectable solutions containing antioxidants, buffers, bacteriostatic agents and solutes, by which the formulation is isotonic with respect to the blood or synovial fluid of the recipient being treated; as well as aqueous and non-aqueous sterile suspensions, which may contain suspension media and thickeners. Formulations may be supplied in single-dose or multi-dose containers, such as sealed ampoules and vials, and stored in a freeze-dried (lyophilized) state so that only the addition of a sterile carrier liquid, such as water for injection, is necessary immediately prior to use. Solutions and suspensions for injection prepared in accordance with the disclosed formulation can be obtained from sterile powders, granules and tablets.

The compounds according to the invention can also be administered as liposome delivery systems such as, for example, small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from various phospholipids such as, for example, cholesterol, stearylamine or phosphatidylcholines.

The compounds of the invention can also be combined with soluble polymers as drug targeting carriers. Such polymers may include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamidophenol, polyhydroxyethylaspartamidophenol or polyethylene oxide-polylysine substituted with palmitoyl radicals. In addition, the compounds according to the invention can be combined with biodegradable polymers which are suitable for providing sustained drug release, such as, for example, polylactic acid, poly-epsilon-caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydroxypyrans, polycyanoacrylates, lactic acids with glycolic acid, polymers such as dextran-methacrylate conjugates, polyphosphoesters, various polysaccharides and polyamines, and poly-ε-caprolactone, albumin, chitosan, collagen or modified gelatin, and cross-linked or amphipathic hydrogel block copolymers.

Suitable for enteral administration (oral or rectal) are, in particular, tablets, dragees, capsules, syrups, juices, drops or suppositories, and suitable for topical administration are ointments, creams, pastes, lotions, gels, sprays, foams, aerosols, solutions (eg solutions in alcohols such as ethanol or isopropanol, acetonitrile, DMF, dimethylacetamide, 1,2-propanediol or mixtures thereof with each other and/or with water) or powders. Liposomal formulations are also particularly suitable for topical application.

In the case of an ointment formulation, the active compound can be used with either a paraffinic or water-miscible cream base. Alternatively, the active compound may be formulated as a cream with an oil-in-water or water-in-oil cream base.

Drugs adapted for transdermal administration can be delivered as independent patches for prolonged, intimate contact with the recipient's epidermis. Thus, for example, the active compound can be delivered from the patch by iontophoresis as outlined in Pharmaccutical Rescarch, 3 (6), 318 (1986).

It goes without saying that, in addition to the components specifically mentioned above, the medicaments according to the invention may also contain other agents customary in the art for a specific type of pharmaceutical formulation.

The invention also relates to a kit (set) consisting of separate packages

a) an effective amount of a compound of formula I and/or physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers thereof, including mixtures thereof in all proportions, and

b) an effective amount of the additional active drug compound.

The kit includes suitable containers such as boxes or packs, individual bottles, bags or ampoules. The kit may include, for example, separate ampoules, each containing an effective amount of a compound of formula I and/or its pharmaceutically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all ratios, and an effective amount of an additional active drug compound in dissolved or lyophilized form.

In addition, drugs according to the invention can be used to provide additive or synergistic effects in the case of certain known therapies and/or can be used to restore the effectiveness of certain existing therapies.

In addition to the compounds according to the invention, pharmaceutical preparations according to the invention may also contain additional active compounds of the drug, for example for use in the treatment of cancer, other anticancer drugs. For the treatment of the other diseases mentioned, the pharmaceutical preparations according to the invention may also, in addition to the compounds according to the invention, contain additional active compounds of the drug, which, as is known to the person skilled in the art, are used for their treatment.

In one general embodiment, methods are provided to enhance an immune response in a host in need thereof. The immune response can be enhanced by decreasing T cell tolerance, including by increasing IFN-γ release, decreasing production or activation of regulatory T cells, or increasing production of antigen-specific memory T cells in the host. In one embodiment, the method includes administering a compound of the present invention to a host in combination or alternately with an antibody. In specific sub-embodiments, the antibody is a therapeutic antibody. In one particular embodiment, a method for enhancing the efficacy of passive antibody therapy is provided, comprising administering a compound of the present invention in combination or alternately with one or more passive antibodies. This method may increase the effectiveness of antibody therapy for the treatment of proliferative disorders such as cancer, or may increase the effectiveness of therapy in the treatment or prevention of infectious diseases. The compound of the present invention can be administered in combination or alternately with antibodies such as, for example, rituximab, herceptin or erbitux.

In another general embodiment, a method is provided for treating or preventing abnormal cell proliferation, comprising administering a compound of the present invention to a host in need thereof, substantially in the absence of another anticancer agent.

In another general embodiment, a method is provided for treating or preventing abnormal cell proliferation in a host in need thereof, comprising administering to the host a first compound of the present invention substantially in combination with a first antineoplastic agent, followed by administering a second A _2A and/or A _2B receptor antagonist. . In one sub-embodiment, the second antagonist is administered substantially in the absence of the other anticancer agent. In another general embodiment, a method is provided for treating or preventing abnormal cell proliferation in a host in need thereof, comprising administering to the host a compound of the present invention substantially in combination with a first anticancer agent, and thereafter administering a second anticancer agent in the absence of an antagonist.

Thus, the treatment of cancer disclosed herein may be carried out either in the form of therapy with a compound of the present invention, or in combination with surgery, radiation, or chemotherapy. Chemotherapy of this type may include the use of one or more active compounds from the following classes of anticancer active compounds:

(i) antiproliferative/antineoplastic/DNA damaging active compounds and combinations thereof that are used in medical oncology, such as alkylating active compounds (eg, cisplatin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulfan, and nitrosoureas); antimetabolites (eg, antifolates such as fluoropyrimidines such as 5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside, hydroxyurea and gemcitabine); antitumor antibiotics (eg anthracyclines such as adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin and mithramycin); antimitotic active compounds (eg vinca alkaloids such as vincristine, vinblastine, vindesine and vinorelbine and taxoids such as taxol and taxotere); topoisomerase inhibitors (eg, epipodophyllotoxins such as etoposide and teniposide, amsacrine, topotecan, irinotecan, and camptothecin) and active compounds affecting cell differentiation (eg, all-trans retinoic acid, 13-cis-retinoic acid, and fenretinide) ;

