Novel sulfonamide derivatives

Abstract

The invention relates to a compound of formula (I), wherein A1 and A2 and R1 to R3 are defined as in the description and in the claims. The compound of formula (I) can be used as a medicament.

Description

[0001] F. Hoffmann-La Roche AG, CH-4070 Basel, Switzerland

[0002] Case: P39772

[0003] Novel sulfonamide derivatives

[0004] The present invention relates to organic compounds useful for therapy and / or prophylaxis in a mammal, and in particular to compounds that are TMEM175 enhancers. The compound of formula (I) is particularly useful in the treatment or prophylaxis of neurodegenerative diseases, fibrotic disorders or inflammatory disorders, more specifically, synucleinopathies, Parkinson’s disease, Dementia with Lewy body disease, REM-sleep behavior disorder, amyotrophic lateral sclerosis and lysosomal storage disorders.

[0005] The invention relates in particular to a compound of formula (I)

[0006]

[0007] wherein

[0008] one of A1and A2is nitrogen and the other one is carbon;

[0009] R1and R2together with the carbon and nitrogen atoms to which they are attached form pyrrolyl, susbtitutued pyrrolyl, pyrrazolyl or substituted pyrrazolyl, wherein substituted pyrrolyl and substituted pyrrazolyl are pyrrolyl and

[0010] DP / 24.09.25 pyrrazolyl substituted with one substituent selected from halogen, alkyl, cycloalkyl, haloalkyl or cycloalkoxy; and

[0011] R3is hydrogen or halogen;

[0012] or a pharmaceutically acceptable salt thereof.

[0013] TMEM175, or Transmembrane Protein 175, is a cation-selective lysosomal ion channel shown to conduct both potassium and proton currents (Hu M, Li P, Wang C, Feng X, Geng Q, Chen W, Marthi M, Zhang W, Gao C, Reid W, Swanson J, Du W, Hume RI, Xu H. Parkinson's disease-risk protein TMEM175 is a proton- activated proton channel in lysosomes. Cell 185: 2292-2308. e20, 2022). TMEM175 contributes to maintenance of the lysosomal membrane potential and stabilization of the lysosomal pH gradient both of which are crucial for proper lysosomal function (Cang, C.; Aranda, K.; Seo, Y.-J.; Gasnier, B.; Ren, D. TMEM175 Is an Organelle K+Channel Regulating Lysosomal Function. Cell 2015, 162, 1101-1112). Lysosomal catabolic activity is dependent upon a variety of enzymes confined to the lysosomal lumen. The majority of these enzymes exhibit pH dependent activity and are optimally functional at an acidic pH between 4.5 to 5.0 (Mellman I. Organelles observed: lysosomes. Science 244: 853-854, 1989. doi:

[0014] 10.1126 / science.244.4906.853). TMEM175 acts as a proton-activated channel that will preferentially expel protons from the lysosomal lumen at pH <4.5. At pH > 5.0, protondependent activation is reduced and the proton flux from the lumen to the cytosol is decreased, helping to maintain the optimal pH range for normal lysosomal function (Hu et al 2022, op. cit.). In mice, knockout of TMEM175 has been shown to reduce the enzymatic degradation of BSA and more specifically, the pH dependent activity of cathepsins B and D are reduced (Wie J, Liu Z, Song H, Tropea TF, Yang L, Wang H, Liang Y, Cang C, Aranda K, Lohmann J, Yang J, Lu B, Chen-Plotkin AS, Luk KC, Ren D. A growth-factor-activated lysosomal K+channel regulates Parkinson’s pathology. Nature 591: 431-437, 2021. doi: 10.1038 / s41586-021-03185-z).

[0015] TMEM175 has been identified as a novel lysosomal ‘leak-like’ potassium channel representing the major K+permeability of lysosomes (Cang et al., 2015, op. cit.).

[0016] TMEM175 is unique among other canonical potassium channels for several reasons: (1) no sequence homology with other tetrameric potassium channels, (2) it possesses a unique structure (Lee C, Guo J, Zeng W, Kim S, She J, Cang C, Ren D, Jiang Y. The lysosomal potassium channel TMEM175 adopts a novel tetrameric architecture. Nature. 2017 Jul 27;547 (7664):472-475; Oh S, Paknejad N, Hite RK. Gating and selectivity mechanisms for the lysosomal K+channel TMEM175. Elife. 2020 Mar 31;9:e53430) and assembles as a homodimer of 2 homologous copies of a six-transmembrane helix domain, where (3) the transmembrane helix 1 and 7 serve as the pore forming helix and (4) it is localized at lysosomal and endosomal membranes. Furthermore, TMEM175 is not blocked by cesium (Cs+) as are the majority of potassium channels, but instead can conduct Cs+with a similar permeability as K+. It has been shown to be blocked by 4-AP. Interestingly, TMEM175 is activated by growth factors via AKT in a kinase independent fashion (Wie et al., 2021, op. cit.).

[0017] In knockout studies, it has been shown that TMEM175 regulates lysosomal membrane potential, pH stability, and organelle fusion via potassium conductance on lysosomal and endosomal membranes (Cang et al, 2015, op. cit.).

[0018] TMEM175 with the M393T risk mutation is thought to be a (partial) loss of function mutation because measured currents are reduced (Wie et al., 2021, op. cit.) and the effect on lysosomal pH resembles that of knockout cells with a more alkaline pH during starvation. Furthermore, lysosomal localization of TMEM175 M393T might be reduced as compared to wildtype TMEM175 (Jinn S, Blauwendraat C, Toolan D, Gretzula CA, Drolet RE, Smith S, Nalls MA, Marcus J, Singleton AB, Stone DJ. Functionalization of the TMEM175 p. M393T variant as a risk factor for Parkinson disease. Hum Mol Genet. 2019 Oct 1;28( 19):3244-3254). In contrast to wildtype TMEM175, M393T overexpression does not reduce PFF-induced phospho a-synuclein (Jinn et al., 2019, op. cit.).

[0019] The Q65P variant is considered as a gain-of-function mutation under stress. When cells are starved, it leads to a reduction in TMEM175 current that is dependent and gated by AKT. In the Q65P mutant, the starvation-induced reduction in K+ current is delayed and early during starvation, the Q65P TMEM175 lysosomes carry a higher current than wildtype TMEM175. Taken together, these data suggest that the Q65P mutation represents a gain-of-function mutation under stress (Wie et al., 2021, op. cit.).

