RU2786361C2 - Method for production of jak inhibitors and their intermediate compounds - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to the production of intermediate compounds for the production of drugs. A method for the production of a compound of the formula J-15 or its salt is proposed, where X is Br; PG¹ is a protective group for carboxylic acid; PG² is an amino protective group, and PG³ is an amino protective group. The method includes: (a) interaction of a compound of the formula J-14 or its salt with hydrazine to obtain the compound of the formula J-15, and (b) optionally, formation of salt of the compound J-15. Compounds of formulas J-14, J-15 and a method for the production of a compound of the formula J-16 are also proposed.

EFFECT: obtainment of intermediate compounds for JAK inhibitors.

20 cl, 3 tbl, 13 ex

Description

State of the art invention

The technical field to which the invention belongs

The invention relates to a method for the preparation of compounds that can be used as intermediates for the preparation of drugs. In particular, the invention relates to the preparation of intermediates for JAK inhibitors.

State of the Art

The JAK family consists of four members: JAK1, JAK2, JAK3, and tyrosine kinase 2 (TYK2). Binding of the cytokine to the JAK-dependent cytokine receptor causes dimerization of the receptor, which results in phosphorylation of tyrosine residues on the JAK kinase, influencing JAK activation. Phosphorylated JAKs, in turn, bind and phosphorylate various STAT proteins, which dimerize, internalize in the cell nucleus, and directly modulate gene transcription, leading, among other things, to downstream effects associated with inflammatory disease. JAKs usually associate with cytokine receptors in pairs as homodimers or heterodimers. Each of the four members of the JAK family is involved in the signaling of at least one of the cytokines associated with inflammation. Therefore, a chemical inhibitor with pan activity against all members of the JAK family can modulate a wide range of pro-inflammatory pathways that contribute to inflammatory lung diseases such as severe asthma, COPD, and chronic lung allograft dysfunction (CLAD). Therefore, it would be desirable to have an efficient process for the production of specific JAK inhibitors.

The essence of the invention

The invention is directed to a process for the preparation of compounds that can be used as intermediates for the preparation of drugs having inhibitory activity against JAK.

Accordingly, in one aspect, the invention provides a process for preparing a compound of formula J-15, or a salt thereof:

where

X is selected from the group consisting of Br, I and Cl;

PG ¹ is a protecting group for a carboxylic acid;

PG ² is an amino protecting group; and

PG ³ is an amino protecting group;

method including:

(a) reacting a compound of formula J-14 :

or its salt with hydrazine to give a compound of formula J-15 and

(b) optionally forming a salt of compound J-15 .

In another aspect, the invention relates to a compound of formula J-14 :

or its salt, where

X is selected from the group consisting of Br, I and Cl;

PG ¹ is a protecting group for a carboxylic acid;

PG ² is an amino protecting group; and

PG ³ is an amino protecting group.

In another aspect, the invention relates to a compound of formula J-15 :

or its salt

where

X is selected from the group consisting of Br, I and Cl;

PG ¹ is a protecting group for a carboxylic acid;

PG ² is an amino protecting group; and

PG ³ is an amino protecting group.

In another aspect, the invention relates to a process for preparing a compound of formula J-16, or a salt thereof:

where

PG ¹ is a protecting group for a carboxylic acid;

PG ² is an amino protecting group;

PG ³ is an amino protecting group;

PG ⁴ is a hydroxyl protecting group; and

R is H or F;

including:

(a) reacting a compound of formula J-13 :

where X is selected from the group consisting of Br, I and Cl; and Y is a leaving group; with a compound of formula J-11 :

in the presence of a base, to give a compound of formula J-14 :

and optionally forming a salt of compound J-14 ;

(b) reacting a compound of formula J-14 or a salt thereof with hydrazine to give a compound of formula J-15:

and optionally forming a salt of compound J-15 ;

(c) reacting a compound of formula J-15 , or a salt thereof, with a compound of formula J-5 , J-6 , or J-7 :

where R ^a and R ^b are each independently selected from C _1-8 alkyl, where R ^a and R ^b may optionally be connected to form a 4-8 membered ring; in the presence of a base, a palladium catalyst and a phosphine ligand to give a compound of formula J-16 , and optionally forming a salt of compound J-16 .

DETAILED DESCRIPTION OF THE INVENTION

In one aspect, the invention relates to a process for preparing a compound of formula J-15 or a salt thereof:

where

X is selected from the group consisting of Br, I and Cl;

PG ¹ is a protecting group for a carboxylic acid;

PG ² is an amino protecting group; and

PG ³ is an amino protecting group;

including:

(a) reacting a compound of formula J-14 :

or its salt with hydrazine to give a compound of formula J-15 and

(b) optionally forming a salt of compound J-15 .

In some aspects, the reaction with hydrazine is carried out at about 60°C. In some aspects, the reaction with hydrazine is carried out at 60°C±20°C.

In some aspects, X is selected from the group consisting of Br, I and Cl; PG ¹ is an alkyl or benzyl group, where the benzyl group is optionally substituted; PG ² is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group; and PG ³ is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group.

In some aspects, X is Br. In some aspects, X is Br, PG ¹ is benzyl, PG ² is t-butoxycarbonyl, and PG ³ is benzyl.

In some aspects, compound J-14, or a salt thereof, is:

(a) reacting a compound of formula J-13 :

where Y is a leaving group, with a compound of formula J-11 :

in the presence of a base, to give J-14 , and

(b) optionally forming a salt of compound J-14 .

In some aspects, Y is Cl.

In another aspect, the invention relates to a compound of formula J-14 :

or its salt, where

X is selected from the group consisting of Br, I and Cl;

PG ¹ is a protecting group for a carboxylic acid;

PG ² is an amino protecting group; and

PG ³ is an amino protecting group.

In some aspects, X is selected from the group consisting of Br, I and Cl; PG ¹ is an alkyl or benzyl group, where the benzyl group is optionally substituted; PG ² is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group; and PG ³ is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group.

In another aspect, the invention relates to a compound of formula I-14 :

or its salt.

In another aspect, the invention relates to a compound of formula J-15 :

or its salt

where

X is selected from the group consisting of Br, I and Cl;

PG ¹ is a protecting group for a carboxylic acid;

PG ² is an amino protecting group; and

PG ³ is an amino protecting group.

In some aspects, X is selected from the group consisting of Br, I and Cl; PG ¹ is an alkyl or benzyl group, where the benzyl group is optionally substituted; PG ² is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group; and PG ³ is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group.

In another aspect, the invention relates to a compound of formula I-15 :

or its salt.

In another aspect, the invention relates to a process for preparing a compound of formula J-16, or a salt thereof:

where

PG ¹ is a protecting group for a carboxylic acid;

PG ² is an amino protecting group;

PG ³ is an amino protecting group;

PG ⁴ is a hydroxyl protecting group; and

R is H or F;

including:

(a) reacting a compound of formula J-13 :

where X is selected from the group consisting of Br, I and Cl; and Y is a leaving group; with a compound of formula J-11 :

in the presence of a base, to give a compound of formula J-14 :

and optionally forming a salt of compound J-14 ;

(b) reacting a compound of formula J-14 or a salt thereof with hydrazine to give a compound of formula J-15:

and optionally forming a salt of compound J-15 ;

(c) reacting a compound of formula J-15, or a salt thereof, with a compound of formula J-5 , J-6 , or J-7 :

where R ^a and R ^b are each independently selected from C _1-8 alkyl, where R ^a and R ^b may optionally be connected to form a 4-8 membered ring; in the presence of a base, a palladium catalyst and a phosphine ligand to give a compound of formula J-16 , and optionally forming a salt of compound J-16 .

In some aspects, step (c) is carried out in the presence of a diboronic reagent. In some aspects, step (c) is carried out in the presence of tetrahydroxydiborone or a diboronic acid ester. In some embodiments, the catalyst is the reaction product of bis(pinacolato)diborone with potassium fluoride hydrofluoride. In some embodiments, the catalyst is prepared by reacting bis(pinacolato)diborone in propan-2-ol with potassium fluoride hydrofluoride in water, followed by filtration and drying of the resulting solids.

In some aspects, the interaction with hydrazine in stage (b) is carried out at 60°C±20°C.

In some aspects, X is selected from the group consisting of Br, I and Cl;

PG ¹ is an alkyl or benzyl group, where the benzyl group is optionally substituted; PG ² is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group; PG ³ is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group; PG ⁴ is selected from the group consisting of a silyl group, an acyl group and an arylmethyl group. In some aspects, X is Br. In some aspects, X is Br, PG ¹ is benzyl, PG ² is t-butoxycarbonyl, PG ³ is benzyl, and PG ⁴ is benzyl.

In some aspects, Y is Cl.

In some aspects, the palladium catalyst and the phosphine ligand of step (c) are bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II).

In the present method, the J-13 compound:

interacts with a compound of formula J-11 :

in the presence of a base, to give a compound of formula J-14 :

Typically, the reaction is carried out in a solvent such as acetonitrile in the presence of an excess (eg 3 to 7 equivalents) of a base such as trimethylamine. Usually one to two J-13 equivalents are used.

In the present method, compound J-14 or a salt thereof is reacted with hydrazine to give a compound of formula J-15:

Typically, the reaction is carried out in a solvent such as THF and an excess of hydrazine is used, for example 2 to 10 equivalents. The reaction is heated at about 60° C. to completion, typically 0.5 to 6 hours.

In the present method, compound J-15, or a salt thereof, is reacted with a compound of formula J-5 , J-6 , or J-7 :

in the presence of a base, a palladium catalyst and a phosphine ligand to give a compound of formula J-16 :

The reaction is typically carried out by contacting J-16 with about 1 to about 1.5 equivalents of J-5, J-6, or J-7 in the presence of a catalytic amount of a palladium catalyst and a phosphine ligand (between about 0.005 and about 0.1 equivalents ) and from about 2 to about 6 base equivalents. Carrying out the reaction in the presence of an additional catalyst increases the yield of product J-16. The additional catalyst may be a diboronic reagent. The additional catalyst may be tetrahydroxydiborone, a diboronic acid ester, or the reaction product of bis(pinacolato)diborone with potassium fluoride hydrofluoride as shown in Preparation 4.

Suitable palladium catalysts include bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II), tris(dibenzylideneacetone)dipalladium(0) (Pd ₂ dba ₃ ), palladium(II) acetate (Pd(OAc) ₂ ) , dichloro(1,1'-bis(diphenylphosphino)ferrocene)dipalladium(II) (Pd(dppf)Cl ₂ ), dichlorobis(triphenylphosphine)-palladium(II) (Pd(PPh ₃ ) ₂ Cl ₂ ), etc. similar, where generally accepted abbreviations are given in brackets. Phosphine ligands used in the present reaction include tricyclohexylphosphine (PCy ₃ ), tricyclohexylphosphinetetrafluoroborate (PCy ₃ HBF ₄ ), 1,1'-bis(diphenylphosphino)ferrocene (dppf), 1,1'-bis(di-tert-butylphosphino) ferrocene, tri(2-furyl)phosphine, 1,3-bis(diphenylphosphino)propane (dppp), 1,5-bis(diphenylphosphino)pentane (dpppe), tri-tert-butylphosphine (P( t -Bu) ₃ ) , and 99,9-dimethyl-4,5-bis(diphenylphosphino)xanthene (xanthos).

