Heterocyclic compounds that can activate STING

Abstract

The present invention relates to compounds of formula (I) that can activate STING (interferon gene stimulating factor). The present invention further relates to pharmaceutical compositions comprising at least one compound of formula (I), and the use of these compounds or pharmaceutical compositions as pharmaceuticals, for example, as pharmaceuticals for treating cancer in dogs or cats, or as vaccine adjuvants. JPEG2026512969000062.jpg6259 formula (I)

Description

[Technical Field]

[0001] The present invention relates to small molecules capable of activating STING (interferon gene stimulating factor), and salts thereof. More specifically, the present invention relates to heterocyclic compounds capable of activating STING. Furthermore, the present invention relates to pharmaceutical compositions and combinations comprising these compounds, and their use as pharmaceuticals. These compounds and pharmaceutical compositions comprising at least one of these compounds may be suitable as pharmaceuticals, for example, as pharmaceuticals for the treatment of cancer, such as cancer in dogs and / or cats, and as vaccine adjuvants, for example, as vaccine adjuvants for use in pigs. Accordingly, the present invention also relates to compounds and pharmaceutical compositions comprising at least one of these compounds for use in the treatment of cancer in cats or dogs. [Background technology]

[0002] STING is a type of pattern recognition receptor (PRRP) that plays a central role in the innate immune system, distinguishing pathogens from host cells by detecting extracellular and intracellular danger signals, including damage-associated molecular patterns (DAMPs) and pathogen-associated molecular patterns (PAMPs). These recognition processes constitute the first line of defense against viral and bacterial infections and malignant cells. However, pathogens and cancer cells have evolved ways to evade recognition by the immune system. Therefore, the goal of immunotherapy is to initiate an antigen-specific immune response or to reactivate existing responses of the immune system in certain cell types against pathogenic invaders or cancerous cells. Within the PRRP family, STING (also known as TMEM173, MPYS, MITA, and ERIS) belongs to the family of nucleic acid sensors and is an adapter for cytosolic DNA signaling. In healthy mammalian cells, DNA is compartmentalized within the nucleus. In pathogenic situations, such as the invasion of DNA-containing pathogens, or in malignant cells, DNA is located in the cytoplasm. Here, STING is crucial for detecting the aforementioned cytosolic DNA and inducing an immune response to pathogenic events. As a member of the innate immune defense mechanism, STING is expressed in almost all cell types, particularly endothelial, epithelial, and immune cells, such as macrophages and dendritic cells. In dogs and cats, the main expression of STING is observed in the spleen, lungs, blood, lymph nodes, and brain (Zhang et al, Microbial Pathogenesis, 113, 202 - 208, 2017; Zhang et al, Veterinary Immunology and Immunopathology 169, 54 - 62, 2016).

[0003] In its basal state, STING exists as a dimer, with its N-terminal domain fixed to the ER and its C-terminal domain in the cytosol. Cyclic dinucleotides (CDNs) produced by the protein cyclic GMP-AMP synthase (cGAS) are the native ligands for STING (Ablasser et al, Nature 498, 380-384, 2013). Binding of CDNs to STING induces conformational changes that enable relocalization from the ER to perinuclear endosomes, following the binding and activation of TANK-binding kinase (TBK1) and interferon regulator 3 (IRF3) (Liu et al, Science 347, Issue 6227, 2630-1-2630-14, 2015). Phosphorylation of transcription factors IRF3 and NF-κB by TBK1 results in the expression of several cytokines, including type I interferon (IFN). Type I IFN production by antigen-presenting cells and other cell types is considered a major event in T cell activation and, consequently, differentiation of antigen-specific effector CD4 and CD8 T cells. The absence of type I IFN has been shown to result in a reduced T cell-dependent immune response to viral infection or tumor cells (Zitvogel et al, Nature Reviews Immunology 15, 405 - 414, 2015). On the other hand, the presence of a type I IFN signature during cancer therapy is associated with an increase in the number of tumor-infiltrating T cells and potentially favorable clinical outcomes (Sistigu et al, Nature Medicine 20, 1301 - 1309, 2014).

[0004] The antitumor effects of type I interferon have been investigated in vitro and in vivo for feline and canine cancers. Based on field studies in which human recombinant interferon was administered to dogs with tumors (lymphoma, fibrosarcoma, osteosarcoma, mycosarcoma, liposarcoma), the delay or prevention of local recurrence and metastasis has been documented (Klotz et al, Veterinary Immunology and Immunopathology 191, 80 - 93, 2017). Recent studies in mice have shown that the efficient secretion of type I IFN in the tumor microenvironment and the induction of a T cell-dependent immune response against cancer cells depend on the presence of STING (Woo et al, Immunity 41, 5, 830 - 842, 2014; Corrales et al, Cell Reports 11, 1018 - 1030, 2015; Deng et al, Immunity 41, 5, 843 - 852, 2014). In several mouse tumor models, STING deletion resulted in reduced type I IFN levels and diminished antitumor activity in the tumor microenvironment, thereby highlighting the importance of type I IFN presence. On the other hand, specific activation of STING resulted in improved antigen-specific T cell immune responses against cancer cells.

[0005] Type I interferons can significantly enhance the anti-tumor immune response by inducing the activation of both adaptive and innate immune cells. Given the importance of type I interferon in several malignancies, including viral infections and cancer therapy, strategies that enable specific activation of STING are therapeutically interesting. STING activation may show synergistic effects with various approved chemotherapeutic agents or other anticancer therapies, such as radiotherapy (Wu et al., Med Res Rev 2020 May;40(3):1117-1141) or infectious disease therapies. Prior art, for example, describes a low molecular weight modifier of STING in International Publication No. 2020 / 075790. [Overview of the Initiative]

[0006] The compounds according to the present invention are novel activators of STING, as demonstrated in an ex vivo system using whole canine blood. In one embodiment, the present invention relates to the following formula (I) [ka] Equation (I) (In the formula, R 1 These are -C1-6-alkyl or -C3-6 cycloalkyl, R 2a It is selected from the group consisting of -H or -C1-6-alkyl, R 2b It is selected from the group consisting of -H, -C1-6-alkyl, -C1-6-alkylene-OH, -C(O)OH, -C(O)O-C1-6-alkyl, and -pyrazolyl-C1-6-alkyl. R 2c is -H or -C1-6-alkyl, R 3 teeth,

[0007] [ka] Selected from the group consisting of (* indicates the attachment point), R 4 is selected from H, -C1-6-alkyl, and C3-6-cycloalkyl, R 5 is selected from H, -C1-6-alkyl, and C3-6-cycloalkyl, R 6 is selected from H, -C1-6-alkyl, and C3-6-cycloalkyl, R 7 is selected from H, -C1-6-alkyl, C3-6-cycloalkyl and -OH, R 8 is (CH2) n where n is an integer from 1 to 3, preferably 1 or 2, X is CH or N, Y is -O-, -S-, -S(O)-, -S(O)2-) relates to a compound of or a pharmaceutically acceptable salt thereof.

[0008] In another aspect, the present invention relates to a pharmaceutical composition comprising at least one compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. In another aspect, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the same for use as a medicament. In another aspect, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising the same for use in the treatment of cancer in cats or dogs.

Mode for Carrying Out the Invention

[0009] The compounds of the present invention have several advantageous properties, such as a particularly good, preferably dog STING, binding affinity for STING from various mammalian species, such as cats, mice, pigs and dogs, and a favorable cellular EC 50 measured by, i.e., showing a favorable cellular activity in dog whole blood. Accordingly, in a further embodiment, the present invention provides novel compounds of formula (I), including salts thereof, that activate STING and thus induce cytokine production in a STING-dependent manner in vitro and / or in vivo, for example in dogs, and that have pharmacological and pharmacokinetic properties suitable for therapeutic use, i.e., pharmaceutically acceptable use. The binding of the compound to the protein can be determined by known methods, such as surface plasmon resonance, scintillation proximity assay, isothermal titration calorimetry, or differential scanning fluorescence assay. In the last test, the melting temperature T m The temperature at which proteins, also known as ligatures, unfold is measured by the change in fluorescence of a dye that binds to the hydrophobic portion of the protein. m The shift correlates with the binding affinity of this small molecule. The high binding affinity of STING agonists is due to a T of >10°C, preferably >13°C, more preferably >15°C. m This is reflected in the shift. When measured by a binding assay, the compounds according to the present invention preferably have a T of >15°C, more preferably >20°C, and even more preferably >25°C, as determined by DSF. m This shows the interaction with canine STING (dSTING) reflected in the shift.

[0010] Since STING has been demonstrated to stimulate the production of type I interferons, such as interferon-beta (IFNb), in bone marrow and dendritic cells, the efficacy of STING agonists can be evaluated in a canine whole blood (cWB) assay using IFNb secretion as a readout. In this assay, freshly collected canine blood is incubated with a STING agonist, and the interferon-beta level in the supernatant is quantified by ELISA. The compounds according to the present invention typically exhibit a cellular EC50 of less than 10 μM, preferably less than 5 μM, more preferably less than 1 μM, and most preferably less than 0.5 μM. According to the present invention, a combination of high dDSF and low dWB is particularly preferred.

[0011] General formula (I) [ka] Equation (I) (In the formula, R 1 These are -C1-6-alkyl or -C3-6 cycloalkyl, R 2a It is selected from the group consisting of -H or -C1-6-alkyl, R 2b It is selected from the group consisting of -H, -C1-6-alkyl, -C1-6-alkylene-OH, -C(O)OH, -C(O)O-C1-6-alkyl, and -pyrazolyl-C1-6-alkyl. R 2c is -H or -C1-6-alkyl, R 3 teeth,

[0012] [ka] Selected from the group consisting of (* indicates the attachment point), R 4 These are selected from H, C1-6-alkyl, and C3-6 cycloalkyl groups. R 5 These are selected from H, C1-6-alkyl, and C3-6 cycloalkyl groups. R 6 These are selected from H, C1-6-alkyl, and C3-6 cycloalkyl groups. R 7 is selected from H, -C1-6-alkyl, C3-6 cycloalkyl, and -OH. R 8 (CH2) n Here, n is an integer from 1 to 3, preferably 1 or 2. X is either CH or N, Y is -O-, -S-, -S(O)-, -S(O)2-. The compounds of the present invention or their pharmaceutically acceptable salts are particularly suitable for the treatment of pathophysiological processes associated with or regulated by STING, especially cancer, for example, cancer in cats or dogs, or for example, as vaccine adjuvants in pigs.

[0013] Accordingly, in another embodiment, the present invention further relates to a pharmaceutical composition for use as a pharmaceutical, comprising a compound of formula (I) as defined herein or a pharmaceutically acceptable salt thereof, or at least one compound of formula (I). Another aspect of the present invention relates to a pharmaceutical composition for use in the treatment of cancer in cats or dogs, comprising a compound of formula (I) as defined herein, a pharmaceutically acceptable salt thereof, or at least one compound of formula (I). Other aspects of the present invention will be immediately apparent to those skilled in the art from the above and below descriptions and examples.

[0014] Terms and definitions used Terms not explicitly defined herein should be given the meanings that a person skilled in the art would give them in light of this disclosure and context. However, where used herein, unless otherwise specified, the following terms shall have the meanings indicated and shall be governed by the following conventions. In the groups, radicals, or parts defined below, the number of carbon atoms is often specified before the group; for example, C 1-6 -Alkyl refers to an alkyl group or radical having 1 to 6 carbon atoms. Generally, for groups such as HO, H2N, (O)S, (O)2S, NC (cyano), HOOC, F3C, etc., a person skilled in the art can understand the attachment point of the group to the molecule from the free valence of the group itself. For combined groups containing two or more subgroups, the attachment point of the group is the last named subgroup, for example, the substituent "aryl-C". 1-3 -Alkylene is an aryl group with C 1-3 - This refers to elements bonded to alkyl groups, the latter of which are bonded to the core or to a group to which a substituent is attached. Where the compounds of this invention are described in both chemical nomenclature and formula form, in the event of any discrepancy, the formula form shall prevail. Asterisks or tildes may be used in subformulas to indicate bonds connected to the defined core molecule.

[0015] For example, the term "3-carboxypropyl group" refers to the following substituents: [ka] (In the formula, the carboxyl group is attached to the third carbon atom of the propyl group.) The terms "1-methylpropyl-", "2,2-dimethylpropyl-", or "cyclopropylmethyl-" groups refer to the following groups: [ka] It represents. A wavy line may be used in a subformula to indicate bonds connected to the core molecule being defined. Alternatively, an asterisk may be used in a subformula to indicate bonds connected to the core molecule being defined.

[0016] 1.1.1.1 The term "replaced" As used herein, the term “substituted” means that one or more hydrogen atoms on a given atom are replaced by a group selected from a defined set of substituents, provided that the substitution does not exceed the normal valence of the given atom and that the substitution results in a stable compound. Similarly, the term “substituted” may be used in relation to a chemical moiety instead of a single atom, for example, “substituted alkyl” or “substituted aryl.” 1.1.1.2 Stereochemistry - Solvates - Hydrates Unless otherwise explicitly stated, throughout this specification and the accompanying claims, a given chemical formula or name includes tautomers and all stereo, optical and geometric isomers (e.g., enantiomers, diastereomers, E / Z isomers, etc.) and their racemates, as well as mixtures of different proportions of distinct enantiomers, mixtures of diastereomers, or any of the aforementioned forms in which such isomers and enantiomers exist, and their solvates, e.g., hydrates, etc. Unless otherwise specified, "pharmaceutically acceptable salts," as defined in more detail below, also include their solvates, such as hydrates.

