EP4788429A1 - Modulators of cystic fibrosis transmembrane conductance regulator - Google Patents

Modulators of cystic fibrosis transmembrane conductance regulator

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Publication number
EP4788429A1
EP4788429A1 EP24794995.1A EP24794995A EP4788429A1 EP 4788429 A1 EP4788429 A1 EP 4788429A1 EP 24794995 A EP24794995 A EP 24794995A EP 4788429 A1 EP4788429 A1 EP 4788429A1
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EP
European Patent Office
Prior art keywords
compounds
substituted
pharmaceutically acceptable
alkyl
tautomers
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP24794995.1A
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German (de)
French (fr)
Inventor
Alexander Russell Abela
Jeremy Clemens
Sara Sabina Hadida Ruah
Thomas Cleveland
Christopher Dean COOK
Timothy Richard Coon
Andrew Nguyen DINH
Fabrice Pierre
Haripada Khatuya
Brian LEON MOSTAJO
Mark Thomas Miller
Somayeh MOTEVALLI
Adela NANO
Oleg Ursu
Johnny Uy
Jinglan Zhou
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Vertex Pharmaceuticals Inc
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Vertex Pharmaceuticals Inc
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Application filed by Vertex Pharmaceuticals Inc filed Critical Vertex Pharmaceuticals Inc
Publication of EP4788429A1 publication Critical patent/EP4788429A1/en
Pending legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/404—Indoles, e.g. pindolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/443—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a five-membered ring with oxygen as a ring hetero atom
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00

Definitions

  • Cystic fibrosis is a recessive genetic disease that affects approximately 92,000 children and adults worldwide. Despite progress in the treatment of CF, there is no cure.
  • CFTR mutations in CFTR endogenously expressed in respiratory epithelia lead to reduced apical anion secretion causing an imbalance in ion and fluid transport. The resulting decrease in anion transport contributes to increased mucus accumulation in the lung and accompanying microbial infections that ultimately cause death in CF patients.
  • CF patients In addition to respiratory disease, CF patients typically suffer from gastrointestinal problems and pancreatic insufficiency that, if left untreated, result in death.
  • the CFTR2 database contains information on only 432 of these identified mutations, with sufficient evidence to define 352 mutations as disease-causing.
  • the most prevalent disease-causing mutation is a deletion of phenylalanine at position 508 of the CFTR amino acid sequence and is commonly referred to as the F508del mutation. This mutation occurs in many of the cases of cystic fibrosis and is associated with severe disease. [0006]
  • the deletion of residue 508 in CFTR prevents the nascent protein from folding correctly. This results in the inability of the mutant protein to exit the endoplasmic reticulum (ER) and traffic to the plasma membrane.
  • ER endoplasmic reticulum
  • the number of CFTR channels for anion transport present in the membrane is far less than observed in cells expressing wild-type CFTR, i.e., CFTR having no mutations.
  • the mutation results in defective channel gating.
  • the reduced number of channels in the membrane and the defective gating lead to reduced anion and fluid transport across epithelia.
  • the channels that are defective because of the F508del mutation are still functional, albeit less functional than wild-type CFTR channels.
  • CFTR is a cAMP/ATP-mediated anion channel that is expressed in a variety of cell types, including absorptive and secretory epithelia cells, where it regulates anion flux across the membrane, as well as the activity of other ion channels and proteins. In epithelial cells, normal functioning of CFTR is critical for the maintenance of electrolyte transport throughout the body, including respiratory and digestive tissue.
  • CFTR is composed of 1480 amino acids that encode a protein which is made up of a tandem repeat of transmembrane domains, each containing six transmembrane helices and a nucleotide binding domain. The two transmembrane domains are linked by a large, polar, regulatory (R)-domain with multiple phosphorylation sites that regulate channel activity and cellular trafficking.
  • R regulatory
  • Chloride transport takes place by the coordinated activity of ENaC and CFTR present on the apical membrane and the Na + -K + -ATPase pump and Cl- channels expressed on the basolateral surface of the cell.
  • One aspect of the disclosure provides novel compounds, including compounds of Formula I, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
  • Formula I encompasses compounds falling within the following structure: tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing (i.e., pharmaceutically acceptable salts of compounds, tautomers, and deuterated derivatives), wherein: Ring A is selected from 7- and 8-membered heterocycles containing 0-2 additional heteroatoms selected from O and N, wherein Ring A is substituted with 0-4 groups selected from: • C 3 -C 6 cycloalkyl (substituted with 0-4 groups selected from C 1 -C 4 alkyl), • C1-C5 alkyl (substituted with 0-4 groups selected from C 3 -C 6 cycloalkyl, halogen, OH, NH2, oxo, and 4- to 6-membered heterocycle); • C 1 -C 4 alkoxy (substituted with 0-4 groups selected from C 3 -C 6 cycloalkyl, hal
  • Formula I also includes compounds of Formula I′: tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing (i.e., pharmaceutically acceptable salts of compounds, tautomers, and deuterated derivatives), wherein: Ring A is selected from 7- and 8- membered heterocycles containing 0-2 additional heteroatoms selected from O and N, wherein Ring A is substituted with 0-2 groups selected from: • C 3 -C 6 cycloalkyl (substituted with 0-2 groups selected from C1-C4 alkyl), • C 1 -C 5 alkyl (substituted with 0-2 groups selected from C 3 -C 6 cycloalkyl, halogen, OH, NH2, oxo, and C4-C6 heterocycle); • C1-C4 alkoxy (substituted with 0-2 groups selected from C 3 -C 6 cycloalkyl, halogen
  • Formula I also includes compounds of Formula Ia: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein all variables are as defined for Formula I′.
  • Formula I also includes compounds of Formula Ib: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein all variables are as defined for Formula I′.
  • Formula I also includes compounds of Formula Ic: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein all variables are as defined for Formula I′.
  • Formula II encompasses compounds falling within the following structure: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein R 4 is selected from: • C 3 -C 6 cycloalkyl (substituted with 0-2 groups selected from C1-C4 alkyl), • C 1 -C 5 alkyl (substituted with 0-2 groups selected from C 3 -C 6 cycloalkyl, halogen, OH, NH 2 , oxo, and C 3 -C 6 heterocycle); • C1-C4 alkoxy (substituted with 0-2 groups selected from C 3 -C 6 cycloalkyl, halogen, OH, NH 2 , and
  • Formula II also includes compounds of Formula IIa: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein R 4 is as defined for Formula II, and wherein all other variables are as defined for Formula I′.
  • Formula II also includes compounds of Formula IIb: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein R 4 is as defined for Formula II, and wherein all other variables are as defined for Formula I′.
  • Formula II also includes compounds of Formula IIc: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein R 4 is as defined for Formula II, and wherein all other variables are as defined for Formula I′.
  • Formula I also includes compounds of Formula IIIa: (IIIa), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein: Y 1 is selected from O and -CH2-; Y 2 is selected from NR 9 and CH; Y 3 is selected from O and CH; R 4 is selected from: C1-C4 alkyl (substituted with 0-3 groups selected from halogen and cyclopropyl); C 1 -C 4 alkoxy; and C3-C4 cycloalkyl; and R 8 is selected from C1-C4 alkyl and C3-C4 cycloalkyl, each of which is substituted with 0-1 trifluoromethyl groups and, if R 9 is present, R 9 is methyl; or R 8 and R 9 are taken together to form a five-membered heterocyclic ring, substituted with 0-2 methyl groups.
  • Formula IIIa Formula IIIa: (IIIa), t
  • Formula I also includes compounds of Formula IIIb: (IIIb), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein: R 4 is selected from: C1-C4 alkyl (substituted with 0-3 groups selected from halogen and cyclopropyl); C1-C4 alkoxy; and C 3 -C 4 cycloalkyl; and R 8 is selected from C3 alkyl and C3-C4 cycloalkyl, each of which is substituted with 0-1 trifluoromethyl groups and R 9 is methyl; or R 8 and R 9 are taken together to form a five-membered heterocyclic ring, substituted with 0-2 methyl groups.
  • R 4 is selected from: C1-C4 alkyl (substituted with 0-3 groups selected from halogen and cyclopropyl); C1-C4 alkoxy; and C 3 -C 4 cycloalky
  • the compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb are chosen from Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
  • compositions comprising at least one compound chosen from the novel compounds disclosed herein, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and at least one pharmaceutically acceptable carrier, which compositions may further include at least one additional active pharmaceutical ingredient.
  • the at least one additional active pharmaceutical ingredient is at least one other CFTR modulator.
  • the at least one other CFTR modulator is selected from CFTR potentiators and CFTR modulators.
  • another aspect of the disclosure provides methods of treating the CFTR- mediated disease cystic fibrosis comprising administering at least one compound chosen from the novel compounds disclosed herein, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and at least one pharmaceutically acceptable carrier, optionally as part of a pharmaceutical composition comprising at least one additional component, to a subject in need thereof.
  • the at least one additional active pharmaceutical ingredient in the methods of treating disclosed herein is at least one other CFTR modulator.
  • the at least one other CFTR modulator is selected from CFTR potentiators and CFTR correctors.
