RU2800072C1 - Bicyclic agonists of the interferon gene stimulator sting - Google Patents

Abstract

FIELD: medicine, therapy.

SUBSTANCE: group of inventions is intended for the treatment of tumors in patients suffering from them. Compounds of formula (I) or (II) are provided, wherein the meanings of the various substituents are defined herein. In another embodiment, a pharmaceutical composition containing the said compounds is provided. Also a method of treating a tumor in a patient using the said compounds is provided, which can be delivered orally for systemic exposure or intratumorally. Also the use of the said compounds for the treatment of a tumor in a patient is provided. Antitumor therapy using these compounds may further include the administration of an effective dose of a drug that targets the checkpoint of the immune response.

EFFECT: group of inventions provides new compounds of formula (I) or (II) having agonistic bioactivity with respect to interferon gene stimulator (STING), which can be used in the treatment of tumors in patients suffering from them.

9 cl, 3 tbl, 4 ex

Description

[0001] This application claims priority under U.S. Provisional Application No. 62/889,679, filed August 21, 2019.

Background of the Invention

[0002] The cGAS-STING signaling pathway plays a key role in the innate immune response that is generated by mammalian host cells to eliminate a variety of DNA and RNA viruses. STING (interferon gene stimulator) is a resident endoplasmic reticulum (ER) signaling protein partially localized to mitochondria-associated membranes that is widely expressed in both immune and non-immune cell types. In response to cyclic dinucleotides (CDNs), including 2'-3' cGAMP, produced in response to cytosolic DNA via cyclic GMP-AMP synthase (cGAS), STING translocates to the perinuclear region, where it rapidly induces the production of type I interferon (IFN) and the pro-inflammatory cytokine TBK1-IRF3-dependently. STING has also been found to bind directly to cytosolic DNA, although the physiological significance of direct DNA detection has not yet been fully characterized.

[0003] Recent work has demonstrated that STING plays key roles in immune responses to tumor cells. Tumor-initiated efficient T cell priming within the tumor microenvironment requires the production of interferon-beta (IFN-β) by resident dendritic cells, and IFN-β expression has been shown to be dependent on activation of the STING pathway (1). Indeed, intratumoral delivery of nucleotide-based STING agonists has been shown to induce marked regression of established tumors in syngeneic mouse models (1). In addition, activation of the STING pathway has been shown to significantly contribute to the antitumor effect of irradiation through an IFN-β-mediated immune response in the irradiated tumor microenvironment.

Brief summary of the invention

[0004] In various embodiments, the present disclosure relates to an interferon gene stimulator (STING) agonist that can be used in the treatment of tumors.

[0005] According to various embodiments, the present disclosure relates to a compound of formula (I) or formula (II), or a pharmaceutically acceptable salt thereof:

Where

X is S, -N=C(R ¹ )- or -C(R ¹ )=C(R ¹ )-;

each R ¹ is independently H, F, Cl, C ₁ -C ₆ alkyl, ethenyl or ethynyl (each of which may be substituted), cyano, alkoxy, haloalkyl or C ₃ -C ₆ cycloalkyl;

or both R ¹ together with the carbon atoms to which they are attached form a fused phenyl;

R is H, alkyl optionally substituted with -((C ₁ -C ₆ alkyl)OC(O)OC ₁ -C ₆ alkyl), or benzyl, where benzyl may be unsubstituted or substituted with methoxy or an acid or ester isostere; And

ring A is a bicyclic fully aromatic or partially reduced heteroaryl ring system containing 3, 4 or 5 N atoms, substituted with 0, 1, 2, 3 or 4 substituents, each independently selected from the group consisting of NH ₂ , cyano, NHC(=O)O-tBu,OH, carboxamido, C ₁ -C ₆ -alkyl, -S(O) ₂ (C ₁ -C ₆ -alkyl), -S(O)(C ₁ -C ₆ -alkyl), alkylnitrile, alkoxy and haloalkyl; with the proviso that the partially reduced heteroaryl ring system may be substituted with an oxo group.

[0006] According to various embodiments, the present disclosure relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof:

Where

X is S, -N=C(R ¹ )- or -C(R ¹ )=C(R ¹ )-;

each R ¹ is independently H, F, Cl, C ₁ -C ₆ alkyl, ethenyl or ethynyl (each of which may be substituted), cyano, alkoxy, haloalkyl or C ₃ -C ₆ cycloalkyl;

or both R ¹ together with the carbon atoms to which they are attached form a fused phenyl;

R is H, alkyl optionally substituted with -((C ₁ -C ₆ alkyl)OC(O)C ₁ -C ₆ alkyl), or benzyl, where benzyl may be unsubstituted or substituted with methoxy or an acid or ester isostere; And

ring A is a bicyclic fully aromatic or partially reduced heteroaryl ring system containing 3, 4 or 5 N atoms, substituted with 0, 1, 2, 3 or 4 substituents, each independently selected from the group consisting of NH ₂ , cyano, NHC(=O)O-tBu,OH, carboxamido, C ₁ -C ₆ -alkyl, -S(O) ₂ (C ₁ -C ₆ -alkyl), -S(O)(C ₁ -C ₆ -alkyl), alkylnitrile, alkoxy and haloalkyl; with the proviso that the partially reduced heteroaryl ring system may be substituted with an oxo group.

More specifically, according to exemplary embodiments, a compound of the present disclosure includes any of the specific compounds shown in Table 1 below.

[0007] According to an embodiment, the present disclosure also relates to a method of stimulating interferon gene expression, comprising administering to a patient an effective dose of an interferon gene stimulator (STING) agonist containing a compound described herein, and a method of treating a tumor in a patient, comprising administering to a patient an effective dose of an interferon gene stimulator (STING) agonist containing a compound of formula (I).

[0008] Additionally, the method according to the present disclosure can be carried out using an effective dose of any of the specific compounds disclosed herein (see, for example, Table 1).

[0009] According to various embodiments, the method of treating a tumor may further comprise administering an effective dose of a compound disclosed herein via oral or intratumoral administration, or both.

[0010] According to various embodiments, the method of treating a tumor may further comprise administering an effective dose of a compound disclosed herein, the administration comprising administering the compound to a patient as an antibody-drug conjugate or in a liposomal dosage form.

[0011] According to various embodiments, the method of treating a tumor may further comprise administering an effective dose of a compound disclosed herein, further comprising administering an effective dose of a drug that targets an immune response checkpoint. For example, a drug that targets an immune response checkpoint may be an anti-PD-L1 antibody, an anti-PD-1 antibody, an anti-CTLA-4 antibody, or an anti-4-1BB antibody.

[0012] According to various embodiments, the method of treating a tumor may further comprise administering an effective dose of a compound disclosed herein, further comprising administering ionizing radiation or administering anti-cancer drugs.

[0013] According to various embodiments, the present disclosure also relates to a pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt thereof disclosed herein and a pharmaceutically acceptable carrier.

Detailed disclosure of the present invention

[0014] There is considerable interest in the development of STING pathway agonists for various immuno-oncological applications. Most importantly, STING pathway agonists have significant potential as part of combination therapies with checkpoint-targeting drugs in patients who fail to respond to checkpoint blockade alone.

[0015] The authors have developed a robust platform for the detection of non-nucleotide small molecular weight STING agonists. The platform was developed using a primary assay using the human THP-1 cell line carrying an IRF-inducible reporter with 5 copies of an IFN signaling element. Preliminary screening studies, including alternative reporter constructs, rodent cell assays, and cGAS and STING knockout cell lines, were used to eliminate luciferase artifacts and to confirm cross-species cross-reactivity in humans and rodents, as well as signaling selectivity. Biochemical assays, including assays for cGAS enzymatic activity and STING protein binding, were used to determine the specific targets of the identified compounds.

[0016] The term "administering treatment" or "treatment" as used herein refers to alleviating symptoms associated with a disorder or disease, or inhibiting further progression or worsening of such symptoms, or preventing or preventing a disease or disorder, or curing a disease or disorder. By analogy, as used herein, an "effective amount" or "therapeutically effective amount" of a compound of the present disclosure refers to an amount of a compound that alleviates or partially alleviates the symptoms associated with a disease or condition, or stops or slows the further progression or worsening of such symptoms, or prevents or prevents the disease or condition. In particular, the term "therapeutically effective amount" refers to an amount that, at the desired dosage and duration, is effective to achieve the desired therapeutic result. A therapeutically effective amount is also the amount at which the beneficial therapeutic effects outweigh any toxic or adverse effects of the compounds of this disclosure.

[0017] The expression "effective amount" used to describe therapy for an individual suffering from a disease refers to the amount or concentration of a compound according to the present disclosure, which is effective to inhibit or otherwise affect STING in the tissues of the individual in which STING is involved in the violation, and such inhibition or other effect occurs in an amount sufficient to obtain a beneficial therapeutic effect.

[0018] Typically, an initial therapeutically effective amount of a compound described herein, or a salt thereof, is in the range of about 0.01 to about 200 mg/kg, or about 0.1 to about 20 mg/kg, of the patient's body weight per day, with a typical starting range being about 0.3 to about 15 mg/kg/day. Oral unit dosage forms, such as tablets and capsules, may contain from about 0.1 mg to about 1000 mg of the compound, or a pharmaceutically acceptable form thereof. In another embodiment, such dosage forms contain from about 50 mg to about 500 mg of the compound, or a pharmaceutically acceptable form thereof. In yet another embodiment, such dosage forms contain from about 25 mg to about 200 mg of the compound, or a pharmaceutically acceptable form thereof. In yet another embodiment, such dosage forms contain from about 10 mg to about 100 mg of the compound, or a pharmaceutically acceptable form thereof. In a further embodiment, such dosage forms contain from about 5 mg to about 50 mg of the compound, or a pharmaceutically acceptable form thereof. According to any of the above embodiments, the dosage form can be administered once a day or twice a day.