(ii) cytostatic active compounds such as antiestrogen (eg, tamoxifen, toremifene, raloxifene, droloxifene, and iodoxifene), estrogen receptor regulators (eg, fulvestrant), antiandrogens (eg, bicalutamide, flutamide, nilutamide, and cyproterone acetate), LHRH antagonists, or LHRH agonists (eg goserelin, leuprorelin and buserelin), progesterones (eg megestrol acetate), aromatase inhibitors (eg anastrozole, letrozole, vorazole and exemestane) and 5α-reductase inhibitors such as finasteride;

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

(iv) inhibitors of growth factor function, eg anti-growth factor antibodies, anti-growth factor receptor antibodies, eg anti-erbb2 antibody trastuzumab [Herceptin™] and anti-erbbl antibody cetuximab [C225]), farnesyl transferase inhibitors, tyrosine kinase inhibitors and inhibitors serine/threonine kinases, e.g. epidermal growth factor family inhibitors (e.g. EGFR family tyrosine kinase inhibitors such as N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazoline-4-amine ( gefitinib, AAD1839), N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazoline-4-amine (erlotinib, OSI-774) and 6-acrylamido-N-(3-chloro-4-fluorophenyl )-7-(3-morpholinopropoxy)quinazoline-4-amine (CU 1033), eg inhibitors of the platelet growth factor family and eg inhibitors of the hepatocyte growth factor family;

(v) anti-angiogenic active compounds such as bevacizumab, angiostatin, endostatin, linomide, batimastat, captopril, cartilage inhibitor, genistein, interleukin 12, lavendustin, medroxyprogesterone acetate, recombinant human platelet factor 4, tecogalan, thrombospondin, TNP-470, anti-VEGF monoclonal antibody, soluble VEGF receptor chimeric protein, anti-VEGF receptor antibodies, anti-PDGF receptor antibodies, integrin inhibitors, tyrosine kinase inhibitors, serine/threonine kinase inhibitors, antisense oligonucleotides, antisense oligodeoxynucleotides, miRNAs, anti-VEGF aptamers, pigment epithelial factor and compounds which have been published in international patent applications WO 97/22596, WO 97/30035, 97/32856 and 98/13354);

(vi) vascular disruptors such as combretastatin A4 and compounds that have been 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 therapies, such as those directed at the targets mentioned above, such as ISIS 2503, antisense therapy directed against the Ras gene;

(viii) gene therapy methods, including, for example, atypical, modified gene replacement methods such as atypical p53 or atypical BRCA1 or BRCA2, GDEPT (Gene Targeted Enzyme Prodrug Therapy) methods, such as methods that use cytosine deaminase, thymidine kinase, or bacterial nitroreductase enzyme, and methods that increase patient tolerance to chemotherapy or radiotherapy, such as multidrug resistance therapy; And

(ix) methods of immunotherapy, including, for example, methods of increasing the immunogenicity of the patient's tumor cells ex vivo and in vivo, such as transfection with cytokines such as interleukin 2, interleukin 4 or granulocyte-macrophage colony-stimulating factor, methods of reducing T-cell anergy, methods using transfected immune cells such as cytokine-transfected dendritic cells, methods using cytokine-transfected tumor cells, and methods using anti-idiotypic antibodies;

(x) chemotherapeutic agents, including, for example, abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, anastrozole, arsenic trioxide, asparaginase, live BCG, bevacizumab, bexarotene, bleomycin, bortezomib, busulfan, calusterone, camptothecin, capecitabine, carboplatin, carmustine, celecoxib, cetuximab, chlorambucil, cinacalcet, cisplatin, cladribine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, darbepoetin alfa, daunomycin, denileukin-diphthitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone, epirubicin, epoestinide, epoetin, exemestane, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, and gemcitabine.

Drugs from Table 1 may preferably, but not exclusively, be combined with compounds of formula I.

Even without additional embodiments, it is expected that a person skilled in the art will be able to use the above description to the widest extent. The preferred embodiments, therefore, are to be considered merely as a descriptive disclosure that does not limit the present invention in any way.

Thus, the following examples are intended to illustrate the invention and do not limit it. Unless otherwise indicated, percentages mean percentages by weight. All temperatures are in degrees Celsius. "Conventional work-up": if necessary, water is added, the pH is adjusted, if necessary, to values between 2 and 10, depending on the structure of the final product, the mixture is extracted with ethyl acetate or dichloromethane, the phases are separated, the organic phase is dried over sodium sulfate, filtered and evaporated, and the product is purified by silica gel chromatography and/or by crystallization.

Rf values are given for silica gel; mass spectrometry: EI (electron impact ionization): M ⁺ , FAB (fast atom bombardment): (M + H) ⁺ , THF (tetrahydrofuran), NMP (N-methylpyrrolidone), DMSO (dimethyl sulfoxide), EA (ethyl acetate), MeOH (methanol), TLC (thin layer chromatography).

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 - Absorbed fraction

IV - Intravenous

LC-MS/MS - Liquid chromatography with tandem mass spectrometry

LLOQ - Lower limit of quantitation

NC - Not rated

ND - Undefined

PEG - Polyethylene glycol

Pgp - Permeability Glycoprotein

PK - Pharmacokinetic (pharmacokinetics)

p / o - Orally (oral drug)

t _1/2 - Half-life

t _max - The time at which the maximum concentration of the drug substance is reached in plasma

UHPLC - Ultra High Performance Liquid Chromatography

V _ss - Volume of distribution (at steady state)

vol./rev. - Volume to volume

Example 1 Examples of Compounds of the Present Invention

The invention particularly relates to the compounds of Table 2 and their physiologically acceptable salts, derivatives, solvates, prodrugs and stereoisomers, including mixtures thereof in all ratios.

Example 2 Preparation of Compounds of the Present Invention and Analytical Methods

All solvents used were commercially available and were used without further purification. Reactions were typically carried out using anhydrous solvents under an inert nitrogen atmosphere. Flash column chromatography was usually performed using Silica gel 60 (particle size 0.035-0.070 mm).

Data from 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. NMR. All deuterated solvents typically contained from 0.03% to 0.05% v/v. tetramethylsilane, which was used as a standard signal (set to δ=0.00 for both ¹ H and ¹³ C).