  • the pharmaceutical compositions of the disclosure comprise at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
  • the pharmaceutical compositions of the disclosure comprise at least one compound chosen from Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
  • compositions comprising at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing may optionally further comprise (a) at least one (i.e., one or more) compound chosen from (R)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-N-(1-(2,3- dihydroxypropyl)-6-fluoro-2-(1-hydroxy-2-methylpropan-2-yl)-1H-indol-5- yl)cyclopropanecarboxamide (tezacaftor), 3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5- yl)cyclopropane carboxamid
  • compositions comprising at least one compound chosen from Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing may optionally further comprise (a) at least one (i.e., one or more) compound chosen from (R)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-N-(1-(2,3- dihydroxypropyl)-6-fluoro-2-(1-hydroxy-2-methylpropan-2-yl)-1H-indol-5- yl)cyclopropanecarboxamide (tezacaftor), 3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5- yl)cyclopropane carboxamido)-3-
  • Another aspect of the disclosure provides methods of treating the CFTR-mediated disease cystic fibrosis comprising administering to a patient in need thereof at least one compound chosen from the novel compounds disclosed herein, pharmaceutically acceptable salts thereof, and deuterated derivatives of any of the foregoing, and optionally further administering one or more additional CFTR modulating agents selected from tezacaftor, ivacaftor, and lumacaftor.
  • compounds of the disclosure e.g., compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), and pharmaceutical compositions comprising those compounds, and optionally further comprising one or more CFTR modulating agents, are used in therapy or in the manufacture of a medicament.
  • the one or more additional CFTR modulating agents are selected from CFTR potentiators.
  • the one or more additional CFTR modulating agents are selected from CFTR correctors. In some embodiments, the one or more additional CFTR modulating agents are selected from tezacaftor, lumacaftor, ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and (6R)-17-amino-12,12- dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5] nonadeca- 1(18),2,4,14,16-pentaen-6-ol; and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing.
  • a further aspect of the disclosure provides intermediates and methods for making the compounds and compositions disclosed herein.
  • “Tezacaftor” as used herein, refers to (R)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-N- (1-(2,3-dihydroxypropyl)-6-fluoro-2-(1-hydroxy-2-methylpropan-2-yl)-1H-indol-5- yl)cyclopropanecarboxamide, which can be depicted with the following structure: .
  • Tezacaftor may be in the form of a deuterated derivative, a pharmaceutically acceptable salt, or a pharmaceutically acceptable salt of a deuterated derivative.
  • Tezacaftor and methods of making and using tezacaftor are disclosed in WO 2010/053471, WO 2011/119984, WO 2011/133751, WO 2011/133951, WO 2015/160787, and US 2009/0131492, each of which is incorporated herein by reference.
  • Ivacaftor refers to N-[2,4-bis(1,1-dimethylethyl)- 5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide, which is depicted by the structure: .
  • Ivacaftor may also be in the form of a deuterated derivative, a pharmaceutically acceptable salt, or a pharmaceutically acceptable salt of a deuterated derivative.
  • Ivacaftor and methods of making and using ivacaftor are disclosed in WO 2006/002421, WO 2007/079139, WO 2010/108162, and WO 2010/019239, each of which is incorporated herein by reference.
  • a deuterated derivative of ivacaftor (deutivacaftor) is employed in the compositions and methods disclosed herein.
  • a chemical name for deutivacaftor is N-(2-(tert-butyl)-5-hydroxy-4-(2-(methyl-d 3 )propan-2-yl-1,1,1,3,3,3-d 6 )phenyl)-4-oxo-1,4- dihydroquinoline-3-carboxamide, as depicted by the structure: .
  • Deutivacaftor may be in the form of a further deuterated derivative, a pharmaceutically acceptable salt, or a pharmaceutically acceptable salt of a deuterated derivative.
  • Deutivacaftor and methods of making and using deutivacaftor are disclosed in WO 2012/158885, WO 2014/078842, and US Patent No.8,865,902, each of which is incorporated herein by reference.
  • “Lumacaftor” as used herein refers to 3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5- yl)cyclopropanecarboxamido)-3-methylpyridin-2-yl)benzoic acid, which is depicted by the chemical structure: .
  • Lumacaftor may be in the form of a deuterated derivative, a pharmaceutically acceptable salt, or a pharmaceutically acceptable salt of a deuterated derivative.
  • Lumacaftor and methods of making and using lumacaftor are disclosed in WO 2007/056341, WO 2009/073757, and WO 2009/076142, each of which is incorporated herein by reference.
  • (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo [12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, deuterated derivatives, and pharmaceutically acceptable salts and methods of making and using (6R)-17-amino-12,12- dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo [12.3.1.12,5]nonadeca- 1(18),2,4,14,16-pentaen-6-ol are described in WO 2022/109573, WO 2023/224924, and WO 2023/224931, each of which is incorporated herein by reference.
  • aliphatic or “aliphatic group,” as used herein, means a straight-chain (i.e., unbranched) or branched, substituted, or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic (also referred to herein as “cycloaliphatic,” “carbocycle,” or “cycloalkyl”), that has a single point of attachment to the rest of the molecule. Unless otherwise specified, aliphatic groups contain 1-20 aliphatic carbon atoms.
  • aliphatic groups contain 1-10 aliphatic carbon atoms. In other embodiments, unless otherwise specified, aliphatic groups contain 1-8 aliphatic carbon atoms. In still other embodiments, unless otherwise specified, aliphatic groups contain 1-6 aliphatic carbon atoms, and in yet other embodiments, unless otherwise specified, aliphatic groups contain 1-4 aliphatic carbon atoms.
  • cycloaliphatic refers to a monocyclic C3-8 hydrocarbon or bicyclic or tricyclic C8-14 hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point of attachment to the rest of the molecule wherein any individual ring in said bicyclic ring system has 3-7 members.
  • Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and hybrids thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, and (cycloalkyl)alkenyl.
  • Suitable cycloaliphatic groups include cycloalkyl, bicyclic cycloalkyl (e.g., decalin), bridged bicycloalkyl such as norbornyl or [2.2.2]bicyclo-octyl, and bridged tricyclic such as adamantyl.
  • the term “unsaturated” means that a moiety has one or more units of unsaturation.
  • the term “pi bond” means a covalent bond formed by the p orbitals of adjacent atoms. Pi bonds exist where there is a multiple bond, i.e., a double or triple bond, between two atoms. For example, a carbon-carbon double bond consists of one pi bond, and a carbon-carbon triple bond consists of two pi bonds.
  • haloalkyl group refers to an alkyl group substituted with one or more halogen atoms, e.g., fluoroalkyl, which refers to an alkyl group substituted with one or more fluorine atoms.
  • one carbon atom of the alkyl group is substituted with one or more halogen atoms.
  • each carbon atom of the alkyl group is substituted with one or more halogen atoms.
  • one or more carbon atoms of the alkyl group is a perhalo carbon atom (i.e., all hydrogen atoms of the alkyl group are substituted by halogen atoms).
  • each carbon atom of the alkyl group is a perhalo carbon atom.
  • fluoroalkyl include —CHF 2 , —CH 2 F, —CF 3 , —CF 2 —, and perhaloalkyl, such as —CF 2 CF 3 .
  • halogen or “halo” means F, Cl, Br, or I.
  • alkoxy refers to an alkyl or cycloalkyl covalently bonded to an oxygen atom. Alkoxy groups may be linear or branched, and substituted or unsubstituted.
  • cycloalkyl refers to a cyclic, bicyclic, tricyclic, or polycyclic non- aromatic hydrocarbon groups having 3 to 12 carbons (such as, for example 3-10 carbons) and may include one or more unsaturated bonds.
  • Cycloalkyl encompass monocyclic, bicyclic, tricyclic, bridged, fused, and spiro rings, including mono spiro and dispiro rings.
  • Non- limiting examples of cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantyl, norbornyl, dispiro[2.0.2.1]heptane, bicyclo[1.1.1]pentane, and spiro[2,3]hexane. Cycloalkyl groups may be substituted or unsubstituted. [0046]
  • aryl is a functional group or substituent derived from an aromatic ring and encompasses monocyclic aromatic rings and bicyclic, tricyclic, and fused ring systems wherein at least one ring in the system is aromatic.
  • aryl group may be optionally substituted with one or more substituents.
  • aryl groups include phenyl, naphthyl, and 1,2,3,4-tetrahydronaphthalenyl.
  • heteroatom means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the quaternized form of any basic nitrogen; and a substitutable nitrogen of a heterocyclic ring, for example, N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR + (as in N-substituted pyrrolidinyl)).
  • heteroatom refers to oxygen, sulfur, and nitrogen.
  • heteroaliphatic means aliphatic groups wherein one or more carbon atoms are independently replaced with one or more heteroatoms, for example, oxygen, sulfur, nitrogen, phosphorus, or silicon. In some embodiments, the heteroatoms are selected from oxygen, sulfur, and nitrogen. Heteroaliphatic groups may be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and include “heterocycle,” “heterocyclyl,” “heterocycloaliphatic,” and “heterocyclic” groups.
  • heteroaryl ring refers to an aromatic ring comprising at least one ring atom that is a heteroatom, such as O, N, S, P, and Si.
  • the heteroaryl ring contains 1-5 heteroatoms selected from oxygen, sulfur, and nitrogen.
  • the heteroaryl ring contains 1-3 heteroatoms selected from oxygen, sulfur, and nitrogen.
  • Heteroaryl groups encompass monocyclic rings and bicyclic, tricyclic, bridged, fused, and spiro ring systems (including monospiro and dispiro rings) having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic, at least one ring in the system contains one or more heteroatoms, and wherein each ring in the system contains three to seven ring members.
  • Heteroaryl groups also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloalkyl, or heterocyclyl rings, where the point of attachment is on the heteroaromatic ring.
  • heteroaryl rings include pyridine, quinoline, indole, indoline, azaindole, pyrrolopyrazine, furopyridine, and furopyrazine.
  • a heteroaryl group may be optionally substituted with one or more substituents.
  • the term “heteroaryl ring” encompasses heteroaryl rings with various oxidation states, such as heteroaryl rings containing N-oxides and sulfoxides.
  • Non-limiting examples of such heteroaryl rings include pyrimidine N-oxides, quinoline N-oxides, thiophene S-oxides, and pyrimidine N-oxides.