Specific coupling bases include potassium fluoride, cesium carbonate and cesium fluoride. Alternatively, sodium carbonate, potassium carbonate, sodium acetate, potassium tert-butoxide, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) or 1,4-diazabicyclo[2.2.2] can be used as the base. octane (DABCO). The reaction is usually carried out in an inert solvent such as tetrahydrofuran, N,N-dimethylformamide, N,N-dimethylacetamide or N-methylpyrrolidone. Suitable mixed solvent systems include tetrahydrofuran and water, tetrahydrofuran and N,N-dimethylformamide, tetrahydrofuran and N-methylpyrrolidone, acetone and water, ethanol and water, and isopropanol and water. The reaction is usually carried out at a temperature of from about 40 to about 120°C for from about 1 to about 20 hours, or until the reaction is essentially complete. Product J-16 is isolated as a solid by conventional means.

Definitions

In describing the present invention, including its various aspects and embodiments, the following terms have the following meanings, unless otherwise indicated.

The term "alkyl" means a monovalent saturated hydrocarbon group, which may be linear or branched, or combinations thereof. Unless otherwise indicated, such alkyl groups typically contain from 1 to 10 carbon atoms. Representative alkyl groups include, for example, methyl (Me), ethyl (Et), n-propyl (n-Pr) or (nPr), isopropyl (i-Pr) or (iPr), n-butyl (n-Bu) or (nBu), sec-butyl, isobutyl, tert-butyl (t-Bu) or (tBu), n-pentyl, n-hexyl, 2,2-dimethylpropyl, 2-methylbutyl, 3-methylbutyl, 2-ethylbutyl, 2 ,2-dimethylpentyl, 2-propylpentyl and the like.

When a certain number of carbon atoms is for a certain term, the number of carbon atoms is indicated before the term. For example, the term "C _1-3 alkyl" means an alkyl group having 1 to 3 carbon atoms, where the carbon atoms are in any chemically acceptable configuration, including linear or branched configurations.

The term "cycloalkyl" means a monovalent saturated carbocyclic group, which may be monocyclic or polycyclic. Unless otherwise indicated, such cycloalkyl groups typically contain from 3 to 10 carbon atoms. Representative cycloalkyl groups include, for example, cyclopropyl (cPr), cyclobutyl (cBu), cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, adamantyl, and the like.

The term "cpropyl" means cyclopropyl.

The term "heterocyclyl", "heterocycle", "heterocyclic", or "heterocyclic ring" means a monovalent saturated or partially unsaturated cyclic non-aromatic group having 3 to 10 ring atoms in common, where the ring contains 2 to 9 ring carbon atoms and 1 to 4 ring heteroatoms selected from nitrogen, oxygen and sulfur. Heterocyclic groups may be monocyclic or polycyclic (ie, fused or bridged). Typical heterocyclyl groups include, for example, pyrrolidinyl, piperidinyl, piperazinyl, imidazolidinyl, morpholinyl, thiomorpholyl, indolin-3-yl, 2-imidazolinyl, tetrahydropyranyl, 1,2,3,4-tetrahydroisoquincholin-2-yl, 7-azanorbornanyl, northropanyl and the like, where the point of attachment is at any available ring carbon or nitrogen atom. Where the point of attachment of a heterocyclic group is clear from the context, such groups may alternatively be referred to as non-valent species, ie, pyrrolidine, piperidine, piperazine, imidazole, tetrahydropyran, and the like.

The term "halogen" means fluorine, chlorine, bromine or iodine.

The term "therapeutically effective amount" means an amount sufficient to effectively treat when administered to a patient in need of treatment.

The term "treatment" or "therapy" means the prevention, improvement, or suppression of a medical condition, disease, or disorder being treated (eg, a respiratory disease) in a patient (in particular, a human); or relieving symptoms of a health condition, disease, or disorder.

The term "pharmaceutically acceptable salt" means a salt that is acceptable for administration to a patient or a mammal, such as a human (eg, salts that have acceptable mammalian safety for a given dosage regimen). Typical pharmaceutically acceptable salts include acetic, ascorbic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, edisyl, fumaric, gentisic, gluconic, glucuronic, glutamic, hippuric, hydrobromic, hydrochloric, isethionic, lactic, lactobionic, maleic, malic, almond, methanesulfonic, mucus, naphthalenesulfonic, naphthalene-1,5-disulfonic, naphthalene-2,6-disulfonic, nicotinic, nitric, orotic, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic and xinafoic acids and the like.

The term "salt thereof" means a compound formed when the hydrogen of an acid is replaced by a cation such as a metal cation or an organic cation and the like. For example, the cation may be the protonated form of the compound, that is, the form in which one or more amino groups have been protonated with an acid. Typically, the salt is a pharmaceutically acceptable salt, although this is not required for salts of intermediates that are not intended to be administered to a patient.

The term "amino protecting group" means a protecting group suitable for preventing undesired reactions of the nitrogen of the amino group. Exemplary amino protecting groups include, but are not limited to, formyl; acyl groups, for example alkanoyl groups such as acetyl and trifluoroacetyl; alkoxycarbonyl groups such as t-butoxycarbonyl (Boc); arylmethoxycarbonyl groups such as benzyloxycarbonyl (Cbz), 9-fluorenylmethoxycarbonyl (Fmoc), p-nitrobenzyloxycarbonyl (PNZ), 2,4-dichlorobenzyloxycarbonyl and 5-benzisoxazolylmethoxycarbonyl; arylmethyl groups such as benzyl (Bn), 4-methoxybenzyl, trityl (Tr) and 1,1-di-(4'-methoxyphenyl)methyl; silyl groups such as trimethylsilyl (TMS), t-butyldimethylsilyl (TBDMS), [2-(trimethylsilyl)ethoxy]methyl (SEM); etc.

The term "carboxylic acid protecting group" means a protecting group suitable for preventing unwanted reactions with a carboxylic acid. Exemplary carboxylic acid protecting groups include, but are not limited to, esters such as methyl, ethyl, t-butyl, benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm), trimethylsilyl (TMS), t -butyldimethylsilyl (TBS, TBDMS), diphenylmethyl (benzhydryl, DPM) and the like.

The term "hydroxyl protecting group" means a protecting group suitable for preventing undesired reactions on a hydroxyl group. Exemplary hydroxyl protecting groups include, but are not limited to, silyl groups, including tri(C _1-6 alkyl) silyl groups such as trimethylsilyl (TMS), triethylsilyl (TES), t-butyldimethylsilyl (TBS), and the like; esters (acyl groups) including C _1-6 alkanoyl groups such as formyl, acetyl and the like; arylmethyl groups such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm), diphenylmethyl (benzhydryl, DPM) and the like.

Numerous protecting groups and their introduction and removal are described in TW Greene and PGM Wuts, Protecting Groups in Organic Synthesis , Third Edition, Wiley, New York

Examples

The following synthetic examples are provided to illustrate the invention and should not be construed in any way as limiting the scope of the invention. In the examples below, the following abbreviations have the following meanings, unless otherwise noted. Abbreviations not listed below have their commonly used meanings.

ACN=acetonitrile

DMF= N, N -dimethylformamide

EtOAc=ethyl acetate

EtOH=ethanol

NaHMDS=sodium bis(trimethylsilyl)amide

MeTHF=2-methyltetrahydrofuran

MTBE=tert-butyl methyl ether

psi inch=pounds per square inch

R t =retention time

Reagents and solvents were purchased from commercial suppliers (Aldrich, Strem Chemicals, Inc., etc.) and used without further purification. The progress of the reaction was monitored by analytical high performance liquid chromatography and mass spectrometry. Endo/exo product ratios were determined by HPLC analysis using the protocols described below. The reaction mixtures were worked up as described specifically for each reaction; usually they were purified by extraction and other purification methods such as crystallization, depending on temperature and solvent, and precipitation. Characterization of the reaction products in the conventional manner was carried out using mass and ¹ H-NMR spectrometry. For NMR measurements, samples were dissolved in a deuterated solvent (DMSO- d ₆ or CDCl ₃ ) and ¹ H-NMR spectra were recorded using Varian Gemini 2000 (400 MHz) equipment under standard measurement conditions. Mass spectrometric identification of compounds was performed using Agilent (Palo Alto, CA) model 1100 LC/MSD equipment.

General HPLC conditions

Column: Zorbax SB-Aq, 5 µm. 4.6×250 mm

Column temperature: 40ºC

Flow rate: 1.0 ml/min

Mobile phases: A=water/ACN (98:2) + 0.1% TFA

B=water/ACN (10:90) + 0.1% TFA,

Injection volume: 10 µl

Detector wavelength: 214 nm

HPLC Method 1

The crude compounds were dissolved in a mixture of water/ACN (50:50) in an amount of approximately 1 mg/ml and analyzed using the following gradient over 20 min (time (min)/% B): 0/10, 2.5/20, 9/ 75, 15/90, 17/90, 18/10, 20/10.

HPLC method 2

Compounds were dissolved in water/ACN (90:10) at approximately 1 mg/mL and analyzed using the following gradient over 30 min (time (min)/% B): 0/10, 13/10, 23/65 , 28/90, 29/90, 30/10.

HPLC Method 3

Compounds were dissolved in water/ACN (90:10) at approximately 1 mg/mL and analyzed using the following gradient over 55 min (time (min)/% B): 0/10, 10/20, 46/75 , 47/90, 50/10, 55/10.

Receipt 1: 4-(benzyloxy)-2-ethylphenyl)trifluoro-λ⁴-borane, potassium salt I-5

(a) 1-(benzyloxy)-3-ethylbenzene (I-2)

To a stirred solution of 3-ethylphenol (I-1) (25.0 g, 204.0 mmol) in ACN (250 ml, 10 vol) was added potassium carbonate (42.0 g, 306 mmol) at room temperature. The resulting reaction mass was stirred at room temperature for 15 minutes, followed by the dropwise addition of benzyl bromide (24.0 ml, 204 mmol). The resulting reaction mixture was stirred for 6 hours at room temperature. After completion of the reaction (TLC monitoring), the resulting reaction mass was poured into water (1.0 L) followed by extraction of the compound with EtOAc (2×2 L). The combined organic layers were washed with cold water, brine and dried over sodium sulfate, filtered and evaporated under reduced pressure. The crude product was then purified by silica gel column chromatography (100-200 M) using 2% EtOAc in hexane as eluents to give the desired product (I-2) as a light yellow oily compound (35.0 g, 81%). ¹ H NMR (400 MHz, chloroform-d) δ 7.46-7.44 (m, 2H), 7.39 (t, J=7.6 Hz, 2H), 7.34-7.31 (m , 1H), 7.21 (t, J=7.6 Hz), 6.86-6.80 (m, 3H), 5.07 (s, 2H), 2.64 (q, J=7, 6 Hz, 2H), 1.24 (t, J=7.6 Hz, 3H).

(b) 4-(benzyloxy)-1-bromo-2-ethylbenzene (I-3)

To an ice-cold stirred solution of 1-(benzyloxy)-3-ethylbenzene (I-2) (35.0 g, 164 mmol) in ACN (525 ml, 15 vol) was added N-bromosuccinimide (32.0 g, 181 mmol) portions over 15 minutes. The resulting reaction mixture was stirred for 1 hour at room temperature. After completion of the reaction (TLC monitoring), the resulting reaction mass was poured into ice water (1.50 L) followed by extraction of the compound with EtOAc (2×1 L). The combined organic layers were washed with water and dried over sodium sulfate, filtered and evaporated under reduced pressure to give the crude product. n-hexane (250 ml) was added to the crude material to form a suspension, followed by filtration through a Schott filter. The mother liquor was evaporated under reduced pressure to give the desired product I-3 as a light yellow oily compound (42.0 g, 87%). ¹ H NMR (400 MHz, chloroform-d) δ 7.52-7.29 (m, 7H), 6.88 (s, 1H), 6.68 (d, J=6.0 Hz, 1H), 5.04 (s, 2H), 2.69 (kv, J=7.6 Hz, 2H), 1.20 (t, J=7.5 Hz, 3H).