[0017] 1.1.1.3 Stereoisomers In general, substantially pure stereoisomers can be obtained according to synthetic principles known to those skilled in the art, for example, by separation of the corresponding mixtures, by using stereochemically pure starting materials, and / or by stereoselective synthesis. Methods for preparing optically active forms, such as by separation of racemic forms, or by synthesis, such as by starting from optically active starting materials and / or by using chiral reagents, are known in the art. The enantiomerically pure compounds or intermediates of the present invention may be prepared by asymmetric synthesis, for example, by preparing suitable diastereomer compounds or intermediates that can be separated by known methods (e.g., by chromatographic separation or crystallization) and subsequent separation, and / or by using chiral reagents, such as chiral starting materials, chiral catalysts or chiral auxiliaries. Furthermore, methods for preparing enantiomerically pure compounds from corresponding racemic mixtures are known to those skilled in the art, such as by chromatographic separation of the corresponding racemic mixture in a chiral stationary phase; or by resolution of the racemic mixture using a suitable resolving agent, such as by diastereomer formation of the racemic compound using an optically active acid or base, subsequent resolution of the salt and release of the desired compound from the salt; or by derivatization of the corresponding racemic compound using an optically active chiral auxiliary, subsequent diastereomer separation and removal of the chiral auxiliary group; or by kinetic resolution of the racemic mixture (e.g., by enzymatic resolution); by enantioselective crystallization from an aggregate of enantiomorphous crystals under suitable conditions; or by (fractional) crystallization from a suitable solvent in the presence of an optically active chiral auxiliary.

[0018] 1.1.1.4 Salt The phrase "pharmaceutically acceptable" is used herein to mean a compound, material, composition, and / or dosage form that is suitable for use in contact with mammalian tissues within the bounds of sound medical judgment, without excessive toxicity, irritation, allergic response, or other problems or complications, and that is commensurate with a reasonable benefit / risk ratio. As used herein, “pharmaceutically acceptable salt” refers to a derivative of a disclosed compound in which the parent compound has been modified by producing an acid or base salt thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, basic residues, e.g., mineral or organic acid salts of amines; and acidic residues, e.g., alkali or organic salts of carboxylic acids. For example, such salts include salts derived from benzenesulfonic acid, benzoic acid, citric acid, ethanesulfonic acid, fumaric acid, gentisic acid, hydrobromic acid, hydrochloric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, 4-methylbenzenesulfonic acid, phosphoric acid, salicylic acid, succinic acid, sulfuric acid, and tartaric acid.

[0019] Further pharmaceutically acceptable salts can be formed using cations derived from ammonia, L-arginine, calcium, 2,2'-iminobisethanol, L-lysine, magnesium, N-methyl-D-glucamine, potassium, sodium, and tris(hydroxymethyl)-aminomethane. The pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing a basic or acidic moiety. Generally, such salts can be prepared by reacting the free acidic or basic form of these compounds with a sufficient amount of a suitable base or acid in water or an organic diluent, such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile, or a mixture thereof. For example, salts of acids other than those described above (e.g., trifluoroacetate), which are useful for purifying or isolating the compounds of the present invention, also constitute part of the present invention.

[0020] 1.1.1.5 Halogen The term halogen refers to fluorine, chlorine, bromine, and iodine. 1.1.1.6 Heteroatoms Heteroatoms can exist at all possible oxidation stages. For example, sulfur can exist as a sulfoxide (RS(O)-R') and a sulfone (-RS(O)2-R'). 1.1.1.7 Alkyl "C 1-n The term "-alkyl" (where n is an integer selected from 2, 3, 4, 5, or 6, preferably 4, 5, or 6) refers to an acyclic, saturated, branched, or linear hydrocarbon group having 1 to n carbon atoms, either alone or in combination with another group. For example, C 1-5The term -alkyl encompasses the groups H3C-, H3C-CH2-, H3C-CH2-CH2-, H3C-CH(CH3)-, H3C-CH2-CH2-CH2-, H3C-CH2-CH(CH3)-, H3C-CH(CH3)-CH2-, H3C-C(CH3)2-, H3C-CH2-CH2-CH2-CH2-, H3C-CH2-CH2-CH(CH3)-, H3C-CH2-CH(CH3)-CH2-, H3C-CH(CH3)-CH2-CH2-, H3C-CH2-C(CH3)2-, H3C-C(CH3)2-CH2-, H3C-CH(CH3)-CH(CH3)-, and H3C-CH2-CH(CH2CH3)-.

[0021] 1.1.1.8 Alkylene "C 1-n The term "-alkylene" (where n is an integer selected from 2, 3, 4, 5, or 6, preferably 4, 5, or 6) refers to an acyclic, saturated, branched, or linear divalent alkyl group containing 1 to n carbon atoms, either alone or in combination with another group. For example, C 1-4 -Alkylenes, please, include -CH2-, -CH2-CH2-, -CH(CH3)-, -CH2-CH2-CH2-, -C(CH3)2-, -CH(CH2CH3)-, -CH(CH3)-CH2-, -CH2-CH(CH3)-, -CH2-CH2-CH2-CH2-, -CH2-CH2-CH(CH3)-, -CH(CH3)-CH2-CH2-, -CH2-CH(CH3)-CH2-, -CH2-C(CH3)2-, -C(CH3)2-CH2-, -CH(CH3)-CH(CH3)-, -CH2-CH(CH2CH3)-, -CH(CH2CH3)-CH2-, -CH(CH2CH2CH3)-, -CH(CH(CH3))2-, and -C(CH3)(CH2CH3)-.

[0022] 1.1.1.9 Alkenil "C 2-m The term "-Alkenil" is "C 2-mThis term is used for alkyl groups (where m is an integer selected from 3, 4, 5, or 6, preferably 4, 5, or 6) where at least two carbon atoms of the group are bonded to each other by a double bond. 1.1.1.10 Alkenylene "C 2-m The term "-alkenylene" is used as "C 2-m The term "-alkylene" (where m is an integer selected from 3, 4, 5, or 6, preferably 4, 5, or 6) is used when at least two carbon atoms of the group are bonded to each other by a double bond. 1.1.1.11 Alkinyl "C 2-m The term "-alkynyl" is "C 2-m This term is used for alkyl groups (where m is an integer selected from 3, 4, 5, or 6, preferably 4, 5, or 6) where at least two carbon atoms of the group are bonded to each other by a triple bond.

[0023] 1.1.1.12 Alkynylene "C 2-m The term "-alkynylene" is used as "C 2-m The term is used for an alkylene group (where m is an integer selected from 3, 4, 5, or 6, preferably 4, 5, or 6) where at least two of the carbon atoms of the group are bonded to each other by a triple bond. 1.1.1.13 Cycloalkyl "C 3-k The term "-cycloalkyl" (where k is an integer selected from 3, 4, 5, 6, 7, or 8, preferably 4, 5, or 6) refers to a cyclic, saturated, unbranched hydrocarbon group having 3 to k carbon atoms, either alone or in combination with another group. For example, C 3-7 - The term cycloalkyl includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. 1.1.1.14 Cycloalkenyl "C 3-kThe term "-cycloalkenyl" (where k is an integer selected from 3, 4, 5, 6, 7, or 8, preferably 4, 5, or 6) refers to a cyclic, unsaturated but non-aromatic, unbranched hydrocarbon group having 3 to k carbon atoms, of which at least two are bonded to each other by double bonds, either alone or in combination with another group. For example, C 3-7 - The term cycloalkenyl includes cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl, cycloheptadienyl, and cycloheptatrienylon.

[0024] 1.1.1.15 Halo-(alkyl, alkylene, or cycloalkyl) The terms “halo” attached to an “alkyl,” “alkylene,” or “cycloalkyl” group (saturated or unsaturated) define alkyl, alkylene, or cycloalkyl groups in which one or more hydrogen atoms are replaced by a halogen atom selected from fluorine, chlorine, or bromine, preferably fluorine and chlorine, and particularly preferably fluorine. Examples include H2FC-, HF2C-, and F3C-. 1.1.1.16 Carbocyclyl The term "carbocyclyl" refers to a monocyclic, bicyclic, or tricyclic ring structure consisting of 3 to 14 carbon atoms, either alone or in combination with another group. The term "carbocyclyl" refers to fully saturated, partially saturated, and aromatic ring systems. The term "carbocyclyl" also encompasses condensed, bridging, and spirocyclic systems.

[0025] [ka]

[0026] 1.1.1.17 Ariel As used herein, the term “aryl” refers to a carbocyclic aromatic monocyclic group containing six carbon atoms, which may be further condensed, either alone or in combination with another group, to a second five-membered or six-membered carbocyclic group that is aromatic, saturated, or unsaturated. Aryls include, but are not limited to, phenyl, indanyl, indenyl, naphthyl, anthracenyl, phenantrenyl, tetrahydronaphthyl, and dihydronaphthyl. 1.1.1.18 Heterocycline The term "heterocyclyl" refers to a saturated or unsaturated monocyclic or polycyclic ring system containing one or more heteroatoms selected from N, O, S, SO, and SO2, and consisting of 3 to 14 ring atoms, which may include an aromatic ring, and in which none of the heteroatoms are part of an aromatic ring. The term "heterocyclyl" is intended to include all possible isomer forms.

[0027] Therefore, the term “heterocyclyl” includes the following exemplary structures (each form is not depicted as a base, as it may be attached to any atom via covalent bonds, as long as the appropriate valence is maintained). [ka] [ka]

[0028] 1.1.1.19 Heteroaryl The term "heteroaryl" means a monocyclic or polycyclic ring system containing one or more heteroatoms selected from N, O, S, SO, or SO2, and comprising at least one aromatic ring consisting of 5 to 14 ring atoms, wherein at least one of the heteroatoms is part of the aromatic ring, and the resulting ring system must be chemically stable. The term "heteroaryl" is intended to include all possible isomer forms. Therefore, the term “heteroaryl” includes the following exemplary structures (each form is not depicted as a base, as it may be attached to any atom via covalent bonds, as long as the appropriate valence is maintained).

[0029] [ka]

[0030] Many of the terms listed above may be used repeatedly in the definitions of formulas or bases, and in each case, independently of each other, they have one of the meanings listed above. The term "bicyclic ring system" refers to a group consisting of two linked cyclic substructures, including spiro rings, condensation rings, and bridging ring systems. Preferred Embodiment A preferred embodiment of the present invention is the following formula (I) [ka] Equation (I) (In the formula, R 1 These are -C1-6-alkyl or -C3-6 cycloalkyl, R 2a It is selected from the group consisting of -H or -C1-6-alkyl, R 2b It is selected from the group consisting of -H, -C1-6-alkyl, -C1-6-alkylene-OH, -C(O)OH, -C(O)O-C1-6-alkyl, and -pyrazolyl-C1-6-alkyl. R 2c is -H or -C1-6-alkyl, R 3 teeth,

[0031] [ka] Selected from the group consisting of (* indicates the attachment point), R 4These are selected from H, C1-6-alkyl, and C3-6 cycloalkyl groups. R 5 These are selected from H, C1-6-alkyl, and C3-6 cycloalkyl groups. R 6 These are selected from H, C1-6-alkyl, and C3-6 cycloalkyl groups. R 7 is selected from H, -C1-6-alkyl, C3-6 cycloalkyl, and -OH. R 8 (CH2) n Here, n is an integer from 1 to 3, preferably 1 or 2. X is either CH or N, and if X is N, then R 7 It is not -OH, Y is -O-, -S-, -S(O)-, -S(O)2-. This relates to the compound or its pharmaceutically acceptable salt.

[0032] Another particularly preferred embodiment is R 1 However, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, which is C1-6-alkyl, preferably methyl. Another particularly preferred embodiment is R 2a However, -C 1-6 -alkyl, preferably methyl, R 2b However, it is -H or -C1-6-alkyl, R 2c The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound is -H or -C1-6-alkyl.

[0033] Another particularly preferred embodiment is R 1 However, it is a C1-6-alkyl group, preferably a methyl group, and R 2a However, -C 1-6 -Alkyl, preferably methyl, R 2b However, it is -H or -C1-6-alkyl, R 2c The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound is -H or -C1-6-alkyl. Another particularly preferred embodiment is R 2a However, it is methyl, and R 2b However, it is -H, and R 2c The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound is -H. Another particularly preferred embodiment is R 1 However, it is a C1-6-alkyl group, preferably a methyl group, and R 2a However, it is methyl, and R 2b However, it is -H, and R 2c The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound is -H. Another particularly preferred embodiment is R 3 but, [ka] This relates to the compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0034] Another particularly preferred embodiment is R 3 but, [ka] (In the formula, R 7 The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, which is a C1-6-alkyl compound, preferably methyl.

[0035] Another particularly preferred embodiment is R 3 but, [ka] And R 1 However, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, which is C1-6-alkyl, preferably methyl.

[0036] Another particularly preferred embodiment is R 3 but, [ka] And R 2a However, -C 1-6-Alkyl, preferably methyl, and R 2b is -H or -C1-6-alkyl, and R 2c is -H or -C1-6-alkyl, relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0037] Another particularly preferred embodiment is where R 3 is

Chemical formula

[0041] Another particularly preferred embodiment is R 3 but, [ka] This relates to the compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0042] Another particularly preferred embodiment is R 3 but, [ka] This relates to the compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0043] Another particularly preferred embodiment is R 3 but, [ka] And R 4 However, it is -H or methyl, preferably -H, and R 5 and R 6 However, independently, it is -H or methyl, preferably R 5 and R 6 The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein both are -H and Y is O. Another particularly preferred embodiment is R 3 but, [ka] And R 1 However, it is methyl, and R 4 However, it is -H or methyl, preferably -H, and R 5 and R 6 However, independently, it is -H or methyl, preferably R 5 and R 6 The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein both are -H and Y is O.