LC-MS analyzes were performed on a SHIMADZU LC-MS instrument including a UFLC 20-AD MS detector LCMS 2020. The column used was Shim-pack XR-ODS, 2.2 μM, 3.0×50 mm. A linear gradient was applied starting at 95% A (A: 0.05% TFA in water) to 100% B (B: 0.05% TFA in acetonitrile) over 2.2 minutes with a total run time of 3.6 minutes . The column temperature was 40°C at a given flow rate of 1.0 ml/min. Using a diode array detector, scanning was performed in the range of 200–400 nm. The mass spectrometer was equipped with an electrospray ion source (ES) operating in positive or negative ion mode. Scanning on a mass spectrometer was performed in the range m/z 90–900 with a scan time of 0.6 s.

1. N-[4-methoxy-7-(morpholin-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-1-(2-methoxyethyl)-1H-pyrazole -4-carboxamide, 4

A. Ethyl 1-(2-methoxyethyl)-1H-pyrazole-4-carboxylate

To a solution of ethyl 1H-pyrazole-4-carboxylate (950 mg, 6.78 mmol) in N,N-dimethylformamide (18 mL) was added 1-bromo-2-methoxyethane (1.14 g, 8.20 mmol), potassium carbonate (1.80 g, 13.8 mmol) at room temperature. The reaction mixture was irradiated with microwave radiation for 2 h at 160°C. When the reaction was complete, the solids were filtered off. The filtrate was then concentrated in vacuo to give ethyl 1-(2-methoxyethyl)-1H-pyrazole-4-carboxylate as a light yellow liquid (1.50 g, crude). MS: m/z=199.2 [M+H] ⁺ .

b. 1-(2-methoxyethyl)-1H-pyrazole-4-carboxylic acid

Water was added to a solution of ethyl 1-(2-methoxyethyl)-1H-pyrazole-4-carboxylate (1.50 g, crude) in tetrahydrofuran (10 ml). LiOH (20 ml, 32 mmol, 1.6 M) at room temperature. The resulting mixture was stirred for 3 hours at room temperature. When the reaction was complete, the mixture was quenched with H ₂ O (10 ml). The resulting mixture was extracted with dichloromethane (100 ml×3) and the organic phases were combined, washed with brine and dried over anhydrous Na ₂ SO ₄ . The solvent was removed under reduced pressure and the residue was purified by flash chromatography, eluting with MeOH in DCM (0%-70% gradient) to give 1-(2-methoxyethyl)-1H-pyrazole-4-carboxylic acid as white solids (600 mg, 52% in 2 steps). MS: m/z=171.2 [M+H] ⁺ .

c. 5-Bromo-4-chloropyridine-3-amine

To a solution of 3-bromo-4-chloro-5-nitropyridine (9.50 g, 40.01 mmol) in EtOH (500 ml) was added _NH4Cl (13.27 g, 248 mmol), water (50 ml), and Fe (22.39 g, 401 mmol) at room temperature. The resulting mixture was stirred for 2 hours at 80°C. When the reaction was complete, the solids were filtered off. The resulting mixture was concentrated under reduced pressure and the residue was purified by flash chromatography, eluting with 0%-60% EtOAc in petroleum ether to give 5-bromo-4-chloropyridine-3-amine as a yellow oil (5.94 g , 72%). MS: m/z=209.2 [M+H] ⁺ .

d. N-[7-bromo-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide

To a solution of 5-bromo-4-chloropyridine-3-amine (2.97 g, 14.3 mmol) in acetone (64 ml) was added benzoylisothiocyanate (4.47 g, 27.4 mmol) at room temperature. The resulting mixture was stirred for 16 hours at 50°C. When the reaction was complete, the solids were collected by filtration to give N-[7-bromo-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide as a yellow solid (3.36 g, 70% ). MS: m/z=333.8 [M+H] ⁺ .

e. N-[7-(morpholin-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide

Morpholine (7 ml), a precatalyst RuPhos 2nd generation (88 mg, 0.11 mmol), RuPhos (104 mg, 0.22 mmol), t-BuOK (380 mg, 3.39 mmol) at room temperature. After three vacuum/nitrogen flash cycles, the reaction mixture was irradiated with microwave radiation for 2 h at 160°C. Solids were filtered off. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography, eluting with EtOAc in petroleum ether (0%-66% gradient) to give N-[7-(morpholin-4-yl)-[1,3] thiazolo[4,5-c]pyridin-2-yl]benzamide as a white solid (99 mg, 29%). MS: m/z=341.0 [M+H] ⁺ .

f. N-[4-bromo-7-(morpholin-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide

At 70°C, to a solution of N-[7-(morpholin-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (653 mg, 1.92 mmol) in N, N-dimethylformamide (55 ml) was added dropwise with a solution of NBS (410 mg, 2.30 mmol) in N,N-dimethylformamide (15 ml) over 4 h with stirring. When the addition was complete, the resulting mixture was stirred for 10 minutes at 70°C. The resulting mixture was concentrated in vacuo to give the crude product, which was washed with hot EA (100 mL x 3) to give N-[4-6rom-7-(morpholin-4-yl)-[1,3]thiazolo[4, 5-c]pyridin-2-yl]benzamide as a light yellow solid (437 mg, 53%). MS: m/z=418.8 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.23-8.15 (m, 2H), 7.72 (s, 1H), 7.60-7.46 (m, 3H), 3.86-3.78 (m, 4H), 3.25-3.18 (m, 4H).

g. 4-methoxy-7-(morpholin-4-yl)-[1,3]thiazolo[4,5-c]pyridine-2-amine

To a solution of N-[4-bromo-7-(morpholin-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (388 mg, 0.94 mmol) in methanol (20 ml) MeONa in MeOH (7.9 ml, 42 mmol, 5.4 M) was added at room temperature. The reaction mixture was irradiated with microwave radiation for 2 h at 140°C. When the reaction was complete, the mixture was quenched with H ₂ O (6 ml). The resulting mixture was extracted with dichloromethane (50 ml×3) and the organic phases were combined, washed with brine and dried over Na ₂ SO ₄ . The solvent was removed under reduced pressure and the residue was purified by flash chromatography, eluting with MeOH in DCM (0%-5% gradient) to give 4-methoxy-7-(morpholin-4-yl)-[1,3] thiazolo[4,5-c]pyridine-2-amine as a light yellow solid (150 mg, 60%). MS: m/z=267.0 [M+H] ⁺ .

h. N-[4-methoxy-7-(morpholin-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-1-(2-methoxyethyl)-1H-pyrazol-4 -carboxamide

