Tetrahydrobenzo-quinoline sulfonamide derivatives useful as IGE modulators

Abstract

The present invention relates to tetrahydrobenzo-isoquinoline sulfonamide derivatives of formula (I), [Formula 1] JPEG2023508150000046.jpg5259 It relates to processes for their preparation, pharmaceutical compositions containing them, and their use in the treatment of disorders caused by IgE (such as allergic responses, non-allergic mast cell responses or certain autoimmune responses), in particular disorders caused by the interaction of IgE with the FcεRI receptor.

Description

[Technical field]

[0001] The present invention relates to tetrahydrobenzo-isoquinoline sulfonamide derivatives of formula (I), processes for their preparation, pharmaceutical compositions containing them and their use in the treatment of disorders caused by IgE, such as allergic responses, non-allergic mast cell responses or certain autoimmune responses, in particular disorders caused by the interaction of IgE with the FcεRI receptor. [Background technology]

[0002] IgE (immunoglobulin E) is a member of the immunoglobulin family and mediates allergic reactions such as asthma, food allergies, type 1 hypersensitivity and the well-known sinusitis.

[0003] IgE is secreted by B cells and expressed on their surface. IgE synthesized by B cells is anchored to the B cell membrane by a transmembrane domain linked to the mature IgE sequence by a short membrane-binding region. IgE also binds to B cells (and monocytes, eosinophils and platelets) via its Fc region to the low affinity IgE receptor (FcεRII). When a mammal is exposed to an allergen, B cells that synthesize IgE that binds to the allergen are clonally expanded. This IgE is then released by B cells into the circulation, where it is bound by B cells (via FcεRII) and by mast cells and basophils via the so-called high affinity receptor (FcεRI) found on the surface of mast cells and basophils. Such mast cells and basophils are thereby sensitized to the allergen. Subsequent exposure to the allergen crosslinks the FcεRI on these cells, thus activating the release of histamine and other factors that are responsible for clinical hypersensitivity and anaphylaxis.

[0004] Currently, allergic diseases, urticaria and asthma are usually treated with one or more of the following drugs: (1) antihistamines and anti-leukotrienes that antagonize the inflammatory mediators histamine and leukotrienes, (2) local or systemic (oral or injectable) corticosteroids or immunosuppressants that suppress a wide range of inflammatory mechanisms, (3) short- or long-acting bronchodilators that relax the smooth muscle of the narrowed airways in asthma, or (4) mast cell stabilizers that inhibit the degranulation of mast cells that is normally triggered by IgE binding at FcεRI, (5) biologics that prevent IgE binding at FcεRI. The use of peptides that modulate IgE binding to FcεRI has also been attempted. As an example, WO 96 / 01643 describes peptides consisting of 4-50 amino acids for treating immediate-type allergic responses.

[0005] However, there remains a need to identify compounds that have therapeutic utility in the treatment or prevention of disorders caused by IgE, particularly disorders caused by the interaction of IgE with the FcεRI receptor. Summary of the Invention

[0006] It has been found that compounds of formula (I) and their pharma- ceutically acceptable salts may be used for this purpose. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0007] The present invention provides compounds of formula (I) and pharma- ceutically acceptable salts thereof. [ka] R1 and R2 each independently represent a group selected from the following:

[0008] hydrogen; or NHC(O)NH-C1-6-alkyl; or NHSO2-C1-6-alkyl; or one or more R1 aNHC(O)NH-heteroaryl optionally substituted with; or amino; C1-6-alkyl; C(O)O-C1-6-alkyl; nitrile; one or more R1 a Heteroaryl optionally substituted with;NH-C1-6-alkyl;NH-C1-6-heterocycloalkyl;NH-C3-9-cycloalkyl; one or more R1 a Heteroaryl optionally substituted with one or more groups selected from NH-heteroaryl optionally substituted with; or C1-6-alkyl; C1-6-hydroxyalkyl; C3-9-hydroxyheterocycloalkyl, one or more R1 a or NHC(O)-C1-6-alkyl; or one or more R1 a NHC(O)-heteroaryl optionally substituted with R1 a represents a group selected from the following: Halogen; nitrile; C1-6-alkyl; C1-6-haloalkyl; C1-6-alkoxy; C1-6-haloalkoxy; C(O)O-C1-6-alkyl; C(O)OH. R3 represents a group selected from the following: R3 a C1-6-alkyl optionally substituted with one or more groups selected from among One or more R3 a C1-3-alkanediyl-C3-6-cycloalkyl optionally substituted with One or more R3 a C1-3-alkanediyl-C3-6-heterocycloalkyl optionally substituted with One or more R3 a C3-6-heterocycloalkyl optionally substituted with; One or more R3 a C3-6-cycloalkyl optionally substituted with. R3 arepresents a group selected from hydrogen; halogen; C1-2-alkyl; hydroxy; C1-2-alkoxy. R4 represents a group selected from the following: One or more R4 a C3-6-cycloalkyl optionally substituted with a group; or one or more R4 a C1-6-alkanediyl-C3-6-cycloalkyl optionally substituted with a group; or one or more R4 a C1-6-alkanediyl-C3-6-heterocycloalkyl optionally substituted with a group. R4 a represents a group selected from hydroxy; halogen; C1-2-alkyl.

[0009] The term "pharmaceutical acceptable salts" according to the present invention includes salts of compounds of formula (I) with pharmaceutical acceptable acids or bases, in particular acid addition salts. The acid addition salt forms of compounds of formula (I) occurring in free form as bases can be obtained by treating the free base with a suitable acid such as inorganic acids, e.g. hydrohalic acids such as hydrochloric acid or hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., or organic acids, e.g. acetic acid, trifluoroacetic acid, oxalic acid, hydroxyacetic acid, propanoic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclamic acid, salicylic acid, p-aminosalicylic acid, pamoic acid, etc.

[0010] The present invention also relates to all stereoisomeric forms, such as enantiomeric and diastereomeric forms of the compounds of formula (I), or mixtures thereof, including all possible mixtures of stereoisomers, such as racemates. In the context of the present invention, reference to one or more compounds is intended to encompass that compound in each of its possible isomeric forms and mixtures thereof, unless a particular isomeric form is specifically mentioned.

[0011] Some of the compounds of formula (I) may exist in tautomeric forms. Although not explicitly shown in the above formula, such forms are intended to be included within the scope of the present invention.

[0012] It should be understood that each individual atom present in formula (I) or any formula shown herein may in fact be present in the form of any of its naturally occurring isotopes, with the most abundant isotope(s) being preferred.Thus, by way of example, each individual hydrogen atom present in formula (I) or any formula shown herein may be present as 1H, 2H (deuterium) or 3H (tritium) atom, preferably 1H.Similarly, by way of example, each individual carbon atom present in formula (I) or any formula shown herein may be present as 12C, 13C or 14C atom, preferably 12C.

[0013] The present invention includes within its scope solvates of the compounds of formula (I) above. Such solvates may be formed using common organic solvents or water.

[0014] The present invention also includes within its scope co-crystals of compounds of formula (I) above. The technical term "co-crystal" is used to describe a situation in which neutral molecular components are present in a crystalline compound in a well-defined stoichiometric ratio. The preparation of pharmaceutical co-crystals allows modifications to be made to the crystalline form of a pharma- ceutical active ingredient, thereby altering its physicochemical properties without compromising its intended biological activity (see Pharmaceutical Salts and Co-crystals, ed. J. Wouters & L. Quere, RSC Publishing, 2012).

[0015] Compounds according to the present invention may exist in different polymorphic forms. Although not explicitly shown in the above formula, such forms are intended to be included within the scope of the present invention.

[0016] The present invention also includes within its scope prodrugs of the compounds of formula (I) above. The term "prodrug" refers to a compound that is metabolized in vivo to a compound of the present invention or a salt thereof. A prodrug can be identified by administering the prodrug to a mammal, such as a rat, mouse, monkey or human, and identifying the compound or a salt thereof, for example, in the blood or urine.

[0017] Within the framework of the present invention Ct-z represents a carbon chain that may have t to z carbon atoms, for example a C1-7 carbon chain that may have 1 to 7 carbon atoms; Alkyl is a saturated, linear or branched aliphatic group, for example, a C1-6-alkyl group represents a linear or branched carbon chain of 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertbutyl, pentyl, hexyl. Alkyl is a group in which one or more hydrogen atoms are replaced by a deuterium atom. 2 H-replaced deuterated groups are included.

