WO2024178219A1 - 1-acyl-3-aminoindazoles for treating cystic fibrosis - Google Patents

Abstract

The invention is directed to 1-acyl-3aminoindazoles and pharmaceutical compositions thereof that either stimulate or inhibit the cystic fibrosis transmembrane conductance regulator (CFTR). The genus of the compounds that stimulates CFTR (formula I) is useful for treating cystic fibrosis. The genus that inhibits CFTR (formula II) is useful for treating secretory diarrhea.

Description

Docket No.2877.037AWO 1-ACYL-3-AMINOINDAZOLES FOR TREATING CYSTIC FIBROSIS Cross Reference to Related Application [0001] This application claims priority of US provisional application 63/486,577, filed February 23, 2023, the disclosure of which is hereby incorporated herein by reference in its entirety. Field of the Invention [0002] The present application relates generally to 1-acyl-3-aminoindazoles that either stimulate or inhibit the cystic fibrosis transmembrane conductance regulator (CFTR). The subgenus of the compounds that stimulates CFTR is useful for treating cystic fibrosis. The subgenus that inhibits CFTR is useful for treating secretory diarrhea. Background Information [0003] The cystic fibrosis transmembrane conductance regulator (CFTR) is an anion channel essential for human health. Widely expressed in epithelial cells, CFTR regulates salt and fluid homeostasis in the lung, intestine, pancreas, reproductive tract, and other organs. Mutations that disrupt CFTR biosynthesis, folding, trafficking, or ion permeation cause cystic fibrosis (CF), a lethal genetic disease without cure. On the other hand, overactivation of CFTR by Vibrio cholerae and enterotoxigenic Escherichia coli leads to secretory diarrhea, the second leading cause of mortality in children under the age of 5. For these reasons, CFTR modulators that either enhance or downregulate its activity have been long pursued as drug candidates. [0004] Whereas no negative CFTR modulators have been advanced to clinic, progress has made in developing positive modulators, including correctors that increase the abundance of CFTR at the cell surface and potentiators that enhance CFTR ion flux. Thus far, one potentiator (ivacaftor) and three correctors (lumacaftor, tezacaftor, and elexacaftor) are available to CF patients. The potentiator ivacaftor, prescribed singly or in combination with correctors, is approved to treat 178 CFTR mutations. While this drug ^{H2932377.1} 1 Docket No.2877.037AWO has had beneficial effects on the health of CF patients, it does retain liabilities. Its pharmacokinetics are far from optimum; it is difficult to formulate due to low water solubility (<0.05 ug/mL); and its bioavailability is highly variable, as 99% of ivacaftor is bound to plasma proteins. Furthermore, the side effects of ivacaftor, including liver disease and cataracts in children, may limit its usage for some patients. [0005] The molecular mechanism of ivacaftor has been well studied. Electrophysiology measurements showed that the drug increases the open probability (Po) of both wild-type (wt) and many mutant CFTRs, suggesting that it acts through an allosteric site. Cryo-EM structures revealed that ivacaftor binds to CFTR inside the membrane, coinciding with a hinge region important for gating. A different potentiator, GLPG1837, representing a distinct chemotype, also binds the same site on CFTR. The potentiator-binding pocket is unique to CFTR compared to other closely related proteins, and is thus an excellent therapeutic target, as molecules binding at this site are less likely to have off-target activities. [0006] Molecular docking, electrophysiology, cryo-EM, and medicinal chemistry were employed to identify novel CFTR ligands. The structure of the CFTR/ivacaftor complex was used to screen a virtual library of diverse chemical scaffolds. The binding site of the lead compound was verified by cryo-EM. Described below are nano-molar affinity potentiators that are chemically unrelated to the known potentiators. Also described for the first time are negative modulators that bind at the potentiator site but inhibit CFTR ion conduction, thus demonstrating the possibility of down-regulating CFTR activity. SUMMARY OF THE INVENTION [0007] The invention is directed to 1-acyl-3aminoindazoles and pharmaceutical compositions thereof that either stimulate or inhibit the cystic fibrosis transmembrane conductance regulator (CFTR). The genus of the compounds that stimulates CFTR is useful for treating cystic fibrosis. The genus that inhibits CFTR is useful for treating secretory diarrhea. ^{H2932377.1} 2 Docket No.2877.037AWO [0008] The present invention relates, in a first aspect, to compounds formula I:

X¹ and X² are chosen independently from N and CH; R¹ is chosen from hydroxymethyl, halogen, amino, methoxy, -CONH₂, and -NHC(=O)H; R² is hydrogen, or taken together with R¹, forms a six-membered fused carbocyclic or heterocyclic ring; R³ is methyl or ethyl; R⁴ is chosen from hydrogen, halogen, and lower-alkyl; and R⁵ is hydrogen, or taken together with R⁴, forms a six-membered fused carbocyclic ring. This subgenus is a positive modulator of CFTR. [0009] In another aspect, the invention relates to formula II: formula II: . wherein

{^H2932377.1} 3 Docket No.2877.037AWO X¹ and X² are chosen independently from N and CH; R³ is methyl or ethyl; R¹⁰ is H or NH2; and R¹³ is H or halogen. This subgenus is a negative modulator of CFTR. [0010] In another aspect, the invention relates to pharmaceutical compositions comprising a pharmaceutically acceptable carrier and a compound as described herein. [0011] In another aspect, the invention relates to a method for treating cystic fibrosis comprising administering a compound of formula I as described herein. [0012] In another aspect, the invention relates to a method for treating secretory diarrhea comprising administering a compound of formula II as described herein. BRIEF DESCRIPTION OF THE DRAWINGS [0013] In the accompanying drawings the drawings are not necessarily complete when viewed without reference to the text, emphasis instead being placed upon illustrating the principles of the invention. [0014] FIG.1 presents a bar graph showing relative stimulation of CFTR for a series of compounds. DETAILED DESCRIPTION OF THE INVENTION [0015] It has been found that compounds of formulas I and II are modulators of CFTR. The invention can thus be divided into two genera. In the first genus, the compounds of formula I are (S) enantiomers at the R³ position. In the second genus, the compounds of formula II are (R) enantiomers at the R³ position. The chirality at R³ in combination with ^{H2932377.1} 4 Docket No.2877.037AWO the substitution pattern at R¹ and R¹⁰ appears to determine whether a 1-acyl-3- aminoindazole is a positive or negative modulator of the CFTR channel. [0016] In an aspect, the disclosure relates to a substantially pure single enantiomer of a compound represented by formula I: , wherein

X¹ and X² are chosen independently from N and CH; R¹ is chosen from hydroxymethyl, halogen, amino, methoxy, -CONH2, and -NHC(=O)H; R² is hydrogen, or taken together with R¹, forms a six-membered fused carbocyclic or heterocyclic ring; R³ is methyl or ethyl; R⁴ is chosen from hydrogen, halogen, and lower-alkyl; and R⁵ is hydrogen, or taken together with R⁴, forms a six-membered fused carbocyclic ring. [0017] In some embodiments, the substantially pure single enantiomer is represented by formula Ia: . ^{H2932377.1}

Docket No.2877.037AWO [0018] In some embodiments, the substantially pure single enantiomer is represented by formula Ib: .

is represented by formula Ic: . [0020] In

enantiomer of a compound represented by formula II: , wherein

X¹ and X² are chosen independently from N and CH; R³ is methyl or ethyl; R¹⁰ is H or NH₂; and {^H2932377.1} 6 Docket No.2877.037AWO R¹³ is H or halogen. [0021] In some embodiments, the substantially pure single enantiomer is represented by formula IIa: . [0022] In some