[0019] The term "pharmaceutically acceptable salts" refers to non-toxic addition salts of organic or inorganic acids and/or bases (see, for example, Lit, et al., Salt Selection for Basic Drugs (1986), Int J. Pharm., 33, 201-217, incorporated herein by reference). Typical pharmaceutically acceptable salts include, for example, alkali metal salts, alkaline earth metal salts, ammonium salts, water-soluble and water-insoluble salts such as acetates, amsonates (4,4-diaminostilbene-2,2-disulfonate), benzenesulfonates, benzonates, bicarbonates, bisulfates, bitartrates, borates, bromide, butyrates, calcium salts, calcium edetates, camsylates, carbonates, chlorides, citrates, clavularates, dihydrochlorides, edetates, edisylates, estolates, esylates, fiunarate, gluceptates, gluconates, glutamates, glycolylarsanilates, hexafluorophosphates, hexylresorcinates, hydrabamines, hydrobromides, hydrochlorides, hydroxynaphthoates, iodides, isothionates, lactates, lactobionates, laurates, malates, maleates, mandelates, mesylates, methyl bromides, methyl nitrates, methyl sulfates, mukats, napsilates, nitrates, N-methylglucamine-ammonium salts, 3-hydroxy-2-naphthoates, oleates, oxalates, palmitates, pamoates (1,1-methene-bis-2-hydroxy-3-naphthoates, ein bonates), pantothenates, phosphates/diphosphates, picrates, polygalacturonates, propionates, p-toluenesulfonates, salicylates, stearates, subacetates, succinates, sulfates, sulfosalicylates, suramates, tannates, tartrates, theoclates, tosylates, triethiodides and valerates. A pharmaceutically acceptable salt may contain more than one charged atom in its structure. In this case, the pharmaceutically acceptable salt may contain a plurality of counterions. Thus, a pharmaceutically acceptable salt may contain one or more charged atoms and/or one or more counterions.

[0020] Standard abbreviations for chemical groups are used, such as those well known in the art; e.g., Me=methyl, Et=ethyl, i-Pr=isopropyl, Bu=butyl, t-Bu=tert-butyl, Ph=phenyl, Bn=benzyl, Ac=acetyl, Bz=benzoyl, etc.

[0021] The term "alkyl" refers to a straight or branched hydrocarbon radical containing from 1 to about 20 carbon atoms. For example, alkyl may contain 1 to 10 carbon atoms, or 1 to 6 carbon atoms. Иллюстративный алкил включает в себя неразветвленные алкильные группы, такие как метил, этил, пропил, бутил, пентил, гексил, гептил, октил, нонил, децил, ундецил, додецил, и т.п., и также включает в себя разветвленные изомеры неразветвленных алкильных групп, например без ограничения -СН(СН ₃ ) ₂ , СН(СН ₃ )(СН ₂ СН ₃ ), -СН(СН ₂ СН ₃ ) ₂ , -С(СН ₃ ) ₃ , -С(СН ₂ СН ₃ ) ₃ , -СН ₂ СН(СН ₃ ) ₂ , -СН ₂ СН(СН ₃ )(СН ₂ СН ₃ ), -СН ₂ СН(СН ₂ СН ₃ )2, -СН ₂ С(СН ₃ ) ₃ , -СН ₂ С(СН ₂ СН ₃ ) ₃ , СН(СН ₃ )СН(СН ₃ )(СН ₂ СН ₃ ), -СН ₂ СН ₂ СН(СН ₃ ) ₂ , -СН ₂ СН ₂ СН(СН ₃ )(СН ₂ СН ₃ ), СН ₂ СН ₂ СН(СН ₂ СН ₃ ) ₂ , -СН ₂ СН ₂ С(СН ₃ ) ₃ , -СН ₂ СН ₂ С(СН ₂ СН ₃ ) ₃ , -СН(СН ₃ )СН ₂ СН(СН ₃ ) ₂ , -СН(СН ₃ )СН(СН ₃ )СН(СН ₃ ) ₂ , и т.п.. Таким образом, алкильные группы включают в себя первичные алкильные группы, вторичные алкильные группы и третичные алкильные группы. The alkyl group may be unsubstituted or optionally substituted with one or more substituents as described herein.

[0022] The term "alkoxy" or "alkoxy" refers to an -O-alkyl group containing the specified number of carbon atoms. For example, a (C ₁ -C ₆ )-alkoxy group includes -O-methyl, -O-ethyl, -O-propyl, -O-isopropyl, -O-butyl, -O-sec-butyl, -O-tert-butyl, -O-pentyl, -O-isopentyl, -O-neopentyl, -O-hexyl, -O-isohexyl, and -O-neohexyl.

[0023] Unless otherwise indicated, the terms "halogen" or "halogen" or "halogen", alone or as part of another substituent, means a fluorine, chlorine, bromine or iodine atom, preferably fluorine, chlorine or bromine.

[0024] A "haloalkyl" group includes monohaloalkyl groups, polyhaloalkyl groups where all halogen atoms may be the same or different, and perhaloalkyl groups where all hydrogen atoms are replaced by the same or different halogen atoms, such as fluorine and/or chlorine atoms. Examples of haloalkyl include trifluoromethyl, 1,1-dichloroethyl, 1,2-dichloroethyl, 1,3-dibromo-3,3-difluoropropyl, perfluorobutyl, and the like.

[0025] Aryl groups are cyclic aromatic hydrocarbons that do not contain heteroatoms in the ring. As is well known in the art, an aromatic compound is a multiple unsaturated ring system that contains 4n+2π electrons, where n is an integer. Thus, aryl groups include, without limitation, phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chryzenyl, biphenylenyl, anthracenyl, and naphthyl groups. In some embodiments, aryl groups contain from about 6 to about 14 carbon atoms in the ring portions of the groups. Aryl groups may be unsubstituted or substituted as defined above. Representative substituted aryl groups may be monosubstituted or may be substituted more than once, such as, without limitation, 2-, 3-, 4-, 5-, or 6-substituted phenyl or 2-8-substituted naphthyl groups, which may be substituted with carbon or non-carbon groups, such as those listed above.

[0026] Heterocyclyl groups or the term "heterocyclyl" includes aromatic and non-aromatic ring compounds containing 3 or more ring atoms, of which one or more ring atoms is a heteroatom, such as, but not limited to, N, O, and S. Thus, heterocyclyl may be cycloheteroalkyl or heteroaryl, or any combination thereof, provided that it is polycyclic. In some embodiments, heterocyclyl groups contain from 3 to about 20 ring atoms, while other similar groups contain from 3 to about 15 ring atoms. The heterocyclyl group, referred to as C ₂ -heterocyclyl, may be a 5-ring with 2 carbon atoms and three heteroatoms, a 6-ring with 2 carbon atoms and four heteroatoms, and the like. By analogy, C ₄ -heterocyclyl can be a 5-ring with one heteroatom, a 6-ring with two heteroatoms, and the like. The number of carbon atoms and the number of heteroatoms add up to the total number of ring atoms. Ring size can also be expressed by the total number of atoms in the ring, for example, a 3-10 membered heterocyclyl group containing both carbon and non-carbon ring atoms. The heterocyclyl ring may also contain one or more double bonds. The heteroaryl ring is an embodiment of the heterocyclyl group. The term "heterocyclyl group" includes groups of fused rings, including groups containing fused aromatic non-aromatic groups. For example, the dioxalanyl ring system and the benzdioxalanyl ring system (methylenedioxyphenyl ring system) are both heterocyclyl groups as used herein. The term also includes polycyclic, eg, bicyclic or tricyclic ring systems containing one or more heteroatoms, such as, but not limited to, quinuclidinyl. The heterocyclyl groups may be unsubstituted or may be substituted.

[0027] Heteroaryl groups are heterocyclic aromatic cyclic compounds containing 5 or more ring atoms, of which one or more atoms is a heteroatom, such as, but not limited to, N, O, and S; for example, heteroaryl rings may contain from 5 to about 8-12 ring atoms. The heteroaryl group is a variety of heterocyclyl groups that have an aromatic electronic structure that is a multiple substituted ring system that contains 4n+2π electrons, where n is an integer. A heteroaryl group, referred to as C ₂ -heteroaryl, may be a 5-ring (i.e., 5-membered ring) with 2 carbon atoms and three heteroatoms, a 6-ring (i.e., 6-membered ring) with 2 carbon atoms and four heteroatoms, and the like. By analogy, C ₄ -heteroaryl can be a 5-ring with one heteroatom, a 6-ring with two heteroatoms, and the like. The number of carbon atoms and the number of heteroatoms add up to the total number of ring atoms. Heteroaryl is meant to include oxidized S or N atoms such as sulfinyl, sulfonyl and N-oxide of the tertiary ring nitrogen. The carbon atom or heteroatom is the point of attachment of the heteroaryl ring structure, resulting in a stable compound. Examples of heteroaryl groups include, without limitation, pyridinyl, pyridazinyl, pyrazinyl, quinaoxalinyl, indolizinyl, benzo[b]thienyl, quinazolinyl, purinyl, indolyl, quinolinyl, pyrimidinyl, pyrrolyl, pyrazolyl, oxazolyl, thiazolyl, thienyl, isoxazolyl, oxathiadiazolyl, isothiazole yl, tetrazolyl, imidazolyl, triazolyl, furanyl, benzofuryl and indolyl. The heteroaryl group may be unsubstituted or may be substituted with one or more substituents as described herein.