[0044] Another particularly preferred embodiment is R 3 but, [ka] And R 2a However, -C 1-6 -Alkyl, preferably methyl, R 2b However, it is -H or -C1-6-alkyl, R 2c However, it is -H or -C1-6-alkyl, R 4 However, it is -H or methyl, preferably -H, and R 5 and R 6 However, independently, it is -H or methyl, preferably R 5 and R 6 The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein both are -H and Y is O.

[0045] Another particularly preferred embodiment is R 3 but, [ka] And R 1 However, it is methyl, and R 2a However, -C 1-6 -Alkyl, preferably methyl, R 2b However, it is -H or -C1-6-alkyl, R 2c However, it is -H or -C1-6-alkyl, R 4 However, it is -H or methyl, preferably -H, and R 5 and R 6 However, independently, it is -H or methyl, preferably R 5 and R 6 The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein both are -H and Y is O.

[0046] Another particularly preferred embodiment is R 1 However, it is methyl, and R 2a However, it is methyl, and R 2b and R 2c However, it is -H, and R3 but, [ka] And R 4 , R 5 and R 6 The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein is -H and Y is O.

[0047] Particularly preferred compounds of the present invention, with the following structure [ka] A compound selected from or a pharmaceutically acceptable salt thereof.

[0048] Further particularly preferred compounds of the present invention are: [ka] [ka] or a pharmaceutically acceptable salt thereof.

[0049] In one embodiment, the present invention relates to a compound of formula (I) in a salt-free form. In another embodiment, the present invention relates to a compound of formula (I) in a pharmaceutically acceptable salt form. Regarding equation (I), the R specified above 1 , R 2a , R 2b , R 2c , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 The definitions of X and Y, and each of them, may be combined with one another. In one embodiment, the present invention relates to a pharmaceutical composition comprising at least one compound according to formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. Compounds of formula (I) or pharmaceutically acceptable salts thereof have been found to be useful in the prevention and / or treatment of diseases and / or conditions in which the regulation of STING is therapeutically beneficial. Accordingly, in another embodiment, the present invention relates to a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof or at least one compound of formula (I) for use as a pharmaceutical. In one embodiment, the present invention relates to a pharmaceutical composition comprising a compound of formula (I), a pharmaceutically acceptable salt thereof or at least one of these compounds for use in the treatment of cancer in cats or dogs.

[0050] In another embodiment, the compound according to the present invention has a temperature of >15°C, more preferably >20°C, and even more preferably >25°C, as determined by DSF. m This shows the interaction with canine STING (dSTING) reflected in the shift. In preferred embodiments, the compounds according to the present invention exhibit interaction with dSTING determined by DSF (dDSF) and induce cytokine secretion in canine whole blood (dWB). In a more preferred embodiment, the compound according to the present invention exhibits a combination of high dDSF and low dWB.

[0051] Treatment method In one embodiment, the present invention relates to the use of a pharmaceutical composition comprising a compound of formula (I), a pharmaceutically acceptable salt thereof, or at least one of these compounds in a method for treating a disease. In particular, compounds of general formula (I) or salts thereof are useful for the prevention and / or treatment of diseases and / or conditions in mammals, such as cats, mice, pigs, and dogs, where the regulation of STING is therapeutically beneficial. Furthermore, due to their activity, the compounds of the present invention are suitable as vaccine adjuvants. Diseases and conditions associated with or regulated by STING include, but are not limited to, inflammatory, allergic, or autoimmune diseases such as allergic rhinitis or asthma, infections, or cancer. Autoimmune diseases include, but are not limited to, systemic lupus erythematosus, psoriasis, insulin-dependent diabetes mellitus (IDDM), dermatomyositis, and Sjögren's syndrome (SS). The compounds of the present invention may be used to treat inflammation of any tissue and organ of the body, including but not limited to musculoskeletal inflammation, vasculitis, neuroinflammation, gastrointestinal inflammation, ocular inflammation, reproductive system inflammation, and other inflammations. Examples of musculoskeletal inflammations that can be treated with the compounds of the present invention include arthritis (e.g., osteoarthritis, rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, acute and chronic infectious arthritis, arthritis associated with gout and pseudogout, and juvenile idiopathic arthritis), tendinitis, synovitis, tenosynovitis, bursitis, connective tissue inflammation (fibromyalgia), epicondylitis, myositis, and osteitis (e.g., Paget's disease, pubic osteitis, and cystic fibrous osteitis). Examples of ocular inflammations that can be treated with the compounds of the present invention include blepharitis, blepharochalasis, conjunctivitis, dacryoadenitis, keratitis, keratoconjunctivitis sicca (dry eyes), scleritis, trichiasis, and uveitis.

[0052] Examples of inflammation of the nervous system that can be treated with the compounds of the present invention include encephalitis, Guillain-Barré syndrome, meningitis, neuromyotonic syndrome, narcolepsy, multiple sclerosis, myelitis, and schizophrenia. Examples of inflammation of the vascular or lymphatic system that can be treated with the compounds of the present invention include arthritis, arthritis, phlebitis, vasculitis, and lymphangitis. Examples of inflammatory conditions of the digestive system that can be treated with the compounds of the present invention include cholangitis, cholecystitis, enteritis, colitis, gastritis, gastroenteritis, inflammatory bowel disease (such as Crohn's disease and ulcerative colitis), ileitis, and proctitis. Examples of inflammatory conditions of the reproductive system that can be treated with the compounds of the present invention include cervicitis, chorioamnionitis, endometritis, epididymitis, omphalitis, oophoritis, orchitis, salpingitis, tubal-ovarian abscess, urethritis, vaginitis, vulvitis, and vulvar pain. The compound may be used to treat autoimmune conditions having inflammatory components. Such conditions include acute disseminated alopecia universalis, Behçet's disease, Chagas disease, chronic fatigue syndrome, autonomic dysphagia, encephalomyelitis, ankylosing spondylitis, aplastic anemia, hidradenitis suppurativa, autoimmune hepatitis, autoimmune oophoritis, celiac disease, Crohn's disease, type 1 diabetes mellitus, giant cell arteritis, Goodpasture syndrome, Graves' disease, Guillain-Barré syndrome, Hashimoto's disease, Henoch-Schönlein purpura, Kawasaki disease, lupus erythematosus, and microscopic This includes microscopic colitis, microscopic polyarteritis, mixed connective tissue disease, multiple sclerosis, myasthenia gravis, ocular clonus myoclonus syndrome, optic neuritis, Oud's thyroiditis, pemphigus, polyarteritis nodosa, polymyalgia, rheumatoid arthritis, Reiter's syndrome, Sjögren's syndrome, temporal arteritis, Wegener's granulomatosis, warm autoimmune hemolytic anemia, interstitial cystitis, Lyme disease, Morphea, psoriasis, sarcoidosis, scleroderma, ulcerative colitis, and vitiligo.

[0053] The compound may be used to treat T-cell-mediated hypersensitivity disorders having inflammatory components. Such conditions include contact hypersensitivity, contact dermatitis (including those caused by poison ivy), urticaria, cutaneous allergies, respiratory allergies (hay fever, allergic rhinitis), and gluten-sensitive enteropathy (celiac disease). Other inflammatory conditions that can be treated with the compound include, for example, appendicitis, dermatitis, dermatomyositis, endocarditis, connective tissue inflammation, gingivitis, glossitis, hepatitis, hidradenitis suppurativa, iritis, laryngitis, mastitis, myocarditis, nephritis, otitis, pancreatitis, parotitis, pericarditis, peritonitis, pharyngitis, pleurisy, pneumonitis, prostatitis, pyelonephritis, and stomatitis, transplant rejection (involving organs such as the kidneys, liver, heart, lungs, pancreas (e.g., islet cells), bone marrow, cornea, small intestine, skin allografts, skin homografts, and heart valve xenografts, serum sickness, and graft-versus-host disease), acute pancreatitis, chronic pancreatitis, acute respiratory distress syndrome, and Sézary's syndrome. This includes syndrome, congenital adrenal hyperplasia, non-suppurative thyroiditis, hypercalcemia associated with cancer, pemphigus, bullous herpetiform dermatitis, severe erythema multiforme, exfoliative dermatitis, seborrheic dermatitis, seasonal or perennial allergic rhinitis, bronchial asthma, contact dermatitis, atopic dermatitis, drug hypersensitivity reactions, allergic conjunctivitis, keratitis, herpes zoster, iritis and iridocyclitis, chorioretinitis, optic neuritis, symptomatic sarcoidosis, chemotherapy for fulminant or disseminated pulmonary tuberculosis, idiopathic thrombocytopenic purpura in adults, secondary thrombocytopenia in adults, acquired (autoimmune) hemolytic anemia, leukemia and lymphoma in adults, acute leukemia in childhood, focal enteritis, autoimmune vasculitis, multiple sclerosis, chronic obstructive pulmonary disease, solid organ transplant rejection, and sepsis.

[0054] Preferred treatments include treatment of transplant rejection, rheumatoid arthritis, psoriatic arthritis, multiple sclerosis, type 1 diabetes, asthma, inflammatory bowel disease, systemic lupus erythematosus, psoriasis, chronic lung disease, and inflammation associated with infectious conditions (e.g., sepsis). In one embodiment, the disease or condition treated with the compound of the present invention is cancer. Examples of cancerous diseases and conditions in which the compound of formula (I), or its salts or solvates, may potentially have beneficial antitumor effects include cancers of the lung, bone, pancreas, skin, brain, head, neck, uterus, ovaries, stomach, colon, colorectal, breast, esophagus, small intestine, intestine, endocrine system, thyroid, parathyroid, adrenal gland, urethra, prostate, penis, testis, ureter, bladder, kidney or liver, and bile duct; urothelial carcinoma; rectal cancer; anal cancer; carcinoma of the fallopian tube, endometrium, cervix, vagina, vulva, renal pelvis, and renal cells; sarcoma; soft tissue sarcoma; myxoma; and rhabdomyocytosis. Myomas; fibromas; lipomas; teratomas; intrahepatic cholangiocarcinomas; hepatoblastomas; angiosarcomas; hemangiomas; hepatocellular carcinomas; fibrosarcomas; chondrosarcomas; myelomas; chronic or acute leukemias; lymphocytic lymphomas; primary CNS lymphomas; CNS neoplasms; spinal axial tumors; squamous cell carcinomas; synovial sarcomas; malignant pleural mesotheliomas; brainstem gliomas; pituitary adenomas; bronchial adenomas; chondrodic hamartomas (hanlartomas); mesotheliomas; Hodgkin's disease, or any combination of one or more of the aforementioned cancers.

[0055] Preferred cancers that can be treated with the compounds according to the present invention include skin, lung, for example, small cell lung cancer, non-small cell lung cancer, liver, pancreas, colon, colorectal, brain, breast, ovary, prostate, kidney, bladder, bile duct, endometrium, thyroid, cervix, stomach, head, neck, sarcoma, soft tissue sarcoma, esophageal, head and neck cancer, rectal and urothelial cancer, and lymphoma. The novel compounds may be used, optionally, in combination with surgical procedures, radiotherapy, or other “cutting-edge” compounds, such as cytostatic or cytotoxic substances, cell proliferation inhibitors, anti-angiogenic substances, steroids, antibodies, nanobodies, cancer targeting agents, viruses including but not limited to oncolytic viruses, or immunogenic cell death inducers, for the prevention, palliative, curative, or semi-curative, short-term or long-term treatment of the aforementioned diseases. The novel compounds may also be used in combination with surgical procedures, radiotherapy, or other "cutting-edge" compounds, such as cell quiescent or cytotoxic substances, cell proliferation inhibitors, anti-angiogenic substances, steroids, antibodies, nanobodies, cancer targeting agents, viruses including but not limited to oncolytic viruses, or immunogenic cell death inducers, by combining different routes of administration of the compounds, e.g., intravenous, intratumoral, subcutaneous, inhalation, or oral, for the prevention, palliative, curative, or semi-curative, short-term or long-term treatment of the aforementioned diseases. For example, partial or complete tumor resection may be combined with the compounds of the present invention. For example, external beam radiotherapy may be combined with the compounds of the present invention. In its role as an adjuvant, in certain embodiments, the compounds and compositions of the present invention may be used as adjuvants in a vaccine-based therapeutic or preventive strategy. Therefore, the compounds of the present invention, or salts thereof, may be used in conjunction with one or more vaccines selected to stimulate an immune response to one or more predetermined antigens. The compounds of the present invention, or salts thereof, may be provided together with or in addition to such vaccines.

[0056] Such vaccines may include inactivated or attenuated bacteria or viruses containing the antigen of interest, purified antigens, recombinant live viruses or bacterial delivery vectors that express and / or secrete the antigen, antigen-presenting cell (APC) vectors containing cells that have been loaded with the antigen or transfected with a composition containing nucleic acids encoding the antigen, liposomal antigen delivery vehicles, or naked nucleic acid vectors encoding the antigen. This list is not intended to be limiting. For example, such vaccines may also include inactivated tumor cells or oncolytic viruses that express and secrete one or more GM-CSF, CCL20, CCL3, IL-12p70, FLT-3 ligands, or cytokines. Therefore, the present invention relates to a compound of general formula (I) for use as a pharmaceutical, for example, as a pharmaceutical for treating cancer in cats or dogs, or for example, as a vaccine adjuvant for use in pigs. In a preferred embodiment, the present invention relates to a pharmaceutical composition for use in the treatment of cancer in cats or dogs, comprising a compound of formula (I), a pharmaceutically acceptable salt thereof, or at least one compound of formula (I).