is represented by formula IIb: . [0023] In some

is represented by formula IIc: . [0024] In some

is N. In some embodiments, X² is CH. In some embodiments, X² is N. In some embodiments, both X¹ and X² may be CH. In some embodiments, both X¹ and X² may be N. In some embodiments, X¹ is N and X² is CH. In some embodiments, X¹ is CH and X² is N. [0025] In some embodiments, R¹ is hydroxymethyl. In some embodiments, R¹ is halogen. In some embodiments, R¹ is fluorine. In some embodiments, R¹ is chlorine. In ^{H2932377.1} 7 Docket No.2877.037AWO some embodiments, R¹ is amino. In some embodiments, R¹ is methoxy. In some embodiments, R¹ is -CONH2. In some embodiments, R¹ is -NHC(=O)H. [0026] In some embodiments, R² is hydrogen. In some embodiments, R², taken together with R¹, forms a six-membered fused carbocyclic ring. In some embodiments, R², taken together with R¹, forms a six-membered fused heterocyclic ring. In some embodiments, R², taken together with R¹, forms a fused pyran ring. [0027] In some embodiments, R³ is methyl. In some embodiments, R³ is ethyl. [0028] In some embodiments, R⁴ is hydrogen. In some embodiments, R⁴ is halogen. In some embodiments, R⁴ is fluorine. In some embodiments, R⁴ is bromine. In some embodiments, R⁴ is lower-alkyl (e.g., methyl, ethyl, n-propyl, isopropyl, and the like). [0029] In some embodiments, R⁵ is hydrogen. In some embodiments, R⁵, taken together with R⁴, forms a six-membered fused carbocyclic ring. In some embodiments, R⁵, taken together with R⁴, forms a fused benzene ring. [0030] In some embodiments, R⁵ is hydrogen and R⁴ is hydrogen. In some embodiments, R⁵ is hydrogen and R⁴ is halogen. In some embodiments, R⁵ is hydrogen and R⁴ is fluorine. In some embodiments, R⁵ is hydrogen and R⁴ is bromine. In some embodiments, R⁵, taken together with R⁴, forms a fused benzene ring. [0031] In some embodiments, R¹⁰ is H. In some embodiments, R¹⁰ is NH₂. [0032] In some embodiments, R¹³ is H. In some embodiments, R¹³ is F. [0033] In some embodiments, R¹¹, R¹², and R¹³ are H. In some embodiments, R¹⁰, R¹¹, R¹², and R¹³ are H. [0034] In an aspect, the disclosure relates to a method for treating cystic fibrosis comprising administering a substantially pure single enantiomer of a compound represented by formula I, formula Ia, formula Ib, or formula Ic, as disclosed above and herein. [0035] In an aspect, the disclosure relates to a method for treating secretory diarrhea comprising administering a substantially pure single enantiomer of a compound represented by formula II, formula IIa, formula IIb, or formula IIc, as disclosed above and herein. [0036] ^{H2932377.1} 8 Docket No.2877.037AWO [0037] It is to be understood that in various embodiments, the pharmaceutical compositions of the present inventions comprise one or more pharmaceutically acceptable excipients, including, but not limited to, one or more binders, bulking agents, buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, diluents, disintegrants, viscosity enhancing or reducing agents, emulsifiers, suspending agents, preservatives, antioxidants, opacifying agents, glidants, processing aids, colorants, sweeteners, taste-masking agents, perfuming agents, flavoring agents, diluents, polishing agents, polymer matrix systems, plasticizers and other known additives to provide an elegant presentation of the drug or aid in the manufacturing of a medicament or pharmaceutical product comprising a composition of the present inventions. Examples of carriers and excipients 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. [0038] In various embodiments, non-limiting examples of excipients include, but are not limited to, corn starch, potato starch, or other starches, gelatin, natural and synthetic gums such as acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethyl cellulose calcium, sodium carboxymethyl cellulose), polyvinyl pyrrolidone, methyl cellulose, pre-gelatinized starch, hydroxypropyl methyl cellulose, (e.g., Nos.2208, 2906, 2910), hydroxypropyl cellulose, titanium dioxide, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrates, kaolin, silicic acid, sorbitol, starch, pre-gelatinized starch, agar-agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, other starches, pre-gelatinized starch, other starches, clays, other algins, other celluloses, gums, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, ^{H2932377.1} 9 Docket No.2877.037AWO talc, hydrogenated vegetable oil (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laureate, agar, a syloid silica gel (AEROSIL200, manufactured by W.R. Grace Co. of Baltimore, MD), a coagulated aerosol of synthetic silica (marketed by Degussa Co. of Plano, TX), CAB-O- SIL (a pyrogenic silicon dioxide product sold by Cabot Co. of Boston, MA), colorants and mixtures thereof. [0039] The terms "subject" or "subject in need thereof" are used interchangeably herein. These terms refer to a patient who has been diagnosed with the underlying disorder to be treated. Ordinarily, the patient will be a human. [0040] As used herein, the terms “treatment” or “treating" are used interchangeably. These terms refer to an approach for obtaining beneficial or desired results including, but not limited to, therapeutic benefit. Therapeutic benefit includes eradication or amelioration of the underlying disorder being treated; it also includes the eradication or amelioration of one or more of the symptoms associated with the underlying disorder such that an improvement is observed in the patient, notwithstanding that the patient may still be afflicted with the underlying disorder. [0041] The term “substituted” refers to the replacement of one or more hydrogen atoms in a specified group with a specified radical. For example, substituted aryl, heterocyclyl etc. refer to aryl or heterocyclyl wherein one or more H atoms in each residue are replaced with halogen, haloalkyl, alkyl, (C_1-8)hydrocarbyl, etc.. [0042] Halogen refers to fluorine, chlorine, bromine, and iodine. [0043] Unless otherwise specified, alkyl is a linear or branched hydrocarbyl. Unless otherwise specified, an unsubstituted alkyl has from 1 to 20 carbon atoms (e.g., 1 to 6 carbon atoms). Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, s-and t-butyl and the like. Lower alkyl refers to a linear or branched hydrocarbyl of 1 to 4 carbons. ^{H2932377.1} 10 Docket No.2877.037AWO [0044] A hydrocarbon or hydrocarbyl (as a substituent) includes alkyl, cycloalkyl, polycycloalkyl, alkenyl, alkynyl, aryl and combinations thereof. Examples include cyclopropylmethyl, benzyl, phenethyl, cyclohexylmethyl, camphoryl and naphthylethyl. Hydrocarbon refers to any substituent comprised of hydrogen and carbon as the only elemental constituents. Cycloalkyl is a subset of hydrocarbyl and includes cyclic hydrocarbon groups of from 3 to 8 carbon atoms. Examples of cycloalkyl groups include c-propyl, c-butyl, c-pentyl, norbornyl and the like. [0045] Unless otherwise specified, the term “carbocycle” or “carbocyclic” refers to a ring system in which the ring atoms are all carbon but of any oxidation state. Thus (C₃- C8) carbocycle refers to both non-aromatic and aromatic systems, including such systems as cyclopropane, benzene and cyclohexene; (C8-C12) carbopolycycle refers to such systems as norbornane, decalin, indane and naphthalene. Carbocycle, if not otherwise limited, refers to monocycles, bicycles and polycycles. [0046] Unless otherwise specified, acyl refers to formyl and to groups of 1, 2, 3, 4, 5, 6, 7 and 8 carbon atoms of a straight, branched, cyclic configuration, saturated, unsaturated and aromatic and combinations thereof, attached to the parent structure through a carbonyl functionality. One or more carbons in the acyl residue may be replaced by nitrogen, oxygen or sulfur as long as the point of attachment to the parent remains at the carbonyl. Examples include formyl, acetyl, benzoyl, propionyl, isobutyryl, t- butoxycarbonyl, benzyloxycarbonyl and the like. Lower-acyl refers to groups containing one to four carbons. The double bonded oxygen, when referred to as a substituent itself is called “oxo”. [0047] “Heterocycle” or “heterocyclic” refers to a cycloalkyl or aryl residue in which from one to four carbons is replaced by a heteroatom selected from the group consisting of N, O and S. The nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. Unless otherwise specified, a heterocycle may be non-aromatic (i.e. aliphatic) or aromatic. Examples of heterocycles ^{H2932377.1} 11 Docket No.2877.037AWO include pyrrolidine, pyrazole, pyrrole, indole, quinoline, isoquinoline, tetrahydroisoquinoline, benzofuran, benzodioxan, benzodioxole (commonly referred to as methylenedioxyphenyl, when occurring as a substituent), tetrazole, morpholine, thiazole, pyridine, pyridazine, pyrimidine, thiophene, furan, oxazole, oxazoline, isoxazole, dioxane, tetrahydrofuran and the like. It is to be noted that heteroaryl is a subset of heterocycle in which the heterocycle is aromatic. Examples of heteroaromatic rings include: furan, benzofuran, isobenzofuran, pyrrole, indole, isoindole, thiophene, benzothiophene, imidazole, benzimidazole, purine, pyrazole, indazole, oxazole, benzoxazole, isoxazole, benzisoxazole, thiazole, benzothiazole, triazole, tetrazole, pyridine, quinoline, isoquinoline, pyrazine, quinoxaline, acridine, pyrimidine, quinazoline, pyridazine, cinnoline, phthalazine, and triazine. Examples of heterocyclyl residues additionally include piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxo- pyrrolidinyl, 2-oxoazepinyl, azepinyl, 4-piperidinyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyrazinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, benzimidazolyl, thiadiazolyl, benzopyranyl, benzothiazolyl, tetrahydrofuryl, tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl, thiamorpholinylsulfoxide, thiamorpholinylsulfone, oxadiazolyl, triazolyl and tetrahydroquinolinyl. [0048] As used herein, and as would be understood by the person of skill in the art, the recitation of “a compound” - unless expressly further limited - is intended to include salts of that compound. Thus, for example, the recitation “a compound of formula I” as applied to Example I1421, would include both the free base and its salt:

of formula I” refers to the compound or a pharmaceutically acceptable salt thereof. The term "pharmaceutically acceptable salt" refers to salts prepared from pharmaceutically ^{H2932377.1} 12 Docket No.2877.037AWO acceptable non-toxic acids or bases including inorganic acids and bases and organic acids and bases. When the compounds of the present invention are basic, as shown in the depiction above in this paragraph, salts may be prepared from pharmaceutically acceptable non-toxic acids including inorganic and organic acids. Suitable pharmaceutically acceptable acid addition salts for the compounds of the present invention include acetic, adipic, alginic, ascorbic, aspartic, benzenesulfonic (besylate), benzoic, boric, butyric, camphoric, camphorsulfonic, carbonic, citric, ethanedisulfonic, ethanesulfonic, ethylenediaminetetraacetic, formic, fumaric, glucoheptonic, gluconic, glutamic, hydrobromic, hydrochloric, hydroiodic, hydroxynaphthoic, isethionic, lactic, lactobionic, laurylsulfonic, maleic, malic, mandelic, methanesulfonic, mucic, naphthylenesulfonic, nitric, oleic, pamoic, pantothenic, phosphoric, pivalic, polygalacturonic, salicylic, stearic, succinic, sulfuric, tannic, tartaric acid, teoclatic, p- toluenesulfonic, and the like. [0050] The term “substantially pure” as used herein in reference to stereochemical purity means that the compositions contain at least 90% by weight of one enantiomer and 10% by weight or less of the other. In a more preferred embodiment the term "substantially pure" means that the composition contains at least 99% by weight of one enantiomer, and 1% or less of the opposite enantiomer. Preparation of Compounds [0051] The following abbreviations are used in the synthetic routes: THF (tetrahydrofuran), MeOH (methanol), DCM (dicholoromethane), DMF (N,N- dimethylformamide), ACN (acetonitrile), EtOH (ethanol), EtOAc (ethyl acetate), IPA (2- propanol), DMSO (dimethyl sulfoxide), MTBE (methyl tert-butyl ether), TEA (triethylamine), DIPEA (N,N-diisopropylethylamine), TMEDA (tetramethylethylenediamine), DMAP (N,N-dimethylpyridin-4-amine), EDCI (N-(3- Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride), HOBt (1- Hydroxybenzotriazole hydrate), HBTU ((2-(1H-benzotriazol-1-yl)-1,1,3,3- tetramethyluronium hexafluorophosphate), T3P (propanephosphonic acid anhydride), TBAI (tetrabutylammonium iodide), LAH (lithium aluminum hydride), TFA (trifluoroacetic acid). ^{H2932377.1} 13 Docket No.2877.037AWO [0052] The syntheses of the 1-acyl-3-aminoindazole derivatives of type 1 were performed as depicted in scheme S1. Starting from the corresponding phenol derivatives, a recently reported protocol comprising deprotonation with NaH and subsequent SN2- reaction with enantiopure (R)- and (S)-3-bromo-2-methyl propanol, (1) respectively, allowed to obtain the enantiopure (R)- and (S)-3-aryloxy-2-methylpropanol derivatives 11a-h and ent11a-d, respectively. Both phenoxy-derivatives 11a/ent11a (1) and the fluoro derivative 11e (2) have been described in the literature. Jones oxidation gave the corresponding carboxylic acid derivatives 12a-h and ent12a-d. It is worthy of note, that the phenoxy derivatives 12a/ent12a (1), the racemic naphthyl derivative 12b (3) and the biphenyl derivative 12g (4) have been previously published. The coupling reactions using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC × HCl) and 1- hydroxy-7-azabenzotriazole (HOAt) with 6-fluoro-1H-indazole-3-amine (13) proceeded with high regioselectivity to give access to the 1-acyl-3-aminoindazole derivatives 1a-h and ent1a-d in 15-53% yield. Investigation of the alcohol derivatives 11a-h / ent11a-d and the target compounds 1a-h / ent1a-d by chiral HPLC proved the high stereospecificity of the synthetic pathway. [0053] The fluoroindazole derivative renouncing the phenoxy moiety 2 was obtained in high yield by reacting isobutyryl chloride with HOAt, followed by the addition of 6- fluoro-1H-indazole-3-amine (13) (scheme S2). [0054] The derivatives modified in position 3 of the indazole scaffold were prepared as displayed in scheme S3.The 3-desamino derivative 3a was synthesized as a racemate starting from 6-fluoro-1H-indazole (14) and the commercially available racemic carboxylic acid derivative rac-12a.6-Fluoro-3-methylindazole (15) gave access to the enantiopure target compound 3b. For the synthesis of the N-methyl derivative 3c, a novel, efficient pathway for the preparation of 3-(N-methylamino)indazole derivatives had to be established. In detail, the diacetyl derivative 16 was prepared from 6-fluoro-1H-indazole- 3-amine (13) using acetylchloride in the presence of pyridine and 4- dimethylaminopyridine (DMAP). Deprotonation with sodium hydride (NaH) and subsequent reaction with methyl iodide (MeI) gave rise to the N-methyl derivative 17, which was deprotected with hydrogen chloride (HCl) in methanol (MeOH). The resulting 6-fluoro-N-methyl-1H-indazol-3-amine (18) was acylated with the (S)-carboxylic acid ^{H2932377.1} 14 Docket No.2877.037AWO derivative 12a to give the target compound 3c. The aza analog 4 was synthesized starting from 6-fluoro-1H-pyrazolo[4,3-b]pyridin-3-amine (20), which was available by a ring closing reaction of the pyridine derivative 19 with hydrazine hydrate according to a previously reported protocol. (5) It is noteworthy, that the cyclization reaction also led to the nucleophilic attack of hydrazine in position 5 of the pyridine nucleus. Thus, the respective aryl hydrazine side product was formed, which had to be separated by preparative HPLC. Coupling with the (S)-carboxylic acid derivative 12a resulted in the formation of target compound 4. [0055] Furthermore, the fluorine in position 6 of the indazole scaffold was replaced by a variety of substituents (scheme S4). The alkoxy substituted compounds 5a,b were synthesized as racemates starting from rac-12a, employing the respective, commercially available 6-alkoxy-indazole derivatives 21 and 22. Benzotriazol-1- yloxytripyrrolidinophosphonium hexafluorophosphate (PyBOP) in presence of HOAt and N,N-diisopropylethylamine (DIPEA) were used for the coupling reaction under microwave promoted conditions. Starting from the buyable 6-trifluoromethyl-, 6-chloro- or 6-aminoindazole analogs 23,24 and 25, respectively, the EDC/HOAt promoted coupling with the (S)-carboxylic acid derivative 12a furnished the respective target compounds 5c, 5d and 5f. [0056] The syntheses of the congeners bearing an aminocarbonyl or a hydroxymethyl group in position 6 are outlined in scheme S5. The primary amide derivative 27 was obtained from the commercially available carboxylic acid derivative 26 by a one pot coupling protocol employing PyBOP in presence of NH4Cl and DIPEA. The carbinol derivative 29 was synthesized by the cyclization reaction of the commercially available o-fluorobenzonitrile precursor 28 with hydrazine hydrate according to an established method.(6) Both scaffolds were coupled with the (S)-carboxylic acid derivative 12a acid to furnish the both target compounds 5f and 5g. Starting from the (R)-carboxylic acid derivative ent-12a, the corresponding enantiomers ent-5f and ent-5g were prepared as well. Furthermore, the (S)-4-fluorophenoxy analog F-5g was obtained in a similar fashion by the acylation with the (S)-carboxylic acid derivative 12e. [0057] The 6-carboxamide and 6-amino derivatives 7a and 7b renouncing the amino group in position 3 were obtained analogously starting from the respective purchasable ^{H2932377.1} 15 Docket No.2877.037AWO precursors 30 and 31 (scheme S6). The enantiomer ent-7a was synthesized employing (R)-carboxylic acid derivative ent-12a. [0058] The N-formyl congeners 6a, 6b and 8 were prepared starting from the corresponding amine derivatives 5e and 7b taking advantage of activated formic acid (scheme S7). [0059] The preparation of the fused derivative 9 (scheme S8) was achieved starting from the alkine derivative 32, performing a gold(I) promoted hydroarylation reaction to give the chromene derivative 33.(7) Since the original cyclization protocol did not work with satisfying yield and purity, we preferred to apply more regioselective conditions employing toluene as the solvent and working at 0 °C, (8) thus leading to a cleaner formation of only 14% regioisomer instead of 25% reported in the literature. The mixture thus obtained was subjected to hydrogenation to give the chroman derivative 34 (and the respective regioisomer). Reaction with N-bromosuccinimide furnished the 6-brominated derivative 35, when chromatography allowed to reduce the amount of the regioisomer to 6%. Transnitrilation reaction employing n-butyllithium and dimethylmalononitrile allowed to cleanly prepare the cyclization precursor 36, (9) which could be subjected to hydrazine hydrate to obtain the fused indazole-amine derivative 37. Acylation with the racemic carboxylic acid derivative rac-12a led to the formation of the target compound 9. [0060] The preparation of the compounds incorporating a modified spacer was performed as shown in scheme S9. Starting from the commercially available carboxylic acids 39-42 or the respective sodium carboxylate 38, coupling with EDC × HCl and HOAt allowed to obtain the target compounds 10a-e. Preparative chiral HPLC gave rise to the separated enantiomers of 10b-d. [0061] Synthesis Schemes ^{H2932377.1} 16 Docket No.2877.037AWO

[0062] Scheme S1. Synthesis of the enantiopure derivatives modified at the phenoxy site, reagents and conditions: (a) 1. NaH, DMF, 0 °C, 30 min, 2. (R)-3-bromo-2- methylpropan-1-ol or (S)-3-bromo-2-methylpropan-1-ol, DMF, rt, 18-31 h, (50-72% crude); (b) 1. CrO₃, H₂SO₄, H₂O, acetone, 0 °C, 3-5 h, 2. iPrOH, 0 °C -> rt (48-64%, crude); (c) 1. EDC × HCl, HOAt, DMF, rt, 2.6-fluoro-1H-indazole-3-amine (13), rt, 1-3 h (15-53%).

renouncing the phenoxy site, reagents and conditions: (a) 1. isobutyryl chloride, HOAt, DIPEA, DMF, 0 °C, 1 h, 2.6-fluoro- 1H-indazole-3-amine (13), microwave irradiation (90%). ^{H2932377.1} 17 Docket No.2877.037AWO

3 or 4, reagents, and conditions: (a) EDC × HCl, HOAt, DMF, rt (3a: 39%, 3b: 57%, 3c: 29%, 4: 31%); (b) AcCl, pyridine, DMAP, 0 °C to rt, 2 h (72%), (c) NaH, MeI, DMF, rt, 1 h (77%); (d) 1.25 M HCl in MeOH, 115 °C, 2 h (100% crude); (e) N₂H₄, EtOH, 70 °C, 17 h (15%).

Docket No.2877.037AWO [0065] Scheme S4. Synthesis of the derivatives with alkoxy substituents in positions 6, reagents and conditions: (a) PyBOP, HOAt, DMF, microwave irradiation (5a: 56%, 5b: 54%); (b) EDC × HCl, HOAt, DMF, rt (5c: 23%, 5d: 54%, 5e: 44%). ;

6/3, reagents and conditions: (a) EDC × HCl, HOAt, DMF, rt (7a: 64%, 7b: 14%). {^H2932377.1} 19 Docket No.2877.037AWO

a formyl group, reagents and conditions: (a) 1. HCO₂H, Ac₂O, THF, 60 °, 2 h, 2.5e, 0 °C, 30 min (42%); (b) 1. HCO2H, Ac2O, THF, 60 °C, 2 h, 2.5e or 7b 0 °C to rt, for 6b: 60 min, for 8: 90 min (6b: 37%, 8: 28%).

[2- biphenyl)di-tert-butylphosphine]gold(I) hexafluoroantimonate, toluene, 0 °C (crude); (b) H2/Pd(OH)2/C, MeOH, rt, 2 h (crude); (c) N-bromosuccinimide, CH3CN, 0 °C (23% over three steps); (d) 1. BuLi, THF, -79 °C.2. dimethylmalononitrile, THF, -79 °C (54%); (e) N2H4, BuOH, 120 °C, 22 h (76%); (f) EDC × HCl, HOAt, DMF, microwave irradiation (61%). ^{H2932377.1} 20 Docket No.2877.037AWO