[0028] By analogy, other aryl (e.g., phenyl) and heteroaryl (e.g., pyridyl) ring systems described herein can be depicted with either clearly labeled double bonds or "cyclic" aryl nomenclature, but the meanings are the same.

[0029] Cycloalkyl groups are groups containing one or more carbocyclic rings, including, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In some embodiments, a cycloalkyl group may contain from 3 to about 8-12 ring atoms, while in other embodiments, the number of ring carbon atoms ranges from 3 to 4, 5, 6, or 7. Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbornyl, adamantyl, bornyl, campenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to decalinyl, etc. Cycloalkyl groups also include rings that are substituted with straight and branched alkyl groups as defined above.

[0030] Cycloalkenyl groups include cycloalkyl groups containing at least one double bond between 2 carbon atoms. Thus, for example, cycloalkenyl groups include, without limitation, cyclohexenyl, cyclopentenyl, and cyclohexadienyl groups. Cycloalkenyl groups may contain from 3 to about 8-12 ring atoms, while in other embodiments, the number of ring carbon atoms ranges from 3 to 5, 6, or 7. Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to, decalinyl, and so on. .p., provided that they contain at least one double bond in the ring. Cycloalkenyl groups also include rings that are substituted with straight and branched alkyl groups as defined above.

[0031] One or more additional substituents on any group described herein are independently selected from the group consisting of R ^A , OR ^A , halo, -N=NR ^A , NR ^A R ^B , -(C ₁ -C ₆ -alkyl)NR ^A R ^B , -C(O)OR ^A , -C(O)NR ^A R ^B , -OC(O)R ^A and -CN. R ^A и R ^B независимо выбраны из группы, состоящей из Н, -CN, -гидрокси, оксо, C ₁ -С ₆ -алкила, C ₁ -С ₆ -алкокси, С ₂ -С ₆ -алкенила, С ₂ -С ₆ -алкилнила, NH ₂ , -S(O) _0-2 -(С ₁ -С ₆ -алкил), -S(O) _0-2 -(С ₆ -С ₁₀ -арил), -С(O)(С ₁ -С ₆ -алкил), -С(O)(С ₃ -С ₁₄ -карбоциклил), -С ₃ -С ₁₄ -карбоциклила, -(С ₁ -С ₆ -алкил)(С ₃ -С ₁₄ -карбоциклил), С ₆ -С ₁₀ -арила, 3-14-членного гетероциклоалкила и -(С ₁ -С ₆ -алкил)-(3-14-членный гетероциклоалкил) (где 1-4 атомов гетероциклоалкила независимо выбраны из N, О и S), и 5-10-членного гетероарила (где 1-4 атома гетероарила независимо выбраны из N, О и S). Каждый алкильный, алкокси, алкенильный, алкинильный, арильный, карбоциклильный, гетероциклоалкильный и гетероарильный фрагмент R ^A и R ^B необязательно замещен одним или несколькими заместителями, выбранными из группы, состоящей из гидрокси, галогена, -NR' ₂ (где каждый R' независимо выбран из группы, состоящей из C ₁ -С ₆ -алкила, С ₂ -С ₆ -алкенила, С ₂ -С ₆ -алкинила, С ₆ -С ₁₀ -арила, 3-14-членного гетероциклоалкила и -(С ₁ -С ₆ -алкил)-(3-14-членный гетероциклоалкил) (где 1-4 атомов кольца независимо выбраны из N, О и S), и 5-10-членного гетероарила (где 1-4 атомов гетероарила независимо выбраны из N, О и S), -NHC(O)(ОС ₁ -С ₆ -алкил), -NO ₂ , -CN, оксо, -С(O)ОН, -С(O)O(С ₁ -С ₆ -алкил), -С ₁ -С ₆ -алкил(С ₁ -С ₆ -алкокси), -C(O)NH ₂ , C ₁ -C ₆ -алкил, -С(O)С ₁ -С ₆ -алкила, -OC ₁ -С ₆ -алкила, -Si(C ₁ -C ₆ -алкил) ₃ , -S(O) _0-2 -(С ₁ -С ₆ -алкил), С ₆ -С ₁₀ -арила, -(С ₁ -С ₆ -алкил)(С ₆ -С ₁₀ -арил), 3-14-членного гетероциклоалкила и -(С ₁ -С ₆ -алкил)-(3-14-членный гетероцикл) (где 1-4 атомов гетероцикла независимо выбраны из N, О и S), и -O(С ₆ -С ₁₄ -арил). Each alkyl, alkenyl, aryl and heterocycloalkyl described above is optionally substituted with one or more substituents selected from the group consisting of hydroxy, -OC ₁ -C ₆ -alkyl, halogen, -NH ₂ , -(C ₁ -C ₆ -alkyl)NH ₂ , -C(O)OH, CN and oxo.

[0032] The compounds described herein may exist in various isomeric forms, including configurational, geometric, and conformational isomers, including, for example, cis or trans conformations. These compounds may also exist in one or more tautomeric forms, including both individual tautomers and mixtures of tautomers. The term "isomer" is intended to encompass all isomeric forms of a compound of the present disclosure, including tautomeric forms of said compound. Compounds according to the present disclosure may exist in open and cyclized forms. In some cases, one or more cyclized forms may be the result of water loss. The specific composition of open and cyclized forms may depend on how the compound was isolated, stored and administered. For example, a compound may exist predominantly in an open form under acidic conditions, but cyclize under neutral conditions. All forms are included in this disclosure.

[0033] The -CO ₂ H substituent can be replaced with bioisosteric substitutions such as:

and the like, where R has the same definition as R ^A defined herein (see, for example, The Practice of Medicinal Chemistry (Academic Press: New York, 1996), p. 203).

[0034] Some of the compounds described herein may contain asymmetric centers, and therefore exist in various enantiomeric and diastereomeric forms. The compound described herein may be in the form of an optical isomer or a diastereoisomer. Accordingly, the present disclosure encompasses the compounds and their uses described herein in the form of their optical isomers, diastereoisomers, and mixtures thereof, including a racemic mixture. Optical isomers of the compounds of the present disclosure can be obtained by known techniques such as asymmetric synthesis, chiral chromatography, pseudo-moving bed technology, or by chemical separation of stereoisomers using optically active resolving agents.

[0035] Unless otherwise indicated, the term "stereoisomer" means one stereoisomer of a compound that is substantially free of other stereoisomers of that compound. Thus, a stereoisomerically pure compound containing one chiral center will be substantially free of the opposite enantiomer of that compound. A stereoisomerically pure compound containing two chiral centers will be substantially free of other diastereoisomers of that compound. A typical stereoisomeric pure compound contains more than about 80 wt. % of one stereoisomer a of the compound and less than about 20 wt. % other stereoisomers of the compound, for example, more than about 90 wt. % of one stereoisomer of the compound and less than about 10 wt. % other stereoisomers of the compound, or more than about 95 wt. % of one stereoisomer of the compound and less than about 5 wt. % other stereoisomers of the compound, or more than about 97 wt. % of one stereoisomer of the compound and less than about 3 wt. % other stereoisomers of the compound, or more than about 99 wt. % of one stereoisomer of the compound and less than about 1 wt. % other stereoisomers of the compound. The stereoisomer described above can be considered as a composition containing two stereoisomers which are present in the respective mass fractions described herein.

[0036] If there is a discrepancy between the depicted structure and the name given to the structure, then the depicted structure shall prevail. In addition, if the stereochemistry of a structure or part of a structure is not indicated, for example by bold or dashed lines, then the structure or part of a structure should be interpreted as encompassing all of its stereoisomers. However, in some cases where there is more than one chiral center, the structures and names may be presented as separate enantiomers to help describe the relative stereochemistry. Specialists in the field of organic synthesis will be aware if the compounds are obtained in the form of individual enantiomers using the methods used to obtain them.

[0037] Unless otherwise indicated, as used herein, the term "compound" is inclusive in the sense that it encompasses a compound or a pharmaceutically acceptable salt, stereoisomer and/or tautomer thereof. Thus, for example, a compound of formula (I) or formula (II) includes a pharmaceutically acceptable salt of the tautomer of said compound.

[0038] Connections

[0039] According to various embodiments, the present disclosure relates to a compound of formula (I) or formula (II), or a pharmaceutically acceptable salt thereof:

[0040] X is S, -N=C(R ¹ )- or -C(R ¹ )=C(R ¹ )-.

[0041] Each R ¹ is independently H, F, Cl, C ₁ -C ₆ alkyl, ethenyl, or ethynyl (each of which may be substituted), cyano, alkoxy, haloalkyl, or C ₃ -C ₆ cycloalkyl.

[0042] In an embodiment, both R ¹ attached to the X-containing ring, together with the carbon atoms to which they are attached, form a fused phenyl.

[0043] R is H, alkyl, optionally substituted -((C ₁ -C ₆ alkyl)OC(O)OC ₁ -C ₆ alkyl), or benzyl, where benzyl may be unsubstituted or substituted with methoxy or an acid or ester isostere.