[0018] Alkanediyl is a group of the general formula C, such as -CH-CH-. n H 2n is a divalent linear or branched saturated hydrocarbon group. Alkylamino refers to one or more alkyl groups substituted on an amino radical. Examples of alkylamino include methylamino, ethylamino, tertbutylamino, and dimethylamino. Hydroxy is an -OH group; Hydroxyalkyl is an alkyl group in which one or more hydrogen atoms are replaced by hydroxy groups; Haloalkyl is an alkyl group in which one or more hydrogen atoms are replaced by halogen atoms; Alkoxy is an -O-alkyl group; Haloalkoxy is an alkoxy group in which one or more hydrogen atoms are replaced by halogen atoms; The halogen atom is a fluorine, chlorine, bromine or iodine atom; Cycloalkyl refers to a monocyclic or bicyclic aliphatic group that is not aromatic and may contain double bonds and contains 3 to 14 atoms, preferably 3 to 9 atoms in the group. As examples of cycloalkyl, mention may be made of cyclopropyl, cyclobutyl, cyclobutenyl, cyclopentyl, cyclohexyl, spiro-undecanyl, spiro[2.2]pentanyl. Heterocycloalkyl refers to a monocyclic or bicyclic saturated group that is not aromatic and may contain a double bond, containing 3 to 14 atoms, preferably 3 to 10 atoms in the group, in which one or more carbon atoms are replaced by an atom selected from nitrogen, oxygen, sulfur. As examples of heterocycloalkyl, mention may be made of aziridinyl, pyrrolidinyl, piperidyl, oxetane, oxa-spiro-undecanyl. Hydroxyheterocycloalkyl is a heterocycloalkyl group in which one or more hydrogen atoms have been replaced by a hydroxy group. Heteroaryl refers to a monocyclic or bicyclic group containing 5 to 14, preferably 5 to 10, atoms, in which at least one ring is aromatic and at least one atom in the group is selected from among nitrogen, oxygen, and sulfur. As examples of heteroaryl groups, triazolyl, furanyl, pyrrolyl, chromanyl, and isoquinolinyl may be mentioned.

[0019] According to one embodiment, the compounds of the invention are selected from compounds of formula (I) When R1 is different from hydrogen, R2 is hydrogen; When R2 is different from hydrogen, R1 is hydrogen; Other substituents are as defined herein above and below.

[0020] According to one embodiment, the compounds of the invention are selected from compounds of formula (I) In the formula, R3 represents a group selected from the following: R3 a C1-6-alkyl optionally substituted with one or more groups selected from among One or more R3 aC1-3-alkanediyl-C3-6-cycloalkyl optionally substituted with One or more R3 a C1-3-alkanediyl-C3-6-heterocycloalkyl optionally substituted with One or more R3 a C3-6-heterocycloalkyl optionally substituted with; One or more R3 a C3-6-cycloalkyl optionally substituted with. R3 a represents a group selected from hydrogen, halogen, C1-2-alkyl, hydroxy, and C1-2-alkoxy. Other substituents are as defined herein above and below.

[0021] According to one embodiment, the compounds of the invention are selected from compounds of formula (I) In the formula, R4 is One or more R4 a C3-6-cycloalkyl optionally substituted with a group; or one or more R4 a C1-6-alkanediyl-C3-6-cycloalkyl optionally substituted with a group; or one or more R4 a C1-6-alkanediyl-C3-6-heterocycloalkyl, optionally substituted with a group, R4 a represents a group selected from hydroxy; halogen; C1-2-alkyl, Other substituents are as defined herein above and below.

[0022] According to one embodiment, the compounds of the invention are selected from compounds of formula (I) In the formula, R4 represents cyclopropyl; Other substituents are as defined herein above and below.

[0023] Another embodiment of the present invention relates to a pharmaceutical composition comprising a detectable amount of a compound of formula (I) or a pharma- ceutically acceptable salt, solvate, or co-crystal thereof in combination with a pharma- ceutically acceptable diluent or carrier.

[0024] In yet another embodiment, the present invention relates to a compound of formula (I), a pharma- ceutically acceptable salt, solvate or co-crystal thereof, for use as a medicament, in particular for use in a method for the treatment or prevention of disorders caused by IgE, including allergies, type 1 hypersensitivity, the well-known sinusitis, urticaria or related conditions, e.g. airway constriction in asthma, local inflammation in eczema, increased mucus secretion in allergic rhinitis, or increased vascular permeability.

[0025] In a further embodiment, the present invention relates to a method for treating or preventing allergies, type 1 hypersensitivity, common sinusitis, urticaria or related conditions comprising the administration of a therapeutically effective amount of a compound of formula (I).

[0026] According to one embodiment, the compound of the invention is selected from: 3-Cyclopropyl-N-(2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 1-[3-cyclopropyl-5-(2-methylpropylsulfamoyl)-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]-3-ethylurea; 1-[3-cyclopropyl-5-(2-methylpropylsulfamoyl)-7,8,9,10-tetrahydrobenzo[h]isoquinolin-10-yl]-3-ethylurea; 3-Cyclopropyl-7-(methanesulfonamido)-N-(2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 3-Cyclopropyl-10-(methanesulfonamido)-N-(2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 3-Cyclopropyl-N-(2-fluoro-2-methylpropyl)-7-[[6-(2-methyltetrazol-5-yl)pyridin-3-yl]amino]-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 3-Cyclopropyl-7-[3-[(2,5-dimethylpyrazol-3-yl)amino]-1,2,4-triazol-4-yl]-N-(2-fluoro-2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 3-Cyclopropyl-N-(2-fluoro-2-methylpropyl)-10-[[6-(2-methyltetrazol-5-yl)pyridin-3-yl]amino]-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; Ethyl 5-amino-1-[3-cyclopropyl-5-[(2-fluoro-2-methylpropyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]imidazole-4-carboxylate; 3-Cyclopropyl-N-(2-fluoro-2-methylpropyl)-7-[[5-(3-methyl-1,2,4-oxadiazol-5-yl)pyridin-3-yl]amino]-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 1-[3-cyclopropyl-5-[(2-fluoro-2-methylpropyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]-3-(2,5-dimethylpyrazol-3-yl)urea; Ethyl 1-[3-cyclopropyl-5-[(2-fluoro-2-methylpropyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]imidazole-4-carboxylate.

[0027] The following examples show how the compounds covered by formula (I) can be synthesized. They are provided for illustrative purposes only and are not intended to, and should not be construed as, limiting the present invention in any manner. Those skilled in the art will understand that routine variations and modifications of the following examples can be made without departing from the spirit or scope of the present invention. EXAMPLES

[0028] The following examples show how compounds covered by formula I can be synthesized. They are provided for illustrative purposes only and are not intended, nor should they be construed, as limiting the present invention in any manner. Those skilled in the art will understand that routine variations and modifications of the following examples can be made without departing from the spirit or scope of the present invention.

[0029] Abbreviation DCM Dichloromethane THF Tetrahydrofuran EtOAc Ethyl acetate MeCN Acetonitrile MeOH Methanol M mass or mole Brine Saturated sodium chloride solution HPLC High Performance Liquid Chromatography LCMS Liquid Chromatography Mass Spectrometry DIPEA N,N-Diisopropylethylamine RT retention time DMF N,N'-Dimethylformamide sat. saturation aq. aqueous solution tBuXPhos Pd G3 [(2-di-tert-butylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'biphenyl)]methanesulfonate palladium(II) IPA Isopropyl Alcohol conc. concentration SCX Biotage® ISOLUTE® SCX-2 Propyl Sulfonic Acid Functionalized Silica TEA Triethylamine AcOH Acetic acid AIBN 2,2'-Azobis(2-methylpropionitrile) Bedford catalyst Chloro(η 2 -P,C-Tris(2,4-di-tert-butylphenyl)phosphite)(tricyclohexylphosphine)palladium(II) NBS N-Bromosuccinimide min