conditions: (a) 1. EDC × HCl, HOAt, DMF, rt, 10 min, 2.6-fluoro-1H-indazole-3-amine (13), rt, 10a and 10b: 3 h, (10a: 27%, 10b: 45%,). [0071] General materials and methods for organic synthesis [0072] Reagents and solvents were purchased in their purest grade from abcr, Acros, Alfa Aesar, Chemspace, Enamine, Manchester Organics, Sigma Aldrich, TCI, VWR and were used without further purification. Microwave assisted (Discover^® microwave oven, CEM Corp.) synthesis was carried out by 20 × irradiating with microwaves (50 W, irradiation time: 10 s). In between each irradiation step, intermittent cooling of the reaction mixture to a temperature of -10°C was achieved by sufficient agitation in an ethanol-ice bath. TLC analyses were performed using Merck 60 F254 aluminum sheets and analyzed by UV light (254 nm). Purification by flash column chromatography was conducted using silica gel 60 (40-63 µm mesh, Merck) and eluents as binary mixtures with the volume ratios indicated. Preparative HPLC was performed on an Agilent 1200 preparative series HPLC system or on an Agilent HPLC 1260 Infinity system combined with an MWD detector and fraction collector, applying a linear gradient and a flow rate as indicated below. As HPLC column, a Zorbax-Eclipse XDB-C8 PrepHT (21.2 mm × 150 mm, 5 µm) was used. For the separation of enantiomers, a Waters preparative HPLC system consisting of the 2545 binary gradient module, 2707 autosampler, 2998 photodiode array detector and the fraction collector III was employed, using a Chiralpak IC column (250 mm × 30 mm, 5 µm), a flow rate of 45 mL/min and an isocratic binary solvent system specified below. Compounds were characterized by NMR spectroscopy, IR spectroscopy, high-resolution mass spectra (HRMS) and purity was assessed by RP- HPLC. All assayed compounds were >95% pure. ESI-mass spectra were recorded using ^{H2932377.1} 21 Docket No.2877.037AWO LC-MS: Thermo Scientific Dionex Ultimate 3000 UHPLC quarternary pump, autosampler and RS-diode array detector, column: Zorbax-Eclipse XDB-C8 analytical column, 3.0 mm × 100 mm, 3.5 μm, flow rate 0.4 mL/min using DAD detection (230 nm; 254 nm), coupled to a Bruker Daltonics Amazon mass spectrometer using ESI as ionization source. High mass accuracy and resolution experiments were performed on a Bruker Daltonics timsTOF Pro spectrometer using electrospray ionization (ESI) as ionization source. NMR spectra were obtained either on a Bruker Avance III 400 or a Bruker Avance III 600 (600 MHz for ¹H and 151 MHz for ¹³C) spectrometer, the latter equipped with a Prodigy nitrogen cooled probe at 297 K, using the deuterated solvents indicated below. For the spectra recorded in organic solvents, the chemical shifts are reported in ppm (δ) relative to TMS. For measurements in D₂O, the water peak was used for calibration. IR spectra were performed on a Jasco FT/IR 4100 spectrometer using a KBr pellet or with substance film on a NaCl crystal plate, as specified. Substance purities were assessed by analytical HPLC (Agilent 1100 analytical series, equipped with a quarternary pump and variable wavelength detector detector; column Zorbax Eclipse XDB-C8 analytical column, 4.6 mm × 150 mm, 5 μm, flow rate 0.5 mL / min, detection wavelengths: 220 nm, 254 nm; system 1: methanol / 0.1% aq. HCOOH, linear gradient: 10% methanol for 3 min, 10% to 100% methanol in 15 min, 100% methanol for 6 min; system 2: acetonitrile / 0.1% aq. HCOOH, linear gradient: 5% acetonitrile for 3 min, 5% to 95% acetonitrile in 15 min, 95% acetonitrile for 6 min; system 3: acetonitrile / 0.1% aq. TFA, linear gradient: 3% to 85% acetonitrile in 26 min, 85% to 95% acetonitrile in 2 min, 95% acetonitrile for 2 min; system 4: acetonitrile / 0.1% aq. TFA, linear gradient: 3% to 25% acetonitrile in 26 min) system 5: acetonitrile / 0.1% aq. HCOOH, linear gradient: 3% to 85% acetonitrile in 26 min, 85% to 95% acetonitrile in 2 min, 95% acetonitrile for 2 min;. Chiral analytical HPLC was run on an AGILENT series 1100 system equipped with a VWD and detection at 254 nm. As chiral column a DAICEL Chiralpak IC column (4.6 mm × 250 mm, 5 µM) was used at 20 °C and a flow rate 1.0 mL / min with the solvent system as indicated. Specific optical rotation values (°× mL × dm^-1 × g^-1) were obtained from a Jasco P2000 polarimeter with the solvents indicated. Melting points were determined in open capillaries using a Büchi 510 melting point apparatus and are given uncorrected. ^{H2932377.1} 22 Docket No.2877.037AWO General experimental procedures [0073] Unless otherwise noted, reactions were performed under nitrogen atmosphere employing dry solvents of commercial quality, used as purchased. All reactions were carried out using a magnetic stirrer with optional aluminum heating block or ice bath for sealed microwave vials and oil or ice bath for round bottom flasks, respectively. Solvents were evaporated by a rotation evaporator with a membrane vacuum pump. Products purified by preparative HPLC using aqueous solvents were lyophilized. [0074] General procedure A (GPA): preparation of 1.26 M Jones reagent To a stirred solution of CrO₃ (126 mg) in 500 µL water at 0 °C was slowly added conc. H2SO4 (126 µL). The volume was adjusted to 1 mL using water. Detailed synthesis procedures [0075] (R)-2-Methyl-3-phenoxypropan-1-ol [(R)-SX224, 11a] (1) To a stirred solution of phenol

sieve (187 µL, 2.13 mmol) in DMF (4 mL) in a flame-dried flask, sodium hydride (60% in mineral oil; 97.4 mg, 2.44 mmol) was added. After stirring the suspension for 30 min at rt, (R)-3-bromo-2- methylpropan-1-ol (213 µL, 2.03 mmol) was added. The mixture was stirred for additional 18 h, then 2N NaOH was added and extraction with tert-butyl methyl ether (3×) was performed. The combined organic layers were washed with water and brine, dried over Na2SO4 and concentrated. After purification by flash-chromatography (isohexane / tert-butyl methyl ether, 7:3) compound (R)-SX22411a (202 mg, 1.22 mmol, 60%) was obtained as colorless liquid. ESI-MS: m/z 189.3 [M+Na]⁺. ¹H-NMR: (400 MHz, CDCl3) δ 1.04 (d, J = 7.0 Hz, 3H), 1.90 (dd, J = 5.7, 5.7 Hz, 1H), 2.21 (m, 1H), 3.73 – 3.70 (m, 2H), 3.93 (dd, J = 9.1, 7.0 Hz, 1H), 3.98 (dd, J = 9.1, 5.3 Hz, 1H), 6.89 – 6.93 (m, 2H), 6.93 – 6.98 (m, 1H), 7.32 – 7.26 (m, 2H). ^{H2932377.1} 23 Docket No.2877.037AWO ¹³C-NMR: (DEPTQ, 101 MHz, CDCl₃) δ 13.6, 35.7, 66.2, 71.2, 114.5, 120.9, 129.5, 158.8. chiral HPLC:isocratic elution with n-hexane / isopropanol, 95:5: tR = 6.6 min, 99% ee (254 nm). [α]D²⁴: +2.4 (c = 0.9, chloroform). The analytical data of this compound are in accordance with those reported in the literature. [0076] (S)-2-Methyl-3-phenoxypropan-1-ol [(S)-SX224, ent-11a] (1)

(S)-2-Methyl-3-phenoxypropan-1-ol [(S)-SX224] ent-11a was synthesized analogously as described for (R)-SX22411a, starting from (S)-3-bromo-2-methylpropan-1-ol. chiral HPLC: isocratic elution with n-hexane / isopropanol, 95:5: tR = 6.9 min, 99% ee (254 nm). [α]_D ²⁵: -2.9 (c = 1.6, chloroform). The analytical data of this compound are in accordance with those reported in the literature. [0077] (R)-2-Methyl-3-(naphthalen-2-yloxy)propan-1-ol [(R)-SX233, 11b] To a stirred solution of 2-

in DMF (4 mL) in a flame-dried flask sodium hydride (60% in mineral oil; 64.0 mg, 1.60 mmol) was added. After stirring the suspension for 30 min at rt, (R)-3-bromo-2-methylpropan-1-ol (138 µL, 1.32 mmol) was added. The mixture was stirred for additional 22 h, then 2N NaOH was added and extraction with tert-butyl methyl ether (3×) was performed. The combined organic layers were washed with water and brine, dried over Na₂SO₄ and concentrated. After purification by flash-chromatography (isohexane / tert-butyl methyl ether, 7:3) compound ^{H2932377.1} 24 Docket No.2877.037AWO (R)-SX23311b (206 mg, 0.95 mmol, 72%) was obtained as pale yellow liquid, which solidified upon standing (white wax). ESI-MS: m/z 238.9 [M+Na]⁺. HR-ESI-MS: m/z [M+H]⁺ calcd.239.1043 for C₁₄H₁₆NaO₂, found 239.1044. ¹H-NMR: (400 MHz, CDCl3) δ 1.09 (d, J = 7.0 Hz, 3H), 2.22 – 2.34 (m, 1H), 3.74 – 3.78 (m, 2H), 4.06 (dd, J = 9.1, 6.7 Hz, 1H), 4.09 (dd, J = 9.1, 5.5 Hz, 1H), 7.18 – 7.10 (m, 2H), 7.34 (ddd, J = 8.1, 6.9, 1.3 Hz, 1H), 7.44 (ddd, J = 8.2, 6.9, 1.3 Hz, 1H), 7.68 – 7.80 (m, 3H). ¹³C-NMR: (DEPTQ, 101 MHz, CDCl3) δ 13.7, 35.7, 66.2, 71.2, 106.7, 118.8, 123.7, 126.4, 126.7, 127.6, 129.0, 129.4, 134.5, 156.7. chiral HPLC: isocratic elution with n-hexane / isopropanol, 98:2: t_R = 17.5 min, >99% ee (254 nm). [α]D²⁴: +1.8 (c = 0.7, chloroform). [0078] (S)-2-Methyl-3-(naphthalen-2-yloxy)propan-1-ol [(S)-SX233, ent-11b]

Methyl-3-(naphthalen-2-yloxy)propan-1-ol [(S)-SX233] ent-11b was synthesized analogously as described for (R)-SX23311b, starting from (S)-3-bromo-2-methylpropan- 1-ol. The analytical data were in accordance. chiral HPLC: isocratic elution with n-hexane / isopropanol, 98:2: tR = 18.2 min, >99% ee (254 nm). [α]D²³: -3.0 (c = 0.8, chloroform). [0079] (S)-1-(3-Amino-6-(hydroxymethyl)-1H-indazol-1-yl)-3-(4-fluorphenoxy)-2- methylpropan-1-one (I1421 F-5g) ^{H2932377.1} 25 Docket No.2877.037AWO To a solution of (S)-