In a 30 ml sealed tube, to a solution of 1-(2-methoxyethyl)-1H-pyrazole-4-carboxylic acid (143 mg, 0.84 mmol) in THF (10 ml) was added HATU (353 mg, 0.93 mmol), 4- methylmorpholine (170 mg, 1.68 mmol) under N ₂ atmosphere. The reaction mixture was stirred for 16 hours at 50° C. and then 4-methoxy-7-(morpholin-4-yl)-[1,3]thiazolo[4,5-c]pyridine-2-amine (75 mg , 0.28 mmol). The resulting solution was stirred for another 16 hours at 90° C. in an oil bath. When the reaction was complete, the reaction mixture was quenched by adding water (10 ml). The resulting solution was extracted with DCM (30 ml x 3). The organic phases were combined, washed with brine and dried over Na ₂ SO ₄ . The solvent was removed under reduced pressure and the residue was purified by preparative HPLC under the following conditions: Column, XBridge Prep C18 OBD Column, 19×150 mm 5 µm; MeCN in water (with 10 mmol/l NH ₄ HCO ₃ ), gradient 22%-40% in 8 min; Detector, UV 254/220 nm to give N-[4-methoxy-7-(morpholin-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-1-(2 -methoxyethyl)-1H-pyrazole-4-carboxamide as a white solid (54 mg, 45%). HPLC: 98.8% pure, RT=4.216 min. MS: m/z=419.1 [M+H] ⁺ . ^1Н NMR (400 MHz, DMSO-d ₆ ) δ 12.87 (s, 1Н), 8.52 (s, 1Н), 8.21 (s, 1Н), 7.64 (s, 1Н), 4.35 (t, J=5.1 Hz, 2Н ), 3.98 (s, 3Н), 3.84-3.76 (m, 4Н), 3.71 (t, J=5.1 Hz, 2Н), 3.25 (s, 3Н), 3.14-3.06 (m, 4Н).

2. N-[4-methoxy-7-(1-methyl-1H-pyrazol-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-4-(methoxymethyl) benzamide, 14

i. N-[7-(1-methyl-1H-pyrazol-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide

1-methyl-4-( tetramethyl-1,3,2-dioxaborolan-2-yl)-1Н-pyrazole (2.19 g, 10.6 mmol), Pd(dppf)Cl2CH2Cl2 (359 mg, 0.44 mmol), sodium carbonate (3.27 g, 30.8 mmol), water (23 ml) at room temperature. After three vacuum/nitrogen flash cycles, the resulting mixture was stirred for 16 h at 80°C. When the reaction was complete, the solids were filtered off. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography eluting with EtOAc in petroleum ether (0%-100% gradient) to give N-[7-(1-methyl-1H-pyrazol-4-yl)- [1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide as an off-white solid (2.54 g, 86%). MS: m/z=336.2 [M+H]+.

j. N-[4-bromo-7-(1-methyl-1H-pyrazol-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide

At 50°C, to a solution of N-[7-(1-methyl-1H-pyrazol-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (960 mg, 2.86 mmol) in N,N-dimethylformamide (150 mL) NBS (662 mg, 3.72 mmol) in N,N-dimethylformamide (30 mL) was added dropwise over 1 h with stirring. The resulting solution was then stirred for 1 hour at 50°C. When the reaction was complete, the mixture was quenched with H2O (100 ml). The resulting mixture was extracted with dichloromethane (150 mL×3) and the organic phases were combined, washed with brine and dried over anhydrous Na2SO4. The solvent was removed under reduced pressure and the residue was purified by flash chromatography eluting with EtOAc in petroleum ether (0%-60% gradient) to give N-[4-bromo-7-(1-methyl-1H-pyrazole-4 -yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide as a white solid (675 mg, 57%). MS: m/z=414.1 [M+H]+. 1H NMR (400 MHz, Chloroform-d) δ 9.92 (s, 1Н), 8.41 (s, 1Н), 8.03 (d, J=7.7 Hz, 2Н), 7.98 (s, 1Н), 7.92 (s, 1Н) , 7.76-7.68 (m, 1H), 7.66-7.58 (m, 2H), 4.07 (s, 3H).

k. 4-methoxy-7-(1-methyl-1H-pyrazol-4-yl)thiazolo[4,5-c]pyridine-2-amine

To a solution of N-[4-bromo-7-(1-methyl-1H-pyrazol-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (338 mg, 0.81 mmol) in methanol (12 ml) was added MeONa in MeOH (5.5 ml, 29.7 mmol, 5.4 M) at room temperature. The reaction mixture was irradiated with microwave radiation for 2 h at 140°C. When the reaction was complete, the mixture was quenched with ice water (20 ml). The resulting mixture was extracted with dichloromethane (50 ml×3) and the organic phases were combined, washed with brine and dried over Na ₂ SO ₄ . The solvent was removed under reduced pressure and the residue was purified by flash chromatography eluting with MeOH in DCM (0%-9% gradient) to give 4-methoxy-7-(1-methyl-1H-pyrazol-4-yl)thiazolo [4,5-c]pyridine-2-amine as a light yellow solid (33 mg, 11%). MS: m/z=382.2 [M+H]+.

l. N-[4-methoxy-7-(1-methyl-1H-pyrazol-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-4-(methoxymethyl)benzamide, 14

To a solution of 4-(methoxymethyl)benzoic acid (53 mg, 0.32 mmol) in tetrahydrofuran (5 mL) was added 4-methylmorpholine (65 mg, 0.64 mmol), HATU (135 mg, 0.35 mmol) at room temperature. The resulting mixture was stirred for 8 hours at 50°C. To the above reaction mixture was added 4-methoxy-7-(1-methyl-1H-pyrazol-4-yl)-[1,3]thiazolo[4,5-c]pyridine-2-amine (28 mg, 0.11 mmol) and the resulting mixture was stirred for 16 hours at 90°C. When the reaction was complete, the mixture was quenched by adding water (10 ml). The resulting mixture was extracted with DCM (30 ml x 3). The organic phases were combined, washed with brine and dried over Na ₂ SO ₄ . The solvent was removed under reduced pressure and the residue was purified by preparative HPLC under the following conditions: Column, XBridge Shield RP18 OBD Column, 19×150 mm 5 µm; MeCN in water (with 10 mmol/l NH ₄ HCO ₃ + 0.1% NH ₃ .H ₂ O), gradient 30%-52% in 7 min; Detector, UV 254/220 nm. As a result, N-[4-methoxy-7-(1-methyl-1H-pyrazol-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-4-(methoxymethyl )benzamide as a white solid (18 mg, 40%). HPLC: 99.4% pure, RT=5.40 min. MS: m/z=410.1 [M+H] ⁺ . 1Н NMR (400 MHz, DMSO-d6) δ 13.24 (s, 1Н), 8.25-8.13 (m, 4Н), 7.94 (s, 1Н), 7.50 (d, J=8.0 Hz, 2Н), 4.53 (s, 2Н), 4.05 (s, 3Н), 3.97 (s, 3Н), 3.35 (s, 3Н).