[0020] Dysfunction in lysosomal activities is linked to various neurodegenerative diseases, including Parkinson's disease, suggesting a potential role for TMEM175 in neurodegenerative disorders (Bahr B. A., Bendiske J. The neuropathogenic contributions of lysosomal dysfunction. J. Neurochem. 2002;83:481-489). Consistent with this hypothesis, TMEM175 has been identified by genome- wide association studies as a genetic risk factor for Parkinson’s disease (PD) (Hopfner F, Mueller SH, Szymczak S, Junge O, Tittmann L, May S, Lohmann K, Grallert H, Lieb W, Strauch K, Miiller-Nurasyid M, Berger K, Schormair B, Winkelmann J, Mollenhauer B, Trenkwalder C, Maetzler W, Berg D, Kasten M, Klein C, Hbglinger GU, Gasser T, Deuschi G, Franke A, Krawczak M, Dempfle A, Kuhlenbaumer G. Rare variants in specific lysosomal genes are associated with Parkinson’s disease. Mov Disord 35: 1245-1248, 2020. doi: 10.1002 / mds.28037), Rapid-eye-movement (REM) sleep behavior disorder (RBD) (Krohn L, et al. Genetic, structural, and functional evidence link TMEM175 to synucleinopathies. Ann. Neurol. 2020;87:139- 153. doi: 10.1002 / ana.25629) and Dementia with Lewy Bodies (Chia R, Sabir MS, Bandres-Ciga S, et al.; American Genome Center. Genome sequencing analysis identifies new loci associated with Lewy body dementia and provides insights into its genetic architecture. Nat Genet. 2021;53(3):294-303). Multiple coding variants of TMEM175 have been identified and one variant, TMEM175 M393T, is associated with an increased risk and earlier onset of Parkinson’s disease (Blauwendraat C., Heilbron K., Vallerga C. L., Bandres-Ciga S., von Coelln R., Pihlstrom L., Simon-Sanchez J., Schulte C., Sharma M., Krohn L. et al. (2019) Parkinson's disease age at onset genome- wide association study: defining heritability, genetic loci, and alpha- synuclein mechanisms. Mov. Disord., 34, 866-875). Functional analysis of M393T indicated that M393T is a partial loss of function allele, suggesting that enhancement of TMEM175 activity could provide a therapeutic benefit in Parkinson’s disease (Jinn S, Drolet RE, Cramer PE, Wong AH, Toolan DM, Gretzula CA, Voleti B, Vassileva G, Disa J, Tadin-Strapps M, Stone DJ. TMEM175 deficiency impairs lysosomal and mitochondrial function and increases alpha-synuclein aggregation. Proc Natl Acad Sci USA 114: 2389-2394, 2017. doi:

[0021] 10.1073 / pnas.1616332114).

[0022] TMEM175 has also been identified as a comorbid gene between Amyotrophic lateral sclerosis (ALS) and Parkinson’s disease (Tian Y, Ma G, Li H, Zeng Y, Zhou S, Wang X, Shan S, Xu Y, Xiong J, Cheng G. Shared Genetics and Comorbid Genes of Amyotrophic Lateral Sclerosis and Parkinson's Disease. Mov Disord. 2023 Oct;38(10):1813-1821. doi: 10.1002 / mds.29572. Epub 2023 Aug 3) and as a shared genetic risk loci among Alzheimer’s disease related dementias, Parkinson’s disease and Amyotrophic lateral sclerosis (Wainberg, M., Andrews, S. J. & Tripathy, S J. Shared genetic risk loci between Alzheimer’s disease and related dementias, Parkinson’s disease, and amyotrophic lateral sclerosis. Alz Res Therapy 15, 113 (2023)). Further, in a proteome-wide association study (PWAS) for Amyotrophic lateral sclerosis, TMEM175 was identified (Ma, Y., Jia, T., Qin, F. et al. Abnormal Brain Protein Abundance and Cross-tissue mRNA Expression in Amyotrophic Lateral Sclerosis. Mol Neurobiol 61, 510-518 (2024)) and lysosomal dysfunction is an important pathogenic disease mechanism in ALS (Root J, Merino P, Nuckols A, Johnson M, Kukar T. Lysosome dysfunction as a cause of neurodegenerative diseases: Lessons from frontotemporal dementia and amyotrophic lateral sclerosis.

[0023] Neurobiol Dis. 2021 Jul;154:105360). Therefore, TMEM175 enhancers may address the underlying disease biology in ALS.

[0024] Lysosomal storage disorders (Platt FM, d'Azzo A, Davidson BL, Neufeld EF, Tifft CJ. Publisher Correction: Lysosomal storage diseases. Nat Rev Dis Primers. 2019 May 17;5( 1):34) are characterized by accumulation of substrates in excess in lysosomes, often resulting from defects in lysosomal function. Since TMEM175 knockout leads to a reduced lysosomal hydrolysis activity and degradation activity (Hu et al 2022, op. cit.), a TMEM175 enhancer may increase lysosomal degradation capacity and thereby reduce pathological storage in lysosomal storage disorders.

[0025] In the present description the term “alkyl”, alone or in combination, signifies a straight-chain or branched-chain alkyl group with 1 to 8 carbon atoms, particularly a straight or branched-chain alkyl group with 1 to 6 carbon atoms and more particularly a straight or branched-chain alkyl group with 1 to 4 carbon atoms. Examples of straightchain and branched-chain Ci-Cs alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert.-butyl, the isomeric pentyls, the isomeric hexyls, the isomeric heptyls and the isomeric octyls, particularly methyl, ethyl, propyl, butyl and pentyl more particularly methyl, ethyl, propyl, isopropyl, isobutyl, tert.-butyl and isopentyl. Particular examples of “alkyl” are methyl and ethyl.

[0026] The term “cycloalkyl”, alone or in combination, signifies a cycloalkyl ring with 3 to 8 carbon atoms and particularly a cycloalkyl ring with 3 to 6 carbon atoms. Examples of cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl, cycloheptyl and cyclooctyl. A particular “cycloalkyl” is cyclopropy.

[0027] The term “oxy”, alone or in combination, signifies the -O- group.

[0028] The term “cycloalkyloxy” or “cycloalkoxy”, alone or in combination, signifies a group of the formula cycloalkyl-O- in which the term "cycloalkyl" has the previously given significance, such as cyclopropoxy and cyclobutoxy. A particular example of “cycloalkoxy” is cyclopropoxy.

[0029] The terms “halogen” or “halo”, alone or in combination, signifies fluorine, chlorine, bromine or iodine and particularly fluorine, chlorine or bromine, more particularly fluorine and chlorine. The term “halo”, in combination with another group, denotes the substitution of said group with at least one halogen, particularly substituted with one to five halogens, particularly one to three halogens, i.e. one, two or three halogens. Particular halogens are fluorine, bromine and chlorine.

[0030] The term “haloalkyl”, alone or in combination, denotes an alkyl group substituted with at least one halogen, particularly substituted with one to five halogens, particularly one to three halogens. Particular “haloalkyl” are difluoromethyl, difluoroethyl and trifluoroethyl.

[0031] The term “pharmaceutically acceptable salts” refers to those salts which retain the biological effectiveness and properties of the free bases or free acids, which are not biologically or otherwise undesirable. The salts are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, particularly hydrochloric acid, and organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcystein. In addition these salts may be prepared from addition of an inorganic base or an organic base to the free acid. Salts derived from an inorganic base include, but are not limited to, the sodium, potassium, lithium, ammonium, calcium, magnesium salts. Salts derived from organic bases include, but are not limited to salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, lysine, arginine, N-ethylpiperidine, piperidine, polyamine resins. The compound of formula (I) can also be present in the form of zwitterions. Particularly preferred pharmaceutically acceptable salts of compounds of formula (I) are the salts of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid and methanesulfonic acid.

[0032] If one of the starting materials or compounds of formula (I) contain one or more functional groups which are not stable or are reactive under the reaction conditions of one or more reaction steps, appropriate protecting groups (as described e.g. in “Protective Groups in Organic Chemistry” by T. W. Greene and P. G. M. Wuts, 3rdEd., 1999, Wiley, New York) can be introduced before the critical step applying methods well known in the art. Such protecting groups can be removed at a later stage of the synthesis using standard methods described in the literature. Examples of protecting groups are tert-butoxycarbonyl (Boc), 9-fluorenylmethyl carbamate (Fmoc), 2-trimethylsilylethyl carbamate (Teoc), carbobenzyloxy (Cbz) and p-methoxybenzyloxycarbonyl (Moz).