[0044] Ring A is a bicyclic fully aromatic or partially reduced heteroaryl ring system containing 3, 4, or 5 N atoms, substituted with 0, 1, 2, 3, or 4 substituents, each independently selected from the group consisting of NH ₂ , cyano, NHC(=O)O-tBu, OH, carboxamido, C ₁ -C ₆ -alkyl, -S(O) ₂ (C ₁ -C ₆ alkyl), -S(O)(C ₁ -C ₆ alkyl), alkylnitrile, alkoxy and haloalkyl. According to various embodiments, the partially reduced heteroaryl ring system may be substituted with an oxo group.

[0045] According to additional embodiments, the present disclosure relates to a compound of formula (I) wherein X is S, -N=C(R ¹ )-, or -C(R ¹ )=C(R ¹ )-. Each R ¹ is independently H, F, Cl, ethenyl or ethynyl (each of which may be substituted), cyano, alkoxy or haloalkyl. R is H, alkyl or benzyl, where the benzyl may be unsubstituted or substituted with methoxy or an acid or ester isostere. Ring A is a bicyclic fully aromatic or partially reduced heteroaryl ring system containing 3, 4, or 5 N atoms, substituted with 0, 1, 2, 3, or 4 substituents, each independently selected from the group consisting of NH ₂ , cyano, NHC(=O)O-tBu,OH, carboxamido, alkylnitrile, alkoxy, and haloalkyl; with the proviso that the partially reduced heteroaryl ring system may be substituted with an oxo group.

[0046] According to other embodiments, the present disclosure relates to a compound of formula (II) described above.

[0047] In various embodiments, optionally in combination with any other embodiment described herein, Ring A is unsubstituted or substituted as described herein and is a ring selected from the group consisting of:

where the wavy line indicates the position of the binding.

[0048] According to additional embodiments, the present disclosure relates to specific examples of compounds of formula (I) and formula (II), and their pharmaceutically acceptable salts, are presented below in Table 1. The compounds are presented with an indication of a score of activity, in part obtained from data from the ISG-LUC activation assay described herein, and physico-chemical characterization data.

[0049] Table 1: Specific compounds and scores. Scores are based on potency and efficacy data (+ = EC ₅₀ >20,000 nM; ++ = potent but less potent and effective than comparator (EC ₅₀ >1000 nM); +++ = comparable potency to comparator (EC ₅₀ < 3000 nM); ++++ = more potent and/or potent than comparator (EC ₅₀ < 900 nM)).

[0050] Related Documents:

[1] Corrales L, Glickman LH, McWhirter SM, Kanne DB, Sivick KE, Katibah GE, Woo SR, Lemmens E, Banda T, Leong JJ, Metchette K, Dubensky TW Jr, Gajewski TF. (2015) Direct Activation of STING in the Tumor Micro environment Leads to Potent and Systemic Tumor Regression and Immunity. CellRep.11: 1018-30.

[2] Deng, L. et al. (2014) STING-Dependent Cytosolic DNA Sensing Promotes Radiation-Induced Type I Interferon-Dependent Antitumor Immunity in Immunogenic Tumors, Immunity. 41:843.

[3] Corrales L, Matson V, Flood B, Spranger S, Gajewski TF. (2017) Innate immune signaling and regulation in cancer immunotherapy. Cell Res. 27:96-108.

[4] Corrales L, McWhirter SM, Dubensky TW Jr, Gajewski TF. (2016) The host STING pathway at the interface of cancer and immunity. J Clin Invest. 126:2404-11.

[0051] Methods of use

[0052] According to an embodiment, the present disclosure also relates to a method for stimulating the expression of interferon genes in a human patient. Said method comprises administering to a patient an effective dose of a compound or a pharmaceutically acceptable salt thereof as described herein.

[0053] According to another embodiment, the present disclosure relates to a method for treating a tumor in a patient. Said method comprises administering to a patient an effective dose of a compound or a pharmaceutically acceptable salt thereof.

[0054] In the context of combination therapies comprising administering a compound of the present disclosure and a drug targeting an immune response checkpoint, or combination therapies for potentiating ionizing radiation-based therapeutic approaches and existing chemotherapy therapies such as DNA damage-based chemotherapy, the STING agonists of the present disclosure may complement and potentiate the effects of these known therapeutic approaches. This builds on recent documents pointing to the key role of STING-dependent micronucleus-mediated tumor clearance using these approaches (see for example:

[5] Mackenzie, K.F., et al., (2017), cGAS surveillance of micronuclei links genome instability to innate immunity, Nature, 548, 461.

[6] Wang, W. et al., (2016), Effector T Cells Abrogate Stroma-Mediated Chemoresistance in Ovarian Cancer, Cell, 165, 1092-1105.

[7] Charlotte E. Ariyan, et al., January 16, 2018; DOI: 10.1158/2326-6066, Robust antitumor responses result from local chemotherapy and CTLA-4 blockade, cancerimmunolres.aacrjournals.org on January 31, 2018.

[8] Chung Kil Song, et al., www.moleculartherapy.org vol. 15 no. 8 aug. 2007, Chemotherapy Enhances CD8+ T Cell-mediated Antitumor Immunity Induced by Vaccination With Vaccinia Virus).

[0055] The compounds of the present disclosure may be used in therapeutic combinations with the administration of an effective dose of a drug that targets an immune response checkpoint. For example, a drug that targets an immune response checkpoint can be an anti-PD-L1 antibody, an anti-PD-1 antibody, an anti-CTLA-4 antibody, or an anti-4-1BB antibody (see, for example:

[9] Ager, CR, et al., (2017) Cancer Immunol Res; 5(8), 676.

[10] Fu, J. et al. (2015) Sci Transl Med. April 15, 2015; 7(283): 283ra52. doi: 10.1126/scitranslmed.aaa4306.

[11] Wang, H., et al. (2017) PNAS, February 14, 2017, vol. 114, no. 7, 1637-1642).

[0056] Pharmaceutical composition

[0057] According to another embodiment, the present disclosure relates to a pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt thereof described herein in combination with a pharmaceutically acceptable carrier or excipient.

[0058] Compositions of the present disclosure may be administered orally, topically, parenterally, by inhalation or spray, or rectally in unit dosage forms. In the context of this document, the term "parenteral" includes subcutaneous injection, intravenous, intramuscular, intrasternal injection or infusion techniques.

[0059] Suitable oral compositions described herein include, without limitation, tablets, lozenges, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion, hard or soft capsules, syrups, or elixirs.

[0060] Compositions according to the present disclosure, which are suitable for oral administration, can be prepared in accordance with any method known in the field of production of pharmaceutical compositions. For example, liquid dosage forms of the compounds of the present disclosure contain one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents, and preservatives for the purpose of providing pharmaceutically acceptable formulations of the compound or a pharmaceutically acceptable salt thereof.

[0061] For tablet formulations, the compound or a pharmaceutically acceptable salt thereof, in admixture with non-toxic pharmaceutically acceptable excipients, is used to manufacture tablets. Examples of such excipients include, without limitation, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, and sodium phosphate; granulating agents and disintegrators, for example corn starch or alginic acid; binders such as starch, gelatin or acacia; and lubricants such as magnesium stearate, stearic acid or talc. Tablets may be uncoated or may be coated by known coating techniques to delay disintegration and absorption in the gastrointestinal tract and therefore provide a delayed therapeutic effect for the desired period of time. For example, a time delay agent such as glyceryl monostearate or glyceryl distearate may be used.

[0062] Oral formulations may be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules, in which the active ingredient is mixed in water or an oily medium, such as peanut oil, liquid paraffin, or olive oil.

[0063] For aqueous suspensions, the compound, or a pharmaceutically acceptable salt thereof, is mixed with excipients suitable to maintain a stable suspension. Examples of such excipients include, but are not limited to, sodium carboxymethylcellulose, methylcellulose, hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth, and acacia gum.

[0064] Oral suspensions may also contain dispersants or humectants, such as a naturally occurring phosphatide, for example, lecithin, or alkylene oxide condensates with fatty acids, for example, polyoxyethylene stearate, and ethylene oxide condensates with long chain aliphatic alcohols, for example, heptadecaethyleneoxycetanol, or ethylene oxide condensates with partial esters derived from fatty acids and hexitol, such as polyoxyethylene sorbitan monooleate, or condensates of ethylene oxide with partial esters derived from fatty acids and hexitan anhydrides, such as polyethylene sorbitan monooleate. Aqueous suspensions may also contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate, one or more colorants, one or more flavors, and one or more sweeteners, such as sucrose or saccharin.

[0065] Oil suspensions can be formulated by suspending the compound or a pharmaceutically acceptable salt thereof in a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or a mineral oil such as liquid paraffin. Oily suspensions may contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol.

[0066] Sweeteners such as those presented above and flavoring agents may be added to provide palatable oral preparations. These compositions may be preserved by the addition of an antioxidant such as ascorbic acid.

[0067] Dispersible powders and granules suitable for the preparation of an aqueous suspension by the addition of water provide the compound or a pharmaceutically acceptable salt thereof in admixture with a dispersant or humectant, a suspending agent, and one or more preservatives. Suitable dispersion aids or humectants and suspension aids are illustrated by those already mentioned above. Additional excipients may also be present, such as sweeteners, flavors and colors.

[0068] The pharmaceutical compositions of the present disclosure may also be in the form of oil-in-water emulsions. The oil phase can be a vegetable oil, such as olive oil or peanut oil, or a mineral oil, such as liquid paraffin, or mixtures thereof. Suitable emulsifiers can be naturally occurring gums such as acacia or gum tragacanth, naturally occurring phosphatides such as soy, lecithin and esters or partial esters derived from fatty acids and hexitol, anhydrides such as sorbitan monooleate, and the condensation reaction products of said partial esters with ethylene oxide such as polyoxyethylene sorbitan. -monoleate. The emulsions may also contain sweetening and flavoring agents.