  • heterocycle is used interchangeably with “heterocyclyl ring,” and refers to a non-aromatic hydrocarbon containing 3 to 12 atoms in a ring (such as, for example 3-10 atoms) comprising at least one ring atom that is a heteroatom, such as O, N, S, P, or Si and may include one or more unsaturated bonds.
  • the heterocyclyl ring contains 1-5 heteroatoms selected from oxygen, sulfur, and nitrogen.
  • the heterocyclyl ring contains 1-3 heteroatoms selected from oxygen, sulfur, and nitrogen.
  • Heterocyclyl rings encompass monocyclic, bicyclic, tricyclic, polycyclic, bridged, fused, and spiro rings, including monospiro and dispiro rings.
  • a C 3 -C 6 heterocycle is a heterocycle selected from, for example: [0052]
  • a C 4 -C 6 heterocycle is a heterocycle selected from, for example: [0053]
  • the bond designation “ ” is intended to reflect the presence of an aromatic (i.e., conjugated) ring system.
  • certain compounds of this disclosure may exist as separate stereoisomers or enantiomers and/or mixtures of those stereoisomers or enantiomers.
  • a “wedge” ( ) or “hash” ( ) bond to a stereogenic atom indicates a chiral center of known absolute stereochemistry (i.e., one stereoisomer).
  • a “wavy” bond ( ) to a stereogenic atom indicates a chiral center of unknown absolute stereochemistry (i.e., one stereoisomer).
  • a “wavy” bond ( ) to a double-bonded carbon indicates a mixture of E/Z isomers.
  • a (“straight”) bond to a stereogenic atom indicates where there is a mixture (e.g., racemic or enriched).
  • certain compounds of this disclosure may exist as separated atropisomers and/or mixtures of those atropisomers, i.e., a subclass of stereoisomers resulting from hindered rotation about single bonds or chirality axis and that can be isolated as separate chemical species.
  • a stereogenic unit that is notated with a (P) or (M) indicates the stereochemical designation of the stereogenic unit under the Cahn-Ingold-Prelog convention (basic terminology of stereochemistry, IUPAC Recommendations 1996, Pure & Appl. Chem., Vol 68, No.12, pp. 2193-2222, 1996).
  • stable refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and preferably their recovery, purification, and use for one or more of the purposes disclosed herein.
  • stable compounds refers to compounds which possess sufficient stability to allow for their manufacture and which maintain the integrity of the compounds for a sufficient period of time to be useful for the purposes detailed herein (e.g., formulation into therapeutic products, intermediates for use in production of therapeutic compounds, isolatable or storable intermediates, and/or treating a disease or condition responsive to therapeutic agents).
  • any atom not specifically designated as a particular isotope is meant to represent any stable isotope of that atom. Unless otherwise stated, when a position is designated specifically as “H” or “hydrogen,” the position is understood to have hydrogen at its natural abundance isotopic composition.
  • the term “derivative” refers to a collection of molecules having a chemical structure identical to a compound of this disclosure, except that one or more atoms of the molecule may have been substituted with another atom. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms.
  • deuterated derivative(s) refers to a compound having the same chemical structure as a reference compound, with one or more hydrogen atoms replaced by a deuterium atom.
  • the one or more hydrogens replaced by deuterium are part of an alkyl group.
  • the one or more hydrogens replaced by deuterium are part of a methyl group.
  • deuterium In chemical structures, deuterium may be represented as “D.” In compounds identified as a deuterated derivative, or containing a “D” substitution, the deuterium atom is present in amounts far exceeding its natural abundance; for example, the compound may have an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom).
  • CFTR means cystic fibrosis transmembrane conductance regulator.
  • modulator refers to a compound that increases the activity of a biological compound or molecule such as a protein.
  • CFTR modulator refers to a compound that increases the activity of CFTR.
  • the increase in activity resulting from a CFTR modulator includes, but is not limited to, compounds that correct, potentiate, stabilize, and/or amplify CFTR.
  • the terms “corrector” and “CFTR corrector” are used interchangeably and refer to a compound that facilitates the processing and trafficking of CFTR to increase the amount of CFTR at the cell surface.
  • the novel compounds disclosed herein are CFTR correctors. Tezacaftor and lumacaftor, and deuterated derivatives and pharmaceutically acceptable salts thereof as referenced herein, are correctors.
  • CFTR correctors include those disclosed in WO 2023/034946, WO 2023/034992, WO 2021/097054, WO 2021/097057, WO 2021/113806, WO 2021/113808, and WO 2021113809, such as SION-109, SION-638, SION-719, and SION-451.
  • Additional CFTR correctors include ABBV-2851, disclosed in WO 2017/009804, ABBV-3748; ABBV-3903; and ABBV-119.
  • the terms “potentiator” and “CFTR potentiator” refer to a compound that increases the channel activity of CFTR protein located at the cell surface, resulting in enhanced ion transport.
  • a description of a combination of compounds that includes a compound of the disclosure e.g., compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing
  • a CFTR potentiator such as, e.g., ivacaftor, deutivacaftor, (6R,12R)-17-amino-12- methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5] nonadeca- 1(18),2,4,14,16-pentaen-6-ol, and (6R)-17-
  • a combination of at least one compound of the disclosure will include a potentiator selected from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19- dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-
  • a potentiator selected from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19- dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-
  • such a combination may also include a CFTR potentiator enhancer.
  • CFTR potentiator enhancer As used herein, the term “CFTR potentiator enhancer,” “CFTR potentiation enhancer,” and “CFTR co-potentiator” are used interchangeably and refer to a compound that enhances CFTR potentiation.
  • compound when referring to a compound of this disclosure, refers to a collection of molecules having an identical chemical structure, except that there may be isotopic variation among the constituent atoms of the molecules.
  • a novel compound of the disclosure refers to a compound chosen from compounds of any one of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
  • a novel compound of the disclosure is any compound disclosed in PCT/US2021/053858 (incorporated herein by reference).
  • Compounds 1 to 139 refers to the selection of one or more of the compounds from a specified group. “Selected from” and “chosen from” may be used interchangeably herein.
  • a reference to “Compounds 1 to 139” herein is intended to represent a reference to each of Compounds 1 through 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139) encompassed by Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc IIIa, and/or IIIb, and disclosed in Tables 2 and 4 herein, individually or as a group.
  • the term “active pharmaceutical ingredient” or “therapeutic agent” (“API”) refers to a biologically active compound.
  • the terms “patient” and “subject” are used interchangeably and refer to an animal, including a human.
  • the terms “effective dose” and “effective amount” are used interchangeably herein and refer to that amount of a compound that produces the desired effect for which it is administered (e.g., improvement in CF or a symptom of CF, or lessening the severity of CF or a symptom of CF).
  • treatment generally mean the improvement in one or more symptoms of CF or lessening the severity of CF or one or more symptoms of CF in a subject.
  • Treatment includes, but is not limited to, the following: increased growth of the subject, increased weight gain, reduction of mucus in the lungs, improved pancreatic and/or liver function, reduction of chest infections, and/or reductions in coughing or shortness of breath.
  • references herein to methods of treatment e.g., methods of treating a CFTR mediated disease or a method of treating cystic fibrosis
  • methods of treatment e.g., methods of treating a CFTR mediated disease or a method of treating cystic fibrosis
  • additional CFTR modulating agents e.g., a compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb
  • Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • deuterated derivatives of those compounds and tautomers e.g., and pharmaceutically acceptable salts of any of the foregoing, optionally in combination with one or more additional CFTR modulating agents
  • references herein to methods of treatment e.g.,
  • references herein to methods of treatment e.g., methods of treating a CFTR mediated disease or a method of treating cystic fibrosis using a pharmaceutical composition of the disclosure (e.g., a pharmaceutical composition comprising at least one compound chosen from compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and optionally further comprising one or more additional CFTR modulating agents) should also be interpreted as references to: - a pharmaceutical composition (e.g., a pharmaceutical composition comprising at least one compound chosen from compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc
  • the term “in combination with,” when referring to two or more compounds, agents, or additional active pharmaceutical ingredients, means the administration of two or more compounds, agents, or active pharmaceutical ingredients to the patient prior to, concurrent with, or subsequent to each other.
  • the terms “about” and “approximately,” when used in connection with amounts, volumes, reaction times, reaction temperatures, etc. mean an acceptable error for a particular value as determined by one of ordinary skill in the art, which depends in part on how the value is measured or determined. In some embodiments, the terms “about” and “approximately” mean within 1, 2, 3, or 4 standard deviations.
  • the terms “about” and “approximately” mean within 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, or 0.05% of a given value or range. In some embodiments, “about” and “approximately” mean within 15% of a given value or range. . In some embodiments, “about” and “approximately” mean within 10% of a given value or range. . In some embodiments, “about” and “approximately” mean within 5% of a given value or range.
  • the symbol appearing immediately before a numerical value has the same meaning as the terms “about” and “approximately.”
  • the term “at least one” refers to one or more.
  • the term “solvent” refers to any liquid in which the product is at least partially soluble (solubility of product >1 g/L).
  • Non-limiting examples of suitable solvents include, for example, water (H2O), methanol (MeOH), methylene chloride or dichloromethane (DCM; CH 2 Cl 2 ), acetonitrile (MeCN; CH 3 CN; ACN), N,N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), methyl acetate (MeOAc), ethyl acetate (EtOAc), isopropyl acetate (IPAc), tert-butyl acetate (t-BuOAc), isopropyl alcohol (IPA), tetrahydrofuran (THF), 2-methyl tetrahydrofuran (2-MeTHF), methyl ethyl ketone (MEK), tert-butanol, diethyl ether (Et 2 O), methyl tert-butyl ether (MTBE), 1,4-dioxane, and N
  • the term “F508del” refers to a mutant CFTR protein which is lacking the amino acid phenylalanine at position 508, or to a mutant CFTR gene which encodes for a CFTR protein lacking the amino acid phenylalanine at position 508.