[0030] LCMS method Method 1: X-Bridge C18 Waters 2.1x20mm, 2.5μm column Column temperature: 40℃ Mobile phase A: 10 mM ammonium formate in water + 0.1% formic acid Mobile phase B: acetonitrile + 5% water + 0.1% formic acid Gradient program: flow rate 1mL / min Time A% B% 0.00 95.00 5.00 1.50 5.00 95.00 2.25 5.00 95.00 2.50 95.00 5.00

[0031] Method 2: Mobile phase A: 0.1% formic acid in water Mobile phase B: 0.1% formic acid in acetonitrile Phenomenex, Kinetex-XB C18, 2.1mmx100mm, 1.7μm column Flow rate: 0.6mL / min Column temperature: 40℃ Injection volume: 1μL Gradient: Time (min): %A %B 0.00 95 5 5.30 0 100 5.80 0 100 5.82 95 5 7.00 95 5 UV 215nM, PDA spectrum 200~400nm, step: 1nm MSD Scan Positive 150-850

[0032] Method 3: X-Bridge C18 Waters 2.1x20mm, 2.5μM column Column temperature: 40℃ Mobile phase A: 10 mM ammonium formate in water + 0.1% formic acid Mobile phase B: acetonitrile + 5% water + 0.1% formic acid Gradient program: flow rate 1mL / min Time A% B% 0.00 95.00 5.00 4.00 5.00 95.00 5.00 5.00 95.00 5.10 95.00 5.00

[0033] Method 4: Stationary phase: X-Bridge C18 Waters 2.1x20mm, 2.5μM column Mobile phase A: 10 mM ammonium formate + 0.1% ammonia solution in water Mobile phase B: Acetonitrile + 5% water + 0.1% ammonia solution Flow rate: 1mL / min Gradient Program: Time A% B% 0.00 95.00 5.00 1.50 5.00 95.00 2.25 5.00 95.00 2.50 95.00 5.00

[0034] Method 5: System: Waters Classic Acquity-QDa, Acquity PDA Stationary phase: Waters Acquity UPLC BEH, C18, 2.1x50mm, 1.7μm Mobile phase A: 10 mM ammonium formate + 0.1% ammonia solution in water Mobile phase B: Acetonitrile + 5% water + 0.1% ammonia solution Flow rate: 0.7mL / min Temperature: 40℃ Gradient Program: Time A% B% 0.00 98.00 2.00 4.00 5.00 95.00 5.00 5.00 95.00 5.10 98.00 2.00

[0035] Method 6: Stationary phase: X-Bridge C18 Waters 2.1x20mm, 2.5μM column Mobile phase A: 10 mM ammonium formate in water + 0.1% formic acid Mobile phase B: acetonitrile + 5% water + 0.1% formic acid Flow rate: Pump 1: 1mL / min, Pump 2: 0.5mL / min Gradient Program: Pump 1: Pump 2: Time A% B% Time A% B% 0.00 95.10 4.90 0.10 5.00 95.00 4.00 5.00 95.00 1.00 5.00 95.00 5.00 5.00 95.00 1.10 95.00 5.00 5.10 95.10 4.90

[0036] Method 7: Stationary phase: X-Bridge C18 Waters 2.1x20mm, 2.5μM column Mobile phase A: 10 mM ammonium formate + 0.1% ammonia solution in water Mobile phase B: Acetonitrile + 5% water + 0.1% ammonia solution Flow rate: 1mL / min Gradient Program: Time A% B% 0.00 95.00 5.00 4.00 5.00 95.00 5.00 5.00 95.00 5.10 95.00 5.00

[0037] Method 8: Stationary phase: X-Bridge C18 Waters 2.1x20mm, 2.5μM column Mobile phase A: 10 mM ammonium formate + 0.1% ammonia solution in water Mobile phase B: Acetonitrile + 5% water + 0.1% ammonia solution Flow rate: Pump 1: 1mL / min, Pump 2: 0.5mL / min Gradient Program: Pump 1: Pump 2: Time A% B% Time A% B% 0.00 95.10 4.90 0.10 5.00 95.00 4.00 5.00 95.00 1.00 5.00 95.00 5.00 5.00 95.00 1.10 95.00 5.00 5.10 95.10 4.90

[0038] Method 9: Stationary phase: X-Bridge C18 Waters 2.1x20mm, 2.5μM column Mobile phase A: 10 mM ammonium formate + 0.1% ammonia solution in water Mobile phase B: Acetonitrile + 5% water + 0.1% ammonia solution Flow rate: 1mL / min Gradient Program: Time A% B% 0.00 95.00 5.00 1.50 5.00 95.00 2.25 5.00 95.00 2.50 95.00 5.00

[0039] Intermediates Intermediate 1 [ka] 2,3,6,7,8,9-Hexahydrocyclopenta[a]naphthalene-1-one 1,2,3,4-Tetrahydronaphthalene (5.17 mL, 37.8 mmol) was added slowly to a suspension of AlCl3 (10.6 g, 79.4 mmol) and prop-2-enoyl chloride (3.4 mL, 41.6 mmol) in DCM (300 mL) at -78 °C. The mixture was then warmed to room temperature overnight. The solution was carefully hydrolyzed on ice and the organic phase was separated. The aqueous phase was extracted twice with DCM and the combined organic fractions were washed with aqueous potassium carbonate and dried over sodium sulfate. The solvent was removed under vacuum and the crude residue was purified by column chromatography eluting with a gradient of 0% to 20% EtOAc in heptane to give the title compound (1.01 g, 12% yield). LCMS [M+H] + 187, RT 1.89 min (Method 1).

[0040] Intermediate 2 [ka] 2-Hydroxyimino-6,7,8,9-tetrahydro-3H-cyclopenta[a]naphthalen-1-one To a solution of intermediate 1 (1.0 g, 5.36 mmol) in ether (13 mL) was added first saturated ethanolic HCl (0.22 mL) and then a 15% ethanolic solution of ethyl nitrite (4.81 mL, 7.62 mmol) dropwise at 0° C. After 30 min at 0° C., the precipitated product was collected by filtration, washed with ether and dried to give the crude title compound (1.15 g, 74% yield), which was used in the next step without further purification. LCMS [M+H] + 216, RT 1.77 min (Method 1).

[0041] Intermediate 3 [ka] 1,3-Dichloro-7,8,9,10-tetrahydrobenzo[h]isoquinoline To a suspension of intermediate 2 (0.86 g, 3.99 mmol) in POCl3 (24 mL) was added PCl5 (940 mg, 4.51 mmol) at 0° C. Then, gaseous HCl was introduced until the solution was saturated and the reaction was stirred at 60° C. for 4 h. Then, PCl5 (316 mg) was added again and stirring was continued at 80° C. for 2 h. The solvent was removed under vacuum and the residue was slowly hydrolyzed by addition of water to give a precipitate, which was collected by filtration. The collected solid was washed with water and dried to give the crude title compound (1 g, 84% yield), which was used in the next step without further purification. LCMS [M+H] + 252, RT2.50 minutes (method 1).

[0042] Intermediate 4 [ka] 3-Chloro-7,8,9,10-tetrahydrobenzo[h]isoquinoline A mixture of intermediate 3 (0.88 g, 3.52 mmol), a solution of red phosphorus (261 mg, 8.45 mmol) in AcOH (4 mL) and HI (57%) (1.59 mL, 12.1 mmol) was refluxed for 8 h. The hot reaction mixture was filtered, evaporated and the residue was dissolved in water and basified by addition of concentrated aqueous NH4OH. The precipitate was collected by filtration, dissolved in DCM, washed with brine, dried over MgSO4 and evaporated. The crude residue obtained was purified by column chromatography eluting with a gradient of 8% to 40% EtOAc in heptane to give the title compound (600 mg, 74% yield). LCMS [M+H] + 218, RT2.03 minutes (method 1).

[0043] Intermediate 5 [ka] 3-Chloro-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonyl chloride A mixture of intermediate 4 (15.0 g, 68.9 mmol) and chlorosulfonic acid (55 mL, 819 mmol) was heated in a sealed tube at 80° C. for 3 h 15 min. The reaction mixture was then diluted with DCM (400 mL) and poured into H2O (400 mL) at 0° C. The phases were separated, the aqueous phase was extracted with DCM (4×100 mL), the organics were combined, dried (Na2SO4), filtered and concentrated in vacuo to give the crude title compound (21.8 g, assumed quantitative), which was used in the next step without further purification. LCMS [M+H] + 353( i Quench with BuNH2), RT 1.33 min (Method 4).