(c) 2-(4-(benzyloxy)-2-ethylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (I-4)

A stirred solution of 4-(benzyloxy)-1-bromo-2-ethylbenzene (I-3) (42.0 g, 144 mmol), bis(pinacolato) diborone (44.0 g, 173 mmol), and potassium acetate (28 g, 288 mmol) in dioxane (440 ml) was degassed by purging N ₂ (g) for 15 min followed by the addition of PdCl ₂ (dppf).DCM complex (11.0 g, 15 mmol). The resulting reaction mixture was heated to 80° C. for 16 hours. After completion of the reaction (TLC monitoring), the reaction mass was filtered through a pad of celite and the mother liquor was evaporated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (100-200M) using 1% EtOAc in hexanes as eluents to give the desired product (I-4) as a light yellow oily compound (32.0 g, 66%). ¹ H NMR (400 MHz, chloroform-d) δ 7.74 (d, J=8.4 Hz, 1H), 7.45-7.36 (m, 5H), 6.84-6.78 (m , 2H), 5.08 (s, 2H), 2.91 (q, J=7.6 Hz), 1.33 (s, 12H), 1.19 (t, J=7.6 Hz, 3H ).

(d) (4-(benzyloxy)-2-ethylphenyl)trifluoro-λ ⁴ -borane, potassium salt (I-5)

To a stirred solution of 2-(4-(benzyloxy)-2-ethylphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (I-4) (20 g, 59.0 mmol), in acetone:methanol (200 ml, 1:1 ratio, 10 vol.), 3M potassium hydrogen fluoride solution (23.0 g, 295 mmol, dissolved in 98.0 ml water) was added. The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (TLC monitoring), the resulting reaction mass was evaporated under reduced pressure. The solid thus obtained was taken up in water (100 ml) and stirred at room temperature for 30 minutes. The resulting reaction mass was filtered through a Schott funnel, washed with n-hexane and dried under reduced pressure to give the desired product (I-5) as a white solid (14.0 g, 74%). ¹ H NMR (400 MHz, chloroform-d) δ 7.43 (d, J=7.2 Hz, 2H), 7.37 (t, J=7.5 Hz, 2H), 7.30 (t, J=7.1Hz, 1H), 7.22(d, J=8.0Hz), 6.58(s, 1H), 6.53(d, J=7.9Hz, 1H), 5 .00 (s, 2H), 2.65 (kv, J=7.5 Hz, 2H), 1.07 (t, J=7.4 Hz, 3H).

Receipt 2: 6-benzyl 5-(tert-butyl) (S)-3-benzyl-3,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5,6-dicarboxylate (I- eleven)

(a) (S)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine-6-carboxylic acid, hydrochloride salt (I-7)

To an ice-cold stirred suspension of L-histidine (I-6) (5.0 kg, 32.14 mol) in water (40 L, 8 vol) was added concentrated hydrochloric acid (3.93 L, 33.75 mol), followed by the addition of formaldehyde (5.50 L, 67.5 mol, 37% aqueous solution) dropwise. The resulting solution was stirred for 30 minutes at the same temperature and then heated at 80°C for 8 hours. The progress of the reaction was monitored by LCMS. Water was removed under reduced pressure to give a crude product, and the resulting crude product was stirred for 2 hours in toluene (20 L). The organics were removed under reduced pressure to remove excess water and the compound was azeotropically dried. The resulting material was then taken up in diethyl ether (20 L) and stirred for 2 hours. The solid was then filtered and air dried to give the desired product (I-7) as an off-white solid (6.50 kg, 85%). ¹ H NMR (400 MHz, D ₂ O) δ 8.69 (s, 1H), 4.56 (d, J=15.4 Hz, 1H), 4.42 (d, J=15.5 Hz, 1H), 4.20 (dd, J=5.5, 5.2 Hz, 1H), 3.42 (dd, J=5.0, 17.0 Hz, 1H), 3.11 (dd, J =10.2, 16.8 Hz, 1H).

(b) (S)-3,5-bis(tert-butoxycarbonyl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine-6-carboxylic acid (I-8)

To an ice-cold stirred solution of (S)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine-6-carboxylic acid di-hydrochloride (I-7) (6.10 kg, 25 .40 mol) in 1,4-dioxane (48 L, 8 vol.) and water (48 L, 8 vol.) were added dropwise triethylamine (12.36 L, 89 mol) followed by the addition of di-tert-butyl dicarbonate ( 18.07 L, 78.74 mol, dissolved in 1,4-dioxane) for 30 minutes. The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (TLC & LCMS monitoring), the yellowish reaction mixture was diluted with water (10 L) and washed successively with diethyl ether (2×10 L) and EtOAc (2×7.50 L). The organic phase was discarded. The aqueous layer was cooled and adjusted to pH ~3 with 6N HCl. HCl solution The aqueous layer was cooled and adjusted to pH ~ 3 with 6N HCl solution. HCl solution; the aqueous phase was extracted with EtOAc (3×10 L). The combined organic layers were washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The oily residue was crystallized from 30% EtOAc:hexanes to give the desired product (I-8) as an off-white solid (5.1 kg, 55%). (m/z): [M+H]+ calculated for C ₁₇ H ₂₅ N ₃ O ₆ 368.18 found 368.21.

(c) 6-benzyl 3,5-di-tert-butyl (S)-6,7-dihydro-3H-imidazo[4,5-c]pyridine-3,5,6(4H)-tricarboxylate (I- nine)

To an ice-cold solution of (S)-3,5-bis(tert-butoxycarbonyl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine-6-carboxylic acid (I-8) ( 5.1 kg, 13.88 mol) in DCM (51 L, 10 vol) were added successively saturated aqueous sodium bicarbonate (41.0 L, 8 vol), tetrabutylammonium iodide (5.13 kg, 13.88 mol) and benzyl bromide (2.47 L, 20.82 mol). The resulting reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitoring by TLC & LCMS), the biphasic solution was separated. The aqueous layer was extracted with DCM (3×10 L). The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product, which was purified by column chromatography through silica gel (100-200M) using 40% EtOAc in hexane eluents to give the desired product (I -9) as a viscous oil (4.50 kg, 72%). (m/z): [M+H]+ calculated for C ₂₄ H ₃₁ N ₃ O ₆ 458.22 found 458.60.

(d) 6-benzyl 5-(tert-butyl) (S)-3,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5,6-dicarboxylate (I-10)

To an ice-cold solution of 6-benzyl 3,5-di-tert-butyl (S)-6,7-dihydro-3H-imidazo[4,5-c]pyridine-3,5,6(4H)-tricarboxylate (I- 9) (4.50 kg, 9.84 mol) in IPA (45 L, 10 vol) ammonium hydroxide (36 L, 8 vol) was added dropwise. The resulting reaction mixture was further stirred at room temperature for the next 16 hours. After completion of the reaction (TLC & LCMS monitoring), the resulting mixture was diluted with water (25 L) followed by extraction with EtOAc (3×20 L). The combined organic layers were washed with brine, dried over sodium sulfate, filtered and concentrated under reduced pressure to give the crude product, which was purified by column chromatography through silica gel (100-200M) using 2% MeOH in DCM eluents to give the desired product (I- 10) as a thick viscous oil (2.70 kg, 77%). (m/z): [M+H]+ calculated for C ₁₉ H ₂₃ N ₃ O ₄ 358.17 found 358.33.

(f) 6-benzyl 5-(tert-butyl) (S)-3-benzyl-3,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5,6-dicarboxylate (I -eleven)

To an ice-cold solution of 6-benzyl 5-(tert-butyl) (S)-3,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5,6-dicarboxylate (I-10) ( 2.70 kg, 7.55 mol) in DCM (32.4 L, 12 vol.) was added aqueous 1N. sodium hydroxide (24.3 L, 9 vol.) followed by the sequential addition of tetrabutylammonium iodide (2.80 kg, 7.55 mol) and benzyl bromide (0.99 L, 8.31 mol). The resulting reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction (monitoring by TLC & LCMS), the biphasic solution was separated. The aqueous layer was extracted with DCM (3×10 L). The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product, which was purified by column chromatography on silica gel (100-200M) using 40% EtOAc in hexane eluents to give the desired product (I -11) as a viscous oil (1.70 kg, 63%). (m/z): [M+H]+ calculated for C ₂₆ H ₂₉ N ₃ O ₄ 448.22 found 448.20.

Preparation 3: 6-benzyl 5-(tert-butyl) (S)-3-benzyl-2-(6-bromo-1H-indazol-3-yl)-3,4,6,7-tetrahydro-5H-imidazo [4,5-c]pyridine-5,6-dicarboxylate (I-15)

(a) 4-bromo-2-fluorobenzoyl chloride (I-13)

To an ice-cold stirred solution of 4-bromo-2-fluorobenzoic acid (I-12) (1.25 kg, 5.71 mol) in DCM (12.5 L, 15 vol) was added dropwise oxalyl chloride (0.98 L, 11.42 mol). The resulting reaction mixture was stirred for 10 minutes at the same temperature. Then DMF (150 ml) was added dropwise to the reaction mixture. The resulting reaction mass was allowed to warm to room temperature and stirred for 2 hours. After completion of the reaction (TLC monitoring), excess oxalyl chloride was removed under reduced pressure under nitrogen to give crude product (I-13) (1.08 kg, 80%), which was used in the next step without further purification.

(b) 6-benzyl 5-(tert-butyl) (S)-3-benzyl-2-(4-bromo-2-fluorobenzoyl)-3,4,6,7-tetrahydro-5H-imidazo[4,5 -c]pyridine-5,6-dicarboxylate (I-14)

To a stirred solution of 6-benzyl 5-(tert-butyl) (S)-3-benzyl-3,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5,6-dicarboxylate (I -11) (1.70 kg, 3.80 mol) triethylamine (2.11 L, 15.2 mol) was added to ACN (13.6 L, 8 vol) followed by 4-bromo-2-fluorobenzoyl chloride (I-13) (1.08 kg, 4.56 mol in 3.4 L ACN, 2 vol) at room temperature. After completion of the addition, the color of the resulting reaction mixture turned brown from light yellow. The resulting reaction mixture was stirred at the same temperature for 30 minutes and the progress of the reaction was monitored by TLC. The resulting reaction mixture was quenched with ice water (10 L), followed by extraction with EtOAc (3×5 L) and the combined organic layers washed with brine. The organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product, which was purified by column chromatography on silica gel (100-200M) using 20% EtOAc in hexane eluents to give the desired product (I-14) (1, 74 kg, 71%). %). (m/z): [M+H]+ calculated for C ₃₃ H ₃₁ BrFN ₃ O ₅ 648.14 found 648.20.

(c) 6-benzyl 5-(tert-butyl) (S)-3-benzyl-2-(6-bromo-1H-indazol-3-yl)-3,4,6,7-tetrahydro-5H-imidazo [4,5-c]pyridine-5,6-dicarboxylate (I-15)

To a stirred solution of 6-benzyl 5-(tert-butyl) (S)-3-benzyl-2-(4-bromo-2-fluorobenzoyl)-3,4,6,7-tetrahydro-5H-imidazo[4,5 -c]pyridine-5,6-dicarboxylate (I-14) (1.74 kg, 2.68 mol) in THF (26.0 L, 15 vol) was added hydrazine hydrate (0.705 L, 13.4 mol ) at room temperature. The resulting reaction mixture was heated at 60°C for 3 hours. After completion of the reaction (TLC monitoring), the resulting reaction mass was poured into ice water (10 L) followed by extraction of the compound with EtOAc (3×10 L). The combined organic layers were washed with brine and dried over sodium sulfate, filtered and evaporated under reduced pressure to give the crude product, which was purified by column chromatography on silica gel (100-200M) using 20% EtOAc in hexane eluents to give the desired product (I -15) as an off-white solid (1.12 kg, 65%). (m/z): [M+H]+ calculated for C ₃₃ H ₃₂ BrN ₅ O ₄ 642.16 found 642.21.