[0033] The compound of formula (I) can contain several asymmetric centers and can be present in the form of optically pure enantiomers, mixtures of enantiomers such as, for example, racemates, mixtures of diastereoisomers, diastereoisomeric racemates or mixtures of diastereoisomeric racemates.

[0034] The term “asymmetric carbon atom” means a carbon atom with four different substituents. According to the Cahn-Ingold-Prelog Convention an asymmetric carbon atom can be of the “R” or “S” configuration.

[0035] The invention further relates to:

[0036] A compound of formula (I) wherein R1and R2together with the carbon and nitrogen atoms to which they are attached form pyrrolyl, susbtitutued pyrrolyl, pyrrazolyl or substituted pyrrazolyl, wherein substituted pyrrolyl and substituted pyrrazolyl are pyrrolyl and pyrrazolyl substituted with one substituent selected from chlorine, bromine, methyl, cyclopropyl, difluoromethyl, difluoroethyl, trifluoroethyl or cyclopropyloxy;

[0037] A compound of formula (I) wherein R1and R2together with the carbon and nitrogen atoms to which they are attached form

[0038]

[0039] A compound of formula (I) wherein R3is hydrogen or fluorine.

[0040] The invention further relates to a compound selected from

[0041] N- [7 -bromo-2- [methoxy(phenyl)methyl] -4-oxo-pyrrolo [2, 1 -f] [ 1,2,4] triazin- 3-yl] -4-methyl-benzenesulfonamide;

[0042] N-[2-[methoxy(phenyl)methyl]-4-oxo-pyrrolo[2,l-f][l,2,4]triazin-3-yl]-4-methyl-benzenesulfonamide;

[0043] N- [2- [methoxy(phenyl)methyl] -7 -methyl-4-oxo-pyrrolo [2, 1 -f] [ 1,2,4] triazin-3-yl] -4-methyl-benzenesulfonamide;

[0044] N- [7 -cyclopropyl-2- [methoxy(phenyl)methyl] -4-oxo-pyrrolo [2, 1 -f] [ 1,2,4] triazin-3-yl] -4-methyl-benzenesulfonamide; and

[0045] N-(8-chloro-2-(methoxy(phenyl)methyl)-4-oxopyrazolo[l,5-a][l,3,5]triazin-3(4H)-yl)-4-methylbenzenesulfonamide;

[0046] or a pharmaceutically acceptable salt thereof.

[0047] The invention relates in particular to a compound of formula (I-R)

[0048]

[0049] wherein A1and A2and R1to R3are as defined above;

[0050] or a pharmaceutically acceptable salt thereof.

[0051] The following abreviations are used in the present description.

[0052] DMF is N, N-dimethylformamide;

[0053] HPLC is high pressure liquid chromatography;

[0054] LiHMDS is lithium bis(trimethylsilyl)amide;

[0055] MeCN is acetonitrile;

[0056] NBS is N-bromo succinimide;

[0057] Pd(dppf)Cl2is 1,1'-bis(di-tert-butylphosphino)ferrocene palladium dichloride; FA is formic acid;

[0058] TCFH is tetramethylchloroformamidinium hexafluorophosphate;

[0059] NMI is N-Methylimidazole;

[0060] RT is room temperature;

[0061] TLC is thin-layer chromatography;

[0062] EDCI is 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide;

[0063] THF is tetrahydrofuran;

[0064] TMSCN is trimethylsilyl cyanide; CMV is cytomegalovirus;

[0065] FBS is fetal bovine serum;

[0066] HEPES is 4-(2-hydroxyethyl)-l -piperazine ethanesulfonic acid;

[0067] NMDG is N-methyl-D-glucamine diatrizoate;

[0068] EGTA is ethylene glycol-bis(beta- aminoethyl ether)-N, N, N’, N’ -tetraacetic acid;

[0069] DPBS is Dulbecco’s phosphate-buffered saline;

[0070] mV is millivolt;

[0071] DCPIB is 4-[(2-Butyl-6,7-dichloro-2-cyclopentyl-2,3-dihydro- 1-oxo- lH-inden-5-yl)oxy]butanoic acid; and

[0072] 4-AP is 4- aminopyridine.

[0073] The compound of formula (I) can be generated using any of the reaction conditions described below, and the decision on which route to use is based on intermediate availability.

[0074] For more information on the general procedures, please refer to the embodiments relating to the process of making the compound of formula (I) as described herein.

[0075]

[0076] Scheme 1: Synthesis of the compound of Formula (I) having formula 11

[0077] In scheme 1, R is hydrogen, halogen and alkyl.

[0078] Treatment of amino-pyrrole carboxamide 1 with acid 2 using EDO in DCM or TCFH, NMI in DMF at RT leads to hydrazide intermediate 3 (scheme 1), which can then undergo cyclization to intermediate 4 by treatment with KOH in water and ethanol at 100°C for 2 hrs. Subsequent treatment with amino diphenylphosphinate in the presence of potassium tert-butoxide in THF or DMF leads to amino intermediate 5, which can then be reacted with a sulfonyl chloride in presence of LiHMDS in THF at -78 °C to afford the desired sulfonamide of formula II.

[0079] TMSCN, BF3·Et2O MeOH, HCl in Et2O rt, o / n 0 °C, o / n 6 7

[0080]

[0081] Scheme 2: Synthesis of the compound of Formula (I) having formula 111

[0082] Treatment of dimethoxytoluene 6 with TMSCN and BF3 in ether at RT leads to the formation of phenylacetonitrile 7, which is then converted to the corresponding phenylethanimidate 8 with MeOH and HCI. Subsequent treatment with amino-pyrazole in presence of NaOAc in acetonitrile affords intermediate 9, which can then be cyclized upon treatment with dimethyl carbonate and NaOAc in ethanol at 80°C to form intermediate 10. Treatment with diphenylphosphinate and potassium tert-butoxide at 0°C leads to amino intermediate 11, which can then be reacted with sulfonyl chloride in the presence of LiHMDS in THF at -78°C to afford the desired sulfonamide of formula III.

[0083] The invention thus also relates to a process for the preparation of a compound of formula (I) as defined above comprising the reaction of a compound of formula (A)

[0084]

[0085] in the presence of para-toluenesulfonyl chloride and a base, wherein A1, A2and R1to R3are as defined above.

[0086] In the process of the invention, the base is advantageously LiHMDS.

[0087] The process of the invention can be carried out in THF. It can advantageously be performed at a temperature of around -78°C.

[0088] The invention further relates to:

[0089] A compound of formula (I), when manufactured according to a process of the invention;

[0090] A compound of formula (I) for use as therapeutically active substance;

[0091] A pharmaceutical composition comprising a compound of formula (I) and a therapeutically inert carrier;

[0092] The use of a compound of formula (I) for the treatment or prophylaxis of synucleinopathies, Parkinson’s disease, Dementia with Lewy body disease, REM-sleep behavior disorder, amyotrophic lateral sclerosis or lysosomal storage disorders;

[0093] The use of a compound of formula (I) for the preparation of a medicament for the treatment or prophylaxis of synucleinopathies, Parkinson’s disease, Dementia with Lewy body disease, REM-sleep behavior disorder, amyotrophic lateral sclerosis or lysosomal storage disorders; A compound of formula (I) for use in the treatment or prophylaxis of synucleinopathies, Parkinson’s disease, Dementia with Lewy body disease, REM-sleep behavior disorder, amyotrophic lateral sclerosis or lysosomal storage disorders; and

[0094] A method for the treatment or prophylaxis of synucleinopathies, Parkinson’s disease, Dementia with Lewy body disease, REM-sleep behavior disorder, amyotrophic lateral sclerosis or lysosomal storage disorders, which method comprises administering an effective amount of a compound of formula (I) to a patient in need thereof.