[0044] Intermediate 6 [ka] 3-Chloro-N-(2-fluoro-2-methyl-propyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide To a stirred solution of intermediate 5 (21.8 g, 68.9 mmol) in DCM (250 mL) at 0° C. was added 2-fluoro-2-methylpropan-1-amine HCl (9.85 g, 77.2 mmol) followed by DIPEA (30 mL, 172 mmol). The reaction was then stirred at room temperature for 15 h. The mixture was diluted with H2O (200 mL), brine (200 mL) and the phases were separated. The aqueous phase was extracted with DCM (2×50 mL) and the combined organics were extracted with H2O (150 mL) then brine (150 mL), dried and concentrated in vacuo. The resulting solid was triturated with a mixture of EtOAc:iso-hexane=8:2 (100 mL) to give the title compound (19.7 g, 73% yield). The filtrate was concentrated under vacuum and purified by column chromatography eluting with 0-50% EtOAc in iso-hexane to give a second crop of the title compound (3.7 g, 12% yield). LCMS [M+H] + 371, RT 1.23 min (Method 4).

[0045] Intermediate 7&8 [ka] tert-Butyl N-[3-chloro-5-[(2-fluoro-2-methyl-propyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-10-yl]carbamate (7) tert-Butyl N-[3-chloro-5-[(2-fluoro-2-methyl-propyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]carbamate (8) To a solution of intermediate 6 (9.10 g, 24.5 mmol) in EtOAc (235 mL) at 50° C., NBS (4.85 g, 27.2 mmol) and 2,2′-azobis(2-methylpropionitrile) (400 mg, 2.44 mmol) were added. The reaction mixture was heated to 90° C. under N2 for 2.5 h. The mixture was concentrated in vacuo, the residue was dissolved in THF (200 mL), purged with NH3 gas for 10 min, and then heated in a sealed vessel at 70° C. for 11 h. The mixture was then concentrated in vacuo and suspended in DCM (200 mL) in an ice bath, to which was then added a solution of di-tert-butyl dicarbonate (7.9 g, 35 mmol) in DCM (30 mL), followed by TEA (9 mL, 64.6 mmol). After 4 hours, additional di-tert-butyl dicarbonate (1.0 g, 4.4 mmol) was added and the reaction was stirred for 3 days. The reaction mixture was then diluted with brine (200 mL), saturated NaHCO3 (200 mL) and the phases were separated. The aqueous layer was extracted with DCM (3x50 mL) and the combined organics were dried and concentrated in vacuo. Purification by column chromatography eluting with 0-20% EtOAc in iso-hexane gave the title compound.

[0046] Intermediate 7 (2.35g, 19% yield); LCMS[M+H] + 486, RT1.28 (Method 4).

[0047] Intermediate 8 (3.53 g, 27% yield); LCMS [M+H] + 486, RT1.23 (Method 4).

[0048] Intermediate 9 [ka] tert-Butyl N-[3-cyclopropyl-5-[(2-fluoro-2-methyl-propyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]carbamate To a suspension of intermediate 8 (3.52 g, 6.52 mmol) in a mixture of toluene (70 mL), 1,4-dioxane (40 mL) and HO (3.5 mL) was added cyclopropylboronic acid (1.87 g, 21.8 mmol) and potassium phosphate tribasic (3.95 g, 18.2 mmol). The mixture was purged with N for 5 min, after which palladium(II) acetate (90 mg, 0.40 mmol) and tricyclohexylphosphonium tetrafluoroborate (400 mg, 1.05 mmol) were added. The resulting orange mixture was purged with N2 for 5 min and then heated at 120°C for 11 h, after which additional palladium(II) acetate (90 mg, 0.40 mmol) and tricyclohexylphosphonium tetrafluoroborate (400 mg, 1.05 mmol) were added and the mixture was purged with N2 for another 5 min and then heated at 120°C for 3 days. The reaction mixture was concentrated in vacuo and the residue was dissolved in EtOAc (400 mL containing a few mL of IPA). Water (150 mL) and brine (200 mL) were added and the phases were separated. The aqueous phase was extracted with EtOAc (100 mL) and the combined organic layers were dried, concentrated in vacuo and triturated with Et2O (150 mL) to give the title compound (2.20 g, 69% yield). LCMS [M+H] + 492, RT 1.25 min (method 4).

[0049] Intermediate 10 [ka] 7-Amino-3-cyclopropyl-N-(2-fluoro-2-methyl-propyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide Hydrochloric acid (4N in dioxane, 6 mL) was added to intermediate 9 (500 mg, 0.98 mmol), followed by MeOH (2 mL) and the resulting solution was stirred at room temperature for 1.5 h. The reaction mixture was concentrated in vacuo and passed through a 10 g SCX cartridge eluting with 7N NH3 in MeOH, then triturated with Et2O (20 mL) to give the title compound (356 mg, 93% yield). LCMS [M+H] + 392, RT 1.32 min (Method 9).

[0050] Intermediate 11 [ka] tert-Butyl N-[3-cyclopropyl-5-[(2-fluoro-2-methyl-propyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-10-yl]carbamate To a suspension of intermediate 7 (2.35 g, 4.84 mmol) in a mixture of toluene (50 mL), 1,4-dioxane (20 mL), and HO (3 mL) was added cyclopropylboronic acid (1.2 g, 14 mmol) and potassium phosphate tribasic (2.6 g, 12 mmol). The mixture was purged with N for 5 min, followed by the addition of palladium(II) acetate (55 mg, 0.24 mmol) and tricyclohexylphosphonium tetrafluoroborate (270 mg, 0.71 mmol). The resulting orange mixture was purged with N for 5 min and then heated at 120° C. for 9 h, after which additional palladium(II) acetate (55 mg, 0.24 mmol) and tricyclohexylphosphonium tetrafluoroborate (270 mg, 0.71 mmol) were added and the mixture was purged with N for an additional 5 min and then heated at 120° C. for 8 h. The reaction mixture was concentrated in vacuo, dissolved in EtOAc (400 mL) and washed with brine (150 mL). The phases were separated and the aqueous phase was extracted with EtOAc (2x100 mL). The combined organics were dried, concentrated in vacuo and triturated with Et2O (100 mL) to give the title compound (1.96 g, 82% yield). LCMS [M+H] + 492, RT 1.29 min (Method 4).

[0051] Intermediate 12 [ka] 10-Amino-3-cyclopropyl-N-(2-fluoro-2-methyl-propyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide Hydrochloric acid (4N in dioxane, 6 mL) was added to intermediate 11 (500 mg, 1.00 mmol), followed by MeOH (2 mL) and the solution was stirred at room temperature for 1 h. The reaction mixture was concentrated in vacuo and passed through an SCX cartridge eluting with 7N NH3 in MeOH, then triturated with Et2O (10 mL) to give the title compound (307 mg, 75% yield). LCMS [M+H] + 392, RT 0.97 min (method 4).

[0052] Intermediate 13 [ka] 3-Chloro-N-isobutyl-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide To a solution of intermediate 5 (475 mg, 1.50 mmol) in DCM (20 mL), 2-methylpropan-1-amine (0.74 mL, 7.51 mmol) was added dropwise and the reaction mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with DCM (50 mL) and washed with water. The organic layer was dried over MgSO4 and concentrated under reduced pressure. The residue was then purified by column chromatography eluting with a gradient of 12% to 60% EtOAc in heptane to give the title compound (497 mg, 92% yield). LCMS [M+H] + 353, RT3.97 minutes (method 2).

[0053] Intermediate 14 & 15 [ka] 7-Bromo-3-chloro-N-isobutyl-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide (14) 10-Bromo-3-chloro-N-isobutyl-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide (15) Intermediate 13 (150 mg, 0.42 mmol) was dissolved in EtOAc (15 mL) and NBS (75.6 mg, 0.42 mmol) was added followed by AIBN (6.98 mg, 0.043 mmol). The heterogeneous mixture was heated to 90° C. for 3 h. The reaction mixture was diluted with EtOAc and washed with Na2S2O3, water and brine. The organic layer was dried over MgSO4 and concentrated under reduced pressure. The crude residue was purified by column chromatography eluting with a gradient of 0% to 50% EtOAc in heptane to give the title compound (145 mg, 56% yield) as a mixture of regioisomers in 71% purity. LCMS [M+H] + 431 / 433, RT 3.33 min (method 3) [Note: both positional isomers co-eluted at the same RT].