3. N-[4-methoxy-7-(1-methyl-1H-pyrazol-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-4-(methoxymethyl) benzamide, 30

m. 4-Chloro-5-iodine-3-nitropyridin-2-ol

To a solution of 4-chloro-3-nitropyridin-2-ol (40.00 g, 229.22 mmol) in CH ₃ CN (500 ml) was added NIS (56.72 g, 252.14 mmol) at room temperature. The resulting mixture was stirred for 12 hours at 80°C. When the reaction was complete, the solvent was removed under reduced pressure. The residue was purified by flash chromatography eluting with EtOAc in PE (0%-100% gradient) to give 4-chloro-5-iodo-3-nitropyridin-2-ol as a yellow solid (46.43 g, 63% ). MS: m/z=301.0 [M+H] ⁺ .

n. 4-Chloro-5-iodine-2-methoxy-3-nitropyridine

Ag ₂ CO ₃ (30.88 g, 112.0 mmol), CH ₃ I (31.81 g, 224.2 mmol) at room temperature. The resulting mixture was stirred for 16 hours at 80°C. When the reaction was complete, the solids were filtered off. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography eluting with EtOAc in PE (0%-20% gradient) to give 4-chloro-5-iodo-2-methoxy-3-nitropyridine as light yellow solids (19.50 g, 91%). ¹ H NMR (400 MHz, DMSO-d6) δ 8.82 (s, 1H), 4.00 (s, 3H).

O. 4-chloro-5-iodine-2-methoxypyridine-3-amine

NH ₄ Cl (61.26 g, 1145 mmol), water (180 ml), Fe ( 53.42 g, 953.85 mmol) at room temperature. The resulting mixture was stirred for 10 hours at 80°C. When the reaction was complete, the solids were filtered off. The filtrate was concentrated under reduced pressure and the residue was diluted in H ₂ O (1.5 L). The resulting solution was extracted with dichloromethane (1.5 l×3) and the combined organic layers were washed with brine, dried over anhydrous Na ₂ SO ₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography eluting with EtOAc in PE (0%-100% gradient) to give 4-chloro-5-iodo-2-methoxypyridine-3-amine as a light yellow solids (17.36 g, 96%). MS: m/z=284.8 [M+H] ⁺ .

R. N-(7-iodo-4-methoxythiazolo[4,5-c]pyridin-2-yl)benzamide

To a solution of 4-chloro-5-iodo-2-methoxypyridine-3-amine (11.13 g, 39.13 mmol) in acetone (293 ml) was added benzoylisothiocyanate (9.58 g, 58.70 mmol). The resulting mixture was stirred for 16 hours at 50°C. When the reaction was complete, the solvent was removed under reduced pressure and the residue was purified by flash chromatography eluting with EtOAc in PE (0%-100% gradient) to give N-(7-iodo-4-methoxythiazolo[4,5- c]pyridin-2-yl)benzamide as a light yellow solid (16.00 g, 99%). MS: m/z=412.0 [M+H] ⁺ .

q. N-(7-(3,6-dihydro-2H-pyran-4-yl)-4-methoxythiazolo[4,5-c]pyridin-2-yl)benzamide

2-( 3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.18 g, 10.40 mmol), Pd(dppf)Cl2CH2Cl2 (566 mg, 0.69 mmol), sodium hydroxide (1.05 g, 26.34 mmol) and water (30 ml) at room temperature. After three vacuum/nitrogen flash cycles, the resulting mixture was stirred for 16 h at 100°C. When the reaction was complete, the solids were filtered off. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography eluting with MeOH in DCM (0%-10% gradient) to give N-(7-(3,6-dihydro-2H-pyran-4-yl) -4-Methoxythiazolo[4,5-c]pyridin-2-yl)benzamide as an off-white solid (702 mg, 28%). MS: m/z=368.2 [M+H] ⁺ .

r. N-(4-methoxy-7-(tetrahydro-2H-pyran-4-yl)thiazolo[4,5-c]pyridin-2-yl)benzamide

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 (702 mg , 1.91 mmol) in MeOH (30 mL) was added Pd/C (10%, 500 mg) under nitrogen atmosphere. After three vacuum/hydrogen flash cycles, the reaction mixture was hydrogenated at 50° C. for 16 hours under a hydrogen atmosphere using a hydrogen balloon. When the reaction was complete, the resulting mixture was filtered through celite. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography eluting with MeOH in DCM (0%-10% gradient) to give N-(4-methoxy-7-(tetrahydro-2H-pyran-4-yl) thiazolo[4,5-c]pyridin-2-yl)benzamide as a white solid (330 mg, 47%). MS: m/z=370.3 [M+H]+.

s. 4-methoxy-7-(tetrahydro-2H-pyran-4-yl)thiazolo[4,5-c]pyridine-2-amine

To a solution of N-[4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]benzamide (330 mg, 0.89 mmol) in methanol (10 ml) was added water (10 ml), sodium hydroxide (357 mg, 8.93 mmol) at room temperature. The resulting mixture was stirred for 16 hours at 90°C. When the reaction was complete, the methanol was removed under reduced pressure. Water (30 ml) was added to the residue and extracted with dichloromethane (30 ml x 3). The combined organic phases were washed with brine, dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography eluting with MeOH in DCM (0%-10% gradient) to give 4-methoxy-7-(tetrahydro-2H-pyran-4-yl)thiazolo[4 ,5-c]pyridine-2-amine as a white solid (106 mg, 42%). MS: m/z=266.0 [M+H] ⁺ .

t. Phenyl (4-methoxy-7-(tetrahydro-2H-pyran-4-yl)thiazolo[4,5-c]pyridin-2-yl)carbamate