(12.4 mg, 0.09 mmol) in DMF (0.5 mL) was added EDC × HCl (17.4 mg, 0.09 mmol). After stirring for 10 min at rt, a solution of I139629 (14.8 mg, 0.09 mmol) in DMF (0.5 mL) was added. After stirring the reaction mixture for 3 h, the solvent was removed by lyophilization and the residue was purified by preparative HPLC using a solvent system of CH3OH / 0.1% aq. HCOOH, a flow rate of 10 mL / min and a gradient of 50% to 80% CH₃OH in 15 min (t_R: 10.2 min, λ = 220 nm) to afford I1421 F-5g (14.2 mg, 41.4 µmol, 45%) as a white solid. ESI-MS: m/z 344.3 [M+H]⁺. HR-ESI-MS: m/z [M+H]⁺ calcd.344.1404 for C18H19FN3O3, found 344.1404. IR (KBr): 3473, 3357, 3314, 1636 cm^-1. m.p.: 136 °C. ¹H-NMR: (600 MHz, DMSO-d₆) δ 1.27 (d, J = 7.3 Hz, 3H), 4.03 (dd, J = 9.1, 5.6 Hz, 1H), 4.09 (ddq, J = 7.9, 5.6, 7.3 Hz, 1H), 4.36 (dd, J = 9.1, 7.9 Hz, 1H), 4.63 (d, J = 5.6 Hz, 2H), 5.36 (t, J = 5.6 Hz, 1H), 6.94 – 6.98 (m, 2H), 7.07 – 7.12 (m, 2H), 7.28 (dd, J = 8.3, 1.1 Hz, 1H), 7.82 (d, J = 8.3 Hz, 1H), 8.26 (brs, 1H). ¹³C-NMR: (DEPTQ, 151 MHz, DMSO-d6) δ 14.0, 37.7, 62.9, 69.7, 112.7, 115.77 (d, J = 12 Hz), 115.82 (d, J = 19 Hz), 119.1, 120.2, 122.4, 139.6, 145.0, 152.9, 154.7, 156.5 (d, J = 239 Hz), 172.1. HPLC: system 3: λ = 220 nm, t_R = 20.4 min, purity: >99%. chiral HPLC: isocratic elution with n-hexane / ethanol, 9:1: tR = 18.7 min, 99% ee (254 nm). [α]D²²: +31.8 (c = 0.5, methanol). [0080] (S)-1-(3-Amino-6-(hydroxymethyl)-1H-indazol-1-yl)-2-methyl-3- phenoxypropan-1-one (I14085g) ^{H2932377.1} 26 Docket No.2877.037AWO

To a solution of (S)-SX225 (12a,15.0 mg, 0.08 mmol) and HOAt (13.6 mg, 0.10 mmol) in DMF (0.5 mL) was added EDC × HCl (19.2 mg, 0.10 mmol). After stirring for 10 min at rt, a solution of I139629 (16.3 mg, 0.10 mmol) in DMF (0.5 mL) was added. After stirring the reaction mixture for 3 h, the solvent was removed by lyophilization and the residue was purified by preparative HPLC using a solvent system of CH₃OH / 0.1% aq. HCOOH, a flow rate of 10 mL / min and a gradient of 50% to 80% CH₃OH in 15 min (t_R: 12.5 min, λ = 220 nm) to afford I14085g (14.4 mg, 44.3 µmol, 53%) as a white solid. ESI-MS: m/z 326.1 [M+H]⁺. HR-ESI-MS: m/z [M+H]⁺ calcd.326.1499 for C₁₈H₁₉N₃O₃, found 326.1499. IR (KBr): 3478, 3357, 1636 cm^-1. m.p.: 132 °C. ¹H-NMR: (600 MHz, DMSO-d₆) δ 1.28 (d, J = 7.0 Hz, 3H), 4.05 (dd, J = 8.8, 5.5 Hz, 1H), 4.11 (ddq, J = 8.0, 5.5, 7.0 Hz, 1H), 4.39 (dd, J = 8.8, 8.0 Hz, 1H), 4.64 (d, J = 5.8 Hz, 2H), 5.37 (t, J = 5.8 Hz, 1H), 6.49 (s, 2H), 6.90 – 6.95 (m, 3H), 7.25 – 7.30 (m, 3H), 7.82 (d, J = 8.0 Hz, 1H), 8.27 (brs, 1H). ¹³C-NMR: (DEPTQ, 151 MHz, DMSO-d₆) δ 14.1, 37.8, 63.0, 69.0, 112.8, 114.5, 119.2, 120.3, 122.4, 129.5, 139.7, 145.0, 152.9, 158.4, 172.2. HPLC: system 3: λ = 220 nm, tR = 20.2 min, purity: 99%. chiral HPLC: isocratic elution with n-hexane /ethanol, 9:1: t_R = 16.7 min, 98% ee (254 nm). [α]D²²: +27.9 (c = 0.5, methanol). [0081] (S)-1-(3-Amino-6-fluoro-1H-indazol-1-yl)-3-(4-fluorophenoxy)-2- methylpropan-1-one [(S)-SX263, 1e] ^{H2932377.1} 27 Docket No.2877.037AWO

To a solution of (S)- mg, (2.5 mL) were added HOAt (13.6 mg, 0.10 mmol) and EDC × HCl (19.2 mg, 0.10 mmol) at rt. The resulting solution was stirred for 10 min at rt and then 6-fluoro-1H-indazol-3-amine (13, 19.6 mg, 0.13 mmol) in DMF (0.5 mL) was added. After stirring the reaction mixture at rt for 1.5 h, the solvent was removed by lyophilization. Purification by preparative HPLC using a solvent system of CH₃OH / 0.1% aq. HCOOH, a flow rate of 10 mL / min and a gradient of 50% to 80% CH3OH in 15 min, 80% to 95% CH3OH in 2 min, 95% CH3OH for 2 min (tR = 16.0 min, λ = 254 nm) afforded (S)-SX2631e (16.9 mg, 51.0 μmol, 51%) as a white solid. ESI-MS: m/z 332.2 [M+H]⁺. HR-ESI-MS: m/z [M+H]⁺ calcd.332.1205 for C17H16F2N3O2, found 332.1203. IR (NaCl): 3463, 3357, 2925, 2849, 1683, 1625, 1509, 1419 cm^-1. m.p.: 148 °C. ¹H-NMR: (600 MHz, DMSO-d6) δ 1.28 (d, J = 6.9 Hz, 3H), 4.02 – 4.11 (m, 2H), 4.33 – 4.40 (m, 1H), 6.62 (s, 2H), 6.93 – 6.99 (m, 2H), 7.06 – 7.13 (m, 2H), 7.26 (ddd, J = 9.0, 8.9, 2.3 Hz, 1H), 7.92 – 7.99 (m, 2H). ¹³C-NMR: (DEPTQ, 151 MHz, DMSO-d6) δ 13.9, 37.7, 69.5, 101.8 (d, J = 28 Hz), 112.2 (d, J = 25 Hz), 115.7 (d, J = 7 Hz), 115.8 (d, J = 8 Hz), 117.0, 122.7 (d, J = 11 Hz), 139.8 (d, J = 13 Hz), 152.7, 155.1 (d, J = 2 Hz), 156.5 (d, J = 236 Hz), 163.2 (d, J = 245 Hz), 172.4. ¹⁹F-NMR: (377 MHz, DMSO-d6) δ -110.5, -123.3. HPLC: system 1: λ = 254 nm, tR = 20.1 min, purity: 97%. chiral HPLC: isocratic elution with n-hexane / isopropanol, 95:5: t_R = 11.4 min, 99% ee (254 nm). [α]D²³: -2.0 (c = 0.9, methanol). ^{H2932377.1} 28 Docket No.2877.037AWO [0082] (S)-1-(3-Amino-6-chloro-1H-indazol-1-yl)-2-methyl-3-phenoxypropan-1-one (I14145d)

To a solution of (S)-2-methyl-3-phenoxypropionic acid (12a, 15.0 mg, 0.08 mmol) and HOAt (11.3 mg, 0.08 mmol) in DMF (1 mL) was added EDC × HCl (15.9 mg, 0.08 mmol). After stirring for 10 min at rt, a solution of 6-chloro-1H-indazol-3-amine (24, 16.7 mg, 0.10 mmol) in DMF (0.5 mL) was added. After stirring the reaction mixture for 4 h, the solvent was removed by lyophilization and the residue was purified by preparative HPLC using a solvent system of CH3OH / 0.1% aq. HCOOH, a flow rate of 10 mL / min and a gradient of 50% to 80% CH₃OH in 15 min, 80% to 95% CH₃OH in 2 min, 95% CH₃OH for 2 min, 95% to 50% CH₃OH in 2 min (t_R: 20.2 min, λ = 220 nm) to afford I14145d (14.7 mg, 44.7 µmol, 54%) as a grey-white resin. ESI-MS: m/z 330.2 [M+H]⁺. HR-ESI-MS: m/z [M+H]⁺ calcd.330.1004 for C₁₇H₁₇ClN₃O₂, found 330.1005. IR (KBr): 3429, 3335, 3247, 3223, 1685 cm^-1. ¹H-NMR: (600 MHz, DMSO-d6) δ 1.27 – 1.39 (m, 3H), 4.03 – 4.11 (m, 2H), 4.35 – 4.41 (m, 1H), 6.65 (s, 2H), 6.90 – 6.95 (m, 3H), 7.24 – 7.30 (m, 2H), 7.43 (dd, J = 8.4, 2.0 Hz, 1H), 7.93 (d, J = 8.4 Hz, 1H), 8.26 (d, J = 2.0 Hz, 1H). ¹³C-NMR: (DEPTQ, 151 MHz, DMSO-d₆) δ 13.9, 37.8, 68.8, 114.5, 114.7, 119.1, 120.7, 122.4, 124.2, 129.5, 139.7, 152.6, 158.3, 172.5. HPLC: system 3: λ = 220 nm, tR = 25.8 min, purity: 96%. chiral HPLC: isocratic elution with n-hexane / isopropanol, 95:5: tR = 13.0 min, 99% ee (254 nm). [α]D²⁰: +45.3 (c = 0.5, chloroform). ^{H2932377.1} 29 Docket No.2877.037AWO [0083] (S)-1-(2-Methyl-3-phenoxypropanoyl)-1H-indazole-6-carboxamide (I14097a)