[0054] Intermediate 16 & 17 [ka] 7-Amino-3-chloro-N-isobutyl-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide (16) 10-Amino-3-chloro-N-isobutyl-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide (17) A mixture of intermediates 14 and 15 (71%, 145 mg, 0.24 mmol) was dissolved in ammonia 7N in MeOH (2 mL) and stirred at room temperature for 16 h. The solvent was removed under vacuum and DCM was added followed by water. The organic layer was separated, dried over Na2SO4 and concentrated under reduced pressure. The crude residue was purified by column chromatography eluting first with a gradient of 5% to 100% EtOAc in heptane to remove non-brominated starting material and then with a gradient of 0% to 10% MeOH in DCM to give the title compound (84 mg, 94% yield) as a mixture of regioisomers. LCMS [M+H] + 368, RT 2.09 and 2.14 min (Method 3).

[0055] Intermediate 18 & 19 [ka] 1-[3-chloro-5-(isobutylsulfamoyl)-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]-3-ethyl-urea (18) 1-[3-chloro-5-(isobutylsulfamoyl)-7,8,9,10-tetrahydrobenzo[h]isoquinolin-10-yl]-3-ethyl-urea (19) Isocyanatoethane (19.8 μL, 0.25 mmol) was added to a mixture of intermediates 16 and 17 (84 mg, 0.22 mmol) in DCM (4 mL). After the solution was stirred at room temperature for 5 h, a precipitate appeared. MeOH (5 mL) was added. The solvent was then removed and the crude product was purified by acidic reverse phase HPLC to give the title compound.

[0056] Intermediate 18 (37 mg, 70% yield). LCMS[M+H] + 439, RT2.63 minutes (method 3).

[0057] Intermediate 19 (24 mg, 44% yield). LCMS [M+H] + 439, RT2.70 minutes (method 3).

[0058] Intermediate 20 & 21 [ka] 3-Chloro-N-isobutyl-7-(methanesulfonamido)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide (20) 3-Chloro-N-isobutyl-10-(methanesulfonamido)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide (21) DIPEA (103.4 μL, 0.58 mmol) was added followed by methanesulfonyl chloride (23 μL, 0.29 mmol) to a mixture of intermediates 16 and 17 (120 mg, 0.29 mmol) in DCM (4 mL). The reaction mixture was stirred at room temperature for 4 h. The reaction mixture was then diluted with DCM and water. The organic layer was separated, dried over Na2SO4 and concentrated under reduced pressure. Purification by reverse phase HPLC (acidic conditions) afforded the title compound.

[0059] Intermediate 20 (65 mg, 98% yield). LCMS [M+H] + 446, RT2.68 minutes (method 3).

[0060] Intermediate 21 (31 mg, 43% yield). LCMS [M+H] + 446, RT2.74 minutes (method 3).

[0061] Intermediate 22 [ka] 1-[3-cyclopropyl-5-[(2-fluoro-2-methyl-propyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]-3-(2,5-dimethylpyrazol-3-yl)thiourea To a vessel charged with 5-isothiocyanato-1,3-dimethyl-pyrazole (25 mg, 0.16 mmol) was added a suspension of intermediate 10 (50 mg, 0.13 mmol) in DCM (1 mL) and THF (2 mL). The resulting reaction mixture was stirred at room temperature for 3 days and then filtered to give the title compound (56 mg, 76% yield). LCMS [M+H] + 545, RT 1.11 min (Method 4).

[0062] Example 1 [ka] 3-Cyclopropyl-N-(2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide A mixture of intermediate 13 (100 mg, 0.28 mmol) and cyclopropylboronic acid (48.6 mg, 0.56 mmol) was dissolved in nitrogen-sparged dioxane (5 mL). A 2M solution of K2CO3 (0.42 mL, 0.85 mmol) was then added, followed by Bedford catalyst (30.2 mg, 0.028 mmol). The reaction mixture was stirred at 100 °C under N2 for 16 h and then concentrated under reduced pressure to remove dioxane. The remaining solution was diluted with DCM (20 mL) and the organic layer was washed with water, dried over MgSO4, and concentrated under reduced pressure. Purification by reverse phase HPLC (basic conditions) afforded the title compound (41 mg, 40% yield).

number

[0063] Example 2 [ka] 1-[3-cyclopropyl-5-(2-methylpropylsulfamoyl)-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]-3-ethylurea A mixture of intermediate 16 (35 mg, 0.08 mmol) and cyclopropylboronic acid (13.7 mg, 0.15 mmol) was dissolved in nitrogen-sparged dioxane (2 mL). A 2M solution of K2CO3 (119.6 μL, 0.23 mmol) was then added, followed by Bedford catalyst (8.5 mg, 0.008 mmol). The mixture was stirred under N2 in a microwave at 120 °C for 2 h. The reaction mixture was concentrated under reduced pressure to remove dioxane. The remaining solution was diluted with DCM (15 mL) and the organic layer was washed with water, dried over MgSO4, and concentrated under reduced pressure. Purification by reverse phase HPLC (acidic conditions) afforded the title compound (4 mg, 11% yield).

number

[0064] Example 3 [ka] 1-[3-cyclopropyl-5-(2-methylpropylsulfamoyl)-7,8,9,10-tetrahydrobenzo[h]isoquinolin-10-yl]-3-ethylurea A mixture of intermediate 17 (20 mg, 0.046 mmol) and cyclopropylboronic acid (7.8 mg, 0.091 mmol) was dissolved in nitrogen-sparged dioxane (2 mL). A 2M solution of K2CO3 (68.3 μL, 0.23 mmol) was added, followed by Bedford catalyst (4.8 mg, 0.005 mmol). The mixture was stirred under nitrogen in a microwave at 120° C. for 2 h. The reaction mixture was concentrated under reduced pressure to remove dioxane. The remaining solution was diluted with DCM (15 mL) and the organic layer was washed with water, dried over MgSO4, and concentrated under reduced pressure. Purification by reverse phase HPLC (acidic conditions) afforded the title compound (6 mg, 30% yield).

number

[0065] Example 4 [ka] 3-Cyclopropyl-7-(methanesulfonamido)-N-(2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide A mixture of intermediate 16 (63 mg, 0.14 mmol) and cyclopropylboronic acid (24.2 mg, 0.28 mmol) was dissolved in nitrogen sparged dioxane (3 mL). A 2M solution of K2CO3 (0.21 mL, 0.42 mmol) was added, followed by Bedford catalyst (15.1 mg, 0.014 mmol). The reaction mixture was concentrated under reduced pressure to remove dioxane. The remaining solution was diluted with DCM (20 mL) and the organic layer was washed with water, dried over MgSO4, and concentrated under reduced pressure. Purification by reverse phase HPLC (acidic conditions) afforded the title compound (27 mg, 41% yield).

number

[0066] Example 5 [ka] 3-Cyclopropyl-10-(methanesulfonamido)-N-(2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide A mixture of intermediate 17 (30 mg, 0.067 mmol) and cyclopropylboronic acid (11.56 mg, 0.13 mmol) was dissolved in nitrogen-sparged dioxane (2 mL). A 2M solution of K2CO3 (0.10 mL, 0.20 mmol) was added, followed by Bedford catalyst (7.1 mg, 0.007 mmol). The mixture was stirred under nitrogen in a microwave at 120° C. for 2 h. The reaction mixture was concentrated under reduced pressure to remove dioxane. The remaining solution was diluted with DCM (15 mL) and the organic layer was washed with water, dried over MgSO4, and concentrated under reduced pressure. Purification by reverse phase HPLC (acidic conditions) afforded the title compound (10 mg, 33% yield).

number

[0067] Example 6 [ka] 3-Cyclopropyl-N-(2-fluoro-2-methylpropyl)-7-[[6-(2-methyltetrazol-5-yl)pyridin-3-yl]amino]-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide To a stirred suspension of intermediate 10 (30 mg, 0.08 mmol) in anhydrous 1,4-dioxane (2.5 mL) at room temperature was added 5-bromo-2-(2-methyltetrazol-5-yl)pyridine (37 mg, 0.15 mmol), sodium tert-butoxide (22 mg, 0.23 mmol) and tBuXPhos Pd G3 (9 mg, 0.01 mmol). The resulting mixture was purged with N2 for 10 min. After an additional 15 min, the reaction mixture was filtered through Celite and concentrated in vacuo. Purification by column chromatography eluting with 0-20% MeOH in EtOAc followed by basic reverse phase column chromatography afforded the title compound (20 mg, 44% yield).