  • the disclosure also provides processes for preparing salts of the compounds of the disclosure.
  • a salt of a compound of this disclosure is formed between an acid and a basic group of the compound, such as an amino functional group, or a base and an acidic group of the compound, such as a carboxyl functional group.
  • the salt is a pharmaceutically acceptable salt.
  • a “pharmaceutically acceptable counterion” is an ionic portion of a salt that is not toxic when released from the salt upon administration to a recipient.
  • a pharmaceutically acceptable salt thereof is disclosed, the amount of the pharmaceutically acceptable salt form of the compound is the amount equivalent to the concentration of the free base of the compound.
  • a “free base” form of a compound does not contain an ionically bonded salt. It is noted that the disclosed amounts of the compounds or their pharmaceutically acceptable salts thereof herein are based upon their free base form.
  • “10 mg of at least one compound chosen from Compound I and pharmaceutically acceptable salts thereof” includes 10 mg of Compound I and a concentration of a pharmaceutically acceptable salt of Compound I equivalent to 10 mg of Compound I.
  • Suitable pharmaceutically acceptable salts are, for example, those disclosed in S. M. Berge, et al. J. Pharmaceutical Sciences, 1977, 66, 1-19.
  • Table 1 of that article provides the following pharmaceutically acceptable salts: Table 1 of S. M.
  • Non-limiting examples of pharmaceutically acceptable acid addition salts include: salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, or perchloric acid; salts formed with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid; and salts formed by using other methods used in the art, such as ion exchange.
  • Non-limiting examples of pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy- ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate
  • CFTR Modulator Compounds [0095] One aspect of the disclosure provides novel compounds, including compounds of Formula I, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
  • Formula I encompasses compounds falling within the following structure: tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing (i.e., salts of compounds, tautomers, and deuterated derivatives), wherein: Ring A is selected from 7- and 8-membered heterocycles containing 0-2 additional heteroatoms selected from O and N, wherein Ring A is substituted with 0-4 groups selected from: • C 3 -C 6 cycloalkyl (substituted with 0-4 groups selected from C1-C4 alkyl); • C 1 -C 5 alkyl (substituted with 0-4 groups selected from C 3 -C 6 cycloalkyl, halogen, OH, NH2, oxo, and 4- to 6-membered heterocycle); • C 1 -C 4 alkoxy (substituted with 0-4 groups selected from C 3 -C 6 cycloalkyl, halogen,
  • Formula I also includes compounds of Formula I′: tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing (i.e., salts of compounds, tautomers, and deuterated derivatives), wherein: Ring A is selected from 7- and 8- membered heterocycles containing 0-2 additional heteroatoms selected from O and N, wherein Ring A is substituted with 0-2 groups selected from: • C 3 -C 6 cycloalkyl (substituted with 0-2 groups selected from C1-C4 alkyl), • C1-C5 alkyl (substituted with 0-2 groups selected from C 3 -C 6 cycloalkyl, halogen, OH, NH 2 , oxo, and C 4 -C 6 heterocycle); • C1-C4 alkoxy (substituted with 0-2 groups selected from C 3 -C 6 cycloalkyl, halogen,
  • Formula I also includes compounds of Formula II: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein R 4 is selected from: • C 3 -C 6 cycloalkyl (substituted with 0-2 groups selected from C 1 -C 4 alkyl), • C 1 -C 5 alkyl (substituted with 0-2 groups selected from C 3 -C 6 cycloalkyl, halogen, OH, NH2, oxo, and C4-C6 heterocycle); • C 1 -C 4 alkoxy (substituted with 0-2 groups selected from C 3 -C 6 cycloalkyl, halogen, OH, NH2, and oxo); • 5- and 6-membered heterocycle; • OH; • NH2; and • oxo; and wherein all other variables are as defined for Formula I′.
  • R 4 is selected from: • C 3 -
  • Formulae I and II also include compounds of Formula IIIb: tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein: R 4 is selected from: C1-C4 alkyl (substituted with 0-3 groups selected from halogen and cyclopropyl); C1-C4 alkoxy; and C 3 -C 4 cycloalkyl; and R 8 is selected from C3 alkyl and C3-C4 cycloalkyl, each of which is substituted with 0-1 trifluoromethyl groups and R 9 is methyl; or R 8 and R 9 are taken together to form a five-membered heterocyclic ring, substituted with 0-2 methyl groups.
  • R 4 is selected from: C1-C4 alkyl (substituted with 0-3 groups selected from halogen and cyclopropyl); C1-C4 alkoxy; and C 3 -C 4 cycloalkyl
  • Ring A is selected from azepane or azocane, wherein one CH 2 group is optionally replaced by an O atom, and wherein Ring A is substituted with 0-2 groups selected from: • C3 or C4 cycloalkyl (substituted with 0-1 methyl groups), • C 1 -C 5 alkyl (substituted with 0-1 groups selected from C 3 -C 4 cycloalkyl); and • C1-C3 alkoxy (substituted with 0-1 groups selected from C3-C4 cycloalkyl); Ring B is a 9-12 membered heteroaryl group containing 2-4 heteroatoms independently selected from N and O, wherein Ring B is substituted with 1-3 groups selected from: • C1-C4 alkyl, substituted with 0-3 groups selected from: o OMe, o halogen, and o OH; • C 3 -C 4 cycloalkyl, substituted with
  • Ring A in the compound, tautomer, deuterated derivative, or salt of Formulae I, I′, Ia, Ib, Ic is selected from: [00109] In some embodiments, Ring A in the compound, tautomer, deuterated derivative, or salt of Formulae I, I′, Ia, Ib, Ic, is selected from: [00110] In some embodiments, Ring B in the compound, tautomer, deuterated derivative, or salt of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb is selected from: Q is selected from C and N; each W is independently selected from CH and N, W 1 is selected from N, CH, and CF; Y is selected from N, NR, O, S, and CR; wherein Ring B contains 1 to 4 nitrogen atoms; R is selected from H and C 1 -C 3 alkyl (substituted with 0-1 groups selected from C 1 -
  • Ring B in the compound, tautomer, deuterated derivative, or salt of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb is selected from:
  • R is selected from H and C1-C3 (alkyl substituted with 0-1 groups selected from C1-C3 alkoxy and OH);
  • R 5 is selected from H, C 1 -C 4 alkyl, and C 3 - and C 4 -cycloalkyl;
  • R 6 is selected from: • H, • C 1 -C 5 alkyl, • C1-C5 haloalkyl, • C 3 and C 4 cycloalkyl, substituted with 0-1 groups selected from: o C1-C4 alkyl, o CH2OCH3, and o C 1 -C 3 haloalkyl; • 5- and 6-membered heterocycle;
  • R 7 is selected from C1-C3 alkyl;
  • R 8 is selected from C 1 -C 4 alkyl; and
  • G is selected from halogens; and wherein R, R 5 and R 6 are not all simultaneously hydrogen, R 5 and R 6 are not simultaneously hydrogen when R is not present, and R 6 is not
  • Ring B in the compound, tautomers, deuterated derivatives, or salts of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb is selected from:
  • Ring B in the compounds, tautomers, deuterated derivatives, or salts of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb is selected from: [00114]
  • the compounds of Formulae I or I′ are chosen from Compounds 1 to 107, tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
  • the compounds of Formula I or I′ are chosen from Table 3. Table 3.
  • the compounds of Formula I or I′ are chosen from Compounds 108 to 139, tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
  • Table 4 Structures of Compounds 108 to 139
  • the compounds of the invention include silicon derivatives of the compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein at least one carbon atom has been replaced with silicon.
  • the at least one carbon atom replaced with silicon may be a non-aromatic carbon. In some embodiments, the at least one carbon atom replaced with silicon may be an aromatic carbon. In certain embodiments, the silicon derivatives of the invention may also have one or more hydrogen atoms replaced with deuterium. In certain embodiments, the silicon derivatives of the invention may also have one or more carbon atoms replaced with germanium.
  • the compounds of the invention include germanium derivatives of the compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein at least one carbon atom has been replaced with germanium.
  • the at least one carbon atom replaced with germanium may be a non-aromatic carbon.
  • the at least one carbon atom replaced with germanium may be an aromatic carbon.
  • the germanium derivatives of the invention may also have one or more hydrogen atoms replaced with deuterium. In certain embodiments, the germanium derivatives of the invention may also have one or more carbon atoms replaced with silicon. [00120] Exemplary embodiments of the invention include, but are not limited to, the following silicon and germanium derivatives:
  • any of the novel compounds disclosed herein such as, for example, a compound chosen from compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, can act as a CFTR modulator, i.e., it modulates CFTR activity in the body. Individuals suffering from a mutation in the gene encoding CFTR may benefit from receiving a CFTR modulator.
  • a CFTR mutation may affect the CFTR quantity, i.e., the number of CFTR channels at the cell surface, or it may impact CFTR function, i.e., the functional ability of each channel to open and transport ions.
  • Mutations affecting CFTR quantity include mutations that cause defective synthesis (Class I defect), mutations that cause defective processing and trafficking (Class II defect), mutations that cause reduced synthesis of CFTR (Class V defect), and mutations that reduce the surface stability of CFTR (Class VI defect).
  • Mutations that affect CFTR function include mutations that cause defective gating (Class III defect) and mutations that cause defective conductance (Class IV defect).
  • Some CFTR mutations exhibit characteristics of multiple classes. Certain mutations in the CFTR gene result in cystic fibrosis.
  • the disclosure provides methods of treating, lessening the severity of, or symptomatically treating cystic fibrosis in a patient comprising administering to the patient an effective amount of any of the novel compounds disclosed herein, such as, for example, a compound chosen from compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, alone or in combination with another active ingredient, such as one or more CFTR modulating agents.