Potassium carbonate (136 mg, 0.98 mmol), phenyl chloroformate (308 mg, 1.97 mmol) and pyridine (78 mg, 0.98 mmol) at room temperature. The resulting mixture was stirred for 8 hours at 50°C. When the reaction was complete, the solvent was removed under reduced pressure. The resulting mixture was diluted with water (30 ml) and extracted with dichloromethane (30 ml×3). The combined organic phases were washed with brine, dried over Na ₂ SO ₄ and filtered. The filtrate was concentrated under reduced pressure to give phenyl (4-methoxy-7-(tetrahydro-2H-pyran-4-yl)thiazolo[4,5-c]pyridin-2-yl)carbamate as an orange oil (88 mg, 70 %), which was used in the next step without further purification. MS: m/z=386.3 [M+H] ⁺ .

u. N-[4-methoxy-7-(1-methyl-1H-pyrazol-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-4-(methoxymethyl)benzamide, thirty

To a solution of phenyl N-[4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]carbamate (88 mg, 0.23 mmol) and 4- methylpiperidin-4-ol (79 mg, 0.68 mmol) in THF (10 mL) DIPEA (177 mg, 1.37 mmol) was added at room temperature. The resulting mixture was stirred for 16 hours at 50°C. When the reaction was complete, the solvent was removed under reduced pressure and the residue was purified by preparative HPLC under the following conditions: column, XBridge Prep OBD C18 Column, 30×150 mm 5 µm; mobile phase, water (with 10 mmol/L NH ₄ HCO ₃ + 0.1% NH ₃ .H ₂ O) and ACN (13.0% ACN to 40.0% in 8 min); detector, UV 220 nm. The title compound 4-hydroxy-N-[4-methoxy-7-(oxan-4-yl)-[1,3]thiazolo[4,5-c]pyridin-2-yl]-4-methylpiperidin-1 α-carboxamide was obtained as a white solid (30 mg, 32%). HPLC: 99.7% pure, RT=3.13 min. MS: m/z=407.3 [M+H]+. 1Н NMR (400 MHz, DMSO-d6) δ 11.45 (s, 1Н), 7.79 (s, 1Н), 4.41 (s, 1Н), 4.02-3.92 (m, 5Н), 3.84 (d, J=13.2 Hz, 2Н), 3.54-3.44 (m, 2Н), 3.29-3.21 (m, 2Н), 3.03-2.84 (m, 1Н), 1.93-1.68 (m, 4Н), 1.56-1.36 (m, 4Н), 1.14 ( s, 3H).

Example 3: Examination of the compounds of the present invention for inhibitory activity against human adenosine receptors in recombinant cells.

The functional activity of human receptors A _2A , A _2B , A ₁ and A ₃ was determined by determining the amount of cAMP, which is a second messenger for adenosine receptors.

To do this, recombinant HEK293 cells expressing human A _2A or A _2B receptors (both coupled to Gs proteins) were seeded in 394-well microtiter plates, test compounds and agonist (NECA) were added. After a 15 minute incubation, HTRF reagents (cAMP dynamic 2, Cis Bio) were added and cAMP levels in the cells were determined using an ENVISION plate reader (Perkin Elmer).

For human A ₁ and A ₃ receptors, recombinant CHO cells expressing either the A ₁ or A ₃ receptor were used. Since both receptors are coupled to Gi proteins, the assay protocol was adapted:

Cells were seeded in 384-well plates, forskolin, test compounds and agonists (CPA for A ₁ - and IB-MECA for A ₃ receptor) were added. After a 30 minute incubation, HTRF reagents (cAMP dynamic 2, Cis Bio) were added and cAMP levels in the cells were determined using an ENVISION plate reader (Perkin Elmer).

The resulting baseline data were normalized to inhibitor control and neutral control (DMSO) and the normalized data were aligned using the GeneData software.

The compounds of the present invention exhibit high selectivity for the A _2A and A _2B adenosine receptors over the A ₁ and A ₃ adenosine receptors (see, for example, data for some examples of compounds of the present invention in Table 4).

In particular, in contrast to the known adenosine A _2A receptor antagonist tosadenant and similar benzothiazole derivatives, the compounds of the present invention surprisingly exhibit dual A _2A /A _2B activity (see Table 4), which is advantageous for the treatment and/or prevention of hyperproliferative and infectious diseases. and disorders as described above, or the compounds of the present invention exhibit at least high A _2A inhibitory activity, along with other unexpected benefits disclosed herein, which result in high efficacy in the treatment and/or prevention of hyperproliferative and infectious diseases and disorders.

A means IC ₅₀ value is < 10 nM, B means IC ₅₀ value is < 100 nM, C means IC ₅₀ value is < 1 μM, D means IC ₅₀ value is > 1 μM.

Example 4 Study of the action of the compounds of the present invention against the endogenous human A _2A receptor

Endogenous functional activity of the Gs-coupled human A _2A receptor was measured in T cells where this receptor is highly expressed. Determination of receptor activity was carried out by determining the amount of cAMP, which is a secondary messenger for adenosine receptors.

In general, human pan T cells were isolated from human PBMCs (MACS pan T cell isolation kit, Miltenyi Biotec) that were obtained from fresh whole blood. T cells were seeded in 384-well microtiter plates and treated with test compounds. After a 10 minute incubation at room temperature, the adenosine A _2A receptor agonist CGS-21680 was added and the plates were incubated for an additional 45 minutes. Finally, HTRF reagents (cAMP Femto Kit, CisBio) were added to the wells and cAMP levels in the cells were determined 1 h later using an ENVISION plate reader (Perkin Elmer).

The resulting baseline data were normalized to inhibitor control and neutral control (DMSO) and the normalized data were leveled using the GeneData Screener software.

The compounds of the present invention demonstrate the ability to inhibit the A _2A receptor expressed in human T cells that are incubated with the adenosine A _2A receptor agonist CGS-21680 (as measured by cAMP quantification), which is advantageous for the treatment and/or prevention of hyperproliferative and infectious diseases and violations as described above. Thus, the compounds of the present invention are surprisingly able to prevent immunosuppression and therefore are able to maintain the inhibition of tumor growth induced by anti-cancer T cells, the reduction or destruction of metastases, and the prevention of neovascularization.

Example 5: Study of the pharmacokinetic properties of the compounds of the present invention in rats and mice

The aim of the study was to obtain information on the pharmacokinetic properties of the compounds of the present invention in female Wistar rats/mice after a single intravenous and oral administration.