number

[0068] Example 7 [ka] 3-Cyclopropyl-7-[3-[(2,5-dimethylpyrazol-3-yl)amino]-1,2,4-triazol-4-yl]-N-(2-fluoro-2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide To a solution of intermediate 22 (55 mg, 0.10 mmol) in DMF (1 mL) was added formic hydrazide (18 mg, 0.30 mmol) and mercuric chloride (52 mg, 0.19 mmol). TEA (40 μL, 0.29 mmol) was then added and the reaction mixture was heated at 80° C. for 4 h. The reaction mixture was then filtered through Celite (20 mL MeCN washings) and concentrated in vacuo. Purification by column chromatography afforded the title compound (10 mg, 19% yield).

number

[0069] Example 8 [ka] 3-Cyclopropyl-N-(2-fluoro-2-methylpropyl)-10-[[6-(2-methyltetrazol-5-yl)pyridin-3-yl]amino]-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide Synthesized in the same manner as in Example 6 using intermediate 12 (30 mg, 0.08 mmol) and equivalent stoichiometric reagents. Purification by column chromatography eluting with 0-20% MeOH in EtOAc, followed by reverse phase column chromatography (basic conditions) afforded the title compound (6 mg, 14% yield).

number

[0070] Example 9 [ka] Ethyl 5-amino-1-[3-cyclopropyl-5-[(2-fluoro-2-methylpropyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]imidazole-4-carboxylate To a solution of ethyl 2-amino-2-cyanoacetate (15 mg, 0.07 mmol, 64% purity) in MeCN (1 mL) was added triethyl orthoformate (13 μL, 0.08 mmol). The resulting solution was heated at 90° C. for 1 h and then cooled to room temperature. Intermediate 10 (20 mg, 0.05 mmol) was then added and after 1 h the reaction mixture was concentrated in vacuo and purified by reverse phase column chromatography (acidic conditions) to give the title compound (9 mg, 31% yield).

number

[0071] Example 10 [ka] 3-Cyclopropyl-N-(2-fluoro-2-methylpropyl)-7-[[5-(3-methyl-1,2,4-oxadiazol-5-yl)pyridin-3-yl]amino]-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide Synthesized in the same manner as in Example 6 using Intermediate 10 (25 mg, 0.06 mmol) and equivalent stoichiometric reagents and heating at 100° C. for 6 h. Purification by column chromatography afforded the title compound (3 mg, 8% yield).

number

[0072] Example 11 [ka] 1-[3-cyclopropyl-5-[(2-fluoro-2-methylpropyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]-3-(2,5-dimethylpyrazol-3-yl)urea To a stirred solution of 1,3-dimethyl-1H-pyrazol-5-amine (8 mg, 0.07 mmol) in MeCN (1 mL) was added TEA (20 μL, 0.14 mmol) and 1,1′-carbonyldiimidazole (11 mg, 0.07 mmol). After 1 h, intermediate 10 (25 mg, 0.07 mmol) was added. After a further 12 h, the reaction mixture was diluted with brine (20 mL), extracted with DCM (3×20 mL) and the combined organics were dried and concentrated in vacuo. Purification by column chromatography eluting with 0-30% MeOH in EtOAc followed by reverse phase column chromatography (acidic conditions) afforded the title compound (3 mg, 9% yield).

number

[0073] Example 12 [ka] Ethyl 1-[3-cyclopropyl-5-[(2-fluoro-2-methylpropyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]imidazole-4-carboxylate To a stirred solution of ethyl (Z)-3-(dimethylamino)-2-isocyano-prop-2-enoate* (10 mg, 0.06 mmol) in 1-butanol (1 mL) was added intermediate 10 (20 mg, 0.05 mmol) and the resulting mixture was heated at 150° C. for 9 h. The mixture was then concentrated in vacuo and purified by column chromatography eluting with 0-20% MeOH in EtOAc followed by reverse phase column chromatography (basic conditions) to give the title compound (4 mg, 15% yield).

number

[0074] In vitro biochemical assays: Protocol for preparation of IgE-Tb reagent 86 nmol of IgE-Fc (N265Q, N371Q) (Young et al., 1995) at 172 μM in 100 mM NaHCO3, pH 9.5 was added to 1 mg of LanthaScreen™ amine-reactive Tb chelate (ThermoFisher catalogue no. PV3583) and incubated for 16 hours at 20° C. The material was then buffer exchanged into phosphate buffered saline (137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM K2HPO4, pH 7.4) and the material was quantified and the extent of Tb conjugation determined by measuring absorbance at 280 nm and 343 nm.

[0075] The integrity of the conjugated material was determined by analytical size exclusion chromatography on a S200 HR 10x300 column (GE Healthcare).Typical conjugation ratio was 4:1=Tb:IgE-Fc. See Young RJ., Owens, RJ., MacKay GA., Chan CMW., Shi J., Hide M., Francis DM., Henry AJ., Sutton BJ., and Gould HJ (1995) Protein Engineering 8:193-199.

[0076] Preparation protocol for sFcεR1α-Y131A-AF488 reagent 400 nmol of FcεR1α (Y131A mutant) (Cook et al., 1997) at 400 μM in 100 mM NaOAc, pH 5.5 was reacted with a final concentration of 1 mM sodium periodate (in 100 mM NaOAc, pH 5.5) for 60 min at 22 °C. Oxidation was quenched by adding 40 μL of ethanediol and incubating for 60 min at 22 °C. The protein was buffer exchanged into conjugation buffer (50 mM NaHCO3, 150 mM NaCl, pH 9.5) and concentrated to 750 μM.

[0077] 175 nmol of protein was added to 1 mg of Alexa Fluor™ 488 hydrazide (Invitrogen) and incubated for 16 hours at 22° C. Sodium cyanoborohydride (100 mM in conjugation buffer) was added to a final concentration of 1 mM and incubated on ice for 60 minutes. The protein was buffer exchanged into phosphate buffered saline (137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 1.8 mM K2HPO4, pH 7.4) and the material was quantified and the extent of Alexa Fluor™ 488 conjugation was determined by measuring absorbance at 280 nm and 495 nm.

[0078] The integrity of the conjugated material was determined by analytical size-exclusion chromatography on a S200 HR 10x300 column (GE Healthcare).Typical conjugation ratios were 2:1 Alexa Fluor™ 488:sFcεR1α. See Cook JPD., Henry AJ., McDonnell JM., Owens RJ., Sutton BJ., and Gould HJ (1997) Biochemistry 36:15579-15588.

[0079] The aim was to measure IgE-Tb binding to the receptor and its inhibition by compounds using an in vitro Fluorescence Resonance Energy Transfer (FRET) assay.

[0080] reagent The FRET reagents used were IgE labeled with terbium (FRET donor) and soluble IgE receptor FcεRIα with the Y131A mutation labeled with Alexa Fluor™ 488 (FRET acceptor). Unlabeled FcεRIα was also used to generate a background control. The assay buffer consisted of 20 mM Tris pH 7.2, 150 mM NaCl and 0.002% Tween, 1% DMSO.

[0081] Assay reaction The assay was performed as follows: each assay reaction was performed in a volume of 25 μl in a 384-well half-volume plate. A 10-point compound serial dilution (3-fold) was generated in DMSO at a concentration of x50 of the final assay concentration (FAC). Compound solutions were then prepared by diluting IgE-Tb 10-fold in assay buffer. For the assay, 5 μl of diluted compound was added to 10 μl, followed by 10 μl of FcεRIα-Y131A-AF488. The FRET reagent FAC was 5 nM IgE-Tb, 25 nM FcεRIα-Y131A-AF488. Typically, the top FAC of the compound in the assay was 10 μM. The final DMSO concentration was 2%. The minimum signal (MIN) was measured by adding 5 μl of 1 μM unlabeled FcεRIα (FAC=200 nM) to the FRET reagent. The maximum FRET signal (MAX) was measured in wells containing the FRET reagent but no compound.