[0069] Syrups and elixirs may be formulated with sweeteners such as glycerol, propylene glycol, sorbitol, or sucrose. Such formulations may also contain a soothing agent, preservatives and flavors, and colorants. The pharmaceutical compositions may be in the form of a sterile injection, an aqueous suspension, or an oily suspension. Such a suspension may be formulated in accordance with the prior art using suitable dispersion aids or the humectants and suspension aids mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be used are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fatty oils have traditionally been used as a solvent or suspension medium. For this purpose, any light fixed oil can be used, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find their use in the preparation of injectables.

[0070] The compound or a pharmaceutically acceptable salt thereof may also be administered in the form of suppositories for rectal administration. Such compositions can be prepared by mixing the compound with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at rectal temperature and will therefore liquefy in the rectum to release the compound. Illustrative excipients include cocoa butter and polyethylene glycols.

[0071] Compositions for parenteral administration are administered in a sterile environment. Depending on the carrier medium used and the concentration of the compound or its pharmaceutically acceptable salt in the formulation, the parenteral formulation may be in the form of a suspension or a solution containing the dissolved compound. Adjuvants such as local anesthetics, preservatives and buffering agents may also be added to parenteral compositions.

[0072] Examples

[0073] Tissue culture. Unless otherwise noted, wild-type THP-l-Lucia ISG (cat. no. thpl-isg) and STING KO (cat. no. thpd-kostg) cells were purchased from Invivogen and maintained in a culture medium consisting of RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum (FBS), 1000 U/ml penicillin, 1000 µg/ml streptomycin, 0.25 µg/ml amphotericin B and 100 µg/ml zeocin.

[0074] Substances that stimulate type 1 interferon. Poly(dA:dT) and 2'3'-cGAMP were purchased from Invivogen and resuspended according to the manufacturer's instructions.

[0075] ISRE-luciferase assay. THP-1 Lucia ISG cells were resuspended in low serum nutrient medium (2% FBS) at a density of 5×10 ⁵ cells per ml and treated with test compound or vehicle (DMSO). In each well of Greiner 384-well plates, 50 μl of cells were seeded and incubated for 24 hours. To evaluate luciferase reporter expression, 30 µl of Quanti-luc detection reagent (Invivogen) was added to each well and luminescence was read using an Envision plate reader (Perkin Elmer) with an integration time of 0.1 second.

[0076] Viability analysis. Cells were resuspended in low serum nutrient medium at a density of 5×10 ⁵ cells per ml and treated with test compound or vehicle (DMSO). In each well of Greiner 384-well plates, 50 μl of cells were seeded and incubated for 24 hours. To evaluate luciferase reporter expression, 30 μl of CellTiter-Glo detection reagent (Promega) was added to each well, and luminescence was determined using an Envision plate reader set to an integration time of 0.1 second.

[0077] Western blot. Cells were solubilized in 1× proteolytic buffer (25 mM HEPES, pH 7.4, 300 mM NaCl, 1.5 mM MgCl ₂ , 1 mM EGTA, 1% P-40, 1% sodium deoxycholate, 2.5 mM sodium pyrophosphate, 1 mM glycerophosphate) with freshly added protease inhibitors and phosphatase (Cell Signaling). Western blotting was performed using Bolt™ 4-12% Bis-Tris gels and Bolt™ Mini Transfer System following the manufacturer's instructions (ThermoFisher Scientific). Anti-STING and γ-tubulin antibodies were purchased from Cell Signaling, diluted with 5% BSA 1× TBS-T buffer (Table 3). Anti-rabbit HRP antibody was diluted containing 5% skimmed milk powder 1x with TBS-T buffer and the luminescence signal was visualized using ChemiDoc Imager (BioRad).

[0078] Real-time semi-quantitative PCR (qPCR). THP-1 cells were resuspended in low serum nutrient medium at a density of 5×10 ⁵ cells per ml and treated with test compound or vehicle (DMSO). 2.5 ml of cells were seeded in each well of a 6-well plate and incubated for 24 hours. RNA was isolated using the RNeasy Plus Mini kit (Qiagen), and 1 μg of purified RNA was reverse transcribed to obtain cDNA (VILO, cat. no. 11755050, ThermoFisher Scientific). Gene expression was assessed using the Taqman primers and probes listed in Table 4 on Taqman Universal Mix II (Cat. #4440038, ThermoFisher) following the manufacturer's instructions. Gene expression was normalized using the delta-delta-Ct method and reported as fold change in expression.

[0079] STING Thermal Shift Analysis (TSA). Human and mouse STING c-terminal domains (CTDs) were expressed and purified as previously described in detail (Ouyang, S., Song, X., Wang, Y., Ru, FL, Shaw, N., Jiang, Y., Niu, F., Zhu, Y., Qiu, W., Parvatiyar, K., et al. (2012). Structural analysis of the STING adaptor protein reveals a hydrophobic dimer interface and mode of cyclic di-GMP binding Immunity 36, 1073-1086). To STING protein (0.22 mg/mL) diluted 1× with Protein Thermal Shift Buffer provided in Protein Thermal Shift Dye Kit (p/n 4461146, ThermoFisher Scientific), test compound or vehicle control was added. Thermal Shift dye was added and a melt curve was recorded following the parameters described for the dye kit. Melting points (Tm) were calculated using the derivatives method using Protein Thermal Shift v1.3 software (Cat. No. 4466038, ThermoFisher Scientific).

[0080] Wild-type STING Binding Assay (Cisbio Cat #64BDSTGPEH). The assay method format was optimized to demonstrate the binding of recombinant 6x His-labeled human STING protein labeled with terbium cryptate to the natural 2'3'cGAMP ligand labeled with d2 (acceptor). After the two dyes approach, excitation of the donor with a flash lamp in the PHERAstar FSX plate reader initiates fluorescence resonance energy transfer (FRET) to the acceptor, which in turn fluoresces at 665 nm. To evaluate the ability of synthetic small molecule STING ligands to bind to human STING, assays were performed in a competitive format. Titration samples of 10 points for each of the synthetic ligands in 5 µl were transferred to a 384-well plate, followed by the addition of 20 µl of assay buffer containing 6x His-labeled human STING protein and labeled 2'3'cGAMP ligand, and incubated for 3 hours at room temperature. PHERAstar baseline values were used to calculate derived IC ₅₀ values (signal inversely proportional to synthetic ligand binding) by curve fitting in Genedata. Percent inhibition was calculated from the ratio of the maximum amount of synthetic compound binding to the maximum binding of unlabeled 2'3' cGAMP, which was used as a control in each assay.

[0081]

[0082]

[0083] Compounds useful in carrying out the method of the present disclosure may be prepared according to the following procedures, in combination with average knowledge and experience in organic synthesis, substituting appropriate reagents apparent to the practitioner.

[0084] Experimental Techniques

[0085] Abbreviations. Использовали следующие сокращения: тетрагидрофуран (THF), дихлорметан (DCM), N,N-диметилформамид (DMF), диметилацетамид (DMA), диметилсульфоксид (DMSO), трифторуксусная кислота (TFA), триэтиламин (TEA), диизопропилэтиламин (DIPE А), (1-циано-2-этокси-2-оксоэтилиденаминоокси)диметил-аминоморфолинокарбения гексафторфосфат (COMU), 1-[бис(диметиламино)метилен]- ¹ Н-1,2,3-триазоло[4,5-b]пиридиний-3-оксида гексафторфосфат, N-[(диметиламино)- ¹ Н-1,2,3-триазоло[4,5-b]пиридин-1-илметилен]-N-метилметанаминия гексафторфосфата N-оксид (HATU).

[0086] General examples for obtaining compounds according to the present disclosure. Starting materials and intermediates for the preparation of compounds of the present disclosure may be prepared by applying or adapting the methods described below, their apparent chemical equivalents, or, for example, as described in literature such as The Science of Synthesis, Volumes 1-8. Editors E.M. Carreira et al. Thieme publishers (2001-2008). Details regarding additional reagents and conditions are also available from structure and reaction searches using commercial computer search engines such as Scifinder (www.cas.org) or Reaxys (www.reaxys.com).

[0087] Part I: Preparation of intermediates

[0088] Scheme 1: Synthesis of Intermediate A:

[0089] Step 1: Synthesis of 3,6-dichloro-N-(4-methoxybenzyl)pyridazin-4-amine: To a solution of 3,4,6-trichloropyridazine (45.0 g, 245 mmol) in THF (450 ml) was added (4-methoxyphenyl)methanamine (101 g, 736 mmol) and then the mixture was stirred at 50°C for 0 .5 hours. The mixture was diluted with water (1000 ml) and extracted with EtOAc (1000 ml x 3). The combined organic layers were washed with brine (300 ml×2), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by column chromatography to give 3,6-dichloro-N-(4-methoxybenzyl)pyridazine-4-amine (70.0 g, 97% purity, 97% yield) as a white solid. LCMS (ESI): m/z 283.9 [M+H] ⁺ .