Preparation 4: 6-benzyl 5-(tert-butyl) (S)-3-benzyl-2-(6-(4-(benzyloxy)-2-ethylphenyl)-1H-indazol-3-yl)-3.4 ,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5,6-dicarboxylate (I-16)

Bis(pinacolato)diborone (250 g, 984 mmol) was charged into a 5 L three-neck single wall flask pre-etched with fluoride chemistry along with propan-2-ol (1882 mL, 2.46E+04 mmol) and the mixture was stirred until complete dissolution. Dissolution was endothermic (-4°C). In a 4 L Erlenmeyer flask pre-etched using fluoride chemistry, potassium fluoride hydrofluoride (538 g, 6891 mmol) was dissolved in water (2306 ml, 1.28E+05 mmol) to form a 3M solution. Dissolution was endothermic (-12°C). The solution was then filtered to remove a small amount of insoluble material from KHF ₂ . After complete dissolution of both solutions, the contents of the Erlenmeyer flask were loaded into a single wall flask in portions over 15 minutes. A moderate exothermic effect (+10°C) was observed. The solution turned into a thick and translucent gray slurry during the addition and the stirring was increased to mix the contents well. The mixture was stirred for 1.5 hours and then filtered through a coarse porous glass funnel (4 L, pre-etched). It took 30-45 minutes to complete the filtration. The clear biphasic filtrate was discarded. The white solids were dried for 10 minutes on the filter (cake cracking observed). The solids were transferred back to a cleaned 5 L three neck single wall flask and resuspended with water (1330 mL, 7.38E+04 mmol). The suspension was stirred for 2 h after which time it formed a transparent homogeneous hydrogel. The solution was stirred for an additional 1 hour, after which the solids/gel were filtered off with a coarse 4 L glass funnel (pre-etched). The solids were allowed to dry on the filter for 30 minutes. The solids were transferred back to a cleaned 5 L three neck single wall flask and resuspended in acetone (1084 mL, 1.48E+04 mmol). The white/grey suspension was stirred for 1 hour and then filtered through a coarse 4 L glass funnel (pre-etched). It took 20 minutes to complete the filtration, and then dried on a funnel for another 1 hour. During this time, the solid particles were stirred from time to time to ensure uniform drying. After drying, a light white powder remained on the filter. The solids were dried for 20 hours at 55° C. under vacuum with a slow release of nitrogen to give a fluffy white solid (collected 200.3 g).

To a stirred solution of 6-benzyl 5-(tert-butyl) (S)-3-benzyl-2-(6-bromo-1H-indazol-3-yl)-3,4,6,7-tetrahydro-5H-imidazo [4,5-c]pyridine-5,6-dicarboxylate (I-15) (10.0 g, 16.0 mmol) in 2-methyltetrahydrofuran (100 ml, 10 vol.) was added (4-(benzyloxy)- 2-ethylphenyl)trifluoro-λ ⁴ -borane, potassium salt (I-5) (8.0 g, 20 mmol) and fluffy white solid obtained above (0.20 g). The resulting reaction mixture was degassed with nitrogen for 30 minutes. To this solution was added a prepared aqueous solution of cesium carbonate (20.0 g, 62.0 mmol in 60 ml of water, 6 vol.). The resulting reaction mixture was further degassed for 15 minutes followed by the addition of bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II) (0.66 g, 0.93 mmol) and the reaction mixture was evacuated and purged nitrogen. The resulting reaction mixture was heated at 110°C for 20 hours. After completion of the reaction (TLC & LCMS monitoring), the resulting reaction mixture was cooled to room temperature and filtered through a Celite pad, then washed with EtOAc (3×0.5 L). The combined organic layers were washed with 1N. sodium hydroxide solution (3×0.5 l). The combined organic layers were then washed with brine and dried over sodium sulfate, filtered, and evaporated under reduced pressure to give the crude product, which was purified by silica gel column chromatography (100-200M) using 20% EtOAc in hexane eluents to give the desired product (I-16) (as a mixture of N-benzyl regioisomers) as a light yellow solid (8.0 g, 66%). (m/z): [M+H]+ calculated for C ₄₈ H ₄₇ N ₅ O ₅ 774.36 found 774.59.

Preparation 5: (S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c ]pyridine-6-carboxylic acid, hydrochloride (I-18)

(a) benzyl (S)-3-benzyl-2-(6-(4-(benzyloxy)-2-ethylphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-3H- imidazo[4,5-c]pyridine-6-carboxylate, hydrochloride (I-17)

6-Benzyl 5-(tert-butyl) (S)-3-benzyl-2-(6-(4-(benzyloxy)-2-ethylphenyl)-1H-indazol-3-yl)-3,4,6, 7-tetrahydro-5H-imidazo[4,5-c]pyridine-5,6-dicarboxylate (I-16) (1.0 g, 1.292 mmol) was dissolved in dioxane (8 ml) and water (1.5 ml) , then a solution of hydrogen chloride, 4 M in dioxane (7 ml, 28.0 mmol) was added and the reaction mixture was stirred at room temperature for 3 hours (reaction progress was monitored by LCMS). The reaction mixture was then frozen and lyophilized, and the crude product (I-17) was used directly in the next reaction (quantitative yield assumed). (m/z): [M+H]+ calculated for C ₄₃ H ₃₉ N ₅ O ₃ 674.31 found 674.3.

(b) (S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c ]pyridine-6-carboxylic acid, hydrochloride (I-18)

Benzyl (S)-3-benzyl-2-(6-(4-(benzyloxy)-2-ethylphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-3H-imidazo[4 ,5-c]pyridine-6-carboxylate, hydrochloride (I-17) (0.918 g, 1.292 mmol) was dissolved in 2-propanol (15 ml), hydrogen chloride solution, 5 M in water (0.258 ml, 1.292 mmol), and water (0.25 ml) at 50°C, then added palladium, 10% wt. on charcoal, 50% water (0.138 g, 0.065 mmol). Then the reaction flask was purged with nitrogen, a hydrogen balloon was attached, and the reaction mixture was stirred at 50°C for 4 days, replenishing the hydrogen balloon if necessary (reaction progress was monitored by LCMS). Then all solids were removed by filtration and the resulting solution was concentrated. The residue was dissolved in ACN/water 1:1, frozen and lyophilized. The resulting powder (I-18) was used without further purification (quantitative yield was assumed). (m/z): [M+H]+ calculated for C ₂₂ H ₂₁ N ₅ O ₃ 404.17 found 404.2.

Preparation 6: (S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-isopropyl-4,5,6,7-tetrahydro-3H-imidazo[4 ,5-c]pyridine-6-carboxylic acid (I-19)

(S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine- 6-carboxylic acid, HCl (I-18) (0.25 g, 0.568 mmol) was suspended in DMF (2.5 ml) and acetone (2.5 ml), then acetic acid (0.098 ml, 1.705 mmol) was added and sodium cyanoborohydride (0.179 g, 2.84 mmol) and the reaction mixture was stirred at room temperature for 24 hours (reaction progress was monitored by LCMS). The reaction mixture was concentrated, then the crude product was purified by reverse phase chromatography (5-70% ACN/water gradient, 50 g C18aq column) to give the TFA salt of the title compound (149 mg, 47% yield). (m/z): [M+H]+ calculated for C ₂₅ H ₂₇ N ₅ O ₃ 446.21 found 446.3.

Receipt 7: (S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-propyl-4,5,6,7-tetrahydro-3H-imidazo[4, 5-c]pyridine-6-carboxylic acid (I-20)

(S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine- 6-carboxylic acid, HCl (I-18) (0.160 g, 0.364 mmol) and propionaldehyde (0.039 ml, 0.546 mmol) were dissolved in methanol (3.0 ml), then sodium cyanoborohydride (0.069 g, 1.091 mmol) was added and the reaction the mixture was stirred at room temperature for 24 hours (the progress of the reaction was followed by LCMS). The reaction mixture was concentrated and the crude product was purified by reverse phase chromatography (5-70% ACN/water gradient, 50 g C18 column) to give the TFA salt of the title compound (78 mg, 38% yield). (m/z): [M+H]+ calculated for C ₂₅ H ₂₇ N ₅ O ₃ 446.21 found 446.3.

Receipt eight: (S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-methyl-4,5,6,7-tetrahydro-3H-imidazo[4,5- c]pyridine-6-carboxylic acid (I-21)

(S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine- 6-carboxylic acid, HCl (I-18) (0.160 g, 0.364 mmol) and formaldehyde solution, 37 wt% in water (0.032 ml, 0.436 mmol) were dissolved in methanol (3.0 ml), then sodium cyanoborohydride ( 0.069 g, 1.091 mmol) and the reaction mixture was stirred at room temperature for 4 hours (reaction progress was monitored by LCMS). Sodium borohydride (14 mg, 0.364 mmol) was added to quench excess formaldehyde, then the reaction mixture was concentrated. The crude product was purified by reverse phase chromatography (5-70% ACN/water gradient, 50 g C18 column) to give the TFA salt of the title compound (110 mg, 57% yield). (m/z): [M+H]+ calculated for C ₂₃ H ₂₃ N ₅ O ₃ 418.18 found 418.2.

Receipt 9: (R)-2-(3-methylpiperazin-1-yl)ethan-1-ol, dihydrochloride (I-24)

(a) tert-butyl (R)-4-(2-hydroxyethyl)-2-methylpiperazine-1-carboxylate (I-23)

(R)-1-boc-2-methyl-piperazine (0.2 g, 0.999 mmol), DIPEA (0.698 ml, 3.99 mmol), and 2-bromoethanol (0.085 ml, 1.198 mmol) were dissolved in DMF (5 ml) and the reaction mixture was stirred at 60°C for 16 hours (the progress of the reaction was followed by LCMS). The reaction mixture was concentrated, then 10 ml of diethyl ether was added to the residue. The solution was sonicated until the gel residue disappeared and a solid formed. The ether solution was then decanted from the solid. The solid was then used directly in the next reaction (quantitative yield assumed). (m/z): [M+H]+ calculated for C ₁₂ H ₂₄ N ₂ O ₃ 245.18 found 245.4.

(b) (R)-2-(3-methylpiperazin-1-yl)ethan-1-ol, dihydrochloride (I-24)

Tert-butyl (R)-4-(2-hydroxyethyl)-2-methylpiperazine-1-carboxylate (0.244 g, 0.999 mmol) was dissolved in dioxane (3.0 ml) and water (0.5 ml), then the solution was added hydrogen chloride, 4 M in dioxane (2.0 ml, 65.8 mmol) and the reaction mixture was stirred at room temperature for 3 hours (the progress of the reaction was followed by LCMS). The reaction mixture was frozen and lyophilized and the resulting product was used without purification (quantitative yield assumed). (m/z): [M+H]+ calculated for C ₇ H ₁₆ N ₂ O 145.13 found 145.4.