[0095] Another embodiment of the invention provides pharmaceutical compositions or medicaments containing the compounds of the invention and a therapeutically inert carrier, diluent or excipient, as well as methods of using the compounds of the invention to prepare such compositions and medicaments. In one example, compounds of formula (I) may be formulated by mixing at ambient temperature at the appropriate pH, and at the desired degree of purity, with physiologically acceptable carriers, i.e., carriers that are non-toxic to recipients at the dosages and concentrations employed into a galenical administration form. The pH of the formulation depends mainly on the particular use and the concentration of compound, but preferably ranges anywhere from about 3 to about 8. In one example, a compound of formula (I) is formulated in an acetate buffer, at pH 5. In another embodiment, the compounds of formula (I) are sterile. The compound may be stored, for example, as a solid or amorphous composition, as a lyophilized formulation or as an aqueous solution.

[0096] Compositions are formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners.

[0097] The compounds of the invention may be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural and intranasal, and, if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.

[0098] The compounds of the present invention may be administered in any convenient administrative form, e.g., tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain components conventional in pharmaceutical preparations, e.g., diluents, carriers, pH modifiers, sweeteners, bulking agents, and further active agents.

[0099] A typical formulation is prepared by mixing a compound of the present invention and a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, e.g., Ansel, Howard C., et al., Ansel’s Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R., et al. Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. The formulations may also include one or more buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present invention or pharmaceutical composition thereof) or aid in the manufacturing of the pharmaceutical product (i.e., medicament).

[0100] The invention will now be illustrated by the following examples which have no limiting character. Examples

[0101] Example 1: N-[7-bromo-2-[methoxy(phenyl)methyl]-4-oxo-pyrrolo[2,l-f][l,2,4]triazin-3-yl]-4-methyl-benzenesulfonamide

[0102]

[0103] To a stirred solution of N-[2-[methoxy(phenyl)methyl]-4-oxo-pyrrolo[2,l-f][l,2,4]triazin-3-yl]-4-methyl-benzenesulfonamide (Example 2, 100 mg, 0.23 mmol, 1.0 equiv) in AcOH (2 mL) were added NBS (42 mg, 0.23 mmol, 1.0 equiv) in portions at 0 °C under nitrogen atmosphere. The resulting mixture was stirred at 0 °C for 1 h under nitrogen atmosphere. The reaction was quenched with Water at room temperature. The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC with the following conditions (Column: Xselect CSH C18 OBD Column 30* 150mm 5μm, n; Mobile Phase A: Water(0.1% formic acid), Mobile Phase B: acetonitrile; Flow rate: 60 mL / min; Gradient: 5% B to 5% B in 1.5 min, 5% B to 46% B in 2 min, 46% to 63% in 12 min; Wave Length: 254nm / 220nm; RT=12.62min) to afford N-[7-bromo-2-[methoxy(phenyl)methyl]-4-oxo-pyrrolo[2, 1 -f] [ 1,2,4]triazin-3-yl]-4-methyl-benzenesulfonamide (22 mg, 18.55% yield) as a white solid. LCMS (ESI): [M-H]+:

[0104] 501.00. ’H NMR (400 MHz, DMSO-d6) δ 11.49 (d, J= 154.1 Hz, 1H), 7.71 (d, J= 8.0 Hz, 2H), 7.39 (d, J= 8.2 Hz, 7H), 7.00 (d, J= 4.6 Hz, 1H), 6.84 - 6.69 (m, 1H), 5.74 (d, J = 11.0 Hz, 1H), 3.39 (s, 3H), 2.41 (s, 3H).

[0105] Example 2: N-[2-[methoxy(phenyl)methyl]-4-oxo-pyrrolo[2,l-f][ l,2,4]triazin-3-yl]-4-methyl-benzenesulfonamide

[0106]

[0107] a) 1 -(2-methoxy-2-phenylacetamido)pyrrole-2-carboxamide

[0108] To a stirred solution of l-aminopyrrole-2-carboxamide (1.0 g, 7.99 mmol, 1.0 equiv) and methoxy (phenyl) acetic acid (2.7 g, 15.98 mmol, 2.0 equiv) in dichloromethane (10 mL) were added EDO (2.5 g, 15.98 mmol, 2.0 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at room temperature for overnight under nitrogen atmosphere. The reaction was quenched with water at room temperature. The resulting mixture was extracted with dichloromethane (2 xlOO mL). The combined organic layers were washed with brine (2 x 100 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with petroleum ether / ethyl acetate (1:1) to afford l-(2-methoxy-2-phenylacetamido)pyrrole-2-carboxamide (600 mg, 27.47% yield) as a white solid. LC-MS (ESI, m / z): [M+ H]+: 274.0

[0109] b) 2- [methoxy (phenyl)methyl] -377-pyrrolo [ 2, 1 - / ] [ 1,2,4] triazin-4-one

[0110] To a stirred solution of l-(2-methoxy-2-phenylacetamido)pyrrole-2-carboxamide (600 mg, 2.19 mmol, 1.0 equiv) and water (4 mL) in EtOH (4 mL) was added KOH (369 mg, 6.58 mmol, 3.0 equiv) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 100 °C for 2 h under nitrogen atmosphere. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, acetonitrile in Water (0.1% formic acid), 10% to 80% gradient in 30 min; detector, UV 254 nm. This resulted in 2-[methoxy(phenyl)methyl]-377-pyrrolo[2,l- / ][l,2,4]triazin-4-one (190 mg, 33.90% yield) as a white solid. LC-MS (ESI, m / z): [M+ H]+: 256.0

[0111] c) 3-amino-2-[methoxy(phenyl)methyl]pyrrolo[2,l- / |[l,2,4]triazin-4-one

[0112] A solution of 2-[methoxy(phenyl)methyl]-377-pyrrolo[2,l- / |[l,2,4]triazin-4-one (170 mg, 0.66 mmol, 1.0 equiv) in THF (2 mL) was treated with t-BuOK (224 mg, 1.99 mmol, 3.0 equiv) at room temperature for 5 min under nitrogen atmosphere followed by the addition of amino diphenylphosphinate (233 mg, 0.99 mmol, 1.5 equiv) in portions at 0 °C. The reaction was quenched with water at room temperature. The resulting mixture was extracted with ethyl acetate (2 x 50 mL). The combined organic layers were washed with brine (2 x 100 mL), dried over anhydrous Mg2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, acetonitrile in Water (0.1% formic acid), 10% to 80% gradient in 30 min; detector, UV 254 nm. This resulted in 3-amino-2-[methoxy(phenyl)methyl]pyrrolo[2,l- / |[l,2,4]triazin-4-one (100 mg, 55.56% yield) as a white solid. LC-MS (ESI, m / z): [M+ H]+: 271.0

[0113] d) N- [2- [methoxy(phenyl)methyl] -4-oxo-pyrrolo [2, 1 -f] [ 1,2,4] triazin-3-yl] -4-methyl-benzenesulfonamide