[0082] The assay was incubated for 2 hours at room temperature, protected from light and evaporation and gently agitated.

[0083] FRET measurements FRET measurements for each well were performed using an Envision plate reader (Perkin Elmer) by exciting at 330 nm and measuring emission at 495 / 520 nm. The FRET ratio was calculated as follows: Illumination at 520 / Illumination at 495x1000

[0084] FRET ratios were used for data analysis.

[0085] Data analysis Z' was calculated as follows (σ=standard deviation, μ=mean): 1-((3xσ MAX )+(3xσ MIN )) / (μ MAX -μ MIN )

[0086] A Z' above 0.5 was considered a good assay.

[0087] Background signal (MIN) was subtracted from all wells. The background subtracted values ​​were used to calculate the percentage inhibition by compound in each test well as follows: 100-Test well FRET ratio / MAX FRET ratio x 100

[0088] Percentage of inhibition was plotted versus compound concentration. IC50 values ​​for each compound were determined using a four-parameter logistic fit model with the XLFIT5 software package.

[0089] The results are as follows. The compounds of the present invention exhibit IC50 values ​​in the range of 19.8 nM to 5098 nM.

[0090] The table below shows the range of IC50 values ​​for each example. [Table 1] The present disclosure includes the following preferred aspects. (1) Compounds of formula (I) and pharma- ceutically acceptable salts thereof, [ka] During the ceremony, R1 and R2 are independent of each other and are as follows: Hydrogen; or NHC(O)NH-C1-6-alkyl; or NHSO 2 -C1-6-alkyl; or One or more R1 a NHC(O)NH-heteroaryl optionally substituted with Amino; C1-6-alkyl; C(O)O-C1-6-alkyl; Nitrile; One or more R1 a Heteroaryl optionally substituted with;NH-C1-6-alkyl;NH-C1-6-heterocycloalkyl;NH-C3-9-cycloalkyl; one or more R1 a heteroaryl optionally substituted with one or more groups selected from NH-heteroaryl optionally substituted with C1-6-alkyl; C1-6-hydroxyalkyl; C3-9-hydroxyheterocycloalkyl, one or more R1 a NH-heteroaryl, optionally substituted with one or more groups selected from heteroaryl, optionally substituted with NHC(O)-C1-6-alkyl; or One or more R1 a NHC(O)-heteroaryl optionally substituted with represents a group selected from R1 a But, Halogen;Nitrile;C1-6-Alkyl;C1-6-Haloalkyl;C1-6-Alkoxy;C1-6-Haloalkoxy;C(O)O-C1-6-Alkyl;C(O)OH represents a group selected from R3 is as follows: R3 a C1-6-alkyl optionally substituted with one or more groups selected from among One or more R3 a C1-3-alkanediyl-C3-6-cycloalkyl optionally substituted with One or more R3 a C1-3-alkanediyl-C3-6-heterocycloalkyl optionally substituted with One or more R3 a C3-6-heterocycloalkyl optionally substituted with; One or more R3 a C3-6-cycloalkyl optionally substituted with represents a group selected from R3 a represents a group selected from hydrogen; halogen; C1-2-alkyl; hydroxy; C1-2-alkoxy, R4 is as follows: One or more R4 a C3-6-cycloalkyl optionally substituted with a group; or one or more R4 a C1-6-alkanediyl-C3-6-cycloalkyl optionally substituted with a group; or one or more R4 a C1-6-alkanediyl-C3-6-heterocycloalkyl optionally substituted with a group represents a group selected from R4 a represents a group selected from hydroxy; halogen; C1-2-alkyl, The compounds of the above formula (I) and pharma- ceutically acceptable salts thereof. (2) R1 and R2 are each independently one of the following: Hydrogen; or C1-6-alkyl; one or more R1 a NH-heteroaryl, optionally substituted with one or more groups selected from heteroaryl, optionally substituted with Amino; C(O)O-C1-6-alkyl; Nitrile; One or more R1 a Heteroaryl optionally substituted with;NH-C1-6-alkyl;NH-C3-9-heterocycloalkyl;NH-C3-9-cycloalkyl; one or more R1a Heteroaryl optionally substituted with one or more groups selected from among NH-heteroaryl optionally substituted with represents a group selected from The compound according to (1). (3) When R1 is different from hydrogen, R2 is hydrogen; When R2 is different from hydrogen, R1 is hydrogen; The compound according to (1) or (2). (4) R3 represents C1-6-alkyl optionally substituted with a fluorine atom; The compound according to any one of (1) to (3). (5) R4 represents cyclopropyl; The compound according to any one of (1) to (4). (6) The following: 3-Cyclopropyl-N-(2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 1-[3-cyclopropyl-5-(2-methylpropylsulfamoyl)-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]-3-ethylurea; 1-[3-cyclopropyl-5-(2-methylpropylsulfamoyl)-7,8,9,10-tetrahydrobenzo[h]isoquinolin-10-yl]-3-ethylurea; 3-Cyclopropyl-7-(methanesulfonamido)-N-(2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 3-Cyclopropyl-10-(methanesulfonamido)-N-(2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 3-Cyclopropyl-N-(2-fluoro-2-methylpropyl)-7-[[6-(2-methyltetrazol-5-yl)pyridin-3-yl]amino]-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 3-Cyclopropyl-7-[3-[(2,5-dimethylpyrazol-3-yl)amino]-1,2,4-triazol-4-yl]-N-(2-fluoro-2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 3-Cyclopropyl-N-(2-fluoro-2-methylpropyl)-10-[[6-(2-methyltetrazol-5-yl)pyridin-3-yl]amino]-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; Ethyl 5-amino-1-[3-cyclopropyl-5-[(2-fluoro-2-methylpropyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]imidazole-4-carboxylate; 3-Cyclopropyl-N-(2-fluoro-2-methylpropyl)-7-[[5-(3-methyl-1,2,4-oxadiazol-5-yl)pyridin-3-yl]amino]-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 1-[3-cyclopropyl-5-[(2-fluoro-2-methylpropyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]-3-(2,5-dimethylpyrazol-3-yl)urea; Ethyl 1-[3-cyclopropyl-5-[(2-fluoro-2-methylpropyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]imidazole-4-carboxylate The compound according to (1), selected from the group consisting of (7) A pharmaceutical composition comprising the compound according to any one of (1) to (6) or a pharma- ceutically acceptable salt thereof. (8) A compound according to any one of (1) to (6) or a pharma- ceutically acceptable salt thereof for use in therapy. (9) The compound according to any one of (1) to (6) or a pharma- ceutically acceptable salt thereof for use in a method for treating or preventing a disorder caused by IgE. (10) The compound according to any one of (1) to (6) or a pharma- ceutically acceptable salt thereof for use in a method for treating or preventing allergy, non-allergic mast cell response, type 1 hypersensitivity, urticaria, or well-known sinusitis. (11) The compound according to any one of (1) to (6) or a pharma- ceutical acceptable salt thereof for use in a method for treating or preventing airway constriction in asthma, local inflammation in eczema, increased mucus secretion in allergic rhinitis, urticaria, or increased vascular permeability. (12) The compound or pharma- ceutical acceptable salt thereof according to any one of (1) to (6) for use in a method for treating or preventing eosinophilic granulomatosis with polyangiitis (also known as "Churg-Strauss syndrome"), aspirin-exacerbated respiratory disease, or cutaneous T-cell lymphoma. (13) A pharmaceutical composition comprising a compound of formula (I) according to any one of (1) to (6) or a pharma- ceutically acceptable salt thereof together with a pharma- ceutically acceptable carrier. (14) Use of the compound according to any one of (1) to (6) or a pharma- ceutically acceptable salt thereof for the manufacture of a pharmaceutical product. (15) Use of the compound according to any one of (1) to (6) or a pharma- ceutical acceptable salt thereof for the manufacture of a pharmaceutical for treating and / or preventing a disorder caused by IgE. (16) Use of a compound according to any one of (1) to (6) or a pharma- ceutically acceptable salt thereof for the manufacture of a medicament for treating and / or preventing allergy, non-allergic mast cell response, type 1 hypersensitivity, urticaria or well-known sinusitis. (17) A method for treating or preventing allergy, non-allergic mast cell response, type 1 hypersensitivity, urticaria, well-known sinusitis, eosinophilic granulomatosis with polyangiitis (also known as "Churg-Strauss syndrome"), aspirin-exacerbated respiratory disease or cutaneous T-cell lymphoma, comprising administering to a patient a therapeutically effective amount of a compound according to any one of (1) to (6) or a pharma- ceutical acceptable salt thereof.