[0090] Step 2: Synthesis of tert-butyl-(3,6-dichloropyridazin-4-yl)(4-methoxybenzyl)-carbamate: Et ₃ N (49.9 g, 492 mmol) was added to a solution of 3,6-dichloro-N-(4-methoxybenzyl)pyridazin-4-amine (70.0 g, 246 mmol) in THF (700 ml) ), DMAP (15.0 g, 123 mmol) and Boc ₂ O (79.2 ml, 345 mmol). After stirring at 25° C. for 0.5 hour, the mixture was diluted with water (1000 ml) and then extracted with EtOAc (1000 ml x 3). The combined organic layers were washed with brine (500 ml×2), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography to give tert-butyl-(3,6-dichloropyridazin-4-yl)(4-methoxybenzyl)carbamate (77 g, 67% yield) as a white solid. LCMS (ESI): m/z 384.1 [M+H] ⁺ .

[0091] Step 3: Synthesis of methyl 5-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-chloropyridazine-3-carboxylate: Et ₃ N (39.7 ml, 285 mmol) and Pd(PPh ₃ ) ₂ Cl (6.95 g, 9.50 mmol). The mixture was stirred at 40°C for 4 hours under pressure of carbon monoxide (50 psi). The mixture was diluted with water (500 ml) and extracted with EtOAc (500 ml x 3). The combined organic layers were washed with brine (300 ml×2), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography to give methyl 5-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-chloropyridazine-3-carboxylate (22.9 g, 47% yield) as a white solid. LCMS (ESI): m/z 408.1 [M+H] ⁺ .

[0092] Step 4: Synthesis of methyl 5-((tert-butoxycarbonyl)amino)-6-chloropyridazine-3-carboxylate: To a solution of 5-((tert-butoxycarbonyl)(4-methoxybenzyl)amino)-6-chloropyridazine-3-carboxylate (15.0 g, 36.7 mmol) in acetonitrile (75.0 ml) and water ( 75.0 ml) CAN (40.3 g, 73.6 mmol) was added at 0° C. and then the mixture was stirred for 0.5 hour. The mixture was diluted with water (200 ml) and extracted with EtOAc (100 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography to give methyl 5-((tert-butoxycarbonyl)amino)-6-chloropyridazine-3-carboxylate (7.20 g, 65% yield) as a white solid. LCMS (ESI): m/z 288.0 [M+H] ⁺ .

[0093] Step 5: Synthesis of methyl 8-((tert-butoxycarbonyl)amino)tetrazolo[1,5-b]pyridazine-6-carboxylate: Sodium azide was added to a solution of 5-((tert-butoxycarbonyl)amino)-6-chloro-pyridazine-3-carboxylate (4.30 g, 13.6 mmol) in DMSO (43.0 ml) (2.65 g, 40.8 mmol). After stirring at 50° C. for 2 hours, the mixture was diluted with water (200 ml) and extracted with EtOAc (200 ml x 3). The combined organic layers were washed with water (200 ml×3) and brine (200 ml×2), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography to give methyl 8-((tert-butoxycarbonyl)amino)tetrazolo[1,5-b]pyridazine-6-carboxylate (2.80 g, 66% yield) as a white solid. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 11.44 (s, 1H), 8.51 (s, 1H), 4.00 (s, 3H), 1.53 (s, 9H). LCMS (ESI): m/z 294.7 [M+H] ⁺ .

[0094] Step b: Synthesis of 8-((tert-butoxycarbonyl)amino)tetrazolo[1,5-b]pyridazine-6-carboxylic acid (A): To a solution of methyl 8-((tert-6utoxycarbonyl)amino)-tetrazolo[1,5-b]pyridazine-6-carboxylate (250 mg, 0.85 mmol) in THF (5 ml) at 0 °C, a solution of lithium hydroxide monohydrate (143 mg, 3.4 mmol) in water (5 ml) was added. The mixture was stirred at 25°C for 4 hours. After completion of the reaction, 3M HCl was added to neutralize the reaction mixture, the precipitate was filtered and washed with cold water (1 ml×2) to give Compound A (191 mg, 80% yield) as a white solid. ¹ H-NMR (400 MHz, DMSO) δ 8.60 (s, 1H), 8.32 (s, 1H), 8.22 (s, 1H), 1.58 (s, 9H).

[0095] Scheme 2: Synthesis of Intermediate B:

[0096] Step 1: Synthesis of methyl 6-chloro-4-((4-methoxybenzyl)amino)pyridazine-3-carboxylate: To a solution of methyl 4,6-dichloropyridazine-3-carboxylate (207 mg, 1 mmol) and Et ₃ N (279 μl, 2 mmol) in DMSO (2 ml) was added 4-methoxy benzylamine (1 44 μl, 1.1 mmol), and the reaction mixture was stirred at room temperature overnight. After completion of the reaction, water (10 ml) was added and a white precipitate formed. The mixture was filtered and washed with water to give methyl 6-chloro-4-((4-methoxybenzyl)amino)-pyridazine-3-carboxylate (257 mg, 84% yield) as a white solid. LCMS (ESI): m/z 308.0 [M+H] ⁺ .

[0097] Step 2: Synthesis of 7-((4-methoxybenzyl)amino)tetrazolo[1,5-b]pyridazine-6-carboxylic acid: To a solution of methyl 6-chloro-4-((4-methoxybenzyl)amino)pyridazine-3-carboxylate (257 mg, 0.84 mmol) in DMSO (3 mL) was added sodium azide (109 mg, 1. 68 mmol) and stirred at 100° C. for 16 hours. After completion of the reaction, water (5 ml) was added and a yellow precipitate formed. The mixture was maintained at 0°C for 10 minutes, filtered and washed with water (2 ml × 2) to obtain a yellow solid. The solid was suspended in water (10 ml) and neutralized by adding HCl (3M). The suspension was filtered to give Compound B (183 mg, 13% yield) as a yellow solid. ¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 8.43 (s, 1H), 7.42-7.32 (m, 2H), 7.22 (s, 1H), 6.96-6.88 (m, 2H), 4.48 (d, J=5.0 Hz, 2H), 3.73 (s, 3H). LCMS (ESI): m/z 300.8 [M+H] ⁺ .

[0098] Scheme 3: Synthesis of Intermediate C:

[0099] Step 1: Synthesis of ethylimidazo[1,2-a]pyrimidine-7-carboxylate: To a suspension of ¹ H-imidazole-2-amine hemisulfate (8.5 g, 64.33 mmol) in EtOH (100 mL) was added sodium methoxide (4.4 g, 82.2 mmol) and the resulting mixture was stirred at 90°C for 30 min. . Then, it was brought to room temperature and a solution of methyl-(E)-4-ethoxy-2-oxobut-3-enoate (10 g, 63.29 mmol) in EtOH (40 ml) was added. The reaction mixture was slowly heated until a condensate appeared and stirred for 16 hours. After completion of the reaction, the reaction mixture was cooled to room temperature and the volatiles were removed under reduced pressure. The residue was purified by column chromatography (0-4% MeOH in DCM) to give ethylimidazo[1,2-a]pyrimidine-7-carboxylate (5.2 g, 43% yield) as an off-white solid. ¹ H-NMR (400 MHz, CDCl ₃ ): δ 8.64-8.61 (m, 1H), 8.06-8.04 (m, 1H), 7.74-7.67 (m, 2H), 4.51 (q, J=7.2 Hz, 2H), 1.47 (t, J=7.2 Hz, 3H). LCMS(ESI): m/z 192.1 [M+H] ⁺ .

[0100] Step 2: Synthesis of ethyl 3-bromimidazo[1,2-a]pyrimidine-7-carboxylate (C): To a suspension of ethylimidazo[1,2-a]pyrimidine-7-carboxylate (5.2 g, 27.2 mmol) in MeOH (52 ml) at 0° C. under nitrogen atmosphere was added KBr (3.24 g, 27.2 mmol) and sodium acetate (3.43 g, 40.8 mmol). Bromine (4.78 g, 29.9 mmol) was added to the mixture over 10-15 minutes. After completion of the reaction, the reaction mixture was quenched by adding 1M Na ₂ SO ₃ aqueous solution (10 MP). Then, the volatiles were removed under reduced pressure, and a saturated solution of NaHCO ₃ (30 ml) was added to the residue. The resulting solid was collected by filtration and purified by column chromatography (0-2%) MeOH in DCM) to give Compound C (5.3 g, 72%) as an off-white solid. ¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 8.97 (m, 1H), 8.18 (s, 1H), 7.71 (d, J=7.2 Hz, 1H), 4.42 (q, J=7.2 Hz, 2H), 3.95 (s, 3H), 1.38 (t, J=7.2 Hz, 3H). LCMS (ESI): m/z 270.2 [M+H] ⁺ .

[0101] Part II: Preparation of exemplary compounds

[0102] All compounds according to the present disclosure was obtained using the methods illustrated below.

[0103] Example 1

[0104] Scheme 4: Synthesis of Compound 1:

[0105] Step 1: Synthesis of methyl 2-(8-((tert-6utoxycaronyl)amino)tetrazolo[1,5-b]pyridazine-6-carboxamido)-4-ethynyl-5-fluorobenzoate: To a solution of compound A (27.8 mg, 0.1 mmol) was added HATU (48 mg, 0.15 mmol), methyl-2-amino-4-ethyne yl-5-fluorobenzoate (19.4 mg, 0.11 mmol) and DIPEA (35 μl, 0.2 mmol), and the reaction mixture was stirred at 50° C. overnight. The crude mixture was purified by preparative HPLC to give methyl 2-(8-((tert-butoxycarbonyl)amino)-tetrazolo[1,5-b]pyridazine-6-carboxamido)-4-ethynyl-5-fluorobenzoate as a white solid (28 mg, 51% yield). ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 12.47 (s, 1H), 11.54 (s, 1H), 8.80 (d, J=6.1 Hz, 1H), 8.67 (s, 1H), 7.93 (d, J=9.2 Hz, 1H), 4.90 (s, 1H), 3.97 (s, 3 H), 1.55 (s, 9H). LCMS (ESI): m/z 456.6 [M+H] ⁺ .