Example 1: (S)-(2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-propyl-4,5,6,7-tetrahydro-3H-imidazo[ 4,5-c]pyridin-6-yl)(4-(2-hydroxyethyl)piperazin-1-yl)methanone

(S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-propyl-4,5,6,7-tetrahydro-3H-imidazo[4,5- c]pyridine-6-carboxylic acid, TFA (I-20) (40 mg, 0.071 mmol), n-(2-hydroxyethyl)piperazine (0.018 ml, 0.143 mmol) and DIPEA (0.025 ml, 0.143 mmol) were dissolved in DMF (1.5 ml), then HATU (40.8 mg, 0.107 mmol) was added and the reaction mixture was stirred at room temperature for 3 hours (reaction progress was monitored by LCMS). Hydrazine (0.011 ml, 0.357 mmol) was added to cleave unwanted by-products, then the solution was stirred at room temperature for 10 minutes. The reaction mixture was then concentrated and the crude product was purified by preparative HPLC (5-70% ACN/water gradient, column C18) to give the TFA salt of the title compound (31 mg, 56% yield). (m/z): [M+H]+ calculated for C ₃₁ H ₃₉ N ₇ O ₃ 558.31 found 558.3. ¹ H NMR (400 MHz, DMSO-d6) δ 13.10 (s, 1H), 12.27 (d, J=48.92 Hz, 1H), 9.40 (s, 1H), 8.28 ( t, J=8.10 Hz, 1H), 7.30 (s, 1H), 7.04 (m, 2H), 6.71 (d, J=2.46 Hz, 1H), 6.64 ( dd, J=2.50, 8.23Hz, 1H), 4.44(t, J=5.31Hz, 1H), 4.11(q, J=5.26, 2H), 3.96 (m, 1H), 3.86-3.52 (m, 6H), 3.49 (q, J=6.01 Hz, 2H), 2.95 (m, 2H), 2.48 (q, J=7.48Hz, 2H), 2.42-2.21(m, 4H), 2.37(t, J=6.20Hz, 2H), 1.42(m, 2H), 1, 00 (t, J=7.52 Hz, 3H), 0.81 (m, 3H).

Example 2: ((S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-propyl-4,5,6,7-tetrahydro-3H-imidazo[ 4,5-c]pyridin-6-yl)((1S,4S)-5-methyl-2,5-diazabicyclo[2.2.1]heptan-2-yl)methanone

(S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-propyl-4,5,6,7-tetrahydro-3H-imidazo[4,5- c]pyridine-6-carboxylic acid, TFA (I-20) (40 mg, 0.071 mmol), (1S,4S)-5-methyl-2,5-diazabicyclo[2.2.1]heptane dihydrobromide (29.4 mg , 0.107 mmol), and DIPEA (0.062 ml, 0.357 mmol) were dissolved in DMF (1.5 ml), then HATU (40.8 mg, 0.107 mmol) was added and the reaction mixture was stirred at room temperature for 3 hours (per stroke reactions were monitored by LCMS). Hydrazine (0.011 ml, 0.357 mmol) was added to cleave unwanted by-products, then the solution was stirred at room temperature for 10 minutes. The reaction mixture was then concentrated and the crude product was purified by preparative HPLC (5-70% ACN/water gradient, column C18) to give the TFA salt of the title compound (32 mg, 59% yield). (m/z): [M+H]+ calculated for C ₃₁ H ₃₇ N ₇ O ₂ 540.30 found 540.3.

Example 3: ((S)-2,4-dimethylpiperazin-1-yl)((S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-methyl -4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridin-6-yl)methanone 3

(a) tert-butyl (S)-4-((S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-methyl-4,5,6 ,7-tetrahydro-3H-imidazo[4,5-c]pyridine-6-carbonyl)-3-methylpiperazine-1-carboxylate (I-25)

(S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-methyl-4,5,6,7-tetrahydro-3H-imidazo[4,5- c]pyridine-6-carboxylic acid, TFA (I-21) (55 mg, 0.103 mmol), (s)-4-n-boc-2-methylpiperazine (41.5 mg, 0.207 mmol), and DIPEA (0.036 ml, 0.207 mmol) was dissolved in DMF (1.5 ml), then HATU (59.0 mg, 0.155 mmol) was added and the reaction mixture was stirred at room temperature for 16 hours (the reaction was followed by LCMS). Hydrazine (0.016 ml, 0.517 mmol) was added to cleave unwanted by-products, then the reaction mixture was stirred at room temperature for 10 minutes. The reaction mixture was then concentrated and the crude product was purified by reverse phase chromatography (5-70% ACN/water gradient, 50 g C18 column) to give the TFA salt of the title compound (54 mg, 72% yield). (m/z): [M+H]+ calculated for C ₃₃ H ₄₁ N ₇ O ₄ 600.33 found 600.3.

(b) ((S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-methyl-4,5,6,7-tetrahydro-3H-imidazo[ 4,5-c]pyridin-6-yl)((S)-2-methylpiperazin-1-yl)methanone (I-26)

Tert-butyl (S)-4-((S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-methyl-4,5,6,7- tetrahydro-3H-imidazo[4,5-c]pyridine-6-carbonyl)-3-methylpiperazine-1-carboxylate, TFA (I-25) (0.126 g, 0.177 mmol) was dissolved in dioxane (1.5 ml) and water (0.3 ml), then a solution of hydrogen chloride, 4 M in dioxane (1.5 ml, 6.00 mmol) was added and the reaction mixture was stirred at room temperature for 2 hours (the reaction was followed by LCMS). The reaction mixture was frozen and lyophilized, and the resulting powder was used directly in the next reaction (quantitative yield was assumed). (m/z): [M+H]+ calculated for C ₂₈ H ₃₃ N ₇ O ₂ 500.27 found 500.3.

(c) ((S)-2,4-dimethylpiperazin-1-yl)((S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-methyl -4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridin-6-yl)methanone

((S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-methyl-4,5,6,7-tetrahydro-3H-imidazo[4,5 -c]pyridin-6-yl)((S)-2-methylpiperazin-1-yl)methanone, dihydrochloride (0.101 g, 0.176 mmol) and formaldehyde solution, 37% wt. in water (0.016 ml, 0.212 mmol) was dissolved in methanol (3.0 ml), then sodium cyanoborohydride (0.055 g, 0.882 mmol) was added and the reaction mixture was stirred at room temperature for 16 hours (reaction progress was monitored by LCMS) . Sodium borohydride (7 mg, 0.176 mmol) was added to quench the remaining formaldehyde. The reaction mixture was concentrated, then the crude product was purified by preparative HPLC (5-60% ACN/water gradient, column C18) to give the TFA salt of the title compound (28 mg, 21% yield). (m/z): [M+H]+ calculated for C ₂₉ H ₃₅ N ₇ O ₂ 514.29 found 514.3.

Example 4: (S)-(2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-isopropyl-4,5,6,7-tetrahydro-3H-imidazo[ 4,5-c]pyridin-6-yl)(4-methyl-1,4-diazepan-1-yl)methanone

(S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-isopropyl-4,5,6,7-tetrahydro-3H-imidazo[4,5- c]pyridine-6-carboxylic acid, TFA (I-19) (50 mg, 0.089 mmol), 1-methylhomopiperazine (0.022 ml, 0.179 mmol), and DIPEA (0.031 ml, 0.179 mmol) were dissolved in DMF (1.5 ml), then HATU (51.0 mg, 0.134 mmol) was added and the reaction mixture was stirred at room temperature for 3 hours (reaction progress was monitored by LCMS). Hydrazine (0.014 mL, 0.447 mmol) was added to cleave unwanted by-products and the solution was stirred at room temperature for 10 minutes. The reaction mixture was then concentrated and the crude product was purified by preparative HPLC (2-70% ACN/water gradient, column C18) to give the TFA salt of the title compound (29 mg, 42% yield). (m/z): [M+H]+ calculated for C ₃₁ H ₃₉ N ₇ O ₂ 542.32 found 542.3. ¹ H NMR (400 MHz, DMSO-d6) δ 13.08 (s, 1H), 12.21 (d, J=29.9 Hz, 1H), 9.40 (s, 1H), 8.27 ( d, J=8.33 Hz, 1H), 7.30 (s, 1H), 7.04 (t, J=8.05, 2H), 6.71 (d, J=2.53 Hz, 1H ), 6.64 (dd, J=2.54, 8.23 Hz, 1H), 4.11 (m, 3H), 3.91-3.52 (m, 6H), 2.97 (m, 1H), 2.91-2.53 (m, 4H), 2.49 (q, J=7.46, 2H), 2.23 (d, J=13.9 Hz, 3H), 1.76 (m, 2H), 1.0 (m, 9H).

Example 5: ((S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-methyl-4,5,6,7-tetrahydro-3H-imidazo[ 4,5-c]pyridin-6-yl)((R)-4-(2-hydroxyethyl)-2-methylpiperazin-1-yl)methanone

(S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-methyl-4,5,6,7-tetrahydro-3H-imidazo[4,5- c]pyridine-6-carboxylic acid, TFA (I-21) (55 mg, 0.103 mmol), (R)-2-(3-methylpiperazin-1-yl)ethan-1-ol, dihydrochloride (I-24) (33.7 mg, 0.155 mmol) and DIPEA (0.090 ml, 0.517 mmol) were dissolved in DMF (1.5 ml), then HATU (59.0 mg, 0.155 mmol) was added and the reaction mixture was stirred at room temperature for 16 hours (reaction progress was monitored by LCMS). Hydrazine (0.016 ml, 0.517 mmol) was added to cleave unwanted by-products, then the reaction mixture was stirred at room temperature for 10 minutes. The reaction mixture was then concentrated and the crude product was purified by preparative HPLC (5-70% ACN/water gradient, column C18) to give the TFA salt of the title compound (22 mg, 28% yield). (m/z): [M+H]+ calculated for C ₃₀ H ₃₇ N ₇ O ₃ 544.30 found 544.3.

Example 6: ((S)-3-(dimethylamino)pyrrolidin-1-yl)((S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5- isopropyl-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridin-6-yl)methanone

(S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-isopropyl-4,5,6,7-tetrahydro-3H-imidazo[4,5- c]pyridine-6-carboxylic acid, TFA (I-19) (50 mg, 0.089 mmol), (S)-(-)-3-(dimethylamino)pyrrolidine (0.023 ml, 0.179 mmol), and DIPEA (0.031 ml , 0.179 mmol) was dissolved in DMF (1.5 ml), then HATU (51.0 mg, 0.134 mmol) was added and the reaction mixture was stirred at room temperature for 3 hours (the reaction was followed by LCMS). Hydrazine (0.014 mL, 0.447 mmol) was added to cleave unwanted by-products and the solution was stirred at room temperature for 10 minutes. The reaction mixture was then concentrated and the crude product was purified by preparative HPLC (5-70% ACN/water gradient, column C18) to give the TFA salt of the title compound (37 mg, 53% yield). (m/z): [M+H]+ calculated for C ₃₁ H ₃₉ N ₇ O ₂ 542.32 found 542.3.