[0057] In one embodiment, the present invention relates to a compound or a pharmaceutically acceptable salt thereof or a pharmaceutical composition for use in the treatment of canine cancer, wherein the canine cancer is selected from osteosarcoma (OSA), oral melanoma, B-cell lymphoma, urothelial carcinoma (UC), angiosarcoma, mast cell tumor, soft tissue sarcoma, squamous cell carcinoma, T-cell lymphoma, mammary gland adenocarcinoma, and anal sac cancer. In another embodiment, the present invention relates to a compound or a pharmaceutically acceptable salt thereof or a pharmaceutical composition for use in the treatment of feline cancer, wherein the feline cancer is selected from B-cell and / or T-cell lymphoma, squamous cell carcinoma, mammary gland adenocarcinoma, mast cell tumor, and injection site sarcoma.

[0058] In a further embodiment, the present invention relates to a method for treating and / or preventing the diseases and conditions described above, comprising administering to a target an effective amount of a pharmaceutical composition comprising a compound of formula (I), a pharmaceutically acceptable salt thereof, or at least one compound of formula (I). In a particularly preferred embodiment, the present invention relates to a method for treating cancer in cats or dogs, comprising administering to a cat or dog an effective amount of a pharmaceutical composition comprising a compound of formula (I), a pharmaceutically acceptable salt thereof, or at least one compound of formula (I). In a more preferred embodiment, the present invention relates to a method for treating canine cancer, comprising administering to a dog an effective amount of a pharmaceutical composition comprising a compound of formula (I), a pharmaceutically acceptable salt thereof, or at least one compound of formula (I), wherein the canine cancer is selected from osteosarcoma (OSA), oral melanoma, B-cell lymphoma, urothelial carcinoma (UC), angiosarcoma, mast cell tumor, soft tissue sarcoma, squamous cell carcinoma, T-cell lymphoma, mammary gland adenocarcinoma, and anal sac cancer.

[0059] In another, more preferred embodiment, the present invention relates to a method for treating feline cancer, comprising administering to a cat an effective amount of a pharmaceutical composition comprising a compound of formula (I), a pharmaceutically acceptable salt thereof, or at least one compound of formula (I), wherein the feline cancer is selected from B-cell and / or T-cell lymphoma, squamous cell carcinoma, mammary gland adenocarcinoma, mast cell tumor, and injection site sarcoma. In a further embodiment, the present invention relates to a compound of general formula (I) for use in the treatment and / or prevention of the above-mentioned cancers, before or after tumor resection and / or radiotherapy. In a further embodiment, the present invention relates to the use of compounds of general formula (I) for the preparation of pharmaceuticals for treating and / or preventing the diseases and conditions described above. In another embodiment, the present invention relates to a method for treating cancer in dogs or cats, comprising administering to a dog or cat in combination with radiotherapy an effective amount of a pharmaceutical composition comprising a compound of formula (I), a pharmaceutically acceptable salt thereof, or at least one compound of formula (I).

[0060] Pharmaceutical composition In another aspect of the present invention, a pharmaceutical composition comprising at least one of the above-mentioned compounds is provided. The pharmaceutical composition may be formulated in a manner suitable for administering a therapeutically effective dose of the compound. Suitable preparations for administering the compound of formula (I) will be apparent to those skilled in the art and include, for example, tablets, pills, capsules, suppositories, lozenges, troches, solutions, syrups, elixirs, sachets, injectable solutions (subcutaneous, intravenous, intramuscular, intraperitoneal, intratumoral and peritumoral), inhalants, injectables, elixirs, emulsions, and powders. Furthermore, the compound according to the present invention may be administered by a targeted delivery platform, for example, an antibody-drug conjugate, a nanobody-drug conjugate, a peptide-drug conjugate, a virus-like particle, or a nanoparticle formulation. Suitable tablets may be obtained, for example, by mixing one or more compounds according to Formula I with known excipients, such as inert diluents, carriers, disintegrants, adjuvants, surfactants, binders, and / or lubricants. For the purposes of this disclosure, the pharmaceutical composition may be administered by a variety of means, including non-parenterally, parenterally, by inhalation spray, topically, nasally, orally, or rectally, as a formulation containing a pharmaceutically acceptable carrier, adjuvant, and vehicle. The pharmaceutical composition of this disclosure may also be administered in the form of a sterile injectable preparation, such as a sterile aqueous or oily suspension for injection.

[0061] Combination therapy The compounds of the present invention may be used alone or in combination with one or more further therapeutic agents. In a further embodiment, the present invention provides a method for treating a disease or condition in which modification of STING is beneficial, comprising administering a therapeutically effective dose of a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, and at least one further therapeutic agent. In a further embodiment, the present invention provides a method for treating inflammation, allergic or autoimmune diseases, infections or cancer, comprising administering a therapeutically effective dose of a combination comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, and at least one further therapeutic agent. The actual pharmacokinetically effective dose or therapeutic dose naturally depends on factors known to those skilled in the art, such as the patient's age and weight, the route of administration, and the severity of the disease. In any case, the combination is administered in a dose and manner that allows for the delivery of a pharmacokinetically effective dose based on the patient's specific condition. In certain embodiments, the compounds and compositions described herein are administered in conjunction with one or more additional compositions, including vaccines; adjuvants; CTLA-4 and PD-1 pathway antagonists, lipids, liposomes, chemotherapeutic agents, immunomodulatory cell lines, cancer targeting agents, immunogenic cell death inducers, and immunomodulators, where immunomodulators can be understood as a general range of activator-modulatory agents and agents that modulate and / or increase the frequency of certain immune cell subtypes.

[0062] The compounds and compositions thereof described herein may be administered before, after, and / or concurrently with additional therapeutic or prophylactic compositions or modalities. The compounds and compositions according to the present invention may be administered via mucosal routes (e.g., oral, sublingual, vaginal, nasal, cervical, etc.), intratumor, intraperitoneal, peritumor, percutaneous, inhalation, or parenteral routes (e.g., subcutaneous, intravenous, intramuscular, intra-arterial, intradermal, intrathecal, and epidural administration), including any combination with one or more additional therapeutic agents. Furthermore, the compounds and compositions according to the present invention may be administered by a targeted delivery platform, including any combination with one or more additional therapeutic agents, for example, such a targeted delivery platform may be an antibody-drug conjugate, a nanobody-drug conjugate, a peptide-drug conjugate, a virus-like particle, or a nanoparticle. Of the possible administration methods, intraperitoneal, intratumoral, peritumoral, subcutaneous, inhalation, or intravenous administration is preferred. The compounds and compositions according to the present invention may also be administered before, after, and / or simultaneously by different combinations of administration methods, including any combination with one or more additional therapeutic agents. For example, intratumoral or peritumoral administration may be performed following inhalation or intravenous administration, or inhalation or intravenous administration may be performed following intratumoral or peritumoral administration. In addition, such administration of the compounds by different routes may be before or after additional therapeutic steps, such as tumor resection or radiotherapy.

[0063] In particularly preferred embodiments, a pharmaceutical composition comprising the compound of the present invention, a pharmaceutically acceptable salt thereof, or at least one compound of the present invention is used in combination with radiotherapy. Simply as an example, the compound of the present invention may be administered after radiotherapy. Furthermore, the compound of the present invention may be administered intravenously after radiotherapy. Furthermore, the compound of the present invention may be administered intravenously after tumor resection. Furthermore, the compound of the present invention may be administered intratumorally after radiotherapy. Furthermore, the compound of the present invention may be administered peritumorally after radiotherapy. Furthermore, the compound of the present invention may be administered by inhalation after tumor resection. Furthermore, the compound of the present invention may be administered intratumorally following intravenous administration, with all administrations occurring after radiotherapy. Furthermore, the compound of the present invention may be administered intravenously following intratumor administration, with all administrations occurring after radiotherapy. Furthermore, the compound of the present invention may be administered peritumorally following intravenous administration, with all administrations occurring after radiotherapy. Furthermore, the compound of the present invention may be administered intravenously following peritumorally, with all administrations occurring after radiotherapy.

[0064] Methods for co-administering additional therapeutic agents are well known in the art. Due to the adjuvant properties of the compounds of the present invention, their use may also be combined with other therapeutic modalities, including other vaccines, adjuvants, antigens, antibodies, and immunomodulators. In addition to the compounds and compositions of the present invention described herein, the compositions or methods of the present invention may further comprise one or more additional substances which, due to their properties, can stimulate or otherwise utilize the immune system to act in response to cancer antigens present on targeted tumor cells. The compounds of the present invention can be used in combination with immune checkpoint inhibitors selected from the group consisting of CTLA-4 pathway antagonists, PD-1 pathway antagonists, Tim-3 pathway antagonists, Vista pathway antagonists, BTLA pathway antagonists, LAG-3 pathway antagonists, or TIGIT pathway antagonists.

[0065] The compounds of the present invention can be used in combination with cancer immune agonists, T cell receptor agonists, or TNF receptor superfamily agonists or antagonists. The compounds of the present invention can be used in combination with therapeutic antibodies or therapeutic nanobodies. In some embodiments, the mechanism of action of the therapeutic antibody is antibody-dependent cell-mediated cytotoxicity (ADCC). In additional embodiments of the methods described herein, the compounds of the present invention are used in combination with chemotherapeutic agents known to those skilled in the art (e.g., small molecule pharmaceutical compounds). Accordingly, the methods further involve administering an effective amount of one or more chemotherapeutic agents to the target as an additional or combination treatment.