  • a compound chosen from compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Com
  • the one or more CFTR modulating agents are selected from ivacaftor, deutivacaftor, lumacaftor, and tezacaftor.
  • the patient has an F508del/minimal function (MF) genotype, F508del/F508del genotype (homozygous for the F508del mutation), F508del/gating genotype, or F508del/residual function (RF) genotype.
  • MF F508del/minimal function
  • F508del/F508del genotype homozygous for the F508del mutation
  • F508del/gating genotype F508del/gating genotype
  • F508del/residual function (RF) genotype F508del/residual function
  • the patient is heterozygous and has one F508del mutation.
  • the patient is homozygous for the N1303K mutation
  • 1 mg to 500 mg of a compound disclosed herein, a tautomer thereof, deuterated derivatives of the compound and tautomer, or a pharmaceutically acceptable salt of any of the foregoing are administered daily.
  • the patient has at least one F508del mutation in the CFTR gene.
  • the patient has a CFTR gene mutation that is responsive to a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt of the invention based on in vitro data.
  • the disclosure also is directed to methods of treatment using isotope-labelled compounds of the afore-mentioned compounds, or pharmaceutically acceptable salts thereof, wherein the formula and variables of such compounds and salts are each and independently as described above or any other embodiments described above, provided that one or more atoms therein have been replaced by an atom or atoms having an atomic mass or mass number which differs from the atomic mass or mass number of the atom which usually occurs naturally (isotope labelled).
  • isotopes which are commercially available and suitable for the disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, for example 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F, and 36 Cl, respectively.
  • the isotope-labelled compounds and salts can be used in a number of beneficial ways. They can be suitable for medicaments and/or various types of assays, such as substrate tissue distribution assays. For example, tritium ( 3 H)- and/or carbon-14 ( 14 C)-labelled compounds are particularly useful for various types of assays, such as substrate tissue distribution assays, due to relatively simple preparation and excellent detectability.
  • deuterium ( 2 H)-labelled ones are therapeutically useful with potential therapeutic advantages over the non- 2 H-labelled compounds.
  • deuterium ( 2 H)-labelled compounds and salts can have higher metabolic stability as compared to those that are not isotope-labelled owing to the kinetic isotope effect described below. Higher metabolic stability translates directly into an increased in vivo half-life or lower dosages, which could be desired.
  • the isotope-labelled compounds and salts can usually be prepared by carrying out the procedures disclosed in the synthesis schemes and the related description, in the example part and in the preparation part in the present text, replacing a non-isotope-labelled reactant by a readily available isotope-labelled reactant.
  • the isotope-labelled compounds and salts are deuterium ( 2 H)- labelled ones.
  • the isotope-labelled compounds and salts are deuterium ( 2 H)-labelled, wherein one or more hydrogen atoms therein have been replaced by deuterium. In chemical structures, deuterium may be represented as “D” or “ 2 H.”
  • deuterium may be represented as “D” or “ 2 H.”
  • concentration of the isotope(s) (e.g., deuterium) incorporated into the isotope- labelled compounds and salt of the disclosure may be defined by the isotopic enrichment factor.
  • the term “isotopic enrichment factor” as used herein means the ratio between the isotopic abundance and the natural abundance of a specified isotope.
  • a substituent in a compound of the disclosure is denoted as deuterium
  • such compound has an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation).
  • One aspect disclosed herein provides methods of treating cystic fibrosis and other CFTR mediated diseases using any of the novel compounds disclosed herein, such as, for example, compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, in combination with at least one additional active pharmaceutical ingredient.
  • At least one additional active pharmaceutical ingredient is selected from mucolytic agents, bronchodilators, antibiotics, anti-infective agents, and anti- inflammatory agents.
  • the additional therapeutic agent is an antibiotic.
  • Exemplary antibiotics useful herein include tobramycin, including tobramycin inhaled powder (TIP), azithromycin, aztreonam, including the aerosolized form of aztreonam, amikacin, including liposomal formulations thereof, ciprofloxacin, including formulations thereof suitable for administration by inhalation, levofloxacin, including aerosolized formulations thereof, and combinations of two antibiotics, e.g., fosfomycin and tobramycin.
  • the additional agent is a mucolyte.
  • mucolytes useful herein include Pulmozyme®.
  • the additional agent is a bronchodilator.
  • Exemplary bronchodilators include albuterol, metaproterenol sulfate, pirbuterol acetate, salmeterol, or terbutaline sulfate.
  • the additional agent is an anti-inflammatory agent, i.e., an agent that can reduce the inflammation in the lungs.
  • Exemplary such agents useful herein include ibuprofen, docosahexaenoic acid (DHA), sildenafil, inhaled glutathione, pioglitazone, hydroxychloroquine, or simvastatin.
  • the additional agent is a nutritional agent.
  • Exemplary nutritional agents include pancrelipase (pancreatic enzyme replacement), including Pancrease®, Pancreacarb®, Ultrase®, or Creon®, Liprotomase® (formerly Trizytek®), Aquadeks®, or glutathione inhalation.
  • the additional nutritional agent is pancrelipase.
  • At least one additional active pharmaceutical ingredient is selected from CFTR modulating agents.
  • the at least one additional active pharmaceutical ingredient is selected from CFTR potentiators.
  • the potentiator is selected from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing.
  • the at least one additional active pharmaceutical ingredient is chosen from CFTR correctors.
  • the correctors are selected from lumacaftor, tezacaftor, and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing.
  • the at least one additional active pharmaceutical ingredient is chosen from (a) tezacaftor, lumacaftor, and deuterated derivatives and pharmaceutically acceptable salts thereof; and/or (b) ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing.
  • the at least one additional active pharmaceutical ingredient is (a) a compound chosen from (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19- dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol and (6R)-17- amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, deuterated derivatives of the compound, and pharmaceutically acceptable salts of any of the foregoing, and may optionally include tezacaftor, deuterated derivatives, or pharmaceutically acceptable salts of any of the foregoing.
  • the combination therapies provided herein comprise (a) a compound selected from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing; and (b) at least one compound selected from tezacaftor, lumacaftor, and deuterated derivatives and pharmaceutically acceptable salts thereof; or (c) at least one compound selected from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5] nonadeca-1(18
  • the combination therapies provided herein comprise (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing; (b) at least one compound selected from tezacaftor and pharmaceutically acceptable salts thereof; and (c) at least one compound selected from ivacaftor, deutivacaftor, and pharmaceutically acceptable salts thereof.
  • the combination therapies provided herein comprise (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing; (b) at least one compound selected from tezacaftor, lumacaftor, and deuterated derivatives and pharmaceutically acceptable salts thereof; and/or (c) at least one compound selected from (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R,12R)-17
  • the combination therapies provided herein comprise (a) a compound chosen from (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa- 3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol and (6R)-17-amino- 12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol, deuterated derivatives of the compound, and pharmaceutically acceptable salts of any of the foregoing, and (b) optionally tezacaftor, deuterated derivatives, or pharmaceutically acceptable salts of any of the foregoing.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • deuterated derivatives of those compounds and tautomers i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers
  • pharmaceutically acceptable salts of any of the foregoing is administered in combination with at least one compound chosen from tezacaftor and pharmaceutically acceptable salts thereof.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • deuterated derivatives of those compounds and tautomers i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers
  • pharmaceutically acceptable salts of any of the foregoing is administered in combination with at least one compound chosen from lumacaftor and pharmaceutically acceptable salts thereof.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • deuterated derivatives of those compounds and tautomers i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers
  • pharmaceutically acceptable salts of any of the foregoing is administered in combination with at least one compound chosen from ivacaftor and pharmaceutically acceptable salts thereof.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • deuterated derivatives of those compounds and tautomers i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers
  • pharmaceutically acceptable salts of any of the foregoing is administered in combination with at least one compound chosen from deutivacaftor and pharmaceutically acceptable salts thereof.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • Compounds 1 to 139 is administered in combination with at least one compound chosen from (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers
  • pharmaceutically acceptable salts of any of the foregoing is administered in combination with at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof and at least one compound chosen from ivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers
  • pharmaceutically acceptable salts of any of the foregoing is administered in combination with at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof and at least one compound chosen from deutivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers
  • pharmaceutically acceptable salts of any of the foregoing is administered in combination with at least one compound chosen from lumacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof and at least one compound chosen from ivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • deuterated derivatives of those compounds and tautomers i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • deuterated derivatives of those compounds and tautomers i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • pharmaceutically acceptable salts of any of the foregoing is administered in combination with at least one compound chosen from lumacaftor and deuterated derivatives and pharmaceutically
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • Compounds 1 to 139 is administered in combination with at least one compound chosen from lumacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof and at least one compound chosen from (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • Compounds 1 to 139 is administered in combination with at (a) a compound chosen from (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol and (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo
  • Each of the compounds of the disclosure (e.g., Compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), independently can be administered once daily, twice daily, or three times daily.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • deuterated derivatives of those compounds and tautomers i.e., deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • deuterated derivatives of those compounds and tautomers i.e., deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers
  • deuterated derivatives of those compounds and tautomers and pharmaceutically acceptable salts of any of the foregoing
  • at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof are administered once daily.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing
  • at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof are administered twice daily.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing
  • at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof are administered once daily and at least one compound chosen from ivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, are administered twice daily.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing
  • at least one compound chosen from lumacaftor and pharmaceutically acceptable salts thereof are administered once daily and at least one compound chosen from ivacaftor and pharmaceutically acceptable salts thereof, are administered twice daily.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing
  • at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof are administered once daily and at least one compound chosen from (6R,12R)-17- amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo [12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15- bis(trifluor
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing
  • at least one compound chosen from lumacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof are administered once daily and at least one compound chosen from (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa- 3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino-12,12- dimethyl-6,15-bis(trifluoromethyl)
  • Compounds of the disclosure e.g., compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), as well as additional CFTR modulator compounds, such as, e.g., tezacaftor, lumacaftor, ivacaftor, deutivacaftor, (6R,12R)- 17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-
  • Such pharmaceutical compositions can be administered once daily or multiple times daily, such as twice daily.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • Compounds 1 to 139 is administered in a first pharmaceutical composition
  • at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a second pharmaceutical composition
  • at least one compound chosen from ivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a third pharmaceutical composition.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • Compounds 1 to 139 is administered in a first pharmaceutical composition
  • at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a second pharmaceutical composition
  • at least one compound chosen from deutivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a third pharmaceutical composition.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in a first pharmaceutical composition; at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a second pharmaceutical composition; at least one compound chosen from (6R,12R)-17-amino-12- methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca- 1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in a first pharmaceutical composition; at least one compound chosen from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino-12,12- dimethyl-6,15-bis(trifluoromethyl)-19-ox
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing
  • a first pharmaceutical composition is administered in a first pharmaceutical composition
  • the second pharmaceutical composition comprises a half of a daily dose of said at least one compound chosen from ivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, and the other half of the daily dose of said at least one compound chosen from ivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a third pharmaceutical composition.