[0114] To a stirred solution of 3-amino-2-[methoxy(phenyl)methyl]pyrrolo[2,l- / |[l,2,4]triazin-4-one (80 mg, 0.29 mmol, 1.0 equiv) and p-toluenesulfonyl chloride (84 mg, 0.44 mmol, 1.5 equiv) in THF (2 mL) was added LiHMDS (1.0 M in THF) (99 mg, 0.59 mmol, 2.0 equiv) in portions at -78 °C under nitrogen atmosphere. The reaction was quenched with sat. NH4CI (aq.) at room temperature. The resulting mixture was extracted with ethyl acetate (2 x 20 mL). The combined organic layers were washed with brine (2 x 10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC with the following conditions (Column: XSelect CSH Fluoro Phenyl 30*150 mm, 5m; Mobile Phase A: Water(0.1% formic acid), Mobile Phase B: acetonitrile; Flow rate: 60 mL / min; Gradient: 30% B to 50% B in 10 min; Wave Length: 254nm / 220nm; RT= 14.2min) to afford N-[2-[methoxy(phenyl)methyl]-4-oxo-pyrrolo[2,l-f][l,2,4]triazin-3-yl]-4-methyl-benzenesulfonamide (16.8 mg, 13.37% yield) as a white solid. LCMS (ESI): [M-H]’: 423.05.1H NMR (400 MHz, DMSO-d6) δ 11.42 (d, J= 150.5 Hz, 1H), 7.70 (d, J= 8.0 Hz, 3H), 7.39 (d, J= 8.7 Hz, 7H), 6.94- 6.83 (m, 1H), 6.57 (s, 1H), 5.72 (s, 1H), 3.36 (s, 3H), 2.41 (s, 3H).

[0115] Example 3: N-[2-[methoxy(phenyl)methyl]-7-methyl-4-oxo-pyrrolo[2,l-f][l,2,4]triazin-3-yl]-4-methyl-benzenesulfonamide

[0116]

[0117] a) methyl l-amino-5-methylpyrrole-2-carboxylate

[0118] A solution of bis(methyl 5-methyl-1H-pyrrole-2-carboxylate) (3.00 g, 10.77 mmol, 1.00 equiv) in DMF (50 mL) was treated with t-BuOK (1.45 g, 12.93 mmol, 1.20 equiv) at 0°C followed by the addition of O-(2,4-dinitrophenyl)hydroxylamine (4.29 g, 21.55 mmol, 2.00 equiv) dropwise at room temperature. The resulting mixture was stirred at room temperature for 2 hours under nitrogen atmosphere. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 20% to 50% gradient in 20 min; detector, UV 254 nm to afford methyl l-amino-5-methylpyrrole-2-carboxylate (2.37 g, 71.31% yield) as a yellow solid. LCMS( ESI) [M + H]+: 155.00

[0119] b) methyl l-(2-methoxy-2-phenylacetamido)-5-methylpyrrole-2-carboxylate

[0120] A solution of methyl l-amino-5-methylpyrrole-2-carboxylate (450.00 mg, 2.91 mmol, 1.00 equiv) in MeCN (5 mL) was treated with methoxy(phenyl)acetic acid (727.57 mg, 4.37 mmol, 1.50 equiv), TCFH (982.77 mg, 3.50 mmol, 1.20 equiv) at room temperature under nitrogen atmosphere followed by the addition of NMI (718.97 mg, 8.75 mmol, 3.00 equiv) dropwise at room temperature. The resulting mixture was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in Water (0.1% FA), 20% to 50% gradient in 20 min; detector, UV 254 nm to afford methyl l-(2-methoxy-2-phenylacetamido)-5-methylpyrrole-2-carboxylate (630 mg, 71.39% yield) as a yellow solid. LCMS( ESI) [M + H]+: 303.00

[0121] c) l-(2-methoxy-2-phenylacetamido)-5-methylpyrrole-2-carboxamide

[0122] A solution of methyl l-(2-methoxy-2-phenylacetamido)-5-methylpyrrole-2-carboxylate (630.00 mg, 2.08 mmol, 1.00 equiv) and Ammonium hydroxide (9% in water) (1 mL) in DMF (6 mL) was stirred at 80°C for 24 hours under nitrogen atmosphere. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in water (0.1% FA), 20% to 50% gradient in 20 min; detector, UV 254 nm to afford l-(2-methoxy-2-phenylacetamido)-5-methylpyrrole-2-carboxamide (210 mg, 35.07% yield) as a white solid. LCMS( ESI) [M + H]+: 288.00

[0123] d) 2-[methoxy(phenyl)methyl]-7-methyl-3H-pyrrolo[2,1-f] [ 1,2,4] triazin-4-one

[0124] A solution of l-(2-methoxy-2-phenylacetamido)-5-methylpyrrole-2-carboxamide (210.00 mg, 0.73 mmol, 1.00 equiv) in EtOH (3 mL) was treated with KOH (82.01 mg, 1.46 mmol, 2.00 equiv) in water (3 mL) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 100°C for 24 hours under nitrogen atmosphere. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in water (0.1% FA), 20% to 50% gradient in 20 min; detector, UV 254 nm to afford 2-[methoxy(phenyl)methyl ]-7-methyl-3H-pyrrolo[2,1-f][1,2,4]triazin-4-one (130 mg, 66.05% yield, 89% purity) as a white solid. LCMS( ESI) [M + H]+: 270.0

[0125] e) 3-amino-2- [methoxy(phenyl)methyl] -7 -methylpyrrolo [2, 1 -f [ 1,2,4] triazin-4-one

[0126] A solution of 2-[methoxy(phenyl)methyl]-7-methyl-377-pyrrolo[2,l- / ][l,2,4]triazin-4-one (130.00 mg, 0.48 mmol, 1.00 equiv) in DMF (5 mL) was treated with amino diphenylphosphinate (225.15 mg, 0.96 mmol, 2.00 equiv) at room temperature under nitrogen atmosphere followed by the addition of t-BuOK (65.00 mg, 0.58 mmol, 1.20 equiv) in portions at room temperature. The resulting mixture was stirred at room temperature for 2 hours under nitrogen atmosphere. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in water (0.1% FA), 20% to 50% gradient in 20 min; detector, UV 254 nm to afford 3-amino-2- [methoxy(phenyl)methyl] -7 -methylpyrrolo [ 2, 1 - / ] [ 1,2,4] triazin-4-one (100 mg, 72.86% yield) as a white solid. LCMS( ESI) [M + H]+: 285.0

[0127] f) N- [2- [methoxy(phenyl)methyl] -7 -methyl-4-oxo-pyrrolo [2, 1 -f] [ 1,2,4] triazin- 3-yl] -4- methyl-benzenesulfonamide

[0128] A solution of 3-amino-2-[methoxy(phenyl)methyl]-7-methylpyrrolo[2,l- / ][l,2,4]triazin-4- one (100.00 mg, 0.35 mmol, 1.00 equiv) in THF (5 mL) was treated with LiHMDS.

[0129] (117.71 mg, 0.70 mmol, 2.00 equiv) at -78 °C for 3 minutes under nitrogen atmosphere followed by the addition of 4-Methylbenzenesulfonyl chloride (100.58 mg, 0.52 mmol, 1.50 equiv) in portions at -78 °C. The reaction was quenched with water at 0 °C. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, MeCN in water (0.1% FA), 20% to 50% gradient in 20 min; detector, UV 254 nm to afford V-{2-[methoxy(phenyl)methyl]-7- methyl-4-oxopyrrolo[2,l- / ][l,2,4]triazin-3-yl}-4-methylbenzenesulfonamide (27.5 mg, 17.83% yield) as a white solid. LCMS (ESI): [M+H]+: 439.15. ’H NMR (300 MHz, DMSO-d6) δ 11.11 (s, 1H), 7.88 - 7.59 (m, 2H), 7.57 - 7.23 (m, 7H), 6.81 (d, 7= 4.2 Hz, 1H), 6.37 (d, J = 4.4 Hz, 1H), 5.75 (s, 1H), 3.39 (s, 3H), 2.42 (s, 6H).