[0066] Additional pharmacologically active substances that may be used together with / in combination with the compound of formula (I)—or its pharmaceutically acceptable salts—(including all individual embodiments or general subsets of compound (I)) or in the medical uses, treatments and / or preventive methods disclosed herein (above and below) include hormones, hormone analogs and antihormones (e.g., tamoxifen, toremifene, raloxifene, fulvestrant, megestrol acetate, flutamide, nilutamide, bicalutamide, aminoglutethimide, cyproterone acetate). , finasteride, buserelin acetate, fludrocortisone, fluoxymesterone, medroxyprogesterone, octreotide); aromatase inhibitors (e.g., anastrozole, letrozole, rialozol, borozole, exemestane, atamestane); LHRH agonists and antagonists (e.g., goserelin acetate, leuprolide); inhibitors of growth factors and / or their corresponding receptors (growth factors include, for example, platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), epidermal growth factor ( EGF), insulin-like growth factor (IGF), human epidermal growth factor (HER, e.g., HER2, HER3, HER4), and / or their corresponding receptors; inhibitors include, for example, (anti) growth factor antibodies, (anti) growth factor receptor antibodies, and tyrosine kinase inhibitors, such as afatinib, dacomitinib, canertinib, neratinib, abitinib, poziotinib, AV412, PF-6274484, HKI357, olmutinib, osimertinib, almonertinib, nazartinib, razertinib, peritinib, erlotinib, gefitinib, and ico This includes tinib, sapitinib, lapatinib, vallitinib, vandetanib, TAK-285, AEE788, BMS599626 / AC-480, GW583340, necitumumab, panitumumab, cetuximab, amivantanab, pertuzumab, trastuzumab, trastuzumab emtansine, or inhibitors of mutant EGFR, HER2 with exon 20 mutations, and hepatocyte growth factor (HGF), c-MET inhibitors, such as emibetuzumab, amivantanab, savolitinib, cabozantinib, and foretinib.Antimetabolites (e.g., methotrexate, larcitrexed, 5-fluorouracil (5-FU), capecitabine, phloxuridine, gemcitabine, mercaptopurine, thioguanine, cladribine, pentostatin, cytarabine (ara) C), combination of fludarabine, trifluridine and tipiracil (=TAS102); antitumor antibiotics (e.g., anthracyclines, e.g., doxorubicin, Doxil (pegylated liposomal doxorubicin hydrochloride), Myoset (non-pegylated liposomal doxorubicin), daunorubicin, epirubicin and idarubicin, mitomycin-C, bleomycin, dactinomycin, plicamycin, streptozocin); platinum derivatives (e.g., cisplatin, oxaliplatin, carboplatin); alkylating agents (e.g., estramustine, mechloretamine, melphalan, chlorambucil, busulfan, dacarbazine, cyclophosphamide, ifosfamide, temozolomide, nitrosourea, e.g., carmustine and lomustine, thiotepa, etc.); antimitotic agents (e.g., vinca alkaloids, e.g., vinblastine, vitamin Ndesine, vinorelbine and vincristine; and taxanes, e.g., paclitaxel, docetaxel, nab-paclitaxel (Abraxane); angiogenesis inhibitors (e.g., tascinimod, bevacizumab), tubulin inhibitors; DNA synthesis inhibitors, PARP inhibitors, topoisomerase inhibitors (e.g., epipodophyllotoxin, e.g., etoposide and etopophos, teniposide, amsacrin, topotecase) Irinotecan, Mitoxantrone); Serine / Threonine Kinase Inhibitors (e.g., PDK1 inhibitors, Raf inhibitors, A-Raf inhibitors, B-Raf inhibitors, C-Raf inhibitors, mTOR Inhibitors (e.g., Rapamycin, Temsirolimus, Everolimus, Ridahololimus, Zotarolimus, Sapanicertib, Trin-1, Dactosilib, GDC-0349, vs-5584; Bistucertib;AZD8055), mTORC1 / 2 inhibitors, PI3K inhibitors, PI3Kα inhibitors (e.g., alpelisib, ceravelisib, GDC-0077, HH-CYH33, AMG511, buparlisib, dactricib, pictilisib, taselicib), dual mTOR / PI3K inhibitors, STK33 inhibitors, AKT inhibitors, PLK1 inhibitors, CDK4 / 6 inhibitors (e.g., palbociclib, ribociclib, abemaciclib, trilaciclib, PF-06873600), aurora kinase inhibitors); tyrosine kinase inhibitors (e.g., PTK2 / FAK inhibitors); protein-protein interaction inhibitors (e.g., IAP inhibitors / SMAC mimetic drugs, MCL-1 (e.g., AZD-5991, AMG-176, AMG-397, S64315, S63845, A- 1210477), MDM2, MDM2 / MDMX); MEK inhibitors (e.g., trametinib, cobimetinib, binimetinib, selumetinib, refametinib); SOS1 inhibitors (i.e., compounds that regulate / inhibit the GEF function of SOS1 by, for example, binding to SOS1 and preventing protein-protein interactions between SOS1 and (mutant) Ras protein, e.g., KRAS; e.g., BAY-293); inhibitors of GDP-loaded or GTP-loaded RAS and / or any variant thereof (i.e., compounds that regulate / inhibit the function of (mutant) RAS protein by, for example, binding to GDP-loaded or GTP-loaded (mutant) RAS protein, e.g., KRAS, NRAS and / or HRAS, preferably KRAS); KRAS Irreversible inhibitors of G12C (AMG-510, MRTX849, ARS-324, GDC-6036); reversible or irreversible binders for GDP-loaded (mutated) KRAS; reversible or irreversible binders for GTP-loaded (mutated) KRAS; ALK inhibitors (e.g., crizotinib, alectinib, entrectinib, brigatinib, ceritinib); ERK inhibitors; FLT3 inhibitors; BRD4 inhibitors; IGF-1R inhibitors; TRAILR2 agonists; Bcl-xL inhibitors; Bcl-2 inhibitors (e.g., venetoclax, ovatoclax, navitoclax, oblimersen); Bcl-2 / Bcl-xL inhibitors; ErbB receptor inhibitors; BCR-ABL inhibitors; ABL inhibitors;Src inhibitors (e.g., dasatinib, ponatinib, bosutinib, vandetanib, KX-01, salakatinib, KX2-391, SU6656, WH-4-023); rapamycin analogs (e.g., everolimus, temsirolimus, ridaflorimus, sirolimus); androgen synthesis inhibitors; androgen receptor inhibitors; DNMT inhibitors; HDAC inhibitors; ANG1 / 2 inhibitors; histone deacetylase inhibitors; IL6 inhibitors; JAK and / or any variant thereof inhibitors; A - Inhibitors of Raf and / or B-Raf and / or C-Raf and / or any variant thereof (encorafenib, dabrafenib, vemurafenib, PLX-8394, RAF-709 (=Example 131 of International Publication No. 2014 / 151616), LXH254, sorafenib, LY-3009120 (=Example 1 of International Publication No. 2013 / 134243), rifirafenib, TAK-632, agerafenib, CCT196969, RO5126766, RAF 265); inhibitors of receptor tyrosine kinases (RTKs) and / or any variants thereof; inhibitors of SHP2 and / or any variants thereof (e.g., SHP099, TNO155, RMC-4550, RMC-4630, IACS-13909); CYP17 inhibitors; radiopharmaceuticals; proteasome inhibitors (e.g., carfilzomib); immunotherapeutic agents, e.g., immune checkpoint inhibitors (e.g., CTLA4, PD1, PD-L1, PD-L2, LAG3, SIRP alpha inhibitors) The body, and TIM3-binding molecules / immunoglobulins (ipilimumab, nivolumab, pembrolizumab, tislerizumab, atezolizumab, avelumab, durvalumab, pizilizumab, PDR-001 (=spartalizumab), AMG-404, ezabenlimab, cintilimab, camrelizumab, tripalimab, tislerizumab); ADCC (antibody-dependent cell-mediated cytotoxicity) enhancers (e.g., anti-CD33 antibody, anti-CD37 antibody, anti-CD20 antibody);This includes, but is not limited to, T cell engagers such as PSMA×CD3, B7H6 / CD3 (disclosed, e.g., in International Publication No. 2021 / 064137), DLL3 / CD3 (disclosed, e.g., in International Publication No. 2019 / 234220), bispecific T cell engagers (BiTE®), e.g., CD3×BCMA, CD3×CD33, CD3×CD19, etc., cancer vaccines, MDM2 inhibitors, oncolytic viruses, and various chemotherapeutic agents such as amifostin, anagrelide, clodronate, filgrastim, interferon, interferon alfa, leucovorin, procarbazine, levamisole, mesna, mitotane, pamidronate, and porfimers. The compounds of the present invention can be used in combination with OX40 agonists, ICOS ligands, CD27 agonists, GITR agonists, and Toll-like receptor agonists.

[0067] In preferred embodiments, additional pharmacologically active substances that may be used with / in combination with the compound of formula (I) - or its pharmaceutically acceptable salts - (including all individual embodiments or general subsets of compound (I)) or in the medical uses, treatments and / or preventive methods disclosed herein (above and below) include checkpoint inhibitors (ipilimumab, nivolumab, pembrolizumab, tislerizumab, atezolizumab, avelumab, durvalumab, pizilizumab, PDR-001 (=spartalizumab), AMG-404, ezabenlimab, cintilimab, This includes camrelizumab, tripalimab, tisrelizumab, taxanes (paclitaxel, docetaxel, nab-paclitaxel (Abraxane)), T cell engagers, e.g., PSMA×CD3, B7H6 / CD3 (e.g., disclosed in International Publication No. 2021 / 604137), DLL3 / CD3 (e.g., disclosed in International Publication No. 2019 / 234220), bispecific T cell engagers (BiTE®), e.g., CD3×BCMA, CD3×CD33, CD3×CD19, etc., cancer vaccines, MDM2 inhibitors, and oncolytic viruses. In additional embodiments of the methods described herein, the compounds of the present invention are used in combination with chemotherapeutic agents and / or additional agents, such as cancer-targeted therapies, for treating the indications described herein. Accordingly, the methods further involve administering an effective amount of one or more cancer-targeted agents to the target as an additional or combination treatment.

[0068] In additional embodiments of the methods described herein, the compounds of the present invention are used in combination with chemotherapeutic agents and / or additional agents and / or additional therapies, such as radiotherapy and / or tumor resection, for treating the indications described herein. In yet another aspect, the present invention relates to a method for treating a disease or condition in a patient that is related to or modulated by STING, comprising the step of administering a therapeutically effective amount of the compound of the present invention to a patient in need of such treatment in combination with a therapeutically effective amount of one or more additional therapeutic agents described herein earlier. The use of the compounds according to the present invention in combination with additional therapeutic agents may be carried out simultaneously or with a time delay. The compounds and one or more additional therapeutic agents according to the present invention may be present together in a single formulation, or separately in two identical or different formulations, for example, as a so-called kit of parts. Accordingly, in a further embodiment, the present invention provides a combination comprising a compound of general formula (I) and at least one further therapeutic agent. A further aspect of the present invention is to provide a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, and at least one further therapeutic agent and one or more pharmaceutically acceptable excipients.

[0069] In a further embodiment, the present invention provides a combination of a compound of formula (I), or a pharmaceutically acceptable salt thereof, and at least one further therapeutic agent for use in therapy. In a further embodiment, the present invention provides a combination of a compound of formula (I), or a pharmaceutically acceptable salt thereof, and at least one further therapeutic agent, for use in the treatment of a disease or condition in which the modification of STING is beneficial. In a further embodiment, the present invention provides a combination of a compound of formula (I), or a pharmaceutically acceptable salt thereof, and at least one further therapeutic agent for use in the treatment of cancer, for example, cancer in dogs or cats. In another aspect, the present invention relates to a pharmaceutical composition comprising the compound according to the present invention and one or more additional therapeutic agents described earlier and later herein, together with one or more inert carriers and / or diluents. Other features and advantages of the present invention will become apparent, for example, from the following more detailed examples illustrating the principles of the present invention.

[0070] chemical synthesis List of abbreviations [Table 1]

[0071] Other features and advantages of the present invention will become apparent from the following more detailed examples, which illustrate the principles of the present invention without limiting their scope. General Unless otherwise stated, all reactions are carried out using commercially available equipment and methods commonly used in chemical laboratories. Starting materials sensitive to air and / or moisture are stored under a protective gas, and the corresponding reactions and associated operations are carried out under a protective gas (nitrogen or argon). Compounds according to the present invention are named in accordance with IUPAC guidelines. Where a compound is represented by both a structural formula and its nomenclature, and there is a discrepancy, the structural formula shall prevail.

[0072] chromatography Thin-layer chromatography is performed on a ready-made TLC plate (containing fluorescent indicator F-254) with silica gel 60 on a Merck glass plate. The Biotage Isolera Four instrument is used for automated preparative NP chromatography with an Interchim Puri Flash column (50 μm, 12-300 g) packed with Millipore silica gel (Granula Silica Si-60A 35-70 μm) or a glass column. Preparative RP HPLC is performed using Waters columns (Sunfire C18, 10 μm, 30 × 100 mm, part number 186003971 or X-Bridge C18, 10 μm, 30 × 100 mm, part number 186003930). Compounds are eluted using different gradients of H2O / acetonitrile or H2O / MeOH with 0.1% TFA added to water, or using different gradients utilizing a basic buffer solution (1 L of water containing 5 mL of ammonium bicarbonate solution (158 g per L of H2O) and 2 mL of ammonia (7 mol / L solution in MeOH)) instead of the water-TFA mixture. HPLC (reaction monitoring) for the analysis of intermediate compounds is performed using columns from Waters and Phenomenex. In each case, the analytical instrument is also equipped with a mass detector.

[0073] HPLC mass spectrometry / UV spectroscopy The retention time / MS-ESI+ for characterizing the example compounds according to the present invention is determined using an HPLC-MS (high-performance liquid chromatography with mass detector), for example, an Agilent HPLC-MS instrument. The compound eluting at the injection peak is given a retention time tR=0. Analytical HPLC method: Acid method HPLC: Agilent 1260 Infinity II MS: Agilent LC / MS (G6125B) Column: Sunfire C18 2.5μm, 3.0×30mm Eluent: A: 0.1% TFA (v / v) in H2O; B: MeCN (HPLC grade) Detection: MS: Positive and negative modes Column temperature: 60℃ Gradient: 0.00 - 0.20 min: 3%B (flow rate 2.2 ml / min) 0.20 - 1.20 min: 3% - 100%B (flow rate 2.2 ml / min) 1.20 - 1.25 min: 100%B (flow rate 3.0 ml / min) 1.25 - 1.40 min: 100%B (flow rate 3.0 ml / min)

[0074] Basic method HPLC: Agilent 1260 Infinity II MS: Agilent LC / MS (G6125B) Column: X - Bridge C18, 2.5μm, 3.0×30 mm Eluent: A: 0.1% NH4OH (v / v) in H2O; B: MeCN (HPLC grade) Detection: MS: positive and negative modes Column temperature: 60°C Gradient: 0.00 - 0.20 min: 3%B (flow rate 2.2 ml / min) 0.20 - 1.20 min: 3% - 100%B (flow rate 2.2 ml / min) 1.20 - 1.25 min: 100%B (flow rate 3.0 ml / min) 1.25 - 1.40 min: 100%B (flow rate 3.0 ml / min)

[0075] Preparative HPLC method: Acidic method HPLC: Agilent 1260 Infinity II MS: Agilent LC / MS (G6125B) Column: Sunfire C18 10μm, 30×30 mm [[ID= forty-five]]Eluent: A: 0.1% TFA (v / v) in H2O; B: MeCN (HPLC grade) Detection: MS: positive and negative modes[[ID= forty-eight]] Flow rate: 50 ml / min Column temperature: 40°C Basic method HPLC: Agilent 1260 Infinity II MS: Agilent LC / MS (G6125B) Column: X-Bridge C18, 10 μm, 30 × 300 mm Eluent: A: 0.1% NH4OH (v / v) in H2O; B: MeCN (HPLC grade) Detection: MS: Positive and negative modes Flow rate: 50ml / min Column temperature: 40℃

[0076] Preparation of the compound according to the present invention The compounds and intermediates according to the present invention are known to those skilled in the art and may be obtained using synthetic methods described in the literature on organic synthesis. These methods are intended as examples of the present invention and do not limit the subject matter and the scope of the claimed compounds to these examples. Preferably, the compounds are obtained in a manner similar to the preparation methods described more fully later herein, particularly as described in the experimental section. In some cases, the order in which the reaction steps are carried out may be changed. Variations of reaction methods known to those skilled in the art but not described in detail herein may also be used. A general process for preparing the compounds according to the present invention will become apparent to those skilled in the art by examining the following scheme. The starting materials may be prepared by methods described in the literature or herein, or by similar or equivalent methods. Any functional groups in the starting materials or intermediates may be protected using conventional protecting groups. These protecting groups may be cleaved again at a suitable stage in the reaction sequence using methods well known to those skilled in the art.

[0077] One method for preparing the compound of formula (I) is illustrated in Scheme I. Indazole B can be synthesized from ortho-methylaniline derivative A. Subsequent iodination yields 3-iodoindazole C. Intermediate D can be obtained, for example, by Chan-Lam coupling using (6-fluoropyridine-3-yl)boronic acid. Conversion to intermediate F can be achieved, for example, by Suzuki coupling with intermediate E. Finally, the compound of formula (I) is synthesized, for example, by aromatic nucleophilic substitution. The product is isolated by conventional means and preferably purified by chromatography.