To a solution of (S)-SX22512a (15.0 mg, 0.08 mmol) and HOAt (13.6 mg, 0.10 mmol) in DMF (0.5 mL) was added EDC × HCl (19.2 mg, 0.10 mmol). After stirring for 10 min at rt, a solution of 1H-indazole-6-carboxamide (30, 16.1 mg, 0.10 mmol) in DMF (0.5 mL) was added. After stirring the reaction mixture for 3 h, the solvent was removed by lyophilization and the residue was purified by preparative HPLC using a solvent system of CH₃OH / 0.1% aq. HCOOH, a flow rate of 10 mL / min and a gradient of 50% to 80% CH3OH in 15 min, 80% to 95% CH3OH in 2 min (tR: 15.5 min, λ = 220 nm) to afford I140912a (17.1 mg, 52.9 µmol, 64%) as a white solid. ESI-MS: m/z 324.1 [M+H]⁺. HR-ESI-MS: m/z [M+H]⁺ calcd.324.1343 for C18H17N3O3, found 324.1345. IR (KBr): 3482, 3437, 3397, 3357, 3197, 1718, 1707, 1662 cm^-1. m.p.: 141 °C. ¹H-NMR: (600 MHz, DMSO-d₆) δ 1.38 (d, J = 7.2 Hz, 3H), 4.21 (dd, J = 10.0, 5.3 Hz, 1H), 4.35 (ddq, J = 6.8, 5.3, 7.2 Hz, 1H), 4.43 (dd, J = 10.0, 6.8 Hz, 1H), 6.90-6.94 (m, 3H), 7.24 – 7.28 (m, 2H), 7.53 (brs, 1H), 7.91 (dd, J = 8.3, 1.6 Hz, 1H), 7.98 (dd, J = 8.3, 0.6 Hz, 1H), 8.24 (brs, 1H), 8.60 (d, J = 1.0 Hz, 1H), 8.85 (ddd, J = 1.6, 1.0, 0.6 Hz, 1H). ¹³C-NMR: (DEPTQ, 151 MHz, DMSO-d6) δ 13.9, 38.2, 68.9, 114.2, 114.4, 120.7, 121.4, 123.9, 129.4, 135.7, 138.2, 140.5, 158.1, 167.6, 174.1. HPLC: system 3: λ = 220 nm, t_R = 21.7 min, purity: 98%. chiral HPLC: isocratic elution with n-hexane / isopropanol, 85:5: tR = 21.6 min, 98% ee (254 nm). [α]_D ²²: +111.4 (c = 0.4, methanol). ^{H2932377.1} 30 Docket No.2877.037AWO [0084] (S)-1-(3-Amino-6-fluoro-1H-indazol-1-yl)-2-methyl-3-(naphthalen-2- yloxy)propan-1-one [(S)-SX240, 1b]

To a solution of compound (S)-SX23712b (25.2 mg, 0.11 mmol) in DMF (2 mL) were added HOAt (15.3 mg, 0.11 mmol) and EDC × HCl (21.3 mg, 0.11 mmol) at rt. The resulting solution was stirred for 10 min at rt and then 6-fluoro-1H-indazole-3-amine (13, 13.8 mg, 91.2 µmol) in DMF (0.5 mL) were added. After stirring the mixture at rt for 1.5 h, the solvent was removed by lyophilization. Purification by preparative HPLC using a solvent system of CH₃OH / 0.1% aq. TFA, a flow rate of 10 mL / min and a gradient of 50% to 80% CH₃OH in 15 min, 80% to 95% CH₃OH in 2 min, 95% CH₃OH for 5 min (tR = 21.0 min, λ = 254 nm) afforded (S)-SX2401b (11.4 mg, 31.3 μmol, 34%) as a white solid. ESI-MS m/z 364.2 [M+H]⁺. HR-ESI-MS: m/z [M+H]⁺ calcd.364.1456 for C21H19FN3O2, found 364.1460. IR (NaCl): 3458, 2924, 2851, 1629, 1424 cm^-1. m.p.: 59 °C. MHz, DMSO-d6) δ 1.34 (d, J = 7.0 Hz, 1H), 4.16 (ddq,

7.0, 5.5 Hz, 1H), 4.21 (dd, J = 9.1, 5.5 Hz, 1H), 4.52 (dd, J = 9.1, 7.8 Hz, 1H), 6.62 (s, 2H), 7.11 (dd, J = 9.0, 2.5 Hz, 1H), 7.26 (ddd, J = 9.0, 8.9, 2.5 Hz, 1H), 7.34 (ddd, J = 8.1, 6.9, 1.2 Hz, 1H), 7.39 (d, J = 2.5 Hz, 1H), 7.45 (ddd, J = 8.1, 6.9, 1.2 Hz, 1H), 7.78 – 7.83 (m, 3H), 7.94 – 7.99 (m, 2H). ¹³C-NMR: (DEPTQ, 151 MHz, DMSO-d₆) δ 14.0, 37.7, 69.0, 101.8 (d, J = 28 Hz), 106.9, 112.2 (d, J = 25 Hz), 117.0, 118.6, 122.7 (d, J = 11 Hz), 123.6, 126.4, 126.7, 127.5, 128.5, 129.3, 134.2, 139.8 (d, J = 13 Hz), 152.7, 156.2, 163.2 (d, J = 245 Hz), 172.5. ^{H2932377.1} 31 Docket No.2877.037AWO ¹⁹F-NMR: (377 MHz, DMSO-d₆) δ -110.5. HPLC: system 1: λ = 254 nm, tR = 20.5 min, purity: 95% chiral HPLC: isocratic elution with n-hexane / isopropanol, 9:1: tR = 9.7 min, >99% ee (254 nm). [α]D²³: -15.8 (c = 0.4, methanol). [0085] (S)-1-(3-Amino-6-fluoro-1H-indazol-1-yl)-3-(4-bromophenoxy)-2- methylpropan-1-one [(S)-SX264, 1f]

To a solution of (S)-SX26112f (19.2 mg, 74.1 μmol) in DMF (2.5 mL) were added HOAt (10.1 mg, 74.1 μmol and EDC × HCl (14.2 mg, 74.1 μmol) at rt. The resulting solution was stirred for 10 min at rt and then 6-fluoro-1H-indazol-3-amine (13, 13.4 mg, 88.9 μmol) in DMF (0.5 mL) was added. After stirring the reaction mixture at rt for 1.5 h, the solvent was removed by lyophilization. Purification by preparative HPLC using a solvent system of CH3OH / 0.1% aq. HCOOH, a flow rate of 10 mL / min and a gradient of 50% to 80% CH₃OH in 15 min, 80% to 95% CH₃OH in 2 min, 95% CH₃OH for 4 min (tR = 18.0 min, λ = 254 nm) afforded (S)-SX2641f (8.22 mg, 21.0 μmol, 28%) as a white solid. ESI-MS: m/z 392.3 [M+H]⁺. HR-ESI-MS: m/z [M+H]⁺ calcd.392.0404 for C₁₇H₁₆BrFN₃O₂, found 392.0402 IR (NaCl): 3466, 3346, 2922, 1680, 1625, 1560, 1423, 1238, 1074 cm^-1. m.p.: 112 °C.

Docket No.2877.037AWO ¹³C-NMR: (DEPTQ, 151 MHz, DMSO-d₆) δ 13.8, 37.6, 69.2, 101.8 (d, J = 28 Hz), 112.1, 112.3 (d, J = 25 Hz), 116.8, 117.0, 122.7 (d, J = 11 Hz), 132.1, 139.8 (d, J = 13 Hz), 152.7, 157.6, 163.2 (d, J = 245 Hz), 172.3. ¹⁹F-NMR: (377 MHz, DMSO-d₆) δ -110.4. HPLC: system 1: λ = 254 nm, tR = 20.8 min, purity: >99%. chiral HPLC: isocratic elution with n-hexane / isopropanol, 95:5: tR = 12.0 min, 98% ee (254 nm). [α]D²⁶: -6.1 (c = 0.3, methanol). [0086] (S)-3-Amino-1-(2-methyl-3-phenoxypropanoyl)-1H-indazole-6-carboxamide (I14245f) To a solution of (S)-

HOAt (9.3 mg, 0.07 mmol) in DMF (0.5 mL) was added EDC × HCl (13.1 mg, 0.09 mmol). After for 10 min at rt, this solution was added to a suspension of 3-amino-1H-indazole-6-carboxamide (12.0 mg, 0.09 mmol) in DMF (0.5 mL). After stirring the reaction mixture for 3 h, the solvent was removed by lyophilization and the residue was purified by preparative HPLC using a solvent system of CH3OH / 0.1% aq. HCOOH, a flow rate of 10 mL / min and a gradient of 10% to 45% CH3OH in 22 min (tR: 11.7 min, λ = 220 nm) to afford I14245f (17.1 mg, 50.6 µmol, 64%) as a white resin. ESI-MS: m/z 339.2 [M+H]⁺. HR-ESI-MS: m/z [M+Na]⁺ calcd.361.1271 for C18H18N4NaO3, found 361.1268. IR (KBr): 3447, 3354, 3310, 3242, 1685 cm^-1. ¹H-NMR: (600 MHz, DMSO-d₆) δ 1.30 (d, J = 7.0 Hz, 3H), 4.08 (dd, J = 9.3, 5.4 Hz, 1H), 4.12 (ddq, J = 7.2, 5.4, 7.0 Hz, 1H), 4.40 (dd, J = 9.3, 7.2 Hz, 1H), 6.63 (brs, 1H), 6.89 – 6.96 (m, 3H), 7.24 – 7.30 (m, 2H), 7.46 (brs, ^{H2932377.1} 33 Docket No.2877.037AWO 1H), 7.81 (dd, J = 8.3, 1.5 Hz, 1H), 7.94 (d, J = 8.3 Hz, 1H), 8.14 (brs, 1H), 8.74 (brs, 1H). ¹³C-NMR: (DEPTQ, 151 MHz, DMSO-d6) δ 14.0, 37.8, 68.9, 114.5, 114.7, 120.4, 120.7, 121.9, 122.9, 129.5, 135.9, 139.0, 152.7, 158.3, 167.8, 172.3. HPLC: system 3: λ = 220 nm, tR = 18.9 min, purity: >99%. [α]D²²: +44.5 (c = 0.25, methanol). [0087] (S)-N-(3-Amino-1-(2-methyl-3-phenoxypropanoyl)-1H-indazol-6-yl)formamide [(S)-SX276, 6a]