Example 7: (S)-(3-(dimethylamino)azetidin-1-yl)(2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-isopropyl-4, 5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridin-6-yl)methanone

(S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-isopropyl-4,5,6,7-tetrahydro-3H-imidazo[4,5- c]pyridine-6-carboxylic acid, TFA (I-19) (50 mg, 0.089 mmol), 3-(dimethylamino)azetidine dihydrochloride (23.20 mg, 0.134 mmol), and DIPEA (0.078 ml, 0.447 mmol) were dissolved in DMF (1.5 ml), then HATU (51.0 mg, 0.134 mmol) was added and the reaction mixture was stirred at room temperature for 3 hours (reaction progress was monitored by LCMS). Hydrazine (0.014 mL, 0.447 mmol) was added to cleave unwanted by-products and the solution was stirred at room temperature for 10 minutes. The reaction mixture was then concentrated and the crude product was purified by preparative HPLC (5-70% ACN/water gradient, column C18) to give the TFA salt of the title compound (25 mg, 37% yield). (m/z): [M+H]+ calculated for C ₃₀ H ₃₇ N ₇ O ₂ 528.30 found 528.3. ¹ H NMR (400 MHz, DMSO-d6) δ 13.09 (s, 1H), 9.40 (s, 1H), 8.27 (d, J=8.31, 1H), 7.30 (s , 1H), 7.04 (m, 2H), 6.71 (d, J=2.54, 1H), 6.64 (dd, J=2.53, 8.26, 1H), 4.26 (m, 1H), 4.06 (m, 2H), 3.82 (m, 2H), 3.64 (m, 2H), 3.03 (m, 2H), 2.74 (m, 2H) , 2.47 (q, J=7.56, 2H), 2.07 (d, J=3.69, 6H), 1.07 (m, 6H), 1.00 (t, J=7, 50, 3H).

Receipt 10: 6-benzyl 5-(tert-butyl) (S)-1-benzyl-2-(6-(4-(benzyloxy)-2-ethyl-5-fluorophenyl)-1H-indazol-3-yl)- 1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5,6-dicarboxylate

To a stirred solution of 6-benzyl 5-(tert-butyl) (S)-1-benzyl-2-(6-bromo-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo [4,5-c]pyridine-5,6-dicarboxylate (1.12 kg, 1.74 mol) in 2-methyltetrahydrofuran (11.2 L, 10 vol) was added ((4-(benzyloxy)-2-ethyl-5-fluorophenyl)trifluoroborane, potassium salt) (0.702 kg, 2.1 mol) and the fluffy white solid isolated in preparation 4 (0.223 kg, 1.04 mol). The resulting reaction mixture was degassed with nitrogen for the next 30 minutes through the dropper inlet. To this solution was added the resulting aqueous solution of Cs₂CO₃ (2.27 kg, 6.96 mol in 7.30 L H₂O, 6 vol.). The resulting reaction mixture was further degassed for a further 15 minutes. Pd(amphos) (0.74 kg, 1.04 mol) was added to the resulting reaction mixture and the reaction mixture was evacuated and purged with nitrogen. The resulting reaction mixture was heated to 90°C for 20 hours. After completion of the reaction, the resulting reaction mixture was cooled to room temperature and filtered through a pad of celite, and washed with ethyl acetate (3×7.5 l). The combined organic layers were washed with 1N NaOH solution (3×3 L). The combined organic layers were washed with brine and dried over Na₂SO₄, filtered and evaporated under reduced pressure to give the crude product, which was purified by silica gel column chromatography (100-200 M) using 20% ethyl acetate in hexane as eluents to give the title product as a mixture of region isomers as not quite white solid (1.10 kg, 80%). (m/z): [M+H]⁺ calculated for C₄₈H₄₆FN_fiveO_five792.92 found 792.34.

Preparation 11: Benzyl (S)-1-benzyl-2-(6-(4-(benzyloxy)-2-ethyl-5-fluorophenyl)-1H-indazol-3-yl)-4,5,6,7- tetrahydro-1H-imidazo[4,5-c]pyridine-6-carboxylate, 2Benzenesulfonic acid

To a solution of 6-benzyl 5-(tert-butyl) (S)-1-benzyl-2-(6-(4-(benzyloxy)-2-ethyl-5-fluorophenyl)-1H-indazol-3-yl)- 1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5,6-dicarboxylate (1.005 g, 1269 mmol) in 2-methyltetrahydrofuran (10.050 ml) and isopropyl acetate (4321.5 ml) benzenesulfonic acid (703 g, 4.442 mmol) was added under nitrogen atmosphere. The resulting reaction mixture was stirred at 50°C for 15 hours. After completion of the reaction, isopropyl acetate (5728.5 ml) was added. The suspension was cooled to 20ºC and kept for 1 hour. The thickened suspension was then filtered under nitrogen pressure. The precipitate was then washed with isopropyl acetate (5000 ml) and dried under nitrogen pressure at 25ºC for 2 hours, followed by drying at 60ºC for 16 hours under high vacuum with a nitrogen purge to give the title compound as an off-white free-flowing solid ( 1210 g, 95% yield). ( m/z ): [M+H] ⁺ calculated for C ₄₃ H ₃₈ FN ₅ O ₃ 692.81 found 692.88.

Preparation 12: Benzyl (S)-1-benzyl-2-(6-(4-(benzyloxy)-2-ethyl-5-fluorophenyl)-1H-indazol-3-yl)-5-isopropyl-4.5, 6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-6-carboxylate

To a suspension of molecular sieves (1.21 kg) in acetone (12.0 L) was added benzyl (S)-1-benzyl-2-(6-(4-(benzyloxy)-2-ethyl-5-fluorophenyl)-1H -indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-6-carboxylate, 2Benzenesulfonic acid (1.20 kg, 1.190 mmol) and acetic acid (26 ml, 446 mmol). The suspension was allowed to mix at 25°C for 30 minutes to obtain a heterogeneous off-white-yellow heterogeneous mixture. Sodium triacetoxyborohydride (492 g, 2.321 mmol) was added to the reaction mixture and stirred at 25° C. for 1 hour. The reaction mixture was filtered through celite and the precipitate was washed with 2-methyltetrahydrofuran (500 ml). The filtrate was diluted with 2-methyltetrahydrofuran (8.0 L) and the solvent was replaced by distillation of acetone. Saturated sodium bicarbonate (3025 ml) was added to wash the solution and the mixture was stirred for 1 hour, stirring was stopped, the layers were separated for 15 minutes, the aqueous layer (pH=7.5) was removed and this time the washing was repeated, leaving 2 hours mixing before separating layers. The organic layer was distilled to 2.0 L, isopropyl acetate (8.0 L) was added and the solvent was replaced by distilling off 2-methyltetrahydrofuran to obtain a suspension. The suspension was stirred at 20° C. for 1 hour, then filtered under nitrogen pressure to give the title compound as an off-white solid (795 g, 91% yield). ( m/z ): [M+H] ⁺ calculated for C ₄₆ H ₄₄ FN ₅ O ₃ 734.89 found 734.96.

Receipt 13: (S)-2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-5-isopropyl-4,5,6,7-tetrahydro-3H- imidazo[4,5-c]pyridine-6-carboxylic acid, 2HCl

To a degassed stirred homogeneous solution of benzyl (S)-1-benzyl-2-(6-(4-(benzyloxy)-2-ethyl-5-fluorophenyl)-1H-indazol-3-yl)-5-isopropyl-4, 5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-6-carboxylate (760 g, 1.036 mmol), propan-2-ol (3800 ml) and 1M hydrogen chloride (aq.) (2589 ml , 2,589 mmol) at 50 ^o C was added 10 wt.% Pd/C, 50 wt.%. H ₂ O (76 g, 35.7 mmol) immediately followed by bubbling hydrogen gas through the reaction mixture for 4 hours. The reaction mixture was filtered through a pad of celite (200 grams). SiliaMetS Thiol (76 g, as a white solid) was added to the clear dark yellow filtrate and stirred at 50 ^° C. for 1 hour to remove residual palladium. SiliaMetS Thiol was filtered through a 0.2 µm filter, obtaining a uniform light yellow filtrate, then SiliaMetS Thiol turned bright orange. Isopropyl acetate (7600 ml) was added to the filtrate and concentrated on a rotary evaporator to approximately 3.0 liters. To the concentrate, then in the form of a thick suspension, was added isopropyl acetate (7600 ml), after which the suspension became free-flowing and filterable. The suspension was filtered and the cake was washed with isopropyl acetate (3000 ml), dried under high vacuum for 1 hour, then further dried under high vacuum with nitrogen purge at 50° C. for 18 hours to give the title compound (472 g, 81% exit). ( m/z ): [M+H] ⁺ calculated for C ₂₅ H ₂₆ FN ₅ O ₃ 464.51 found 464.58.

Example 8: (S)-(3-(dimethylamino)azetidin-1-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-5- isopropyl-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridin-6-yl)methanone

To a stirred solution of (S)-2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-5-isopropyl-4,5,6,7-tetrahydro-3H -imidazo[4,5-c]pyridine-6-carboxylic acid, 2HCl (470 g, 876 mmol) and N,N-dimethylazetidine-3-amine, 2HCl (197 g, 1139 mmol) in N,N-dimethylacetamide ( 2436 ml) cooled to -20°C, DIPEA (413 ml, 2366 mmol) was added over at least 15 minutes (exothermic addition caused the batch temperature to rise to -9.1°C). The batch was again cooled to -15ºC and HCTU (453 g, 1095 mmol) was added. The mixture was heated to 20°C for 1 hour and kept for another hour. To the resulting reaction mixture was added isopropyl acetate (5.0 L) and 1M HCl (2.0 L) and the mixture was stirred for 15 minutes, the layers were separated to extract impurities in the layer of isopropyl acetate. The aqueous layer containing the product was extracted three times with isopropyl acetate (5.0 L each). After the 4th extraction, the aqueous layer was added to 2-methyltetrahydrofuran, then saturated sodium bicarbonate solution (~2.2 L, to adjust the pH=8) was stirred for 15 minutes, the layers were separated, and the aqueous layer was discarded. The organic layer was solvent exchanged with acetonitrile and stirred to a final volume of 2.35 L, whereupon the product precipitated out of solution as amorphous filterable solids. The slurry was then filtered under nitrogen pressure to give 345 grams of crude product. The crude product (345 g) was dissolved in methanol (1035 L) with stirring at 55° C. and held for 15 hours to crystallize the product from solution. The suspension was cooled to 10ºC and kept at this temperature with stirring for 2 hours. The thickened slurry was filtered under nitrogen pressure for 2 hours at 20°C followed by drying under high vacuum with a nitrogen purge at 65°C for 18 hours to give the title compound as a free-flowing off-white to white solid (253 g, 53% yield). ( m/z ): [M+H] ⁺ calculated for C ₃₀ H ₃₆ FN ₇ O ₂ 546.66 found 546.73.

Receipt 14: (S)-5-ethyl-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-3H-imidazo[4, 5-c]pyridine-6-carboxylic acid

(S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine- 6-carboxylic acid, HCl (0.100 g, 0.227 mmol) ( I-18 ) and acetaldehyde (0.019 ml, 0.341 mmol) were dissolved in methanol (3.0 ml), then sodium cyanoborohydride (0.057 g, 0.909 mmol) was added and the reaction the mixture was stirred at room temperature for 16 hours (the progress of the reaction was followed by LCMS). Sodium borohydride (9 mg, 0.227 mmol) was added to quench any remaining acetaldehyde, then the reaction mixture was concentrated. The crude product was then purified by reverse phase chromatography (5-70% ACN/water gradient, 40 g C18 column) to give the TFA salt of the title compound (62 mg, 50% yield). (m/z): [M+H]+ calculated for C ₂₄ H ₂₅ N ₅ O ₃ 432.20 found 432.1.