[0078] Scheme I: [ka]

[0079] Preparation of intermediates Intermediate 1 1-(6-fluoropyridine-3-yl)-3-iodo-7-methyl-1H-indazole [ka] To a stirred reaction mixture of 3-iodo-7-methyl-1H-indazole (16 g; 62 mmol) in DCM (200 ml), copper acetate (16.889 g; 93 mmol), pyridine (9.796 g; 124 mmol), and 6-fluoropyridine-3-boronic acid (14.851 g, 105 mmol) were added. The reaction mixture was then stirred at room temperature for 72 hours. The reaction mixture was filtered through Celite, the filtrate was concentrated, and the crude product was purified using silica chromatography (siRNA:hexane) to obtain the title compound (13 g). 1H NMR (400 MHz, DMSO-d6) δ ppm 2.09 (s, 3 H), 7.25 (m, 1 H), 7.34 (m, 1 H), 7.42 (m, 2 H), 8.28 (ddd, J=8.65, 7.07, 2.79 Hz, 1 H), 8.53 (d, J=2.15 Hz, 1 H).

[0080] Intermediate 2 1-(6-fluoropyridine-3-yl)-2',7-dimethyl-1H,2'H-3,4'-biindazole [ka] A mixture of 1-(6-fluoropyridine-3-yl)-3-iodo-7-methyl-1H-indazole (intermediate 1, 1.0 g, 2.8 mmol), 2-methyl-4-(tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole (1.1 g, 4.2 mmol), 1,1'-bis-(diphenylphosphino)ferrocene]dichloropalladium(II) (0.21 g, 0.28 mmol), and Na2CO3 (0.9 g, 8.5 mmol) was stirred in dioxane (20 ml) and water (5 ml) under an argon atmosphere at 110°C for 2 hours. At room temperature, water (100 ml) was added, and the mixture was extracted with SiO2. The combined organic layer was dried over MgSO4, concentrated under vacuum, and the product was purified by RP HPLC to obtain the title compound (0.8 g). [Examples]

[0081] Example 1 2-[(3R)-1-(5-{2”,7-dimethyl-1H,2”H-[3,4”-biindazole]-1-yl}pyridine-2-yl)pyrrolidine-3-yl]acetic acid [ka] To a solution of intermediate 2 (80 mg, 0.22 mmol) in anhydrous NMP (2 ml), methyl(R)-pyrrolidine-3-acetate HCl (106 mg, 0.56 mmol) was added, followed by the addition of DIPEA (145 mg, 1.12 mmol), and the mixture was heated at 140°C for 7 hours. The reaction product was allowed to cool, and NaOH (aqueous solution) (4 M, 300 μl, 1.2 mmol) was added, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was acidified with TFA and purified by preparative HPLC (acid method) to obtain the title compound (87 mg). 1H NMR (400 MHz, DMSO-d6) δ ppm 1.76 (dq, J=12.29, 8.45 Hz, 1 H), 2.21 (s, 3 H), 2.34 (m, 1 H), 2.48 (m, 2 H), 2.67 (m, 1 H), 3.20 (dd, J=10.65, 7.73 Hz, 1 H), 3.51 (m, 1 H), 3.66 (m, 1 H), 3.81 (dd, J=10.58, 7.29 Hz, 1 H), 4.06 (br d, J=31.05 Hz, 1 H), 4.20 (s, 3 H), 6.78 (br d, J=9.12 Hz, 1 H), 7.25 (m, 2 H), 7.42 (dd, J=8.62, 6.97 Hz, 1 H), 7.67 (d, J=8.62 Hz, 1 H), 7.77 (d, J=6.72 Hz, 1 H), 7.94 (dd, J=9.13, 2.28 Hz, 1 H), 8.09 (d, J=7.10 Hz, 1 H), 8.41 (d, J=2.53 Hz, 1 H), 8.61 (s, 1 H) HPLC (acidic): Rt = 0.802 min ((M+H)+ 481.2)

[0082] Example 2 2-[(3S)-1-(5-{2”,7-dimethyl-1H,2”H-[3,4”-biindazole]-1-yl}pyridine-2-yl)pyrrolidine-3-yl]acetic acid [ka] Using the method described for Example 1, the title compound (19.6 mg) was obtained using intermediate 2 (50 mg) and methyl(S)-pyrrolidine-3-acetate HCl (65 mg). 1H NMR (400 MHz, DMSO-d6) δ ppm 1.75 (dq, J=12.32, 8.40 Hz, 1 H), 2.20 (s, 4 H), 2.47 (d, J=7.35 Hz, 2 H), 2.65 (dquin, J=14.93, 7.36, 7.36, 7.36, 7.36 Hz, 1 H), 3.17 (dd, J=10.58, 7.67 Hz, 1 H), 3.48 (m, 1 H), 3.63 (m, 1 H), 3.79 (dd, J=10.52, 7.35 Hz, 1 H), 4.20 (s, 3 H), 6.64 (d, J=9.00 Hz, 1 H), 7.24 (m, 1 H), 7.42 (dd, J=8.62, 6.97 Hz, 1 H), 7.66 (d, J=8.62 Hz, 1 H), 7.77 (d, J=6.59 Hz, 1 H), 7.82 (dd, J=8.93, 2.60 Hz, 1 H), 8.09 (dd, J=7.60, 1.01 Hz, 1 H), 8.37 (d, J=2.53 Hz, 1 H), 8.61 (s, 1 H) HPLC (acidic): Rt = 0.865 min ((M+H)+ 467)

[0083] Example 3 2-[4-(5-{2',7-dimethyl-1H,2'H-[3,4'-biindazole]-1-yl}pyridine-2-yl)morpholine-2-yl]acetic acid [ka] Using the method described for Example 1, the title compound (57.6 mg) was obtained after preparative HPLC using intermediate 2 (50 mg) and methyl 2-(morpholine-2-yl) acetate HCl (72 mg). HPLC (acidic): Rt=0.947 min ((M+H)+483.2)

[0084] Example 4 2-(5-{2”,7-dimethyl-1H,2”H-[3,4”-biindazole]-1-yl}pyridine-2-yl)-2-azabicyclo[2.2.1]heptan-5-carboxylic acid [ka] Using the method described for Example 1, the title compound (69 mg) was obtained after preparative HPLC using intermediate 2 (50 mg) and methyl 2-azabicyclo[2.2.1]heptane-5-carboxylate (57 mg). HPLC (acidic): Rt=0.811 min ((M+H)+479.2)

[0085] Example 5 (1S,4R,5R)-2-(5-{2”,7-dimethyl-1H,2”H-[3,4”-biindazole]-1-yl}pyridine-2-yl)-2-azabicyclo[2.2.1]heptane-5-carboxylic acid and (1R,4S,5S)-2-(5-{2',7-dimethyl-1H,2'H-[3,4'-biindazole]-1-yl}pyridine-2-yl)-2-azabicyclo[2.2.1]heptane-5-carboxylic acid

[0086] [ka] Using the method described for Example 1, the title compound (56 mg) was obtained using intermediate 2 (50 mg) and methyl (1S,4R,5R)-2-azabicyclo[2.2.1]heptane-5-carboxylate (57 mg). 1H NMR (400 MHz, DMSO-d6) δ ppm 1.68 (m, 1 H), 1.76 (m, 1 H), 1.99 (m, 3 H), 2.22 (s, 5 H), 2.66 (m, 1 H), 2.93 (br s, 2 H), 3.25 (br d, J=9.63 Hz, 2 H), 3.52 (dd, J=9.82, 3.49 Hz, 1 H), 4.20 (s, 3 H), 4.70 (br s, 1 H), 6.79 (br d, J=7.10 Hz, 2 H), 7.25 (m, 3 H), 7.42 (dd, J=8.62, 6.97 Hz, 2 H), 7.66 (d, J=8.74 Hz, 2 H), 7.76 (d, J=6.59 Hz, 2 H), 7.91 (dd, J=8.93, 2.09 Hz, 2 H), 8.08 (d, J=7.10 Hz, 2 H), 8.39 (d, J=2.53 Hz, 2 H), 8.61 (s, 2 H) HPLC (acidic): Rt = 0.811 min ((M+H)+ 479.2)

[0087] Example 6 2-(5-{2',7-dimethyl-1H,2'H-[3,4'-biindazole]-1-yl}pyridine-2-yl)-2-azabicyclo[2.2.2]octane-5-carboxylic acid (A mixture of stereoisomers) [ka] Using the method described for Example 1, the title compound (42 mg) was obtained after preparative HPLC using intermediate 2 (50 mg) and 2-azabicyclo[2.2.2]octane-5-carboxylic acid (0.35 mmol). HPLC (acidic): Rt=0.839 min ((M+H)+493.2)

[0088] Examples 7-10 Example 6 was subjected to chiral preparative HPLC separation under the following conditions to obtain the following stereoisomers. (1S,4R,5R)-2-(5-{2',7-dimethyl-1H,2'H-[3,4'-biindazole]-1-yl}pyridine-2-yl)-2-azabicyclo[2.2.2]octane-5-carboxylic acid (single stereoisomer) (1R,4S,5S)-2-(5-{2',7-dimethyl-1H,2'H-[3,4'-biindazole]-1-yl}pyridine-2-yl)-2-azabicyclo[2.2.2]octane-5-carboxylic acid (single stereoisomer) (1S,4R,5S)-2-(5-{2',7-dimethyl-1H,2'H-[3,4'-biindazole]-1-yl}pyridine-2-yl)-2-azabicyclo[2.2.2]octane-5-carboxylic acid (single stereoisomer) (1R,4S,5R)-2-(5-{2',7-dimethyl-1H,2'H-[3,4'-biindazole]-1-yl}pyridine-2-yl)-2-azabicyclo[2.2.2]octane-5-carboxylic acid (single stereoisomer) Chiral preparative HPLC HPLC: Sepiatec PrepSFC50 Column: CHIRAL ART (registered trademark) Cellulose-SJ_10×250mm_5μm Eluent:A:80%scCO2;B:20%MeOH+20mM NH3 Detection: UV: 220nm Column temperature: 40℃ Flow rate: 15ml / min Gradient: Isocratic

[0089] Example 11 1-[(1R,4R)-5-(5-{2”,7-dimethyl-1H,2”H-[3,4”-biindazole]-1-yl}pyridine-2-yl)-2,5-diazabicyclo[2.2.1]heptan-2-yl]ethane-1-one (single stereoisomer) [ka] A mixture of intermediate 2 (50 mg, 0.14 mmol), (1R,4R)-2,5-diazabicyclo[2.2.1]heptane (30 mg, 0.31 mmol), and DIPEA (90.4 mg, 0.7 mmol) in anhydrous NMP (2 ml) was heated at 140°C for 15 hours. The mixture was allowed to cool to room temperature, and acetic anhydride (57 mg, 0.56 mmol) and DIPEA (90 mg, 0.7 mmol) were added, and the mixture was stirred at room temperature for 1 hour. The product was purified by preparative HPLC (acidic) to obtain the title compound (20.5 mg). HPLC (acidic): Rt=0.859 min ((M+H)+478.2)

[0090] Example 12 1-[(1S,4S)-5-(5-{2”,7-dimethyl-1H,2”H-[3,4”-biindazole]-1-yl}pyridine-2-yl)-2,5-diazabicyclo[2.2.1]heptan-2-yl]ethane-1-one (single stereoisomer) [ka] Using the method described for Example 11, the title compound was obtained after preparative HPLC using intermediate 2 and (1S,4S)-2,5-diazabicyclo[2.2.1]heptane.

[0091] Example 13 1-[5-(5-{2',7-dimethyl-1H,2'H-[3,4'-biindazole]-1-yl}pyridine-2-yl)-2,5-diazabicyclo[2.2.2]octan-2-yl]ethane-1-one (a mixture of stereoisomers) [ka] Using the method described for Example 11, the title compound (23 mg) was obtained after preparative HPLC using intermediate 2 (50 mg) and 2,5-diazabicyclo[2.2.2]octane dihydrochloride (68 mg). HPLC (acidic): Rt=0.896 min ((M+H)+492.2)

[0092] Example 14 - 15 Example 13 was subjected to chiral preparative HPLC separation using the following conditions to obtain the following stereoisomers. 1 - [(1R,4R) - 5 - (5 - {2’,7 - dimethyl - 1H,2’H - [3,4’ - biindazole] - 1 - yl}pyridin - 2 - yl) - 2,5 - diazabicyclo[2.2.2]octan - 2 - yl]ethan - 1 - one (single stereoisomer) 1 - [(1S,4S) - 5 - (5 - {2’,7 - dimethyl - 1H,2’H - [3,4’ - biindazole] - 1 - yl}pyridin - 2 - yl) - 2,5 - diazabicyclo[2.2.2]octan - 2 - yl]ethan - 1 - one (single stereoisomer) Chiral preparative HPLC HPLC: Sepiatec PrepSFC50 Column: CHIRAL ART (registered trademark) Amylose_10×250mm_5μm Eluent: A: 60% scCO2; B: 40% MeOH + 20 mM NH3 Detection: UV: 220 nm Column temperature: 40 °C Flow rate: 15 ml / min Gradient: Isocratic

[0093] Example 14 1 - [(1R,4R) - 5 - (5 - {2’,7 - dimethyl - 1H,2’H - [3,4’ - biindazole] - 1 - yl}pyridin - 2 - yl) - 2,5 - diazabicyclo[2.2.2]octan - 2 - yl]ethan - 1 - one (single stereoisomer)

Chem.

[0094] Example 15 1 - [(1S,4S) - 5 - (5 - {2’,7 - dimethyl - 1H,2’H - [3,4’ - biindazole] - 1 - yl}pyridin - 2 - yl) - 2,5 - diazabicyclo[2.2.2]octan - 2 - yl]ethan - 1 - one (single stereoisomer)

Chem.