Means and methods:

Animal Experiments (Intravital Phase)

Female Wistar rats/mice (n=6) were given either a single intravenous (bolus) injection or oral (gavage) administration of the test compound. Doses of 0.2 and 10 mg/kg (based on compound) were administered intravenously and orally, respectively, as a solution in DMSO (0.2%)/PEG 200 (40%)/water for intravenous administration and as a suspension in Methocel ( 0.5%)/Tween 20 (0.25%) in water for oral dosing. Subsequent blood samples were taken sublingually with isoflurane inhalation in 3 animals per route of administration at 0.1 (i.v. only), 0.25 (oral only), 0.5, 1, 2, 4, 6, and 24 h and further processed to obtain plasma. Also, urine and feces samples from 3 rats per route of administration were collected over a period of time from 0 to 24 hours and pooled for analysis.

Bioanalytics:

Compound concentrations in plasma, faeces were quantified using the UHPLC method together with mass spectrometric 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 device coupled with an API 5500 Q-trap AB Sciex Mass Spectrometer. mm) using a mobile phase gradient with 0.1% formic acid and acetonitrile as eluents. Detection of compounds was carried out using multiple reaction monitoring in positive ionization mode. An internal standard (20 μl) was added to plasma samples and the analyte was extracted from the matrix using tert-butyl methyl ether (tBME). The organic phase was evaporated to dryness in a stream of nitrogen. The residue was dissolved in acetonitrile/0.1% formic acid for LC-MS/MS analysis. Fecal samples were homogenized with 4 times their volume of ethanol/water (4:1, v/v). An internal standard was added to aliquots of water-ethanol extracts, diluted with acetonitrile/water (1:1, v/v) and directly injected into the LC-MS/MS system.

Pharmacokinetic evaluation:

Pharmacokinetic parameters C _max and t _max were taken from the observed data. Area under the curve (AUC), clearance (CL), volume (V), half-life (t _1/2 ), F and all dose-normalized values were calculated using dedicated software "DDS-TOX". "DDS-TOX" values were evaluated for several compounds and shown to be comparable to those provided by the validated WinNonLin software. AUC values were calculated by non-compartmental analysis using the linear-log trapezoidal method. Numerical data for mean plasma concentrations and derived pharmacokinetic parameters have been rounded to 3 significant figures for presentation. Data on oral bioavailability and excretion - expressed as % dose - are displayed using 2 significant figures.

In contrast to the known adenosine A _2A receptor antagonist tosadenant and similar benzothiazole derivatives, the compounds of the present invention unexpectedly show better pharmacokinetic properties in a mouse as an animal model relevant to cancer (see Table 6), which is preferable for treatment and/or prevention of hyperproliferative and infectious diseases and disorders, as described above.

Example 6 Study of the effect of the compounds of the present invention on mouse T cells

Preparation:

Adenosine (Ado) in the tumor microenvironment can inhibit T cell activity by signaling through A _2A receptors and inhibit cytokine secretion by T cells. _A2A -specific agonists such as CGS-21680 do a similar job of inhibiting T cell secretion of cytokines in vitro and in vivo. Potential A _2A antagonists or A _2A /A _2B dual antagonists may rescue T cells from this inhibition. In this case, we describe an in vitro system that we set up using pan-T cells from mouse spleen to screen potential A _2A antagonists or A _2A /A _2B dual antagonists for their activity. The described method involves the use of pre-coated CD3/CD28 beads to stimulate pan-T cells purified from mouse splenocytes, in combination with the addition of an A _2A agonist along with potential A _2A antagonists or dual A _2A /A _2B antagonists to assess the potentiation of T cytokine production. -cells.

Analysis description:

Briefly, mouse pan T cells are purified from BALB/c mouse spleens using the Pan T cell isolation kit Mouse II (MACS Miltenyi biotech Cat# Order No. 130-095-130) according to the protocol. manufacturer. Purified T cells are seeded in Nunc™ 96-well polystyrene round bottom microplates in RPMI with 10% heat-inactivated fetal bovine serum. Cells are kept at 37° C. for 1 hour and then activated with CD3/CD28 pre-coated beads (Dynabeads™ Mouse T-Activator CD3/CD28; Cat# 11456D). After 30 minutes, the cells are treated with various doses of test(s) antagonist(s). Cells are incubated for an additional 30 min at 37°C prior to treatment with A _2A agonist CGS-21680 (1 μM) or neutral control (DMSO). After a 24 hour incubation, IL-2 levels in the supernatants and after a 48 hour incubation, IFN-γ levels in the supernatants are measured by ELISA according to the manufacturer's protocol (R&D systems Cat# DY402 (IL-2); DY485 (IFN-γ) ). Once the concentrations are calculated, the difference in cytokine concentration between the DMSO control and the single agonist control (referred to as Δ) is calculated, and the percentage rescue at each antagonist concentration is calculated using Microsoft Excel. These percent cytokine rescue in a dose dependent manner by the antagonist are plotted in the GraphPad Prism software and the IC ₅₀ value calculated.

In contrast to the known adenosine A _2A receptor antagonist tosadenant, the compounds of the present invention demonstrate that they are able to rescue T cells from inhibition and are able to prevent suppression of cytokine secretion induced by adenosine or A _2A -specific agonists such as CGS-2168 (see Table 7), which is preferred for the treatment and/or prevention of hyperproliferative and infectious diseases and disorders as described above.

Thus, the compounds of the present invention are surprisingly able to prevent immunosuppression and therefore are able to maintain the inhibition of tumor growth induced by anti-cancer T cells, the reduction or destruction of metastases, and the prevention of neovascularization.

Example 7: Injection vials

A solution of 100 g of the compound of the present invention and 5 g of disodium hydrogen phosphate in 3 l of bidistilled water is adjusted to pH 6.5 with 2N sodium chloride. hydrochloric acid, filtered under sterile conditions, transferred to injection vials, lyophilized under sterile conditions and sealed under sterile conditions. Each injection vial contains 5 mg of a compound of the present invention.

Example 8: Mortar

A solution is prepared from 1 g of the compound of the present invention, 9.38 g of NaH ₂ PO ₄ 2 H ₂ O, 28.48 g of Na ₂ HPO ₄ ⋅12H ₂ O and 0.1 g of benzalkonium chloride in 940 ml of bidistilled water. The pH value is adjusted to 6.8, the solution is adjusted to 1 liter and sterilized by irradiation.

Example 9: Ampoules

A solution of 1 kg of a compound of the present invention in 60 L of bidistilled water is filtered under sterile conditions, transferred into ampoules, lyophilized under sterile conditions and sealed under sterile conditions. Each ampoule contains 10 mg of a compound of the present invention.