Example 9: (S)-(3-(dimethylamino)azetidin-1-yl)(5-ethyl-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-4, 5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridin-6-yl)methanone

(S)-5-ethyl-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-3H-imidazo[4,5- c]pyridine-6-carboxylic acid, TFA (30 mg, 0.055 mmol), 3-(dimethylamino)azetidine dihydrochloride (14.28 mg, 0.082 mmol), and DIPEA (0.048 ml, 0.275 mmol) were dissolved in DMF (1. 50 ml), then HATU (31.4 mg, 0.082 mmol) was added and the reaction mixture was stirred at room temperature for 1 hour (reaction progress was monitored by LCMS). Hydrazine (5.18 μl, 0.165 mmol) was added to cleave unwanted by-products, then the solution was stirred at room temperature for 10 minutes. The reaction mixture was then concentrated and the crude product was purified by preparative HPLC (5-60% ACN/water gradient, column C18) to give the TFA salt of the title compound (25 mg, 63% yield). (m/z): [M+H]+ calculated for C ₂₉ H ₃₅ N ₇ O ₂ 514.29 found 514.2.

Example 10: (S)-(3-(dimethylamino)-3-methylazetidin-1-yl)(5-ethyl-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl) -4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridin-6-yl)methanone

(S)-5-ethyl-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-3H-imidazo[4,5- c]pyridine-6-carboxylic acid, TFA (30 mg, 0.055 mmol), N,N,3-trimethylazetidine-3-amine hydrochloride (12.43 mg, 0.082 mmol), and DIPEA (0.048 ml, 0.275 mmol) were dissolved in DMF (1.50 ml), then HATU (31.4 mg, 0.082 mmol) was added and the reaction mixture was stirred at room temperature for 1 hour (reaction progress was monitored by LCMS). Hydrazine (5.18 μl, 0.165 mmol) was added to cleave unwanted by-products, then the solution was stirred at room temperature for 10 minutes. The reaction mixture was then concentrated and the crude product was purified by preparative HPLC (5-60% ACN/water gradient, column C18) to give the TFA salt of the title compound (25 mg, 62% yield). (m/z): [M+H]+ calculated for C ₃₀ H ₃₇ N ₇ O ₂ 528.30 found 528.2.

Example 11: (S)-(3-(dimethylamino)-3-methylazetidin-1-yl)(2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-isopropyl -4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridin-6-yl)methanone

(S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-isopropyl-4,5,6,7-tetrahydro-3H-imidazo[4,5- c]pyridine-6-carboxylic acid (40 mg, 0.090 mmol) ( I-19 ) N,N,3-trimethylazetidine-3-amine hydrochloride (20.29 mg, 0.135 mmol), and DIPEA (0.047 ml, 0.269 mmol ) was dissolved in DMF (1.50 ml), then HATU (51.2 mg, 0.135 mmol) was added and the reaction mixture was stirred at room temperature for 16 hours (reaction progress was monitored by LCMS). Hydrazine (8.45 μl, 0.269 mmol) was added to cleave unwanted by-products, then the solution was stirred at room temperature for 10 minutes. The reaction mixture was then concentrated and the crude product was purified by preparative HPLC (5-60% ACN/water gradient, column C18) to give the TFA salt of the title compound (26 mg, 38% yield). (m/z): [M+H]+ calculated for C ₃₁ H ₃₉ N ₇ O ₂ 542.32 found 542.2.

Example 12: (S)-(3-(dimethylamino)azetidin-1-yl)(2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-propyl-4, 5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridin-6-yl)methanone

(S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-propyl-4,5,6,7-tetrahydro-3H-imidazo[4,5- c]pyridine-6-carboxylic acid, TFA (30 mg, 0.054 mmol), ( I-20 ) 3-(dimethylamino)azetidine dihydrochloride (13.92 mg, 0.080 mmol), and DIPEA (0.047 ml, 0.268 mmol) were dissolved in DMF (1.50 ml), then HATU (30.6 mg, 0.080 mmol) was added and the reaction mixture was stirred at room temperature for 1 hour (reaction progress was monitored by LCMS). Hydrazine (5.05 μl, 0.161 mmol) was added to cleave unwanted by-products, then the solution was stirred at room temperature for 10 minutes. The reaction mixture was then concentrated and the crude product was purified by preparative HPLC (5-60% ACN/water gradient, column C18) to give the TFA salt of the title compound (26 mg, 63% yield). (m/z): [M+H]+ calculated for C ₃₀ H ₃₇ N ₇ O ₂ 528.30 found 528.2.

Example 13: (S)-(3-(dimethylamino)-3-methylazetidin-1-yl)(2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-propyl -4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridin-6-yl)methanone

(S)-2-(6-(2-ethyl-4-hydroxyphenyl)-1H-indazol-3-yl)-5-propyl-4,5,6,7-tetrahydro-3H-imidazo[4,5- c]pyridine-6-carboxylic acid, TFA (30 mg, 0.054 mmol), ( I-20 ) N,N,3-trimethylazetidine-3-amine hydrochloride (12.12 mg, 0.080 mmol), and DIPEA (0.047 ml , 0.268 mmol) was dissolved in DMF (1.50 ml), then HATU (30.6 mg, 0.080 mmol) was added and the reaction mixture was stirred at room temperature for 1 hour (the reaction was followed by LCMS). Hydrazine (5.05 μl, 0.161 mmol) was added to cleave unwanted by-products, then the solution was stirred at room temperature for 10 minutes. The reaction mixture was then concentrated and the crude product was purified by preparative HPLC (5-60% ACN/water gradient, column C18) to give the TFA salt of the title compound (18 mg, 44% yield). (m/z): [M+H]+ calculated for C ₃₁ H ₃₉ N ₇ O ₂ 542.32 found 542.2.

Biological analyzes

Compounds of the present disclosure are characterized in one or more of the following biological assays.

Assay 1: JAK kinase biochemical assays

A panel of four LanthaScreen JAK biochemical assays (JAK1, 2, 3 and Tyk2) were run in total kinase reaction buffer (50 mM HEPES, pH 7.5, 0.01% Brij-35, 10 mM MgCl ₂ and 1 mM EGTA). Recombinant GST-tagged JAK enzymes and GFP-tagged STAT1 peptide substrate were obtained from Life Technologies.

Serially diluted compounds were pre-incubated with each of the four JAK enzymes and substrate in white 384-well microplates (Corning) at ambient temperature for 1 hour. ATP was then added to initiate kinase reactions in a total volume of 10 μl with 1% DMSO. Final enzyme concentrations for JAK1, 2, 3, and Tyk2 were 4.2 nM, 0.1 nM, 1 nM, and 0.25 nM, respectively; the respective ATP Km concentrations used were 25 μM, 3 μM, 1.6 μM and 10 μM; while the substrate concentration was 200 nM for all four assays. Kinase reactions were allowed to proceed for 1 hour at ambient temperature, after which 10 μl of EDTA preparation (final concentration 10 mM) and Tb-anti-pSTAT1 (pTyr701) antibody (Life Technologies, final concentration 2 nM) were added in TR dilution buffer -FRET (Life Technologies). The plates were left to incubate at ambient temperature for 1 hour before being read on an EnVision reader (Perkin Elmer). Emissivity signals (520 nm/495 nm) were recorded and used to calculate percent inhibition values based on DMSO and background controls.

For dose-response analysis, percent inhibition data was plotted against compound concentrations and IC ₅₀ values were determined using a 4-parameter robust fitting model with Prism software (GraphPad Software). The results were expressed as pIC ₅₀ (negative log IC ₅₀ ) and then converted to pK _i (negative log dissociation constant Ki) using the Cheng-Prusoff equation.

Test compounds having a lower K _i value or a higher pK _i value in each of the four JAK assays show greater inhibition of JAK activity.

Assay 2: Inhibition of IL-2 stimulated pSTAT5 in Tall-1 T cells

The potency of test compounds in inhibiting interleukin-2 (IL-2) stimulated STAT5 phosphorylation was measured in the Tall-1 human T cell line (DSMZ) using AlphaLisa. Because IL-2 signals through JAK1/3, this assay provides a measure of JAK1/3 cellular activity.

Phosphorylated STAT5 was measured using the AlphaLISA SureFire Ultra pSTAT5 (Tyr694/699) kit (PerkinElmer).

Human T cells from the Tall-1 cell line were cultured in a humid chamber at 37°C, 5% CO ₂ in RPMI (Life Technologies) supplemented with 15% heat-inactivated fetal bovine serum (FBS, Life Technologies), 2 mM glutamax (Life Technologies), 25 mM HEPES (Life Technologies) and 1X Pen/Strep (Life Technologies). Compounds were serially diluted in DMSO and sonicated into empty wells. Assay medium (DMEM without phenol red (Life Technologies) supplemented with 10% FBS (ATCC)) was dispensed (4 μl/well) and the plates were shaken at 900 rpm for 10 minutes. Cells were plated at 45,000 cells/well in assay medium (4 μl/well) and incubated at 37°C, 5% CO ₂ for 1 hour followed by addition of IL-2 (R&D Systems; final concentration 300 ng/ml) into prewarmed assay medium (4 µl) for 30 minutes. Following cytokine stimulation, cells were lysed with 6 μl of 3x AlphaLisa lysis buffer (PerkinElmer) containing 1x PhosStop and Complete tablets (Roche). The lysate was shaken at 900 rpm for 10 minutes at room temperature (room temp.). Phosphorylated STAT5 was measured using the pSTAT5 AlphaLisa kit (PerkinElmer). The freshly prepared acceptor bead mixture was dispensed onto the lysate (5 μl) under filtered green light <100 lux. The plates were shaken at 900 rpm for 2 minutes, centrifuged briefly and incubated for 2 hours at room temperature in the dark. Donor beads were dispensed (5 μl) under <100 lux filtered green light. The plates were shaken at 900 rpm for 2 minutes, centrifuged briefly and incubated overnight at room temperature in the dark. Luminescence was measured with excitation at 689 nm and emission at 570 nm using an EnVision plate reader (PerkinElmer) in <100 lux green filtered light.

To determine the inhibitory ability of the test compounds in response to IL-2, the average emission intensity of the beads associated with pSTAT5 was measured in a human T cell line. IC ₅₀ values were determined from analysis of signal intensity inhibition curves versus compound concentration. Data were expressed as pIC ₅₀ values (negative decimal logarithm of IC ₅₀ ) (mean ± standard deviation).

Results of in vitro analysis

Compounds of the invention were tested in four JAK enzyme assays; JAK1, JAK2, JAK3 and Tyk2, as well as the analysis of the activity of the BEAS-2B cells described above.

Table 1

Example number JAK1
pK _i JAK2
pK _i JAK3pK _i Tyk2pK _i Tall-1
pIC50 one 10.2 10.3 9.8 9.0 8.8 2 10.3 10.2 9.8 8.9 8.4 3 10.2 10.4 10.2 9.3 8.8 4 10.1 10.4 10.1 9.0 8.6 five 10.3 10.5 10.1 9.2 8.8 6 10.2 10.4 10.0 9.0 8.7 7 10.2 10.5 10.2 9.1 8.6 eight 10.4 10.8 10.1 9.5 8.8 nine 10.3 10.7 10.3 9.2 8.7 10 10.4 10.6 10.2 9.1 8.6 eleven 10.0 10.8 9.9 9.1 8.7 12 10.5 10.6 10.3 9.0 8.7 13 10.3 10.5 9.8 9.0 8.6

Analysis 3: Rat (mouse) model of IL-13 induced pSTAT6 induction in lung tissue.

IL-13 is an important cytokine underlying the pathophysiology of asthma (Kudlacz et al. Eur. J. Pharmacol , 2008 , 582,154-161 ). IL-13 binds to cell surface receptors that activate members of the Janus kinase (JAK) family, which then phosphorylate STAT6 and subsequently activate further transcriptional pathways. In the model described, a dose of IL-13 was delivered locally to the lungs of mice to induce STAT6 (pSTAT6) phosphorylation, which was then measured as an end point.