[0095] Example 16 2-(5-{2',7-dimethyl-1H,2'H-[3,4'-biindazole]-1-yl}pyridine-2-yl)-2-azabicyclo[2.2.1]heptane-5-carboxylic acid (mixture of stereoisomers) [ka] Using the method described for Example 1, the title compound (55 mg) was obtained after preparative HPLC (acidic) using intermediate 2 (50 mg) and methyl 2-azabicyclo[2.2.1]heptane-5-carboxylate (57.1 mg, 0.35 mmol). HPLC (acidic): Rt=0.846 min ((M+H)+479.0)

[0096] Example 17 (1S,4R,5R)-2-(5-{2',7-dimethyl-1H,2'H-[3,4'-biindazole]-1-yl}pyridine-2-yl)-2-azabicyclo[2.2.1]heptane-5-carboxylic acid and (1R,4S,5S)-2-(5-{2',7-dimethyl-1H,2'H-[3,4'-biindazole]-1-yl}pyridine-2-yl)-2-azabicyclo[2.2.1]heptane-5-carboxylic acid (mixture of stereoisomers)

[0097] [ka] Using the method described for Example 1, the title compound (55 mg) was obtained after preparative HPLC (acidic) using intermediate 2 (200 mg) and cis-methyl 2-azabicyclo[2.2.1]heptane-5-carboxylate (215 mg, 1.12 mmol). HPLC (acidic): Rt=0.859 min ((M+H)+479.2)

[0098] Examples 18-19 Example 16 was subjected to chiral preparative HPLC separation using the following conditions to obtain the following stereoisomers. (1R,4S,5S)-2-(5-{2’,7-dimethyl-1H,2’H-[3,4’-biindazol]-1-yl}pyridin-2-yl)-2-azabicyclo[2.2.1]heptane-5-carboxylic acid (single stereoisomer) (1S,4R,5R)-2-(5-{2’,7-dimethyl-1H,2’H-[3,4’-biindazol]-1-yl}pyridin-2-yl)-2-azabicyclo[2.2.1]heptane-5-carboxylic acid (single stereoisomer) Chiral preparative HPLC HPLC: Sepiatec PrepSFC100 Column: CHIRAL ART (registered trademark) Amylose-C_neo_20×250nm_5μm Eluent: A: 60% scCO2; B: 40% MeOH + 20 mM NH3 Detection: UV: 220 nm Column temperature: 40 °C Flow rate: 60 ml / min Gradient: Isocratic

[0099] Example 18 (1R,4S,5S)-2-(5-{2’,7-dimethyl-1H,2’H-[3,4’-biindazol]-1-yl}pyridin-2-yl)-2-azabicyclo[2.2.1]heptane-5-carboxylic acid (single stereoisomer)

Chemical formula

[0100] Example 19 (1S,4R,5R)-2-(5-{2',7-dimethyl-1H,2'H-[3,4'-biindazole]-1-yl}pyridine-2-yl)-2-azabicyclo[2.2.1]heptane-5-carboxylic acid (single stereoisomer) [ka] 1H NMR (400 MHz, DMSO-d6) δ ppm 1.72 (m, 2 H), 1.99 (m, 2 H), 2.23 (s, 3 H), 2.66 (dd, J=8.43, 5.64 Hz, 1 H), 2.94 (br s, 1 H), 3.26 (br d, J=9.63 Hz, 1 H), 3.53 (dd, J=9.76, 3.42 Hz, 1 H), 4.20 (s, 3 H), 4.71 (br s, 1 H), 6.82 (br d, J=7.10 Hz, 1 H), 7.25 (m, 2 H), 7.42 (dd, J=8.62, 6.97 Hz, 1 H), 7.67 (d, J=8.62 Hz, 1 H), 7.76 (d, J=6.72 Hz, 1 H), 7.93 (dd, J=9.06, 2.22 Hz, 1 H), 8.09 (d, J=7.35 Hz, 1 H), 8.40 (d, J=2.53 Hz, 1 H), 8.61 (s, 1 H).

[0101] Pharmacological activity Biological examples The compounds of this disclosure were tested in the following two assays: a canine differential scanning fluorescence assay and a canine whole blood assay. Representative results for the compounds of the present invention are summarized in Tables 1 and 2 below. Further assays for testing the compounds are also described below.

[0102] Canine Differential Scanning Fluorescence (DSF) Assay Production and purification of canine STING (dSTING) protein The protein used in the biophysical experiments was recombinant canine STING protein containing a cytosolic cGAMP-binding ectodomain. A codon-optimized DNA sequence encoding amino acid residues 149-375 of canine STING (for expression in Escherichia coli) was synthesized by GeneArt (Regensburg, Germany) and inserted into the pET17b E. coli expression vector. The protein construct encodes the tobacco etch virus protease (TEV) cleavage site and the STING gene sequence described above, following an N-terminal 8×His tag. The obtained protein sequence (SEQ ID NO: 1) of the canine STING protein used is shown below. His-TEV-dSTING MHHHHHHHHENLYFQSGEKRNFNVAHGLAWSYFIGYLRLILPGLPARIQALHNNMLQGIGSHRLHILFPLDCGVPDDLSVVDPNIRFLYELPQQSANRAGIKRRVYTNSVYELLEKGQPAGI CVLEYATPLQTLFAMSQDGRAGFSREDRLEQAKLFCRTLEDILADAPELQNNCRLIVYQEPAEGSSFSLSQEILRHLRQEEREVTMGSMDTSIVPTSSTLSQEPNLFISGLEQPLPLRTDIF

[0103] For the expression of recombinant canine STING described above, constructs were transformed into E. coli BL21 DE3 strain and grown in LB medium at 15°C in a shaking flask. Expression was induced by adding isopropyl β-D-1-thiogalactopyranoside to a final concentration of 1 mM, and the culture was shaken overnight. The cell pellet was centrifuged and stored at -70°C until use. Following cell thawing in lysis buffer (20 mM TRIS-HCl, pH 8, 500 mM NaCl, 1 mM DTT, 0.5 mg / ml lysozyme, Complete protease inhibitor (Roche) and DNase (Roche)), the protein was purified by metal affinity purification using Ni-NTA resin and elution buffer consisting of 20 mM TRIS-HCl, pH 8, 500 mM NaCl, 1 mM DTT, and 300 mM imidazole. His tag cleavage using TEV protease was performed during overnight dialysis in size exclusion buffer (20 mM TRIS-HCl, pH 8, 100 mM NaCl, 1 mM DTT). For further purification of the target protein, the flow-through was subjected to size exclusion chromatography using a reverse nickel affinity column. Peak fractions were collected and concentrated to 5 mg / mL.

[0104] Biophysical assay - Determination of increased stability of canine STING protein against thermal denaturation, differential scanning fluorescence (DSF) The binding affinity of the compound of the present invention was demonstrated using a thermal shift assay to measure the stability of a suitable protein material from canine STING against thermal denaturation in the presence of the compound. In this assay, the protein unfolding temperature is monitored in the presence of a fluorescent dye that exhibits affinity for hydrophobic amino acids of the protein that are buried in the folded state and gradually exposed during unfolding. The dye fluorescence is quenched in an aqueous environment and increases when the dye associates with the hydrophobic portion of the unfolding protein. A plot of fluorescence intensity as a function of temperature typically shows a sigmoid curve (differential scanning fluorescence measurement) interpreted by a two-state model of protein unfolding. The inflection point of the curve represents the protein's "melting" temperature (Tm), which is numerically calculated using the Boltzmann equation.

[0105] The thermal stability of canine STING protein was measured in an assay buffer containing 20 mM Tris and 150 mM NaCl at pH 7.5. This assay was performed on a CFX384 real-time system (Bio-Rad) using a 384-well qPCR plate (catalog #781358, BRAND) and Microseal® "B" adhesive seals for PCR plates (catalog #MSB-1001, BIO-RAD). A DMSO stock solution of SYPRO orange (SIGMA S5692-500UL) was prepared. The compound stock solution (10 mM in DMSO) was diluted 1:2 with DMSO to an intermediate compound concentration of 5 mM, and then further diluted 1:40 with assay buffer to obtain a compound concentration of 125 μM and 2.5% DMSO. The fluorescent dye stock solution (5000×SYPRO Orange) was then mixed with the target protein and buffer to obtain a concentration of 15 μM protein and 25×SYPRO Orange. 2 μl of this protein-dye mixture was added to 8 μl of compound solution. The final volume was 10 μL. 3–6 well positions were used as negative controls (protein containing 2% DMSO). Plates were prepared for dual measurements and centrifuged at 1000 g for 2 minutes. 160 cycles at 0.5°C were used for the measurements (temperature gradient 15 sec / cycle, 15°C–95°C).

[0106] The final assay concentrations for compound characterization were 100 μM compound, 3 μM target protein, 5 × SYPRO Orange, and 2% DMSO in 10 μl. All dispensing steps were performed using a HamiltonStar pipetting robot (Hamilton). The dissociation curves were processed using Bio-Rad CFX Manager. The peak type was set to "negative". Compound codes for screening were assigned to the plate layout. The TM measurements from two replicates were averaged, and the standard deviation was calculated. If the standard deviation (SD) > 1.5°C, the measurement was repeated. The ΔTm value was generated by subtracting the melting point (Tm) obtained for the STING protein alone from the T value obtained for the protein incubated with the ligand.

[0107] Canine whole blood assay To detect STING activation in a physiological environment, canine whole blood (dWB) was stimulated with the cyclic dinucleotide cGAMP or a test compound. Pathway activity was monitored by measuring IFNb production. The compounds were supplied as a 10 mM DMSO solution, which was diluted and transferred to a 384-well assay plate (Greiner #781182) pre-filled with 10 μl of 1 × HBSS (10 × HBSS (+Ca / +Mg), #14065-049, Gibco) per well using an Echo acoustic dispenser. Typically, eight concentrations were used, with the highest concentration being 10 μM in the final assay volume, followed by a dilution step of approximately 1:4. The DMSO concentration was set to 0.1% in the final assay volume. The 384-well assay plate contained 20 test compounds as well as DMSO in the control and cGAMP standard wells. Canine whole blood was collected as sodium citrate blood (e.g., 3.8% in Sarstedt Monovette) and kept overnight at 4°C until use in the assay. 80 μl whole blood samples were transferred to each well of a 384-well assay plate filled with compound / 1×HBSS. The blood plate was kept at room temperature for 60 minutes, then covered with a lid but not sealed, and continuously shaken at 450 rpm. The 10×cGAMP assay solution was diluted with 1×HBSS from a 2 mM stock solution at room temperature immediately before use. 10 μl of 10×cGAMP / HBSS was added to the high control wells, while only HBSS was added to all compound and low control wells. After covering the assay plate with a lid, the blood plate was kept in an incubator at 37°C for 4 hours without shaking. For the detection of IFNb in canine plasma, a canine interferon beta ELISA kit (Biotrend #SEA222Ca) was used. The whole blood assay plate was centrifuged at 1000 g for 10 minutes at 8°C. 40 μl of supernatant was transferred from a 384-well whole blood plate to a corresponding 96-well ELISA plate pre-filled with 60 μl of assay diluent in each well, using a 96-well pipetting robot. The plate was sealed with a microplate seal and incubated again at 4°C overnight. The ELISA plate was allowed to return to room temperature and then incubated at 37°C for 1 hour. A working solution of detection reagent A was prepared by diluting detection reagent A with assay reagent A at a ratio of 1:100.Next, the liquid was removed from the 96-well ELISA plate, and 100 μL of detection reagent A working solution was added to each well. The ELISA plate was covered with a plate sealer and incubated at 37°C for 1 hour. 1× wash buffer was prepared by diluting 30× wash buffer concentrate with H2O. Detection reagent B working solution was prepared by diluting detection reagent B with assay reagent B in a 1:100 ratio. The ELISA plate was washed three times with 350 μL of wash buffer, then inverted and wiped with absorbent paper to remove all liquid. 100 μL of detection reagent B working solution was added to each well of the ELISA plate, then the ELISA plate was covered with a plate sealer and incubated at 37°C for 30 minutes. After incubation, the ELISA assay plate was washed five times with 350 μL of wash buffer, then inverted again and wiped with absorbent paper to remove all remaining liquid. 90 μL of TMB substrate was added to each well of an ELISA plate. The ELISA plate was then covered with a plate sealer and incubated at 37°C for 15 minutes. The reaction was stopped by adding 50 μL of stop solution, and the absorbance was immediately measured at 450 nm.

[0108] Data evaluation and calculation: For data evaluation and calculation, the control percentage for each well was calculated based on the mean values ​​of the high control (cGAMP-stimulated control) and low control (unstimulated control) using the following standard four-parameter logistic regression equation. [y=(ad) / (1+(x / c)^b)+d] a = low value, d = high price, x=concentration M, c=EC50 M, b = slope

[0109] Canine liver microsome (dLM) assay The metabolic degradation of the test compound was assayed at 37°C using pooled liver microsomes from dogs (beagles). Each time point contained a final incubation volume of 100 μl, consisting of TRIS buffer pH 7.6 (0.1 M), magnesium chloride (5 mM), microsomal protein (1 mg / ml), and the test compound at a final concentration of 1 μM. Following a short pre-incubation period at 37°C, the reaction was initiated by adding beta-nicotinamide adenine dinucleotide phosphate, reduced form (NADPH, 1 mM), and terminated after different time points by transferring aliquots to the solvent. In addition, NADPH-independent degradation was monitored in NADPH-free incubation and terminated at the final time point. The residual percentage of the test compound after NADPH-independent incubation is reflected in parameter c (control) (metabolic stability). Quenched incubations were pelletized by centrifugation (10000 g, 5 min). Aliquots of the supernatant are assayed for the amount of the parent compound by LC-MS / MS. The half-life (t1 / 2 INVITRO) is determined by the slope of the semi-logarithmic plot of the concentration-time profile. Intrinsic clearance (CL_INTRINSIC) is calculated by considering the amount of protein during incubation. CL_INTRINSIC [μl / min / mg protein] = (Ln2 / (half-life [min] * protein content [mg / ml])) * 1000. For better cross-species comparisons, the predicted clearance in each species is expressed as a percentage of hepatic blood flow [QH%]. Generally, high cross-species stability of the compound (corresponding to a low QH%) is desirable.