  • At least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing
  • at least one compound chosen from tezacaftor and pharmaceutically acceptable salts thereof, and at least one compound chosen from ivacaftor, deutivacaftor, and pharmaceutically acceptable salts thereof are administered in a first pharmaceutical composition.
  • the first pharmaceutical composition is administered to the patient twice daily. In some embodiments, the first pharmaceutical composition is administered once daily. In some embodiments, the first pharmaceutical composition is administered once daily and, when the first composition comprises ivacaftor, a second composition comprising only ivacaftor is administered once daily.
  • any suitable pharmaceutical compositions can be used for compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, tezacaftor, ivacaftor, deutivacaftor, lumacaftor and tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
  • Some exemplary pharmaceutical compositions for tezacaftor and its pharmaceutically acceptable salts can be found in WO 2011/119984 and WO 2014/014841, all of which is incorporated herein by reference.
  • Some exemplary pharmaceutical compositions for ivacaftor and its pharmaceutically acceptable salts can be found in WO 2007/134279, WO 2010/019239, WO 2011/019413, WO 2012/027731, and WO 2013/130669, and some exemplary pharmaceutical compositions for deutivacaftor and its pharmaceutically acceptable salts can be found in US 8,865,902, US 9,181,192, US 9,512,079, WO 2017/053455, and WO 2018/080591, all of which are incorporated herein by reference.
  • this disclosure provides a method of treating cystic fibrosis comprising the step of administering a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt disclosed herein (e.g., compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), or a pharmaceutical composition disclosed herein (e.g., pharmaceutical compositions comprising compounds of any of Formulae I, I′, I
  • the method of treating cystic fibrosis further comprises administering at least one additional active therapeutic agent.
  • this disclosure provides use of a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt disclosed herein (e.g., compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), or a pharmaceutical composition disclosed herein (e.g., pharmaceutical compositions comprising compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 139
  • the medicament is formulated to be administered with at least one additional active therapeutic agent.
  • the medicament comprises at least one additional active therapeutic agent.
  • this disclosure provides a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt disclosed herein (e.g., compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), or a pharmaceutical composition disclosed herein (e.g., pharmaceutical compositions comprising compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.
  • this disclosure provides a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt disclosed herein (e.g., compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), or a pharmaceutical composition disclosed herein (e.g., pharmaceutical compositions comprising compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutical
  • the treatment further comprises administering at least one additional active therapeutic agent.
  • this disclosure provides the use of a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt disclosed herein (e.g., compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), or a pharmaceutical composition disclosed herein (e.g., pharmaceutical compositions comprising compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds
  • this disclosure provides the use of a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt disclosed herein (e.g., compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), or a pharmaceutical composition disclosed herein (e.g., pharmaceutical compositions comprising compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers
  • the use further comprises use of at least one additional active therapeutic agent.
  • the at least one additional active therapeutic agent to be administered is selected from mucolytic agents, bronchodilators, antibiotics, anti-infective agents, anti-inflammatory agents, nutritional agents, and CFTR modulators.
  • the at least one additional active therapeutic agent to be administered is selected from CFTR potentiators.
  • the at least one CFTR potentiator to be administered is selected from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)- 13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17- amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing.
  • the at least one additional active therapeutic agent to be administered is selected from CFTR correctors.
  • the at least one CFTR corrector to be administered is selected from lumacaftor, tezacaftor, and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing.
  • the at least one additional active therapeutic agent to be administered is (a) at least one compound selected from tezacaftor, lumacaftor, and deuterated derivatives and pharmaceutically acceptable salts thereof; and (b) at least one compound selected from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)- 13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17- amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts of any of
  • the at least one additional active therapeutic agent to be administered is (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol, and optionally tezacaftor.
  • the at least one additional active therapeutic agent to be administered is (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and optionally tezacaftor.
  • compositions [00174] Another aspect of the disclosure provides a pharmaceutical composition comprising at least one novel compound of the disclosure (e.g., a compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), and at least one pharmaceutically acceptable carrier.
  • a novel compound of the disclosure e.g., a compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb
  • Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof i.e., Compounds 1 to
  • the disclosure provides pharmaceutical compositions comprising at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, in combination with at least one additional active pharmaceutical ingredient.
  • the at least one additional active pharmaceutical ingredient is a CFTR modulator.
  • the at least one additional active pharmaceutical ingredient is a CFTR corrector.
  • the at least one additional active pharmaceutical ingredient is a CFTR potentiator.
  • the pharmaceutical composition comprises at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and at least two additional active pharmaceutical ingredients, one of which is a CFTR corrector and one of which is a CFTR potentiator.
  • the disclosure provides a pharmaceutical composition
  • a pharmaceutical composition comprising (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, (b) at least one compound chosen from tezacaftor and pharmaceutically acceptable salts thereof, and (c) at least one pharmaceutically acceptable carrier.
  • the disclosure provides a pharmaceutical composition
  • a pharmaceutical composition comprising (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, (b) at least one compound chosen from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19- dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino- 12,12-dimethyl-6,15-bis(trifluorol,
  • the disclosure provides a pharmaceutical composition
  • a pharmaceutical composition comprising (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, (b) at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, (c) at least one compound chosen from ivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, and (d) at least one pharmaceutically acceptable carrier.
  • Compounds 1 to 139 i.e., Compounds 1 to 107 and/or Compounds 108 to 139
  • tautomers thereof deuterated derivatives of those compounds and tautomers
  • the disclosure provides a pharmaceutical composition
  • a pharmaceutical composition comprising (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, (b) at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, (c) at least one compound chosen from deutivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, and (d) at least one pharmaceutically acceptable carrier.
  • the disclosure provides a pharmaceutical composition
  • a pharmaceutical composition comprising (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, (a) at least one compound chosen from (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino-12,12- dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-te
  • the disclosure provides a pharmaceutical composition
  • a pharmaceutical composition comprising (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, (b) at least one compound chosen from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19- dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino- 12,12-dimethyl-6,15-bis(trifluorol,
  • any pharmaceutical composition disclosed herein may comprise at least one pharmaceutically acceptable carrier.
  • the at least one pharmaceutically acceptable carrier is chosen from pharmaceutically acceptable vehicles and pharmaceutically acceptable adjuvants.
  • the at least one pharmaceutically acceptable is chosen from pharmaceutically acceptable fillers, disintegrants, surfactants, binders, and lubricants.
  • the pharmaceutical compositions described herein are useful for treating cystic fibrosis and other CFTR mediated diseases.
  • pharmaceutical compositions disclosed herein may optionally further comprise at least one pharmaceutically acceptable carrier.
  • the at least one pharmaceutically acceptable carrier may be chosen from adjuvants and vehicles.
  • the at least one pharmaceutically acceptable carrier includes any and all solvents, diluents, other liquid vehicles, dispersion aids, suspension aids, surface active agents, isotonic agents, thickening agents, emulsifying agents, preservatives, solid binders, and lubricants, as suited to the particular dosage form desired.
  • Non-limiting examples of suitable pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffer substances (such as phosphates, glycine, sorbic acid, and potassium sorbate), partial glyceride mixtures of saturated vegetable fatty acids, water, salts, and electrolytes (such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, and zinc salts), colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, wool fat, sugars (such as lactose, glucose and sucrose), starches (such as corn starch and potato starch), cellulose and its derivatives (such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate), powdered tragacanth, malt,
  • NMR (1D & 2D) spectra were also recorded on a Bruker AVNEO 400 MHz spectrometer operating at 400 MHz and 100 MHz respectively equipped with a 5 mm multinuclear Iprobe.
  • NMR spectra were also recorded on a Varian Mercury NMR instrument at 300 MHz for 1 H using a 45 degree pulse angle, a spectral width of 4800 Hz and 28860 points of acquisition. FID were zero-filled to 32k points and a line broadening of 0.3Hz was applied before Fourier transform.
  • 19 F NMR spectra were recorded at 282 MHz using a 30 degree pulse angle, a spectral width of 100 kHz and 59202 points were acquired.
  • FID were zero-filled to 64k points and a line broadening of 0.5 Hz was applied before Fourier transform.
  • NMR spectra were also recorded on a Bruker Avance III HD NMR instrument at 400 MHz for 1 H using a 30 degree pulse angle, a spectral width of 8000 Hz and 128k points of acquisition. FID were zero-filled to 256k points and a line broadening of 0.3 Hz was applied before Fourier transform.