[0130] Example 4: N-[7-cyclopropyl-2-[methoxy(phenyl)methyl]-4-oxo-pyrrolo[2,l- f][l,2,4]triazin-3-yl]-4-methyl-benzenesulfonamide

[0131]

[0132] To a stirred solution of N-[7-bromo-2-[methoxy(phenyl)methyl]-4-oxo-pyrrolo[2,l-f][l, 2, 4]triazin-3-yl]-4-methyl-benzenesulfonam (Example 1, 100 mg, 0.20 mmol, 1.0 equiv) and cyclopropylboronic acid (27 mg, 0.30 mmol, 1.5 equiv) in dioxane (1 mL) and H2O (0.2 mL) were added Pd(dppf)Cl2(15 mg, 0.02 mmol, 0.1 equiv) and K2CO3(55 mg, 0.40 mmol, 2.0 equiv) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 80 °C for 3 h under nitrogen atmosphere. The reaction was quenched with water at room temperature. The resulting mixture was extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with brine (3x10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, acetonitrile in water (0.1% formic acid), 10% to 50% gradient in 10 min; detector, UV 254 nm. This resulted in N-[7-cyclopropyl-2- [methoxy(phenyl)methyl] -4-oxo-pyrrolo [2, 1 -f] [ 1,2,4] triazin-3-yl] -4-methyl-benzenesulfonamide (37.9 mg, 41.07% yield) as a white solid. LC-MS (ESI, m / z):

[0133] [M+ H]+: 465.15. ’H NMR (400 MHz, Acetonitrile-d3) δ 8.43 (s, 1H), 7.80 - 7.65 (m, 2H), 7.38 (dd, J = 16.1, 9.8 Hz, 7H), 6.71 (d, J= 4.4 Hz, 1H), 6.10 (d, J= 4.3 Hz, 1H), 5.71 (s, 1H), 3.42 (s, 3H), 2.43 (s, 3H), 2.27 (s, 1H), 0.95 (d, J= 95.3 Hz, 4H).

[0134] Example 5: N-(8-chloro-2-(methoxy(phenyl)methyl)-4-oxopyrazolo[l,5-a][l,3,5]triazin-3(4H)-yl)-4-methylbenzenesulfonamide

[0135]

[0136] a) 2-methoxy-2-phenylacetonitrile

[0137] A solution of a, a-dimethoxytoluene (3 g, 19.712 mmol, 1 equiv) and TMSCN (2.93 g, 29.568 mmol, 1.5 equiv) in Et2O (15 mL) was added with BF3(1.60 g, 23.654 mmol, 1.2 equiv) at rt. The mixture was stirred for overnight at rt under nitrogen. The resulting mixture was extracted with water (20 mL) and EtOAc (3 x 10 mL). The combined organic layers were washed with water (2 x 10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The organic layer was purified by Prep-TLC (petroleum ether / ethyl acetate = 1.5:1) Rf= 0.5 to afford 2-methoxy-2-phenylacetonitrile (2 g, 68.94% yield) as a yellow oil.

[0138] b) methyl 2-methoxy-2-phenylethanimidate hydrochloride

[0139] A solution of 2-methoxy-2-phenylacetonitrile (2 g, 13.589 mmol, 1 equiv) in MeOH (10 mL) was added with hydrogen chloride (2.0 M in diethyl ether) (1.49 g, 40.767 mmol, 3 equiv) at 0 °C under nitrogen atmosphere. The mixture was stirred for 3 hours at 0 °C. The resulting mixture was extracted with water (10 mL) and EtOAc (3 x 8 mL). The combined organic layers were washed with water (2 x 8 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give the crude product methyl 2-methoxy-2-phenylethanimidate hydrochloride (2 g, crude) as a white solid.

[0140] LCMS (ESI) [M + H]+: 180.1.

[0141] c) (Z)-A'-(4-chloro-2H-pyrazol-3-yl) -2-methoxy-2-phenylethanimidamide

[0142] A solution of methyl 2-methoxy-2-phenylethanimidate hydrochloride (2 g, 9.273 mmol, 1 equiv) and 4-chloro-277-pyrazol-3-amine (1.31 g, 11.128 mmol, 1.2 equiv) in MeCN (20 mL) was added with AcONa (2.28 g, 27.819 mmol, 3 equiv) at 0 °C. The mixture was stirred for 3 hours at rt. The resulting mixture was extracted with water (20 mL) and EtOAc (3 x 10 mL). The combined organic layers were washed with water (1 x 10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to give (Z)-A'-(4-chloro-2H-pyrazol-3-yl) -2-methoxy-2-phenylethanimidamide (2 g, crude) as a yellow oil. LCMS (ESI) [M + H]+: 265.2

[0143] d) 8-chloro-2-[methoxy(phenyl)methyl]-3H-pyrazolo[1,5-a][1,3,5]triazin-4-one

[0144] A solution of (Z)-A'-(4-chloro-2H-pyrazol-3-yl)-2-methoxy-2-phenylethanimidamide (2 g, 7.55 mmol, 1 equiv) and dimethyl carbonate (1.02 g, 11.33 mmol, 1.5 equiv) in EtOH (15 mL) was added with sodium ethanolate (1.03 g, 15.110 mmol, 2 equiv) at 0°C. The mixture was stirred for 4 hours at 80 °C under nitrogen atmosphere. The resulting mixture was extracted with water (10 mL) and EtOAc (3 x 8 mL). The combined organic layers were washed with water (10 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (petroleum ether / ethyl acetate =1.5:1) R f =0.4 to afford 8-chloro-2-[methoxy(phenyl)methyl]-377-pyrazolo[l,5-a] [1,3,5] triazin-4-one (600 mg, 27.32% yield) as a white solid. LCMS (ESI) [M + H]+: 291.1.

[0145] e) 3-amino-8-chloro-2-[methoxy(phenyl)methyl] pyrazolo[ 1,5-a] [1,3,5] triazin-4-one

[0146] A solution of 8-chloro-2-[methoxy(phenyl)methyl]-377-pyrazolo[l,5-a] [1,3,5] triazin-4-one (500 mg, 1.720 mmol, 1 equiv) in THF (10 mL) was added with t-BuOK (289.5 mg, 2.580 mmol, 1.5 equiv) at 0 °C, 30 min later amino diphenylphosphinate (802.2 mg, 3.440 mmol, 2 equiv) was added at 0 °C. The mixture was stirred 1 hour at rt. The resulting mixture was filtered, the filter cake was washed with EtOAc (3 x 10 mL). The filtrate was concentrated under reduced pressure. The organic layer was purified by silica gel column chromatography, eluted with petroleum ether / ethyl acetate = (2:3) to afford 3-amino-8-chloro-2-[methoxy(phenyl)methyl] pyrazolo[l,5-a] [1,3,5] triazin-4-one (350 mg, 66.56% yield) as a white solid. LCMS (ESI) [M + H]+: 306.1

[0147] f) N-(8-chloro-2-(methoxy(phenyl)methyl)-4-oxopyrazolo[l,5-a][l,3,5]triazin-3(4H)-yl)-4-methylbenzenesulfonamide

[0148] A solution of 3-amino-8-chloro-2-[methoxy(phenyl)methyl] pyrazolo[l,5-a] [1,3,5] triazin-4-one (350 mg, 1.145 mmol, 1 equiv) and p-toluenesulfonyl chloride (261.9 mg, 1.374 mmol, 1.2 equiv) in THF (5 mL) was added with LiHMDS (383.1 mg, 2.290 mmol, 2 equiv) at -78 °C. The mixture was stirred for 1 hour at -78 °C. The reaction was quenched with NH4CI (4 mL) at 0 °C. The aqueous layer was extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with water (3 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The organic layer was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, water (FA) in MeCN, 20% to 50% gradient in 15 min; detector, UV 254 nm to afford N-(8-chloro-2-(methoxy(phenyl)methyl)-4-oxopyrazolo[l,5-a][l,3,5]triazin-3(4H)-yl)-4-methylbenzenesulfonamide (170 mg, 32.3% yield) as a white solid. LCMS (ESI) [M + H]+: 460.1.