[0106] Step 2: Synthesis of lithium 2-(8-aminotetrazolo[1,5-b]pyridazine-6-carboxamido)-4-ethynyl-5-fluorobenzoate (1): 8 mg, 0.061 mmol) in CH ₂ Cl ₂ (0.8 ml) was added TFA (0.4 ml), and the reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction, the mixture was concentrated under reduced pressure. Then, the crude mixture was dissolved in THF (2 ml), an aqueous solution of lithium hydroxide (310 μl, 0.31 mmol, 1M) was added and stirred at 50°C for 6 hours. After completion of the reaction, THF was removed under reduced pressure, and the residue was suspended in water (1 ml). The suspension was filtered and washed with cold water (0.5 ml x 2) and acetonitrile (0.5 ml) to give compound 1 (lithium salt, 14.5 mg, 68% yield) as a white solid. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 15.71 (s, 1H), 8.82 (dd, J=7.0, 1.8 Hz, 1H), 8.33 (s, 2H), 7.76 (dd, J=10.4, 2.0 Hz, 1H), 7.15 (dd, J=2.2, 1.0 Hz, 1H), 4.54 (d, J=2.0 Hz, 1H). LCMS (ESI): m/z 342.46 [M+H] ⁺ .

[0107] The following compounds were synthesized using procedures similar to those described above for compound 1: 5, 6, 7, 8, 9, 10, 11, 14, 15, 16, 17, 18, 19, 21, 22, 24, 25, 26, 28, 29, 30, 31, 32, 33, 34, 35 , 36, 38, 39, 40,41,42,43, 45, 47, 50, 55, 56, 58, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 79, 80, 77, 78, 81, 82, 83, 84, 85 and 86 etc.

[0108] Example 2

[0109] Scheme 5: Synthesis of Compound 2:

[0110] Synthesis of 2-(8-aminotetrazolo[1,5-b]pyridazin-6-yl)-7-ethynyl-6-fluoro-4H-benzo[d][1,3]oxazin-4-one (2): ionyl chloride was removed under vacuum. The solid was dissolved in 1 ml anhydrous acetonitrile and a solution of DIPEA (20 μl, 0.12 mmol) in 1 ml anhydrous acetonitrile was added at room temperature. After stirring for 30 minutes, the resulting precipitate was isolated and washed with acetonitrile to give the product (15 mg, 77% yield). ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 8.49 (s, 2H), 8.11 (d, J=8.4 Hz, 1H), 8.03 (d, J=6.3 Hz, 1H), 7.35 (s, 1H), 5.04 (s, 1H). MS-ESI: m/z 324.49 observed (M+H) ⁺

[0111] Compounds 44, 48 and 49 were prepared using a procedure similar to that described for the preparation of compound 2.

[0112] Example 3

[0113] Scheme 6: Synthesis of Compound 3:

[0114] Step 1: Synthesis of 3-bromimidazo[1,2-a]pyrimidine-7-carboxylic acid: An aqueous solution of lithium hydroxide (0.327 g, 7.8 mmol, 2 ml water) was added to a solution of Compound C (1.4 g, 5.2 mmol) in THF (8 ml) at room temperature. The reaction mixture was stirred at the same temperature for 1 hour. After completion of the reaction, the solvent was removed under reduced pressure, and the aqueous layers were acidified by adding 2N HCl at 0°C. The resulting solid was collected by filtration, washed with cold water (3 mL×2) and dried to give 3-bromimidazo[1,2-a]pyrimidine-7-carboxylic acid (1.0 g, 79% yield) as an off-white solid. ¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 9.07 (d, J=6.8 Hz, 1H), 8.29 (s, 1H), 7.65 (d, J=6.8 Hz, 1H). LCMS (ESI): m/z 243.06 [M+H] ⁺ .

[0115] Step 2: Synthesis of methyl 2-(3-bromimidazo[1,2-a]pyrimidine-7-carboxamido)-4,5-dichlorobenzoate: DIPEA (3 .8 g, 29.2 mmol) and HATU (3.3 g, 8.7 mmol). The mixture was stirred for 10 minutes after which methyl 2-amino-4,5-dichlorobenzoate (1.9 g, 8.7 mmol) was added. The reaction mixture was stirred at 80°C for 5 hours After completion of the reaction, the reaction mixture was cooled to room temperature, and was added a saturated solution of NaHCO ₃ (30 ml). The resulting solid was collected by filtration and purified by column chromatography to give methyl 2-(3-bromimidazo[1,2-a]pyrimidine-7-carboxamido)-4,5-dichlorobenzoate (1.0 g, 39% yield) as an off-white solid. ¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 13.09 (s, 1H), 9.17 (s, 1H), 8.59 (d, J=6.8, 1H), 8.22 (s, 1H), 7.92 (d, J=6.8, 1H), 7.76 (s, 1H), 3.89 (s, 3H). LCMS (ESI): m/z 441.0 [M-H] ^- .

[0116] Step 3: Synthesis of methyl 4,5-dichloro-2-(3-cyanoimidazo[1,2-a]pyrimidine-7-carboxamido)benzoate 3: To a solution of methyl 2-(3-bromimidazo[1,2-a]pyrimidine-7-carboxamido)-4,5-dichlorobenzoate (0.220 g, 0.5 mmol) in DMF (4 mL) ) CuCN (0.157 g, 1.75 mmol) was added and the mixture was irradiated in a microwave reactor at 140° C. for 2 h (1 h + 1 h intervals). The reaction mixture was diluted with EtOAc (25 ml), washed with saturated NaHCO ₃ solution (10 ml), water (10 ml × 2), brine (10 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (0-30% EtOAc in PE) to give 0.11 g of crude product, which was further purified by preparative HPLC to give compound 3 (11 mg, 6% yield) as an off-white solid. ¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 12.79 (s, 1H), 9.28 (d, J=6.8 Hz, 1H), 9.04 (s, 1H), 8.95 (s, 1H), 8.21 (s, 1H), 7.89 (d, J=6.8 Hz, 1H), 4.00 (s, 3H ). LCMS (ESI): m/z 388.1 [M-H] ^- .

[0117] Compounds 13, 46, 52 and 75 were prepared using a procedure similar to that described for compound 3.

[0118] Example 4

[0119] Scheme 7: Synthesis of Compound 4:

[0120] Стадия 1: Синтез метил-3-(1-(тетрагидро-2Н-пиран-2-ил)- ¹ Н-пиразол-4-ил)имидазо[1,2-а]пиримидин-7-карбоксилата: Смесь 1-(тетрагидро-2Н-пиран-2-ил)-4-(4,4,5,5-тетраметил-1,3,2-диоксаборолан-2-ил)- ¹ Н-пиразола (413 мг, 1,48 ммоль, 1,5 экв), метил-3-йодимидазо[1,2-а]пиримидин-7-карбоксилат (0,3 г, 989 мкмоль, 1 экв), K ₃ PO ₄ (315 мг, 1,48 ммоль, 1,5 экв) и SPhos Pd G3 (154 мг, 197 мкмоль, 0,2 экв) в THF (3 мл) трижды дегазировали и продували N ₂ . The mixture was stirred at 60°C for 2 hours under N ₂ atmosphere. The crude reaction mixture was purified by reverse phase Combi-flash (with 0.1% FA added) to give methyl 3-(1-(tetrahydro-2H-pyran-2-yl) ^-1H -pyrazol-4-yl)imidazo[1,2-a]pyrimidine-7-carboxylate (120 mg, 366 µmol, 31% yield) as a yellow solid. LCMS (ESI): m/z 328.1 [M+H] ⁺ .

[0121] Step 2: Synthesis of 3-(1-(tetrahydro-2H-pyran-2-yl) ^-1H -pyrazol-4-yl)imidazo[1,2-a]pyrimidine-7-carboxylic acid: To a mixture of methyl-3-(1-(tetrahydro-2H-pyran-2-yl) ^-1H -pyrazol-4-yl)imidazo[1,2-a]pyrimidine-7- carboxylate (100 mg, 305 µmol, 1 eq) in THF (0.5 ml) and H ₂ O (0.5 ml) LiOH⋅H ₂ O (12.8 mg, 305 µmol, 1 eq) was added at 20°C. The mixture was stirred at 20°C for 0.5 hour and then concentrated to give 3-(1-(tetrahydro-2H-pyran-2-yl) ^-1H -pyrazol-4-yl)imidazo[1,2-a]pyrimidine-7-carboxylic acid (85 mg, crude) as a yellow solid. LCMS (ESI): m/z 314.1 [M+H] ⁺ .