  • 19 F NMR spectra were recorded at 377 MHz using a 30 deg pulse angle, a spectral width of 89286 Hz and 128k points were acquired. FID were zero-filled to 256k points and a line broadening of 0.3 Hz was applied before Fourier transform.
  • Mobile phase A H2O (0.05 % CF3CO2H).
  • Mobile phase B CH3CN (0.035 % CF3CO2H).
  • LC method B Acquity UPLC BEH C 18 column (30 ⁇ 2.1 mm, 1.7 ⁇ m particle) made by Waters (pn: 186002349), and a dual gradient run from 1-99% mobile phase B over 1.0 minute.
  • Mobile phase A H 2 O (0.05 % CF 3 CO 2 H).
  • Mobile phase B CH 3 CN (0.035 % CF 3 CO 2 H).
  • LC method C Reversed phase UPLC using an Acquity UPLC BEH C18 column (50 ⁇ 2.1 mm, 1.7 ⁇ m particle) made by Waters (pn: 186002350), and a dual gradient run from 30- 99% mobile phase B over 2.9 minutes.
  • Mobile phase A H2O (0.05 % CF3CO2H).
  • Mobile phase B CH3CN (0.035 % CF3CO2H).
  • LC method D Merckmillipore Chromolith SpeedROD C 18 column (50 x 4.6 mm) and a dual gradient run from 5 – 100% mobile phase B over 12 minutes.
  • Mobile phase A water (0.1 % CF 3 CO 2 H).
  • Mobile phase B acetonitrile (0.1 % CF 3 CO 2 H).
  • LC method E Merckmillipore Chromolith SpeedROD C 18 column (50 x 4.6 mm) and a dual gradient run from 5 – 100% mobile phase B over 6 minutes.
  • Mobile phase A water (0.1 % CF3CO2H).
  • Mobile phase B acetonitrile (0.1 % CF3CO2H).
  • LC method F Kinetex Polar C 18 3.0 x 50 mm 2.6 ⁇ m, 6 min, 5-95% I in H 2 O (0.1% Formic Acid) 1.2 mL/min.
  • LC method G Acquity UPLC BEH C18 column (50 ⁇ 2.1 mm, 1.7 ⁇ m particle) made by Waters (pn: 186002350), and a dual gradient run from 1-30% mobile phase B over 2.9 minutes.
  • Mobile phase A H2O (0.05 % CF3CO2H).
  • Mobile phase B CH3CN (0.035 % CF3CO2H).
  • LC method H water Cortex 2.7 ⁇ C18 (3.0 mm x 50 mm), Temp: 55 o C; Flow: 1.2 mL/min; mobile phase: 100% water with 0.1% trifluoroacetic (TFA) acid then 100% acetonitrile with 0.1% TFA, gradient run from 5%-100% mobile phase B over 4 min, with stay at 100% B for 0.5 min, equilibration to 5% B over 1.5 min.
  • LC method I UPLC Luna C 18 (2) 50 x 3 mm 3 ⁇ m. Run: 2.5 min.
  • LC method K Xbridge C184.6 x 75 mm, 5 ⁇ m, Initial Gradient at 95% NH 4 HCO 3 /5% MeCN 6 min run with 1 min equilibration gradient 0 to 3 min at 95% MeCN and hold for 3 minutes. Flow 1.5 mL/min.
  • LC method L Luna C183.0 x 50 mm 3.0 ⁇ M, Temp: 45 o C, Flow: 2.0 mL/min, Run Time: 3 minutes.
  • Mobile Phase Initial 95% H 2 O (0.1% Formic Acid) and 5% CH 3 CN (0.1% FA) linear gradient to 95% CH 3 CN (0.1% FA) for 2.0 min then hold at 95% CH 3 CN (0.1% FA) for 1.0 min.
  • LC method M Poroshell 120 EC-C183.0 x 50 mm 2.7 ⁇ M, Temp:45 o C, Flow: 1,5 mL/min, RunTime: 3 min.
  • Mobile phase conditions Initial.95% H 2 O (0.1% Formic Acid) and 5% CH3CN (0.1% FA) linear gradient to 95% CH3CN (0.1% FA) for 1.5 min then hold at 95% CH3CN (0.1% FA) for 1.5 min.
  • LC method N Reversed phase UPLC using an Acquity UPLC BEH C 18 column (50 ⁇ 2.1 mm, 1.7 ⁇ m particle) made by Waters (pn: 186002350), and a dual gradient run from 1- 99% mobile phase B over 4.5 minutes.
  • Mobile phase A H2O (0.05 % TFA).
  • Mobile phase B acetonitrile (0.035 % TFA).
  • Flow rate 1.2 mL/min
  • injection volume 1.5 ⁇ L
  • column temperature 60 °C.
  • LC method O Kinetex Polar C182.6 ⁇ m, 50 x 3.0 mm. Temperature: 45 o C, Flow: 1.2 mL/min, Run time: 3 min.
  • LC method Q Luna C 18 (2) 50 x 3 mm, 3 ⁇ m. Temperature: 45 o C, Flow: 1.5 mL/min, Run time: 3.5 min. Mobile phase conditions: Initial 50% H2O 0.1% FA / 50% MeCN 0.1% FA, linear gradient to 100% MeCN 0.1% FA over 1.5 min then hold for 2 minute at 100% CH 3 CN 0.1% FA. ESI Positive.
  • LC method R Luna C182.0 x 50mm 3 ⁇ m Gradient from 50%H2O + 0.1% FA / 50% MeCN + 0.1% FA to 95% MeCN + 0.1% in 1.5 min and hold for 1.5 minutes. Column temperature: 45 o C Runtime: 3 min. ESI Positive.
  • LC method S Luna C18(2) 50 X 3 mm, 3 ⁇ m. Temperature: 45 o C, Flow: 1.5 mL/min, Run time: 3.5 min. Mobile phase conditions: Initial 95% H2O 0.1% FA / 5% MeCN 0.1% FA, linear gradient to 95% MeCN 0.1% FA over 1.3 min then hold for 2.2 minutes at 95% CH 3 CN 0.1% FA. ESI Positive.
  • LC method T Luna C18(2) 50 X 3 mm, 3 ⁇ m. Temperature: 45 o C, Flow: 1.5 mL/min, Run time: 3 min.
  • LC method X Xbridge C184.6 x 50 mm 3.5 ⁇ m. Temperature: 45 o C, Flow: 2 mL/min, Run time: 3 min. Mobile phase conditions: Initial 95% water 10 mM NH4HCO3 / 5% acetonitrile then linear gradient to 95% acetonitrile for 1.5 min then hold for 1.5 min at 95% acetonitrile.
  • LC method Y Xbridge C 18 4.6 x 50 mm 3.5 ⁇ m. Temperature: 45 o C, Flow: 2 mL/min, Run time: 4 min.
  • LC method 1C Waters Acquity BEH C 18 1.7 ⁇ m 2.1 x 100 mm. Column temp: 40 o C, flow rate: 0.5 mL/min. Eluent A: water 0.1 % FA, eluent B: acetonitrile. Conditions: 95% A 5% B, linear gradient to 5% A 95% B for 3 min, hold at 5% A 95% B for 2 min. Total run time: 5 min. [00220] LC method 1D: Luna C 18 (2) 50 x 3 mm, 3 ⁇ m. Temperature: 45 o C, Flow: 1.5 mL/min, Run time: 4 min.
  • Step 2 tert-Butyl N-tert-butoxycarbonyl-N-[4-chloro-6-(2,6-dimethylphenyl)pyrimidin- 2-yl]carbamate. [00223] All solvents were degassed prior to use.
  • Step 4 4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-amine
  • 4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-amine (hydrochloride salt) (166 g, 614.5 mmol) and 4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-amine (hydrochloride salt) (30 g, 111.0 mmol) were suspended in DCM (2.5 L), treated with NaOH (725 mL of 1 M, 725.0 mmol) and stirred at room temperature for 1 hour. The mixture was transferred into a separatory funnel and left standing over night.
  • the DCM phase was separated and the aqueous phase with insoluble material was extracted twice more with DCM (2 x 500mL).
  • the combined brown DCM phases were stirred over magnesium sulfate and charcoal for 1 hour, filtered and the yellow solution concentrated to a volume of ⁇ 500 mL.
  • the solution was diluted with heptane (750 mL) and DCM was removed under reduced pressure at 60 °C to give a cream suspension. It was stirred at room temperature for 1 hour, filtered, washed with cold heptane and dried to give 4-chloro-6- (2,6-dimethylphenyl)pyrimidin-2-amine (157 g, 91%) as a cream solid.
  • Step 5 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid
  • 4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-amine 235 g, 985.5 mmol was dissolved in MeTHF (2.3 L) and cooled in an ice bath under stirring and nitrogen.
  • methyl 3-chlorosulfonylbenzoate (347 g, 1.479 mol) was added in one portion (seems slightly endothermic) and to the cold pale-yellow solution a solution of 2-methyl-butan-2-ol (Lithium salt) (875 mL of 3.1 M, 2.712 mol) (in heptane) was added dropwise over 1.25 hour (exothermic, internal temperature from 0 to 10 °C). The ice bath was removed, and the greenish solution was stirred for 4 hours at room temperature.
  • 2-methyl-butan-2-ol Lithium salt
  • the phases were separated and the NaOH phase was washed twice with MeTHF (2 x 500 mL) and the combined organic phases were extracted once with 2 M NaOH (1 x 250 mL).
  • the combined NaOH phases were combined, stirred in an ice bath and slowly acidified by addition of HCl (416 mL of 36 %w/w, 4.929 mol) while keeping the internal temperature between 10 and 20 °C.