Claims

1. Compounds of formula (I) and pharma- ceutically acceptable salts thereof, 【Chemistry 1】 During the ceremony, R1 and R2 are independently selected from the following: Hydrogen; or NHC(O)NH-C1-6-alkyl; or N.H.S.O. ２ -C1-6-alkyl; or One or more R1 ａ NHC(O)NH-heteroaryl optionally substituted with Amino; C1-6-alkyl; C(O)O-C1-6-alkyl; nitrile; one or more R1 ａ Heteroaryl optionally substituted with; NH-C1-6-alkyl; NH-C1-6-heterocycloalkyl; NH-C3-9-cycloalkyl; one or more R1 ａ heteroaryl optionally substituted with one or more groups selected from NH-heteroaryl optionally substituted with C1-6-alkyl; C1-6-hydroxyalkyl; C3-9-hydroxyheterocycloalkyl; one or more R1 ａ NH-heteroaryl, optionally substituted with one or more groups selected from heteroaryl, optionally substituted with NHC(O)-C1-6-alkyl; or One or more R1 ａ NHC(O)-heteroaryl optionally substituted with represents a group selected from R1 ａ But, Halogen; nitrile; C1-6-alkyl; C1-6-haloalkyl; C1-6-alkoxy; C1-6-haloalkoxy; C(O)O-C1-6-alkyl; C(O)OH; represents a group selected from R3 is as follows: R3 ａ C1-6-alkyl optionally substituted with one or more groups selected from among One or more R ａ C1-3-alkanediyl-C3-6-cycloalkyl optionally substituted with One or more R ａ C1-3-alkanediyl-C3-6-heterocycloalkyl optionally substituted with One or more R ａ C3-6-heterocycloalkyl optionally substituted with One or more R ａ C3-6-cycloalkyl optionally substituted with represents a group selected from R3 ａ is hydrogen; halogen; C1-2-alkyl; hydroxy; C1-2-alkoxy; represents a group selected from R4 is the following: One or more R4 ａ C3-6-cycloalkyl optionally substituted with a group; or one or more R4 ａ C1-6-alkanediyl-C3-6-cycloalkyl optionally substituted with a group; or one or more R4 ａ C1-6-alkanediyl-C3-6-heterocycloalkyl optionally substituted with a group represents a group selected from R4 ａ represents a group selected from hydroxy; halogen; C1-2-alkyl, The compound of the above formula (I) and its pharma- ceutically acceptable salts.

2. R1 and R2 are independently selected from the following: Hydrogen; or C1-6-alkyl; one or more R1 ａ NH-heteroaryl, optionally substituted with one or more groups selected from heteroaryl, optionally substituted with Amino; C(O)O-C1-6-alkyl; nitrile; one or more R1 ａ Heteroaryl optionally substituted with; NH-C1-6-alkyl; NH-C3-9-heterocycloalkyl; NH-C3-9-cycloalkyl; one or more R1 ａ Heteroaryl optionally substituted with one or more groups selected from among NH-heteroaryl optionally substituted with represents a group selected from The compound of claim 1.

3. When R1 is different from hydrogen, R2 is hydrogen; When R2 is different from hydrogen, R1 is hydrogen; 3. A compound according to claim 1 or 2.

4. R3 represents C1-6-alkyl optionally substituted with a fluorine atom; The compound according to any one of claims 1 to 3.

5. R4 represents cyclopropyl; The compound according to any one of claims 1 to 4.

6. below, 3-cyclopropyl-N-(2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 1-[3-cyclopropyl-5-(2-methylpropylsulfamoyl)-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]-3-ethylurea; 1-[3-cyclopropyl-5-(2-methylpropylsulfamoyl)-7,8,9,10-tetrahydrobenzo[h]isoquinolin-10-yl]-3-ethylurea; 3-cyclopropyl-7-(methanesulfonamido)-N-(2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 3-cyclopropyl-10-(methanesulfonamido)-N-(2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 3-cyclopropyl-N-(2-fluoro-2-methylpropyl)-7-[[6-(2-methyltetrazol-5-yl)pyridin-3-yl]amino]-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 3-cyclopropyl-7-[3-[(2,5-dimethylpyrazol-3-yl)amino]-1,2,4-triazol-4-yl]-N-(2-fluoro-2-methylpropyl)-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 3-cyclopropyl-N-(2-fluoro-2-methylpropyl)-10-[[6-(2-methyltetrazol-5-yl)pyridin-3-yl]amino]-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; Ethyl 5-amino-1-[3-cyclopropyl-5-[(2-fluoro-2-methylpropyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]imidazole-4-carboxylate; 3-cyclopropyl-N-(2-fluoro-2-methylpropyl)-7-[[5-(3-methyl-1,2,4-oxadiazol-5-yl)pyridin-3-yl]amino]-7,8,9,10-tetrahydrobenzo[h]isoquinoline-5-sulfonamide; 1-[3-cyclopropyl-5-[(2-fluoro-2-methylpropyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]-3-(2,5-dimethylpyrazol-3-yl)urea; or Ethyl 1-[3-cyclopropyl-5-[(2-fluoro-2-methylpropyl)sulfamoyl]-7,8,9,10-tetrahydrobenzo[h]isoquinolin-7-yl]imidazole-4-carboxylate 2. The compound of claim 1 selected from:

7. A pharmaceutical composition comprising the compound according to any one of claims 1 to 6 or a pharma- ceutically acceptable salt thereof.

8. A pharmaceutical composition for use in therapy, comprising a compound according to any one of claims 1 to 6 or a pharma- ceutically acceptable salt thereof.

9. A pharmaceutical composition for use in a method for treating or preventing a disorder caused by IgE, comprising a compound according to any one of claims 1 to 6 or a pharma- ceutically acceptable salt thereof.

10. A pharmaceutical composition for use in a method for treating or preventing allergy, non-allergic mast cell response, type 1 hypersensitivity, urticaria, or sinusitis, comprising a compound according to any one of claims 1 to 6 or a pharma- ceutically acceptable salt thereof.

11. A pharmaceutical composition for use in a method for treating or preventing airway constriction in asthma, local inflammation in eczema, increased mucus secretion in allergic rhinitis, urticaria, or increased vascular permeability, comprising a compound according to any one of claims 1 to 6 or a pharma- ceutically acceptable salt thereof.

12. A pharmaceutical composition for use in a method for treating or preventing Eosinophilic Granulomatosis with Polyangiitis (Churg-Strauss Syndrome), Aspirin-Aggravated Respiratory Disease, or Cutaneous T-Cell Lymphoma, comprising a compound according to any one of claims 1 to 6 or a pharma-ceutical acceptable salt thereof.

13. A pharmaceutical composition comprising a compound of formula (I) according to any one of claims 1 to 6, or a pharma- ceutically acceptable salt thereof, together with a pharma- ceutically acceptable carrier.

14. Use of a compound according to any one of claims 1 to 6 or a pharma- ceutically acceptable salt thereof for the manufacture of a medicament.

15. Use of a compound according to any one of claims 1 to 6 or a pharma- ceutically acceptable salt thereof for the manufacture of a medicament for the treatment and / or prevention of disorders caused by IgE.

16. Use of a compound according to any one of claims 1 to 6 or a pharma- ceutically acceptable salt thereof for the manufacture of a medicament for the treatment and / or prevention of allergy, non-allergic mast cell response, type 1 hypersensitivity, urticaria or sinusitis.

17. A pharmaceutical composition comprising the compound according to any one of claims 1 to 6 or a pharma- ceutical acceptable salt thereof, A pharmaceutical composition for use in a method for treating or preventing allergy, non-allergic mast cell response, type 1 hypersensitivity, urticaria, sinusitis, eosinophilic granulomatosis with polyangiitis (Churg-Strauss syndrome), aspirin-exacerbated respiratory disease or cutaneous T-cell lymphoma, comprising administering to a patient a therapeutically effective amount of the compound or a pharma- ceutical acceptable salt thereof.

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