[0110] Mouse liver microsome (mLM) assay Metabolic degradation of the test compound was assayed at 37°C using pooled liver microsomes from (male / female) mice (CD1). Each time point contained a final incubation volume of 100 μl, consisting of TRIS buffer pH 7.6 (0.1 M), magnesium chloride (5 mM), microsomal protein (0.5 mg / ml), and the test compound at a final concentration of 1 μM. Following a short pre-incubation period at 37°C, the reaction was initiated by adding reduced beta-nicotinamide adenine dinucleotide phosphate (NADPH, 1 mM) and terminated after different time points by transferring aliquots to the solvent. In addition, NADPH-independent degradation was monitored in NADPH-free incubation and terminated at the final time point. The residual [%] of the test compound after NADPH-independent incubation is reflected in parameter c (control) (metabolic stability). Quenched incubations were pelletized by centrifugation (10000 g, 5 min). Aliquots of the supernatant are assayed for the amount of the parent compound by LC-MS / MS. The half-life (t1 / 2 INVITRO) is determined by the slope of the semi-logarithmic plot of the concentration-time profile. Intrinsic clearance (CL_INTRINSIC) is calculated by considering the amount of protein during incubation. CL_INTRINSIC [μl / min / mg protein] = (Ln2 / (half-life [min] * protein content [mg / ml])) * 1000. For better cross-species comparisons, the predicted clearance in each species is expressed as a percentage of hepatic blood flow [QH%]. Generally, high cross-species stability of the compound (corresponding to a low QH%) is desirable.

[0111] Canine hepatocyte (dHep) assay The metabolic degradation of the test compound is assayed in a suspension of canine liver cells. Incubation: Thaw the cryopreserved canine hepatocytes and incubate them in a suitable buffer system (KHB buffer or similar buffer or standard cell culture medium) containing 50% fetal bovine serum. Following an acclimation period (15 - 30 minutes) in an incubator (37°C, 5 - 10% CO2, 85 - 95% humidity), add the test compound to the hepatocyte suspension (pH 7.4, typical cell density is approximately 1 million cells / mL; final concentration of test compound is 1 μM, final DMSO concentration < 0.05 v / v%). Incubate the cells for up to 6 hours and take samples at six different time points. Then, quench the samples with acetonitrile and pellet them by centrifugation. Next, analyze the remaining amount of the parent compound in the supernatant by HPLC - MS / MS.

[0112] Calculation: Calculate the elimination rate constant (ke) using the slope of the linear regression of the natural logarithm of [substrate remaining % or substrate concentration] versus time [hours]. ke = (-1) * slope ke = elimination rate constant [1 / hour] Calculate the half - life from the elimination rate constant. t 1 / 2 = ln(2) / ke t 1 / 2 = half - life [hours] Calculation of intrinsic in vitro hepatic clearance: CL_int_in vitro = ke * 1000 / (CD * 60) CL_int_in vitro = intrinsic hepatic clearance, in vitro [μl / min / 1 million cells] CD = cell density [1 million cells / mL] Note: The above equation is only valid when [substrate] << Km.

[0113] The calculated in vitro hepatic intrinsic clearance can be scaled up to the intrinsic in vivo hepatic clearance and used to predict the hepatic in vivo blood clearance (CL_ws) using a liver model (well - stirred model). CL_int_invivo=(CL_int_in vitro*H*L) / 1000 CL_int_invivo = Intrinsic hepatic clearance, in vivo [mL / min / kg] H = hepatocellularity [1 million cells / g liver] L = Liver factor [g / kg body weight] CL_ws=CL_int_invivo*Q / (CL_int_invivo+Q) CL = Liver clearance, estimated in vivo value [mL / min / kg] Q=hepatic blood flow [mL / min / kg] ws=Wellstard QH%=CL_ws*100 / Q QH% = Clearance expressed as a percentage of hepatic blood flow. CONC: Cell concentration during incubation time (10^6 / ml) T_LAST: End point of use (time)

[0114] Mouse hepatocyte (mHep) assay The metabolic degradation of the test compound is assayed in a suspension of mouse liver cells. Incubation: Cryopreserved mouse hepatocytes are incubated in a suitable buffer system (KHB buffer or similar buffer or standard cell culture medium) containing 50% seed serum. Following an acclimatization period (15-30 minutes) in an incubator (37°C, 5-10% CO2, 85-95% humidity), the test compound is added to a hepatocyte suspension (pH 7.4, typical cell density approximately 1 million cells / mL; final concentration of test compound 1 μM, final DMSO concentration <0.05 v / v%). Cells are incubated for up to 6 hours, and samples are collected at six different time points. Samples are then quenched with acetonitrile and pelletized by centrifugation. The residual amount of the parent compound in the supernatant is then analyzed by HPLC-MS / MS. Calculation: The elimination rate constant (ke) is calculated using the slope of a linear regression from the natural logarithm [substrate residual % or substrate concentration] versus time [hours]. ke = (-1) * slope ke = Disappearance rate constant [1 / hour] Calculate the half-life from the disappearance rate constant. t 1 / 2 = ln(2) / ke t 1 / 2 = Half-life [hour]

[0115] Calculation of intrinsic in vitro hepatic clearance: CL_int_in vitro = ke * 1000 / (CD * 60) CL_int_in vitro = Intrinsic hepatic clearance, in vitro [μl / min / 1 million cells] CD = Cell density [1 million cells / mL] Note: The above equation holds only when [substrate] << Km.

[0116] The calculated in vitro hepatic intrinsic clearance can be scaled up to the intrinsic in vivo hepatic clearance and used to predict the hepatic in vivo blood clearance (CL_ws) by using a liver model (Wellstead model). CL_int_invivo = (CL_int_in vitro * H * L) / 1000 CL_int_invivo = Intrinsic hepatic clearance, in vivo [mL / min / kg] H = Hepatocyte occupancy [1 million cells / g liver] L = Liver factor [g / kg body weight] CL_ws = CL_int_invivo * Q / (CL_int_invivo + Q) CL = Estimated value of hepatic clearance, in vivo [mL / min / kg] Q = Hepatic blood flow [mL / min / kg] ws = Wellstead QH% = CL_ws * 100 / Q QH% = Clearance expressed as a percentage of hepatic blood flow CONC: Cell concentration at incubation time (10^6 / ml) T_LAST: Terminal time point used (hour) Solid liver cells, mouse: 120 × 10 e6 cells / g liver Liver factor, mouse: 55 g / kg body weight Blood flow rate, mouse: 90 ml / (min × kg).

[0117] Permeability assay (MDCK-PGP) This assay provides information on the potential for compounds to cross the blood-brain barrier. Permeability measurements across a monolayer of polarized, confluent MDCK-MDR1 cells grown on a permeable filter support are used as an in vitro absorption model. The apparent permeability coefficient (PE) of compounds passing through the MDCK-MDR1 cell monolayer is measured in both apical-to-basal (AB) and basal-to-apical (BA) transport directions (pH 7.4, 37°C). AB permeability (PEAB) represents drug absorption from blood to brain, and BA permeability (PEBA) represents drug efflux from brain to blood. These are due to both passive permeability and active transport mechanisms, primarily mediated by overexpressed human MDR1 P-gp, via efflux and uptake transporters expressed on MDCK-MDR1 cells. Compounds are assigned a permeability / absorption class by comparing AB permeability to that of a reference compound with known in vitro permeability and oral absorption in humans. Identical or similar permeability in both transport directions suggests passive permeability, while directional permeability indicates an additional active transport mechanism. Higher PEBA than PEAB suggests the involvement of active efflux mediated by MDR1 P-gp. Active transport is concentration-dependent and saturates.

[0118] MDCK-MDR1 cells (1-2 × 10^5 cells / 1 cm^2 area) were seeded on a filter insert (Costar transwell polycarbonate or PET filter, pore size 0.4 μm) and cultured for 7 days (DMEM). Subsequently, MDR1 expression was enhanced by culturing the cells with 5 mM sodium butyrate in complete medium for 2 days. Dissolve the compound in a suitable solvent (DMSO, etc., 1-20 mM stock solution). The stock solution is diluted with HTP-4 buffer (128.13 mM NaCl, 5.36 mM KCl, 1 mM MgSO4, 1.8 mM CaCl2, 4.17 mM NaHCO3, 1.19 mM Na2HPO4 × 7H2O, 0.41 mM NaH2PO4 × H2O, 15 mM HEPES, 20 mM glucose, 0.25% BSA, pH 7.4) to prepare the transport solution (0.1–300 μM compound, final DMSO <= 0.5%). The transport solution (TL) is applied to the apical or basal donor side to measure AB or BA permeability (3 filter repeats). The receiver side contains the same buffer as the donor side. Samples are collected from the donor at the start and end of the experiment, as well as from the receiver side at various time intervals for up to 2 hours, for concentration measurement by HPLC-MS / MS or scintillation counting. The sampled receiver volume is replaced with a new receiver solution.

[0119] result [Table 2]

[0120] [Table 3]

[0121] As demonstrated by this embodiment, when measured by a binding assay, the compounds according to the present invention have a T temperature of >15°C, more preferably >20°C, and even more preferably >25°C, as determined by DSF. mIt exhibits interaction with canine STING (dSTING), which is reflected in the shift. Furthermore, the compounds according to the present invention induce cytokine secretion in canine whole blood (dWB). According to the present invention, a combination of high dDSF and low dWB is particularly preferred.

Claims

1. The following formula (I) Equation (I) (In the formula, R 1 is, -C 1 - 6 -Alkyl or -C 3 - 6 It is a cycloalkyl, R 2a is -H or -C 1 - 6 - Selected from the group consisting of alkyl groups, R 2b is selected from the group consisting of -H, -C 1 - 6 -alkyl, -C 1 - 6 -alkylene-OH, -C(O)OH, -C(O)O-C 1 - 6 -alkyl and -pyrazolyl-C 1 - 6 -alkyl R 2c is -H or -C 1 - 6 -It is alkyl, R 3 teeth, Selected from the group consisting of (* indicates the attachment point), R 4 H, -C 1 - 6 - Alkyl and C 3 - 6 Selected from cycloalkyl groups, R 5 H, -C 1 - 6 - Alkyl and C 3 - 6 Selected from cycloalkyl groups, R 6 H, -C 1 - 6 - Alkyl and C 3 - 6 Selected from cycloalkyl groups, R 7 H, -C 1 - 6 - Alkyl, C 3 - 6 Selected from cycloalkyl and -OH, R 8 is, (CH 2 ) n Here, n is an integer from 1 to 3, preferably 1 or 2. X is CH or N, and Y is -O-, -S-, -S(O)-, -S(O) 2 -is) A compound of or a pharmaceutically acceptable salt thereof.

2. R 1 However, -C 1 - 6 - Alkyl, preferably methyl, The compound according to claim 1 or a pharmaceutically acceptable salt thereof.

3. R 2a However, -C 1-6 - Alkyl, preferably methyl, R 2b However, -H or -C 1 - 6 -It is alkyl, R 2c However, -H or -C 1 - 6 - It is alkyl, The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof.

4. R 2a However, it is methyl, R 2b However, it is -H, R 2c However, it is -H. A compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof.

5. R 3 but, (In the formula, R 7 is, -C 1 - 6 - Alkyl, preferably methyl) A compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof.

6. R 3 but, The compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof.

7. R 3 but, And, R 4 However, it is -H or methyl, preferably -H. R 5 and R 6 However, independently, it is -H or methyl, preferably R 5 and R 6 However, both are -H, Y is O, A compound according to any one of claims 1 to 4 or 6, or a pharmaceutically acceptable salt thereof.

8. R 1 However, it is methyl, R 2a However, it is methyl, R 2b and R 2c However, it is -H, R 3 but, And, R 4 , R 5 and R 6 However, it is -H, Y is O, A compound according to any one of claims 1 to 4 or 6, or a pharmaceutically acceptable salt thereof.

9. The following structure A compound selected from or a pharmaceutically acceptable salt thereof.

10. The following structure A compound selected from or a pharmaceutically acceptable salt thereof.

11. A pharmaceutical composition comprising at least one compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

12. A compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 11, for use as a pharmaceutical.

13. A compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 11, for use in the treatment of cancer in cats or dogs.

14. A compound or a pharmaceutically acceptable salt thereof or pharmaceutical composition used according to claim 13, wherein the cancer in dogs is selected from osteosarcoma (OSA), oral melanoma, B-cell lymphoma, urothelial carcinoma (UC), angiosarcoma, mast cell tumor, soft tissue sarcoma, squamous cell carcinoma, T-cell lymphoma, mammary adenocarcinoma and anal sac cancer, or the cancer in cats is selected from B-cell and / or T-cell lymphoma, squamous cell carcinoma, mammary adenocarcinoma, mast cell tumor and injection site sarcoma.

15. A compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition, used in combination with radiotherapy, according to the use described in claim 13 or 14.

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