[0149] 1H NMR (400 MHz, Acetonitrile-d₃) δ 7.96 (s, 1H), 7.80 - 7.72 (m, 2H), 7.47 - 7.38 (m, 2H), 7.37 (ddd, J = 7.1, 4.7, 3.3 Hz, 5H), 5.73 (d, J = 1.4 Hz, 1H), 3.38 (s, 3H), 2.44 (s, 3H).

[0150] Example 6: Biological experiments

[0151] a) The potency of exemplary compounds of formula (I) was determined using thallium fluorescence. Cell Culture

[0152] Flip-IN Hek293-Trex or Hek293 Trex cells stably expressing TMEM175 under a tetracycline inducible CMV promoter were used for these studies. Cells were maintained in DMEM-HG supplemented with heat inactivated 10% FBS (Gemini), 2mM sodium pyruvate, lOmM Hepes, 50ug / ml Hygromycin and 5ug / ml Blasticidin at 37°C in 5% CO₂. All cells were induced with 0.2ug / ml or 2ug / ml Doxycycline 18-24 hour prior to thallium assay.

[0153] Assay solution and compound preparation

[0154] The assay solution consisted of (in mM): 132 Na-Gluconate, 10 K-Gluconate, 1.8 Ca-Gluconate, 0.8 Mg-Gluconate, 10 HEPES, pH 7.4. Test agents were dissolved in DMSO to give 10 mM stocks. 10-point dose response curves were constructed using the Labcyte Echo 550 acoustic dispenser to generate final test concentrations of 30, 15, 7.5, 3.75, 1.88, 0.94, 0.47, 0.23, 0.12, 0.06 uM.

[0155] Assay Protocol

[0156] All thallium fluorescence studies were performed using the Molecular Devices FLIPR TETRA™ fluorescence platform. Plates seeded with Hek-293FlipIN Trex cells expressing TMEM175 channels were loaded with lOpl of a thallium sensitive AM-ester fluorescent dye made up in IX Hank’s buffer + 20mM HEPES. The dye loading buffer contained an extracellular fluorescence quenching agent allowing the dye to remain in the plate throughout the assay. Cells were incubated with dye for 45-60 minutes at room temperature, protected from light.

[0157] After the dye loading incubation time, cell plates and assay plates were loaded into the FLIPR TETRA instrument. The FLIPR program measured a 20 second baseline, after which the embedded 384- well pipetting head added lOul of 2X test agent or controls made up in chloride free assay buffer containing ImM thallium sulfate. Changes in fluorescence (measured at 490nm excitation and 520nm emission) were measured for a 3-minute period.

[0158] Data Analysis

[0159] Percent activation was calculated from changes in fluorescent response using the following equation:

[0160] % activation = ((RFU test agent - RFU buffer) / (RFU positive control - RFU buffer))* 100 EC50 values were calculated using IDBS ActivityBase software using a 4-parameter logistic fit. Fit criteria were defined as follows: Fit minimum values were allowed to float between -10% to 5%, Fit maximum values were allowed to float between 40% and 120%. The data are reported in Table 1 (first column).

[0161] b) The potency of exemplary compounds of formula (I) was determined using the SyncroPatch 384 (Nanion) high throughput electrophysiology platform.

[0162] Cell Culture

[0163] Flip-IN Hek293-Trex or Hek293 Trex cells stably expressing TMEM175 under a tetracycline inducible CMV promoter were used for these studies. Cells were maintained in DMEM-HG supplemented with heat inactivated 10% FBS (Gemini), 2mM sodium pyruvate, lOmM Hepes, 50ug / ml Hygromycin and 5ug / ml Blasticidin at 37°C in 5% CO₂. Cells were cultured in T-225 flasks (Nunc) for 3-4 days to reach 85-95% confluence prior to electrophysiological recording and cells were seeded in PDL coated plates 24hours prior to testing. All cells were induced with 0.2ug / ml or 2ug / ml Doxycycline 18-24hour prior to electrophysiological recording or fluorescent assays, respectively.

[0164] Solutions

[0165] Solutions were of the following composition:

[0166] Extracellular recording solution (in mM): 105 NaCl, 40 NMDG, 4 KC1, 5 CaCl₂, 1 MgCl₂, 10 HEPES, pH 7.4. Seal enhancer solution (in mM): 90 NaCl, 3 KC1, 35 CaCl₂, 10 MgCl₂, 10 HEPES, pH 7.4. Intracellular recording solution (in mM): 150 CsF, 1 Mg- ATP, 10 EGTA, 10 HEPES, 5 MgCl₂, 0.01 Escin, pH 7.2.

[0167] Cell Preparation

[0168] Cells were harvested for electrophysiological recording upon reaching appropriate confluence. Cells were first washed in DPBS (Hyclone, Cat #SH30028.03) and then 2 ml of Accutase (MP Biomedicals #1000449) was added at 28 °C until -90% of cells were suspended. F12 HAM’s media + 1 mM L-glutamine (Hyclone, SH30026.02) was then added to the flask to dilute the accutase. Cells were then triturated until a single cell suspension was achieved, a cell count was performed, and cells were centrifuged for 2 min. at 1,000 rpm. Media was then aspirated, and cells were resuspended in Earle’s balanced salt solution to a concentration of 0.75 X 106 cells / ml and allowed to recover for 25 min at 10°C.

[0169] SyncroPatch Recording At the beginning of each assay, 20 pl of cell suspension was dispensed into each well of a multi-hole 384- well SyncroPatch chip by the onboard pipettor. Cell sealing was initiated, and seal enhancer solution was added to facilitate seal formation. Upon completion of sealing, cells were washed 3 times with extracellular recording solution and the assay voltage protocol was started. Human TMEM175 channels were evaluated using a voltage protocol where cells were voltage-clamped at a holding potential of 0 mV. Ionic currents were evoked every 5 sec with a 200 msec voltage ramp from -100 mV to 60 mV, followed by series of 100 msec voltage steps to -80, 0 mV and +40 mV. The current amplitude at the 100 msec 0 mV step was used to for analysis. Baseline current was assessed for 4 minutes prior to the addition of test compound for 2.5 min. This was followed by addition of the positive control, 10 pM DCPIB for an additional 2.5 min. The experiment was completed with addition of 2 mM 4-AP to fully block the channel.

[0170] Data Analysis

[0171] Data were collected on the SyncroPatch platform using PatchControl software (Nanion) and processed and analyzed using DataControl Software (Nanion). The average leak-subtracted current was determined for control, test compound and DCPIB conditions by subtracting the remaining current following block by 4-AP. Percent activation of test compound was determined from the leak-corrected current using the following equation:

[0172] % Activation = ((Test compoundCorr -ControlCorr) / (DCPIBCorr - ControlCorr))*100

[0173] The % activation data were then plotted as a function of compound concentration and fit will a Hill equation to derive EC50. The data are reported in Table 1 (second column).