[0122] Стадия 3: Синтез метил-5-фтор-2-(3-(1-(тетрагидро-2Н-пиран-2-ил)- ¹ Н-пиразол-4-ил)имидазо[1,2-а]пиримидин-7-карбоксамидо)-4-((триметилсилил)этанил)бензоата: К смеси 3-(1-(тетрагидро-2Н-пиран-2-ил)- ¹ Н-пиразол-4-ил)имидазо[1,2-а]пиримидин-7-карбоновой кислоты (50 мг, 159 мкмоль, 1 экв) и метил-2-амино-5-фтор-4-((триметилсилил)этанил)бензоата (42,3 мг, 159 мкмоль, 1 экв) в пиридине (0,5 мл) при 15°С добавляли POCl ₃ (73,4 мг, 478 мкмоль, 44,4 мкл, 3 экв). The mixture was stirred at 15°C for 30 min, and then diluted with H ₂ O (20 ml) and was extracted with EA (10 ml × 3). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC to afford methyl 5-fluoro-2-(3-(1-(tetrahydro-2H-pyran-2-yl) ^-1H -pyrazol-4-yl)imidazo[1,2-a]pyrimidine-7-carboxamido)-4-((trimethylsilyl)ethanyl)-benzoate (75 mg, 133 µmol, 74% yield after two steps) as yellow solid. ¹ H-ЯМР (400 МГц, DMSO-d ₆ ) δ=12,63 (с, 1H), 9,21 (д, J=7,2 Гц, 1Н), 8,91 (д, J=6,8 Гц, 1H), 8,61 (с, 1H), 8,28 (с, 1H), 8,13 (с, 1H), 7,88 (д, J=10,0 Гц, 1H), 7,75 (д, J=7,2 Гц, 1H), 5,51 (дд, J=2,0, 10,0 Гц, 1H), 4,05-3,95 (м, 4Н), 3,70 - 3,65 (м, 1H), 2,24 - 2,12 (м, 1Н), 2,03 - 1,93 (м, 2Н), 1,78 - 1,66 (м, 1Н), 1,62 - 1,53 (м, 2Н), 0,31 - 0,27 (м, 9Н).

[0123] Step 4: Synthesis of methyl 2-(3-( ^1H -pyrazol-4-yl)imidazo[1,2-a]pyrimidine-7-carboxamido)-5-fluoro-4-((trimethylsilyl)ethanyl)benzoate: To a mixture of methyl 5-fluoro-2-(3-(1-(tetrahydro-2H-pyran-2-yl) ^-1H -pyrazol-4-yl)im dazo[1,2-a]pyrimidine-7-carboxamido)-4-((trimethylsilyl)ethanyl)benzoate (15 mg, 26.7 µmol, 1 eq) in DCM (1 ml) at 15° C. TFA (462 mg, 4.05 mmol, 0.3 ml, 151 eq) was added in one portion. The mixture was stirred at 15° C. for 15 minutes and then concentrated to give a residue. The residue was purified by preparative HPLC (column: Shim-pack C18 150×25×10 μm; mobile phase: [water (0.225% FA)-ACN]; V%: 62%-82%, 10 min) to give methyl-2-(3-( ^1H -pyrazol-4-yl)imidazo[1,2-a]pyrimidine-7-carboxamido)-5- fluoro-4-((trimethylsilyl)ethanyl)benzoate (8 mg, 16.7 µmol, 62% yield) as a yellow solid. LCMS (ESI): m/z 411.0 [M+H] ⁺ .

[0124] Step 5: Synthesis of 2-(3-( ^1H -pyrazol-4-yl)imidazo[1,2-a]pyrimidine-7-carboxamido)-4-ethynyl-5-fluorobenzoic acid (4): To a mixture of methyl 2-(3-( ^1H -pyrazol-4-yl)imidazo[1,2-a]pyrimidine-7-carboxamido)-5-fluoro-4-( (trimethylsilyl)ethanyl)benzoate (15 mg, 31.4 µmol, 1 eq) in THF (0.1 ml) and H ₂ O (0.1 ml) at 15° C. LiOH⋅H ₂ O (1.45 mg, 34.62 µmol, 1.1 eq) was added in one portion. The mixture was stirred at 15° C. for 30 minutes. The mixture was quenched to pH=6 with 1N HCl and then concentrated in vacuo to remove. The precipitate was filtered and the filter cake dried under vacuum to give 2-(3-( ^1H -pyrazol-4-yl)imidazo[1,2-a]pyrimidine-7-carboxamido)-4-ethynyl-5-fluorobenzoic acid (4.28 mg, 10.9 µmol, 34% yield, 100% purity) as a yellow solid. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ=12.95 (s, 1H), 9.21-9.19 (d, J=7.2 Hz, 1H), 9.01-8.99 (d, J=6.4 Hz, 1H), 8.36-8.33 (m, 3H), 7.90-7.87 (d, J=10.0 Hz) , 1H), 7.77-7.75 (d, J=7.2 Hz, 1H), 4.83 (s, 1H). LCMS (ESI): m/z 391.3 [M+H] ⁺ .

[0125] Compounds 51, 59 and 61 were prepared using a procedure similar to that described for the preparation of compound 4.

[0126] While the present disclosure has been described and exemplified in greater detail for those skilled in the art to make and use, various alternatives, modifications, and improvements will be apparent to those skilled in the art without departing from the spirit and scope of the claims.

[0127] All patents and publications referenced herein are herein incorporated by reference equally as if each individual publication were specifically and individually identified as being incorporated herein by reference in its entirety.

Claims (32)

1. Compound of formula (I) (I) Where X is S, -N=C(R ¹ )- or -C(R ¹ )=C(R ¹ )-; each R ¹ is independently H, F, Cl, C ₁ -C ₆ alkyl, ethenyl which may be substituted with F or C ₁ -C ₆ alkyl, ethynyl which may be substituted with C ₁ -C ₆ alkyl, cyano, C ₁ -C ₆ alkoxy, or C ₃ -C ₆ cycloalkyl; where at least one R ¹ is F, Cl, C ₁ -C ₆ alkyl, optionally substituted ethenyl or ethynyl, cyano, C ₁ -C ₆ alkoxy or C ₃ -C ₆ cycloalkyl; or both R ¹ together with the carbon atoms to which they are attached form a fused phenyl; R represents H or C ₁ -C ₆ -alkyl; And ring A is a bicyclic fully aromatic or partially reduced heteroaryl ring system containing 3, 4 or 5 N atoms substituted with 0, 1 or 2 substituents each independently selected from the group consisting of NH ₂ , cyano, NHC(=O)O-tBu,OH, carboxamido, C ₁ -C ₆ -alkyl, -S(O) ₂ (C ₁ -C ₆ -alkyl) and -S(O)(C ₁ -C ₆ -alkyl); with the proviso that the partially reduced heteroaryl ring system may be substituted with an oxo group; or a pharmaceutically acceptable salt thereof, or formula (II) Where each R ¹ is independently H, F, Cl, ethenyl which may be substituted by F or C ₁ -C ₆ alkyl, ethynyl which may be substituted by C ₁ -C ₆ alkyl, cyano, C ₁ -C ₆ alkoxy or C ₃ -C ₆ cycloalkyl; where at least one R ¹ represents F, Cl, optionally substituted ethenyl or ethynyl, cyano, C ₁ -C ₆ -alkoxy or C ₃ -C ₆ -cycloalkyl; or both R ¹ together with the carbon atoms to which they are attached form a fused phenyl; And ring A is a bicyclic fully aromatic or partially reduced heteroaryl ring system containing 3, 4 or 5 N atoms substituted with 0, 1 or 2 substituents each independently selected from the group consisting of NH ₂ , cyano, NHC(=O)O-tBu, OH, carboxamido, C ₁ -C ₆ -alkyl, -S(O) ₂ (C ₁ -C ₆ -alkyl) and -S(O)(C ₁ -C ₆ alkyl); with the proviso that the partially reduced heteroaryl ring system may be substituted with an oxo group; or a pharmaceutically acceptable salt thereof. 2. The compound according to claim 1, where the compound is a compound of formula (I), where X is S, -N=C(R ¹ )- or -C(R ¹ )=C(R ¹ )-; each R ¹ is independently H, F, Cl, ethenyl which may be substituted by F or C ₁ -C ₆ alkyl, ethynyl which may be substituted by C ₁ -C ₆ alkyl, cyano or C ₁ -C ₆ alkoxy; R represents H or C ₁ -C ₆ -alkyl; And ring A is a bicyclic fully aromatic or partially reduced heteroaryl ring system containing 3, 4 or 5 N atoms substituted with 0, 1 or 2 substituents each independently selected from the group consisting of NH ₂ , cyano, NHC(=O)O-tBu,OH and carboxamido; with the proviso that the partially reduced heteroaryl ring system may be substituted with an oxo group. 3. A compound according to claim 1, wherein the compound is a compound of formula (II). 4. Connection to any of paragraphs. 1-3, where ring A is unsubstituted or substituted and is a ring selected from the group consisting of: , , , , , , , , , , , , , , , And , where the wavy line indicates the binding position. 5. Compound selected from the following table: 1 2 3 4 5 48 6 49 7 50 8 51 9 52 10 53 eleven 54 12 55 13 56 14 57 15 58 16 59 17 60 18 61 19 62 20 63 21 64 22 65 23 66 24 67 25 68 26 69 27 70 28 71 29 72 thirty 73 31 74 32 75 33 76 34 77 35 78 36 79 37 80 38 81 39 82 40 83 41 84 42 85 43 86 44 87 45 88 46 47 89 90 91 6. A method of treating a tumor in a patient, comprising administering to the patient an effective dose of a compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs. 1-5. 7. The method of claim. 6, where the introduction includes oral or intratumoral administration, or both. 8. Pharmaceutical composition for the treatment of tumors, containing an effective amount of a compound or a pharmaceutically acceptable salt according to any one of paragraphs. 1-5 and a pharmaceutically acceptable carrier. 9. The use of a compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs. 1-5 for treating a tumor in a patient.