A mixture of formic acid (15.1 µL, 18.5 mg, 401 µmol) and acetic anhydride (25.2 µL, 27.3 mg, 268 µmol) was stirred at 60 °C for 2 h. After cooling to rt, the mixed anhydride was added dropwise to a solution of compound (S)-SX2705e (16.6 mg, 53.5 mol) in anhydrous THF (1 mL) at 0 °C. After stirring the reaction mixture for 30 min at rt, the mixture was immediately treated with CH3OH / water (1:2) in order to hydrolyze the excess of mixed anhydride and then lyophilization was performed. Purification by preparative HPLC using a solvent system of CH3OH / 0.1% aq. HCOOH, a flow rate of 12 mL / min and a gradient of 50% to 75% CH3OH in 12.5 min, 75% to 95% CH3OH in 0.5 min, 95% CH₃OH for 1 min (t_R = 9.7 min, λ = 254 nm) afforded (S)-SX2766a (7.64 mg, 22.6 μmol, 42%) as a white solid. ESI-MS: m/z 339.2 [M+H]⁺. HR-ESI-MS: m/z [M+H]⁺ calcd.339.1452 for C18H19N4O3, found 339.1452. IR (NaCl): 3442, 3336, 2925, 1683, 1618, 1436, 1235 cm^-1. m.p.: 77 °C. ¹H-NMR: (600 MHz, DMSO-d6, two sets of resonances were observed, rotamers, assignment was confirmed by 2D spectroscopy) δ 1.28 (d, J = 6.8 Hz, 3H), ^{H2932377.1} 34 Docket No.2877.037AWO 4.05 (dd, J = 8.8, 5.5 Hz, 1H), 4.07 (ddq, J = 7.4, 6.8, 5.5 Hz 1H), 4.38 (dd, J = 8.8, 7.4 Hz, 1H), 6.47 (s, 1.6 H), 6.49 (s, 0.4 H), 6.90 – 6.96 (m, 3H), 7.24 – 7.29 (m, 2 H and 0.2 H), 7.50 (dd, J = 8.5, 1.8 Hz, 0.8 H), 7.81 (d, J = 8.4 Hz, 0.8 H), 7.82 (d, J = 8.6 Hz, 0.8 H),, 8.04 (d, J = 1.9 Hz, 0.2 H), 8.34 (d, J = 1.8 Hz, 0.8 H), 8.68 (d, J = 1.7 Hz, 0.8 H), 8.89 (d, J = 11.0 Hz, 0.2 H), 10.43 (d, J = 11.0 Hz, 0.2 H), 10.48 (d, J = 1.9 Hz, 0.8 H). ¹³C-NMR: (DEPTQ, 151 MHz, DMSO-d6, two sets of resonances were observed, rotamers) δ 14.0 (minor), 14.0 (major), 37.7, 68.9 (minor), 68.9 (major), 103.4 (minor), 105.3 (major), 113.8 (minor), 114.5, 115.7 (major), 116.1 (major), 116.3 (minor), 120.6, 121.1 (major), 121.8 (minor), 129.5, 139.7 (major), 139.9 (major), 140.1 (minor), 140.2 (minor), 152.7 (major), 152.8 (minor), 158.3, 159.8 (major), 162.6 (minor), 172.1 (major), 172.2 (minor). HPLC: system 2: λ = 254 nm, tR = 16.1 min, purity: >99%. chiral HPLC: isocratic elution with n-hexane / isopropanol, 9:1: tR = 40.7 min, >99% ee (254 nm). [α]_D ²⁶: +78.1 (c = 0.3, methanol). [0088] (S)-1-(3,6-Diamino-1H-indazol-1-yl)-2-methyl-3-phenoxypropan-1-one [(S)- SX270, 5e]

To a solution of (S)-SX22512a (34.8 mg, 0.19 mmol) in DMF (2.5 mL) were added HOAt (25.9 mg, 0.19 mmol) and EDC × HCl (36.4 mg, 0.19 mmol) at rt. The resulting solution was stirred for 10 min at rt and then 1H-indazole-3,6-diamine (25, 34.3 mg, 0.23 mmol) in DMF (0.5 mL) was added. After stirring the reaction mixture at rt for 2 h, ^{H2932377.1} 35 Docket No.2877.037AWO the solvent was removed by lyophilization. Purification by preparative HPLC using a solvent system of CH3OH / 0.1% aq. HCOOH, a flow rate of 12 mL / min and a gradient of 50% to 71% CH3OH in 11 min, 71% to 95% CH3OH in 2 min, 95% CH3OH for 2 min (t_R = 8.0 min, λ = 254 nm) afforded (S)-SX2705e (25.7 mg, 82.9 μmol, 44%) as an off- white solid. ESI-MS: m/z 311.1 [M+H]⁺. HR-ESI-MS: m/z [M+H]⁺ calcd.311.1503 for C₁₇H₁₉N₄O₂, found 311.1505. IR (NaCl): 3452, 3353, 2925, 1676, 1625, 1409, 1238 cm^-1. m.p.: 75 °C. ¹H-NMR: (600 MHz, DMSO-d₆) δ 1.25 (d, J = 6.9 Hz, 3H), 4.00 (dd, J = 8.7, 5.6 Hz, 1H), 4.04 (ddq, J = 7.4, 6.9, 5.6 Hz, 1H), 4.36 (dd, J = 8.7, 7.4 Hz, 1H), 5.66 (s, 2H), 6.15 (s, 2H), 6.55 (dd, J = 8.5, 1.9 Hz, 1H), 6.90 – 6.95 (m, 3H), 7.25 – 7.29 (m, 2H), 7.42 (d, J = 1.9 Hz, 1H), 7.46 (d, J = 8.5 Hz, 1H). ¹³C-NMR: (DEPTQ, 151 MHz, DMSO-d6) δ 14.1, 37.6, 69.0, 98.1, 110.3, 112.0, 114.5, 120.6, 121.0, 129.5, 141.6, 151.2, 153.1, 158.4, 171.7. HPLC: system 1: λ = 254 nm, t_R = 17.3 min, purity: 98%. chiral HPLC: isocratic elution with n-hexane / isopropanol + 0.1% ethylene diamine, 9:1: tR = 41.4 min, >99% ee (254 nm). [α]_D ²⁷: +64.3 (c = 0.5, methanol). [0089] 1-(3-Amino-6-fluoro-1H-indazol-1-yl)-2-(phenoxymethyl)butan-1-one (KD6 10b)

To a solution of 2-(phenoxymethyl)butanoic acid (39, 61.9 mg, 0.32 mmol) and HOAt (43.2 mg, 0.32 mmol) in DMF (1 mL) was added EDC × HCl (61.0 mg, 0.32 mmol). ^{H2932377.1} 36 Docket No.2877.037AWO After stirring for 10 min at rt, a solution of 6-fluoro-1H-indazol-3-amine (13, 40.6 mg, 0.27 mmol) in DMF (0.5 mL) was added. After stirring the reaction mixture for 3 h, the solvent was removed by lyophilization and the residue was purified by preparative HPLC using a solvent system of CH₃OH / 0.1% aq. HCOOH, a flow rate of 10 mL / min and a gradient of 50% to 80% CH3OH in 15 min, 80% to 95% CH3OH in 2 min, 95% CH3OH for 4 min (tR: 19.7 min, λ = 254 nm) to afford KD610b (45.5 mg, 0.14 mol, 45%) as a white solid. The enantiomers were separated by performing chiral preparative HPLC (isocratic solvent system: n-hexane / isopropanol, 98:2) to afford the two enantiomers KD6- 10b-ent1 (tR: 16.5 min) and KD6- 10b-ent2 (tR: 25.1 min). ESI-MS: m/z 328.2 [M+H]⁺. HR-ESI-MS: m/z [M+H]⁺ calcd.328.1456 for C₁₈H₁₉FN₃O₂, found 328.1456. IR (KBr): 3437, 3340, 3231, 1688, 1642, 1624 cm^-1. m.p.: 97 °C. ¹H-NMR: (600 MHz, DMSO-d₆) δ 0.93 (dd, J = 7.7, 7.3 Hz, 3 H), 1.77 (ddq, J = 14.5, 8.5, 7.3 Hz, 1H), 1.82 (ddq, J = 14.5, 7.7, 5.5 Hz, 1H), 4.05 (dddd, J = 8.5, 8.0, 5.8, 5.5 Hz, 1H), 4.13 (dd, J = 9.3, 5.5 Hz, 1H), 4.37 (dd, J = 9.3, 8.0 Hz, 1H), 6.60 (s, 2H), 6.93 – 6.91 (m, 3H), 7.28 – 7.24 (m, 3H), 7.95 (dd, J = 8.6, 5.3, 1H), 7.98 (dd, J = 9.8, 2.3 Hz). ¹³C-NMR: (DEPTQ, 151 MHz, DMSO-d6) δ 11.2, 21.7, 44.0, 67.7, 101.8 (d, J = 28 Hz), 112.2 (d, J = 24 Hz), 114.4, 117.0, 120.6, 122.6 (d, J = 11 Hz), 129.4, 139.6 (d, J = 19 Hz), 152.5, 158.2, 163.1 (d, J = 245 Hz), 171.9. HPLC: system 5: λ = 220 nm, for both enantiomers tR: 25.9 min, KD610b-ent1: purity: >99%, KD610b-ent2: purity: >99%. chiral HPLC: isocratic elution with n-hexane / isopropanol, 98:2: KD610b-ent1: tR = 17.2 min, >99% ee (254 nm); KD610b-ent2: tR = 23.2 min, 98% ee (254 nm). [α]_D ²⁵: KD6 10b-ent1: +23.6 (c = 0.4, methanol); KD610b-ent2: -23.6 (c = 0.2, methanol). [0090] (R,S)-1-(3-Amino-6,7-dihydropyrano[3,2-f]indazol-1(5H)-yl)-2-methyl-3- phenoxypropan-1-one (I13809) ^{H2932377.1} 37 Docket No.2877.037AWO