[0174] TMEM175 TMEM175

[0175] FLIPR EP

[0176] Examples HUMAN HUMAN

[0177] EC50 EC50

[0178] (uM) (uM)

[0179] 1 0.62 0.13

[0180] 2 1.44 0.42

[0181] 3 0.45 0.22

[0182] 4 0.44 0.31

[0183]

[0184] 5 0.92

[0185] Table 1

[0186] Example A Film coated tablets containing the following ingredients can be manufactured in a conventional manner:

[0187] Ingredients Per tablet

[0188] Kernel:

[0189] Compound of formula (I) 10.0 mg 200.0 mg

[0190] Microcrystalline cellulose 23.5 mg 43.5 mg

[0191] Lactose hydrous 60.0 mg 70.0 mg

[0192] Povidone K30 12.5 mg 15.0 mg

[0193] Sodium starch glycolate 12.5 mg 17.0 mg

[0194] Magnesium stearate 1.5 mg 4.5 mg

[0195] (Kernel Weight) 120.0 mg 350.0 mg

[0196] Film Coat:

[0197] Hydroxypropyl methyl cellulose 3.5 mg 7.0 mg

[0198] Polyethylene glycol 6000 0.8 mg 1.6 mg

[0199] Talc 1.3 mg 2.6 mg

[0200] Iron oxide (yellow) 0.8 mg 1.6 mg

[0201] Titan dioxide 0.8 mg 1.6 mg

[0202]

[0203] The active ingredient is sieved and mixed with microcrystalline cellulose and the mixture is granulated with a solution of polyvinylpyrrolidone in water. The granulate is then mixed with sodium starch glycolate and magnesium stearate and compressed to yield kernels of 120 or 350 mg respectively. The kernels are lacquered with an aq. solution / suspension of the above mentioned film coat.

[0204] Example B Capsules containing the following ingredients can be manufactured in a conventional manner:

[0205] Ingredients Per capsule

[0206] Compound of formula (I) 25.0 mg

[0207] Lactose 150.0 mg

[0208] Maize starch 20.0 mg

[0209] Talc 5.0 mg

[0210]

[0211] The components are sieved and mixed and filled into capsules of size 2.

[0212] Example C

[0213] Injection solutions can have the following composition:

[0214] Compound of formula (I) 3.0 mg

[0215] Polyethylene glycol 400 150.0 mg

[0216] Acetic acid q.s. ad pH 5.0

[0217] Water for injection solutions ad 1.0 ml

[0218]

[0219] The active ingredient is dissolved in a mixture of Polyethylene glycol 400 and water for injection (part). The pH is adjusted to 5.0 by addition of acetic acid. The volume is adjusted to 1.0 ml by addition of the residual amount of water. The solution is filtered, filled into vials using an appropriate overage and sterilized.

Claims

Claims1. A compound of formula (I)whereinone of A1and A2is nitrogen and the other one is carbon;R1and R2together with the carbon and nitrogen atoms to which they are attached form pyrrolyl, susbtitutued pyrrolyl, pyrrazolyl or substituted pyrrazolyl, wherein substituted pyrrolyl and substituted pyrrazolyl are pyrrolyl and pyrrazolyl substituted with one substituent selected from halogen, alkyl, cycloalkyl, haloalkyl or cycloalkoxy; andR3is hydrogen or halogen;or a pharmaceutically acceptable salt thereof.

2. A compound according to claim 1, wherein R1and R2together with the carbon and nitrogen atoms to which they are attached form pyrrolyl, susbtitutued pyrrolyl, pyrrazolyl or substituted pyrrazolyl, wherein substituted pyrrolyl and substituted pyrrazolyl are pyrrolyl and pyrrazolyl substituted with one substituent selected from chlorine, bromine, methyl, cyclopropyl, difluoromethyl, difluoroethyl, trifluoroethyl or cyclopropyloxy.

3. A compound according to claim 1 or 2, wherein R1and R2together with the carbon and nitrogen atoms to which they are attached form4. A compound according to any one of claims 1 to 3, wherein R3is hydrogen or fluorine.

5. A compound according to any one of claims 1 to 4 selected fromN-[7-bromo-2-[methoxy(phenyl)methyl]-4-oxo-pyrrolo[2,l-f][l,2,4]triazin-3-yl]-4- methyl-benzenesulfonamide;N-[2-[methoxy(phenyl)methyl]-4-oxo-pyrrolo[2,l-f][l,2,4]triazin-3-yl]-4-methyl- benzenesulfonamide;N- [2- [methoxy(phenyl)methyl] -7 -methyl-4-oxo-pyrrolo [2, 1 -f] [ 1,2,4] triazin-3-yl] -4- methyl-benzenesulfonamide;N- [7 -cyclopropyl-2- [methoxy(phenyl)methyl] -4-oxo-pyrrolo [2, 1 -f] [ 1,2,4] triazin-3- yl] -4-methyl-benzenesulfonamide; andN-(8-chloro-2-(methoxy(phenyl)methyl)-4-oxopyrazolo[l,5-a][l,3,5]triazin-3(4H)- yl)-4-methylbenzenesulfonamide.

6. A compound according to any one of claims 1 to 5 selected fromN-[7-bromo-2-[methoxy(phenyl)methyl]-4-oxo-pyrrolo[2,l-f][l,2,4]triazin-3-yl]-4- methyl-benzenesulfonamide;N- [2- [methoxy(phenyl)methyl] -7 -methyl-4-oxo-pyrrolo [2, 1 -f] [ 1,2,4] triazin-3-yl] -4- methyl-benzenesulfonamide; andN- [7 -cyclopropyl-2- [methoxy(phenyl)methyl] -4-oxo-pyrrolo [2, 1 -f] [ 1,2,4] triazin-3- yl]-4-methyl-benzenesulfonamide;or a pharmaceutically acceptable salt thereof.

7. A a process for the preparation of a compoundof formula (I) as defined in any one of claims 1 to 6, comprising the reaction of a compound of formula (A)in the presence of para-toluenesulfonyl chloride and a base, wherein A1, A2and R1to R3are as defined in any one of claims 1 to 4.

8. A compound according to any one of claims 1 to 6, when manufactured according to a process of claim 6.

9. A compound according to any one of claims 1 to 6 for use as therapeutically active substance.

10. A pharmaceutical composition comprising a compound in accordance with any one of claims 1 to 6 and a therapeutically inert carrier.

11. The use of a compound according to any one of claims 1 to 6 for the treatment or prophylaxis of synucleinopathies, Parkinson’s disease, Dementia with Lewy body disease, REM-sleep behavior disorder, amyotrophic lateral sclerosis or lysosomal storage disorders.

12. The use of a compound according to any one of claims 1 to 6 for the preparation of a medicament for the treatment or prophylaxis of synucleinopathies, Parkinson’s disease, Dementia with Lewy body disease, REM-sleep behavior disorder, amyotrophic lateral sclerosis or lysosomal storage disorders.

13. A compound according to any one of claims 1 to 6 for use in the treatment or prophylaxis of synucleinopathies, Parkinson’s disease, Dementia with Lewy body disease, REM-sleep behavior disorder, amyotrophic lateral sclerosis or lysosomal storage disorders.

14. A method for the treatment or prophylaxis of synucleinopathies, Parkinson’s disease, Dementia with Lewy body disease, REM-sleep behavior disorder, amyotrophic lateral sclerosis or lysosomal storage disorders, which method comprisesadministering an effective amount of a compound as defined in any one of claims 1 to 6 to a patient in need thereof.

15. The invention as hereinbefore described.***

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