Claims (68)

1. Compound of formula I,

Where R ¹ is one of the following structures:

R ² is one of the following structures:

R ³ is

R ⁴ is H, methyl, ethyl, F, Br or Cl, and its stereoisomers. 2. Connection according to claim 1, where R ¹ is phenyl, methylpyrazole or dihydropyran, and R ² , R ³ and R ⁴ have the meanings specified in paragraph 1, and its stereoisomers. 3. Connection according to claim 1 or 2, where R ³ is OMe, and R ¹ , R ² and R ⁴ have the meanings specified in paragraph 1, and its stereoisomers. 4. Connection according to one or more of the preceding claims, where R ¹ is phenyl, methylpyrazole or dihydropyran, R ³ is OMe, and R ² and R ⁴ have the meanings specified in paragraph 1, and its stereoisomers. 5. Connection according to one or more of the preceding claims, where R ⁴ is H, and where R ¹ , R ² and R ³ have the meanings specified in paragraph 1, and its stereoisomers. 6. A compound selected from the group consisting of the following compounds:

and their stereoisomers. 7. A process for the preparation of a compound of formula I, which is characterized in that 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 using 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 urea formation conditions to give a compound of formula I,

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, 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 substantially basic conditions, and a compound of formula VIII is reacted with a compound of formula IX under standard amidation or urea formation conditions to give a compound of formula I,

c) a compound of formula XII is iodinated 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 obtain 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 urea formation conditions to give the compound formula I,

d) the base of the compound of formula I is converted to one of its salts by treatment with an acid, or e) the acid of the compound of formula I is converted into one of its salts by treatment with a base. 8. Pharmaceutical preparation as an inhibitor of the adenosine receptor A _2A and/or A _2B containing an effective amount of at least one compound according to any one of paragraphs. 1-6 and/or its stereoisomers and additionally containing fillers and/or excipients. 9. Drug as an inhibitor of the adenosine receptor A _2A and/or A _2B containing at least one compound according to any one of paragraphs. 1-6 and/or one of its stereoisomers, for use in the treatment and/or prevention of physiological and/or pathophysiological conditions that are associated with A _2A and/or A _2B adenosine receptors. 10. Drug as an inhibitor of the adenosine receptor A _2A and/or A _2B containing at least one compound according to any one of paragraphs. 1-6 and/or one of its stereoisomers, for use in the treatment and/or prevention of physiological and/or pathophysiological conditions that are associated with adenosine receptors A _2A and/or A _2B selected from the group consisting of hyperproliferative and infectious diseases and violations. 11. The medicament of claim 10 wherein the hyperproliferative disease or disorder is a malignant neoplasm. 12. The drug according to claim 11, where the malignant neoplasm is selected from the group consisting of acute and chronic lymphocytic leukemia, acute granulocytic leukemia, adrenal cortex cancer, bladder cancer, brain cancer, breast cancer, cervical cancer, cervical hyperplasia uterus, choriocarcinoma, chronic granulocytic leukemia, chronic lymphocytic leukemia, colon cancer, endometrial cancer, esophageal cancer, essential thrombocytosis, urogenital carcinoma, glioma, glioblastoma, hairy cell leukemia, head and neck carcinoma, Hodgkin's disease, Kaposi's sarcoma, lung carcinoma, lymphoma , malignant carcinoid tumor, malignant hypercalcemia, malignant melanoma, malignant pancreatic insulinoma, medullary thyroid carcinoma, melanoma, multiple myeloma, mycosis fungoides, myeloid and lymphocytic leukemia, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, osteosarcoma, ovarian carcinoma, pancreatic carcinoma, polycythemia vera, primary brain carcinoma, primary macroglobulinemia, prostate cancer, renal cell carcinoma, rhabdomyosarcoma, skin cancer, small cell lung cancer, soft tissue sarcoma, squamous cell carcinoma, gastric cancer, cancer testicular, thyroid cancer, and Wilms tumor. 13. The drug according to claim 10, wherein the hyperproliferative disease or disorder is selected from the group consisting of age-related macular degeneration, Crohn's disease, cirrhosis, chronic inflammation-related disorders, proliferative diabetic retinopathy, proliferative vitreoretinopathy, retrolental fibroplasia, granulomatosis, immune hyperproliferation associated with organ or tissue transplantation, and an immunoproliferative disease or disorder selected from the group consisting of inflammatory bowel disease, psoriasis, rheumatoid arthritis, systemic lupus erythematosus (SLE), vascular hyperproliferation secondary to retinal hypoxia, and vasculitis. 14. The drug according to claim 10, where the infectious disease or disorder is selected from the group consisting of a) virus-induced infectious diseases that are caused by retroviruses, hepadnaviruses, herpesviruses, flaviviruses and/or adenoviruses, where retroviruses are selected from lentiviruses or oncoretroviruses, where lentiviruses are selected from the group consisting of HIV-1, HIV-2, FIV, BIV, SIV viruses , SHIV, CAEV, VMV and EIAV, and oncoretroviruses are selected from the group consisting of HTLV-I, HTLV-II and BLV, hepadnaviruses are selected from the group consisting of HBV, GSHV and WHV, herpesviruses are selected from the group consisting of HSV I, HSV II, EBV, VZV, HCMV or HHV 8, and flaviviruses are selected from the group consisting of HCV, West Nile virus and yellow fever virus, b) bacterial infectious diseases that are caused by gram-positive bacteria, where gram-positive bacteria are selected from the group consisting of methicillin-sensitive and methicillin-resistant staphylococci (including Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus saprophyticus and coagulase-negative staphylococcus), Staphylococcus aureus with intermediate sensitivity to glycopeptides (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), Streptococci group G (GGS) and viridans streptococci), enterococci (including vancomycin-susceptible and vancomycin-resistant strains such as Enterococcus faecalis and Enterococcus faecium), Clostridium difficile, Iisteria monocytogenes, C orynebacterium jeikeium, Chlamydia spp. (including C.pneumoniae) and Mycobacterium tuberculosis, c) bacterial infectious diseases that are caused by gram-negative bacteria, where the gram-negative bacteria are selected from the group consisting of the genus Enterobacteriacae, including Escherichia spp. (including Escherichia coli), Klebsiella spp., Enterobacter spp., Citrobacter spp., Serratia spp., Proteus spp., Providencia spp., Salmonella spp., Shigella spp., Pseudomonas genus (including P. aeruginosa), Moraxella spp. (including M. catarrhalis), Haemophilus spp. and Neisseria spp., 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.