To a solution of (R,S)-2- (rac-12a, 57.1 mg, 0.32 mmol), PyBOP (165 mg, 0.32 mmol) and HOAt (60.0 mg, 0.48 mmol) in DMF (1.5 mL) was added DIPEA (55 µL, 0.44 mmol) and then a solution of I137937 (50.0 mg, 0.26 mmol) in DMF (0.5 mL) at rt. Microwave irradiation (described in the general part) in a sealed tube was performed, then the solvent was removed by lyophilization and the residue was purified by preparative HPLC using a solvent system of CH3CN / 0.1% aq. TFA, a flow rate of 10 mL / min and a gradient of 40% to 65% CH₃CN in 19 min, 65% to 95% CH3CN in 2 min (tR: 19.0 min, λ = 220 nm) to afford I13809 (56.3 mg, 0.16 mmol, 61%) as a white solid. ESI-MS: m/z 352.1 [M+H]⁺. HR-ESI-MS: m/z [M+H]⁺ calcd.352.1656 for C20H22N3O3, found 352.1660. IR (KBr): 3424, 3353, 1647 cm^-1. m.p.: 185 °C. ¹H-NMR: (600 MHz, DMSO-d₆) δ 1.24 – 1.27 (m, 3H), 1.95 (tt, J = 6.3, 5.1 Hz, 1H), 2.86 (t, J = 6.3 Hz, 2H), 4.00 – 4.07 (m, 2H), 4.20 (t, J = 5.1 Hz, 2H), 4.33 – 4.39 (m, 1H), 6.37 (s, 2H), 6.90 – 6.95 (m, 3H), 7.25 – 7.29 (m, 2H), 7.55 (s, 1H), 7.57 (s, 1H). ¹³C-NMR: (DEPTQ, 151 MHz, DMSO-d6) δ 13.9, 21.5, 24.7, 37.5, 66.4, 68.8, 101.8, 113.8, 114.4, 119.7, 120.5, 121.1, 129.4, 138.8, 152.7, 156.4, 158.2, 171.7. HPLC: system 3: λ = 220 nm, t_R = 24.5 min, purity: >99%. [0091] (R,S)-1-(3-Amino-6-methoxy-1H-indazol-1-yl)-2-methyl-3-phenoxypropan-1- one (IK195, 5a) ^{H2932377.1} 38 Docket No.2877.037AWO

To a solution of (R,S)-2- (rac-12, 66.0 mg, 0.37 mmol), PyBOP (194 mg, 0.37 mmol) and HOAt (76 mg, 0.56 mmol) in anhydrous DMF (3 mL) was added DIPEA (64 µL, 0.37 mmol) and then a solution of 6-methoxy-1H-indazol-3- amine (21, 50.0 mg, 0.26 mmol) in DMF (0.5 mL) at rt. Microwave irradiation (described in the general part) in a sealed tube was performed, then the solvent was removed by lyophilization and the residue was purified by preparative HPLC using a solvent system of CH₃CN / 0.1% aq. TFA, a flow rate of 20 mL / min and a gradient of 40% CH₃CN for 1 min, 40% to 65% CH3CN in 10 min (tR: 10.2 min, λ = 254 nm) to afford IK1955a (56.3 mg, 0.17 mmol, 56%) as a white solid. ESI-MS: m/z 326.2 [M+H]⁺. HR-ESI-MS: m/z [M+H]⁺ calcd.326.1499 for C₁₈H₂₀N₃O₃, found 326.1503. IR (KBr): 3446, 3339, 3229, 1676 cm^-1. m.p.: 160-163 °C. ¹H-NMR: (400 MHz, DMSO-d₆) δ 1.28 (d, J = 6.8 Hz, 3H), 3.83 (s, 3H), 4.00 – 4.14 (m, 2H), 4.34 – 4.43 (m, 1H), 6.46 (s, 2H), 6.89 – 6.95 (m, 3H), 6.69 (dd, J = 8.7, 2.2 Hz, 1H), 7.24 – 7.31 (m, 2H), 7.77 (d, J = 8.7 Hz, 1H), 7.79 (d, J = 2.2 Hz, 1H). ¹³C-NMR: (DEPTQ, 151 MHz, DMSO-d6) δ 14.0, 37.7, 55.5, 68.9, 98.3, 113.2, 114.0, 114.4, 120.6, 121.5, 129.5, 140.9, 152.9, 158.3, 161.2, 172.3. HPLC: system 3: λ = 220 nm, t_R = 24.6 min, purity: >99%. [0092] (R)-3-Amino-1-(2-methyl-3-phenoxypropanoyl)-1H-indazole-6-carboxamide (I1422/23 ent-5f) ^{H2932377.1} 39 Docket No.2877.037AWO

I1422/23 ent-5f was as for I14245f, starting from (R)-SX225 ent-12a. The analytical data were in accordance. HPLC: system 3: λ = 220 nm, tR = 18.9 min, purity: 98%. [α]D²²: -44.5 (c = 0.7, methanol). [0093] (R)-1-(2-Methyl-3-phenoxypropanoyl)-1H-indazole-6-carboxamide (I1412 ent- 7a)

I1412 ent-7a was synthesized analogously as described for I14097a, starting from (R)- SX225 ent-12a. The analytical data were in accordance. HPLC: system 3: λ = 220 nm, tR = 21.7 min, purity: 99%. chiral HPLC: isocratic elution with n-hexane / isopropanol, 85:5: t_R = 22.7 min, 98% ee (254 nm). [α]D²²: -111.0 (c = 0.65, methanol). [0094] (S)-1-(3-Amino-6-fluoro-1H-pyrazolo[4,3-b]pyridin-1-yl)-2-methyl-3- phenoxypropan-1-one [BBG3] ^{H2932377.1} 40 Docket No.2877.037AWO BBG3 is synthesized 3-phenoxypropionic

acid and 3-amino-6- was tested as described below and found to exhibit a relative stimulation of approximately 1.6. [0095] (S)-1-(3-amino-6-(hydroxymethyl)-1H-pyrazolo[3,4-b]pyridin-1-yl)-2-methyl- 3-phenoxypropan-1-one [PEH1] PEH1 is synthesized

3-phenoxypropionic acid and 3-amino-6-(hydroxymethyl)-1H-pyrazolo[3,4-b]pyridine. Biological Screening [0096] The molecular mechanism of ivacaftor has been well studied. Published electrophysiology measurements have shown that ivacaftor increases the open probability (Po) of both wild-type (wt) and many mutant CFTRs, suggesting that it acts through an allosteric site. Cryo-EM structures revealed that ivacaftor binds to CFTR inside the membrane, coinciding with a hinge region important for gating. A different potentiator, GLPG1837, representing a distinct chemotype, also binds the same site on CFTR. GLPG1837, a known potent and reversible CFTR potentiator, with EC50s of 3 nM and 339 nM for F508del and G551D CFTR, respectively, was used as positive control. ^{H2932377.1} 41 Docket No.2877.037AWO [0097] Compounds were analyzed by electrophysiology, using inside-out membrane patches containing fully phosphorylated wild type (WT) CFTR channels, and the results were compared to those of GLPG1837. M embrane patches containing WT CFTR were fully phosphorylated by PKA in the presence of 3 mM ATP. The fold stimulation is defined as the ratio of the current in the presence and absence of added compound. Individual measurements were shown as dots. The results are shown in Figure 1. Compounds of the formula I are shown to be potentiators of CFTR flux, whereas compounds of formula II are inhibitors of CFTR flux. Compounds of the formula I would therefore be expected to be useful in the treatment of CF, whereas compounds of formula II would be expected to be useful in the treatment of secretory diarrhea. [0098] We also examined whether I1421 increases the channel activity of mutant CFTR identified in patents. Ten CF-causing mutants, distributed at different positions in CFTR, were cloned and tested. The predominant mutation in cystic fibrosis, ΔF508, is defective in both folding and gating. Newly synthesized ΔF508 CFTR is largely retained in endoplasmic reticulum; and the few channels that reach to the plasma membrane exhibit little activity. The presence of I1421 strongly increased the current of ΔF508, indicating that similar to Ivacaftor and GLPG1837, it can be combined with correctors to rescue ΔF508. About 4% of CF patients carry the mutation G551D, which is expressed on the cell surface but with severe gating defects. In inside-membrane patches, I1421 increased activity of G551D by 25 fold. For the other eight mutations tested, the potentiation activity of I1421 is comparable to that of GLPG1837. These data indicate that I1421 is a strong potentiator that allosterically activates a wide range of CF-causing mutants. ^{H2932377.1} 42

Claims

Docket No.2877.037AWO CLAIMS 1. A substantially pure single enantiomer of a compound represented by formula I:

X¹ and X² are chosen independently from N and CH; R¹ is chosen from hydroxymethyl, halogen, amino, methoxy, -CONH2, and -NHC(=O)H; R² is hydrogen, or taken together with R¹, forms a six-membered fused carbocyclic or heterocyclic ring; R³ is methyl or ethyl; R⁴ is chosen from hydrogen, halogen, and lower-alkyl; and R⁵ is hydrogen, or taken together with R⁴, forms a six-membered fused carbocyclic ring. 2. A substantially pure single enantiomer of a compound according to claim 1 represented by formula Ia: . {^H2932377.1}

Docket No.2877.037AWO 3. A substantially pure single enantiomer of a compound according to claim 1 represented by formula Ib: .

4. A substantially pure single enantiomer of a compound according to claim 1 represented by formula Ic: .

5. A compound according to any of claims 1 to 4 wherein R⁵ is hydrogen and R⁴ is halogen or R⁵ taken together with R⁴, forms a fused benzene ring. 6. A compound according to any of claims 1 to 4 wherein R² is hydrogen. 7. A compound according to any of claims 1 to 4 wherein R², taken together with R¹, forms a fused pyran ring. ^{H2932377.1} 44 Docket No.2877.037AWO 8. A method for treating cystic fibrosis comprising administering a compound according to any of claims 1 to 4. 9. A substantially pure single enantiomer of a compound represented by formula II: . wherein

X¹ and X² are chosen independently from N and CH; R³ is methyl or ethyl; R¹⁰ is H or NH₂; and R¹³ is H or halogen. 10. A substantially pure single enantiomer of a compound according to claim 9 represented by formula IIa: .

11. A substantially pure single enantiomer of a compound according to claim 9 represented by formula IIb: ^{H2932377.1} 45 Docket No.2877.037AWO .

12. A substantially pure single enantiomer of a compound according to claim 9 represented by formula IIc: .

13. A compound according to any of claims 9 to 12 wherein R¹³ is H. 14. A compound according to claim 13 wherein R¹⁰ is H. 15. A compound according to claim 13 wherein R¹⁰ is NH2. 16. A method for treating secretory diarrhea comprising administering a compound according to any of claims 9 to 12. ^{H2932377.1} 46