Balb/c adult mice from Harlan were used in the assay. On the day of the study, animals were lightly anesthetized with isoflurane and administered with either vehicle or test compound (1 mg/ml, total volume 50 μl over several breaths) via oral aspiration. After dosing, the animals were placed in the lateral prone position and observed for complete recovery from anesthesia before returning to their home cage. Four hours later, animals were briefly anesthetized again and either vehicle or IL-13 (total delivered dose 0.03 µg, total volume 50 µl) was administered by oral aspiration, followed by recovery from anesthesia and returned to their home cage. One hour after vehicle or IL-13 administration, whole blood and lungs were collected for both detection of pSTAT6 in lung homogenates using the Perkin Elmer AlphaLISA® SureFire® Ultra™ HV p-STAT6 assay kit (Tyr641) and total drug concentration assay both in the lungs and in plasma. Blood samples were centrifuged (Eppendorf centrifuge, 5804R) for 4 minutes at approximately 12,000 rpm at 4° C. to collect plasma. Lungs were washed in DPBS (Dulbecco's Phosphate Buffered Saline), dried, frozen, weighed and homogenized at a 1:3 dilution in 0.1% formic acid in water by HPLC. Plasma and lung test compound levels were determined by LC-MS analysis against analytical standards plotted against a standard curve in a test matrix. The lung-to-plasma ratio was defined as the ratio of lung concentration in ng/g to plasma concentration in ng/ml after 5 hours.

Activity in the model is confirmed by a decrease in the level of pSTAT6 present in the lungs of treated animals after 5 hours compared to vehicle-treated control animals with IL-13. The difference between vehicle-treated IL-13 control animals and vehicle-treated control animals dictated 0% and 100% inhibitory effect, respectively, in any given experiment. Compounds tested in this assay exhibited inhibition of STAT6 phosphorylation 5 hours after IL-13 administration, as indicated below.

Table 2: Observed pSTAT6 Inhibition and Plasma/Lung Effects

Compound Lung concentration (ng/g) after 5 hours Plasma concentration (ng / ml) after 5 hours The ratio of the content in the lungs and plasma after 5 hours pSTAT6 inhibition after 5 h one 13000±5720 10.3±3.9 1262 67 2 23450±12528 17.5±25 1340 72 3 6330±1131 15.4±7 411 76 4 17350±6625 22.0±23 788 67 five 5445±1862 13.0±3.7 418 73 6 11600±4682 11.3±5.7 1026 37 7 10155±1979 24.0±16.2 423 75 eight 22138±5547 58±17 381 51 nine 7750±1652 23±6 339 72 10 5130±2205 24±6 216 73 eleven 15000±3349 55±21 271 72 12 6940±4248 25±9 281 55 13 7465±3084 23±2.5 330 66

Observation of a significant concentration of test compounds in mouse lungs confirmed that the observed inhibition of IL-13 induced pSTAT6 induction was a result of the activity of the test compound. The lung to plasma ratio at 5 hours indicated that compounds 1 to 6 showed significantly greater lung exposure than mouse plasma exposure.

Assay 4: Inhibition of TSLP-induced TARC release in human peripheral blood mononuclear cells

Thymic stromal lymphopoietin (TSLP) and thymus-activated regulated chemokine (TARC) are overexpressed in the airways in asthma and correlate with disease severity. In the lungs, TSLP can be secreted by bronchial epithelial cells in response to allergens and viral infections. TSLP signals through the IL-7Rα/TSLPR heterodimer have been found in a wide range of tissues and cell types, including epithelial cells, endothelial cells, neutrophils, macrophages, and mast cells. Binding of TSLP to its receptor causes a conformational change that activates JAK1 and JAK2 to phosphorylate various transcription factors, including STAT3 and STAT5. In immune cells, this triggers a cascade of intracellular events that lead to cell proliferation, anti-apoptosis, dendritic cell migration, and production of Th2 cytokines and chemokines. In peripheral blood mononuclear cells (PBMCs), TSLP has a pro-inflammatory effect by activating myeloid dendritic cells to recruit and stimulate T cells, a process mediated by the TARC chemoattractant.

In this assay, it was shown that TSLP stimulation induces the release of TARC from PBMC, and that this response is attenuated in a dose-dependent manner upon treatment with the compound. The efficacy of test compounds was measured by inhibition of TARC release.

Aliquots of PBMC (previously isolated from whole blood and frozen in aliquots at -80°C) from 3-5 donors were thawed at 37°C and added dropwise to 40 ml of pre-warmed, sterile-filtered, complete RPMI medium in a 50 ml Falcon tube . Cells were pelleted and resuspended in complete medium at 2.24×10 ⁶ cells/ml. Cells were seeded at 85 μl (190,000 cells) per well in a 96 well tissue cultured flat bottom plate. The cells were allowed to rest for 1 hour at 37°C with 5% CO ₂ .

Compounds were obtained as 10 mM stock solutions in DMSO. 3.7-fold serial dilutions were performed to give 9 concentrations of test compound in DMSO at 300X the final assay test concentration. 150-fold intermediate dilutions were made in complete medium to give the compound at 2X the final assay test concentration with 0.2% DMSO. After a 1 hour rest period, 95 μl of 2X compound was added to each well of PBMC for a final assay concentration range of 33.33 μM to 0.95 μM. 95 μl of 0.2% DMSO in complete medium was added to untreated control wells. Cells were pre-treated with the compound for 1 hour at 37°C with 5% CO ₂ before stimulation.

Recombinant human TSLP protein was reconstituted at 10 μg/ml in sterile DPBS with 0.1% BSA and stored in aliquots at -20°C. Just prior to use, an aliquot was thawed and prepared at 20X the final concentration of the assay in complete medium. 10 μl of 20X TSLP was added to each well of PBMC for a final assay concentration of 10 ng/ml. 10 μl of complete medium was added to unstimulated control wells. Cells were stimulated in the presence of the compound for 48 hours at 37° C. with 5% CO ₂ .

After stimulation, cell culture supernatants were collected and TARC levels were determined by enzyme-linked immunosorbent assay (ELISA) using the Human CCL17/TARC Quantikine ELISA kit (R&D Systems #DDN00) according to the manufacturer's instructions.

For dose effect analysis, log [test compound (M)] was plotted against percentage response values for each donor, and IC50 values were determined using non-linear regression analysis with GraphPad Prism Software using a 4-parameter sigmoidal dose-response algorithm. response with a variable slope. Data are expressed as mean pIC ₅₀ values (negative decimal logarithm of IC ₅₀ ) calculated from individual donor pIC ₅₀ values and rounded to one decimal place. The inhibition efficiency values are shown in Table 3.

Table 3: Efficacy (pIC ₅₀ ) values of test compounds for inhibiting TSLP-induced TARC release in human peripheral blood mononuclear cells

Compound pIC50 ± standard deviation. one 7.1±0.1 2 6.8±0.2 3 7.3±0.2 4 6.8±0.1 five 7.5±0.6 6 6.8±0.2 7 7.2±0.1 eight 7.0±0.1

While the present invention has been described with reference to specific embodiments thereof, one skilled in the art will appreciate that various changes may be made and equivalents may be substituted without departing from the true spirit and scope of the invention. In addition, many modifications can be made to adapt the particular situation, substance, composition in question, method, process step or steps for the object, spirit and scope of the present invention. All such modifications are intended to be within the scope of the appended claims. In addition, all publications, patents and patent documents cited above in this context, are incorporated into this description in their entirety by reference, as if each of them were included individually by reference.

Claims (89)

1. Method for preparing a compound of formula J-15 or a salt thereof:

where X is Br; PG ¹ is a protecting group for a carboxylic acid; PG ² is an amino protecting group; and PG ³ is an amino protecting group; including: (a) reacting a compound of formula J-14:

or its salt with hydrazine to give a compound of formula J-15 and (b) optionally forming a salt of compound J-15. 2. The method according to p. 1, where the reaction with hydrazine is carried out at 60±20°C. 3. The method according to any one of paragraphs. 1 or 2 where PG ¹ is an alkyl or benzyl group, where the benzyl group is optionally substituted; PG ² is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group; and PG ³ is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group. 4. The method according to any one of paragraphs. 1 or 2, where PG ¹ is benzyl, PG ² is t-butoxycarbonyl and PG ³ is benzyl. 5. The method according to any one of paragraphs. 1-4, where the compound J-14 or its salt is obtained: (a) reacting a compound of formula J-13:

where Y is a leaving group, with a compound of formula J-11:

in the presence of a base to give J-14, and (b) optionally forming a salt of compound J-14. 6. The method according to p. 5, where Y represents Cl. 7. Compound of formula J-14:

or its salt, where X is Br; PG ¹ is a protecting group for a carboxylic acid; PG ² is an amino protecting group; and PG ³ is an amino protecting group. 8. The compound according to claim 7 or its salt, where PG ¹ is an alkyl or benzyl group, where the benzyl group is optionally substituted; PG ² is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group; and PG ³ is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group. 9. The compound according to claim 7, having the formula I-14:

or its salt. 10. Compound of formula J-15:

or its salt, where X is Br; PG ¹ is a protecting group for a carboxylic acid; PG ² is an amino protecting group; and PG ³ is an amino protecting group. 11. The compound according to claim 10 or its salt, where PG ¹ is an alkyl or benzyl group, where the benzyl group is optionally substituted; PG ² is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group; and PG ³ is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group. 12. The compound according to claim 10, having the formula I-15:

or its salt. 13. Method for preparing a compound of formula J-16 or a salt thereof:

where PG ¹ is a protecting group for a carboxylic acid; PG ² is an amino protecting group; PG ³ is an amino protecting group; PG ⁴ is a hydroxyl protecting group; and R is H or F; including: (a) reacting a compound of formula J-13:

where X is Br and Y is a leaving group; with a compound of formula J-11:

in the presence of a base, to give a compound of formula J-14:

and optionally forming a salt of compound J-14; (b) reacting a compound of formula J-14 or a salt thereof with hydrazine to give a compound of formula J-15:

and optionally forming a salt of compound J-15; (c) reacting a compound of formula J-15 or a salt thereof with a compound of formula J-5:

J-5 in the presence of a base, a palladium catalyst and a phosphine ligand to give a compound of formula J-16, and optionally forming a salt of compound J-16. 14. The method of claim 13 wherein step (c) is carried out in the presence of a diboronic reagent or catalyst. 15. The process of claim 14, wherein step (c) is carried out in the presence of tetrahydroxydiborone, a diboronic acid ester, or the reaction product of bis(pinacolato)diborone with potassium fluoride hydrofluoride. 16. The method according to any one of paragraphs. 13-15, where the interaction with hydrazine in step (b) is carried out at 60±20°C. 17. The method according to any one of paragraphs. 13-16, where PG ¹ is an alkyl or benzyl group, where the benzyl group is optionally substituted; PG ² is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group; PG ³ is selected from the group consisting of an acyl group, an alkoxycarbonyl group, an arylmethyl group, and a silyl group; PG ⁴ is selected from the group consisting of a silyl group, an acyl group and an arylmethyl group. 18. The method according to any one of paragraphs. 13-16, where PG ¹ is benzyl, PG ² is t-butoxycarbonyl, PG ³ is benzyl, and PG ⁴ is benzyl. 19. The method according to any one of paragraphs. 13-18, where Y is Cl. 20. The method according to any one of paragraphs. 13-19, wherein the palladium catalyst and the phosphine ligand of step (c) are bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium (II).