  • HCl 416 mL of 36 %w/w, 4.929 mol
  • the final pH was adjusted to 2-3 by addition of solid citric acid.
  • the formed yellow tacky suspension was stirred at room temperature overnight to give a cream crisp suspension.
  • the solid was collected by filtration, washed with plenty of water and sucked dry for 3 hours.
  • Example 2 Preparation of 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin-2- yl]sulfamoyl]benzoic acid
  • Step 1 tert-Butyl N-tert-butoxycarbonyl-N-(4,6-dichloro-5-methyl-pyrimidin-2- yl)carbamate
  • tert-butoxycarbonyl tert-butyl carbonate 159.92 g, 168.34 mL, 710.77 mmol
  • DMAP 3.96 g, 32.090 mmol
  • Step 2 tert-Butyl N-tert-butoxycarbonyl-N-[4-chloro-6-(2,6-dimethylphenyl)-5- methyl-pyrimidin-2-yl]carbamate
  • tert-butyl N-tert-butoxycarbonyl-N-(4,6-dichloro-5-methyl- pyrimidin-2-yl)carbamate 120.85 g, 319.50 mmol
  • DME 850 mL
  • water 120 mL
  • 2,6-dimethylphenyl)boronic acid 57.5 g, 383.38 mmol
  • cesium carbonate 271 g, 831.75 mmol
  • Step 3 4-Chloro-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin-2-amine [00229] To a solution of tert-butyl N-tert-butoxycarbonyl-N-[4-chloro-6-(2,6- dimethylphenyl)-5-methyl-pyrimidin-2-yl]carbamate (100.71 g, 224.82 mmol) in DCM (500 mL) was added HCl (200 mL of 4 M, 800.00 mmol) in dioxane. The solution was stirred at room temperature overnight before being concentrated in vacuum. The residue was then basified with sodium bicarbonate (500 mL) and extracted with ethyl acetate (1 L).
  • Step 4 Methyl 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin-2- yl]sulfamoyl]benzoate [00230] To a solution of 4-chloro-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin-2-amine (35 g, 141.29 mmol) in THF (400 mL) at 0 °C was added methyl 3-chlorosulfonylbenzoate (50 g, 213.08 mmol).
  • Lithium tert-amoxide (46.428 g, 159 mL of 40 %w/w, 197.40 mmol) was added to the solution dropwise keeping the temperature below 5 °C.
  • the solution was allowed to warm to room temperature while it stirred for 3 hours.
  • the solution was acidified with 1 M HCl (200 mL) and extracted with ethyl acetate (3 x 200 mL). The organic layer was washed with brine (300 mL) and dried over sodium sulfate.
  • Step 5 3-[[4-Chloro-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin-2- yl]sulfamoyl]benzoic acid [00231] To a solution of methyl 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin- 2-yl]sulfamoyl]benzoate (59.51 g, 133.45 mmol) in THF (500 mL) was added an aqueous solution of NaOH (300 mL of 2 M, 600.00 mmol) and the mixture was stirred for 2 hours at room temperature.
  • Example 3 Preparation of 3-[[4-Chloro-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2- yl]sulfamoyl]benzoic acid
  • Step 1 2-Amino-5-fluoro-pyrimidine-4,6-diol
  • Step 3 tert-Butyl N-(4,6-dichloro-5-fluoro-pyrimidin-2-yl)carbamate
  • 4 a stirring solution of 4,6-dichloro-5-fluoro-pyrimidin-2-amine (5.153 g, 27.495 mmol) and Boc anhydride (8.43 g, 38.626 mmol) in anhydrous THF (100 mL) at -78 °C under nitrogen was dropwise added a solution of LiHMDS (50 mL of 1.3 M, 65.000 mmol) in THF. After the addition was complete, the reaction mixture was stirred at this temperature for 2 h.
  • LiHMDS 50 mL of 1.3 M, 65.000 mmol
  • the reaction was quenched cold with saturated aqueous ammonium chloride (20 mL) and allowed to warm up to room temperature.
  • the product was extracted with ethyl acetate (3 x 30 mL).
  • the combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate and concentrated.
  • the crude was purified by silica gel chromatography using 0 – 15% ethyl acetate in hexane to afford tert-butyl N-(4,6-dichloro-5-fluoro-pyrimidin-2- yl)carbamate (6.632 g, 86%) as a white solid.
  • Step 5 4-Chloro-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2-amine [00236] To a stirring solution of tert-butyl N-[4-chloro-6-(2,6-dimethylphenyl)-5-fluoro- pyrimidin-2-yl]carbamate (2.43 g, 6.4238 mmol) in DCM (23 mL) at room temperature was added a solution of HCl (6.5 mL of 4 M, 26.000 mmol) in 1,4-dioxane. The reaction mixture was stirred at this temperature for 2 h. The reaction mixture was evaporated to dryness.
  • the obtained white solid was resuspended in saturated aqueous sodium bicarbonate (100 mL) and stirred at room temperature for 15 minutes.
  • the product was extracted with ethyl acetate (3 x 100 mL).
  • the combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate and concentrated.
  • the crude was purified by silica gel chromatography using 0 – 15% hexanes – ethyl acetate to afford 4-chloro-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2- amine (1.42 g, 83%) as white solid.
  • reaction mixture was stirred at this temperature for 1 h.
  • the reaction was quenched cold with 1 M aqueous HCl (180 mL) and then allowed to warm up to room temperature. Volatiles were removed under vacuum and the product was extracted with ethyl acetate (3 x 100 mL). The combined organic layers were washed with brine (60 mL), dried over anhydrous sodium sulfate, and concentrated.
  • the crude was purified by silica gel chromatography using 0 – 20% hexanes – ethyl acetate.
  • Step 7 3-[[4-Chloro-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2- yl]sulfamoyl]benzoic acid [00238] To a stirring solution of methyl 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-fluoro- pyrimidin-2-yl]sulfamoyl]benzoate (490 mg, 1.0892 mmol) in THF (9 mL) at room temperature was added an aqueous solution of NaOH (4.5 mL of 1 M, 4.5000 mmol). The reaction mixture was stirred at this temperature for 2 h.
  • Step 2 tert-Butyl N-tert-butoxycarbonyl-N-[4-chloro-6-(2,6-dimethylphenyl)-5- methoxy-pyrimidin-2-yl]carbamate
  • tert-butyl N-tert-butoxycarbonyl-N-(4,6-dichloro-5-methoxy- pyrimidin-2-yl)carbamate 31.58 g, 80.101 mmol
  • DME 225 mL
  • water 31 mL
  • 2,6-dimethylphenyl)boronic acid (16.5 g, 110.01 mmol
  • cesium carbonate 68 g, 208.71 mmol
  • Step 3 4-Chloro-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2-amine [00241] To a solution of tert-butyl N-tert-butoxycarbonyl-N-[4-chloro-6-(2,6- dimethylphenyl)-5-methoxy-pyrimidin-2-yl]carbamate (50.35 g, 108.52 mmol) in DCM (500 mL) was added HCl (100 mL of 4 M, 400.00 mmol) in dioxane. The solution was stirred at room temperature overnight before being concentrated under vacuum.
  • Step 4 Methyl 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2- yl]sulfamoyl]benzoate [00242] To a solution of 4-chloro-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2-amine (223 mg, 0.8456 mmol) in THF (6 mL) at 0°C was added methyl 3-chlorosulfonylbenzoate (496 mg, 2.1137 mmol).
  • Lithium tert-amoxide (584.00 mg, 2 mL of 40 %w/w, 2.4830 mmol) was added to the solution dropwise keeping the temperature below 5 °C. The solution was allowed to warm to room temperature while it stirred for 3 hours. The solution was acidified with 1 M HCl (5 mL) and extracted with ethyl acetate (20 mL). The organic layer was washed with brine (20 mL) and dried over sodium sulfate.
  • Step 5 3-[[4-Chloro-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2- yl]sulfamoyl]benzoic acid [00243] To a solution of methyl 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2- yl]sulfamoyl]benzoate (386 mg, 0.8356 mmol) in THF (10 mL) was added an aqueous solution of NaOH (5 mL of 1 M, 5.0000 mmol) and stirred for 1 hour at room temperature.
  • Example 5 Preparation of 6-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]pyridine-2-carboxylic acid
  • Step 1 Methyl 6-benzylsulfanylpyridine-2-carboxylate
  • phenylmethanethiol 28.408 g, 26.800 mL, 228.72 mmol
  • NaH 11.200 g, 60 %w/w, 280.03 mmol
  • Step 2 Methyl 6-chlorosulfonylpyridine-2-carboxylate [00245] A solution of methyl 6-benzylsulfanylpyridine-2-carboxylate (121.62 g, 431.47 mmol) in DCM (950 mL) and DI water (300 mL) was cooled in a -1 – 0 °C ice bath and, with vigorous stirring, sulfuryl chloride (228.14 g, 140 mL, 1.6396 mol) was added dropwise while the temperature was maintained below 5 °C. After the addition, the organic phase was separated, washed with DI water (2 x 500 mL), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum.
  • Step 3 Methyl 6-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]pyridine-2-carboxylate [00246] A solution of 4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-amine (16.63 g, 71.161 mmol) and methyl 6-chlorosulfonylpyridine-2-carboxylate (16.8 g, 71.294 mmol) dissolved in anhydrous THF (680 mL) was cooled to – 78 °C.
  • Lithium bis(trimethylsilyl)amide (143 mL of 1 M, 143.00 mmol) in solution in THF was added dropwise. The mixture was allowed to warm up to 0 °C slowly and then 1M aqueous HCl (146 mL) was added, followed by DI water (680 mL). The THF was evaporated, and the aqueous phase was extracted with chloroform (3 x 250 mL). The combined organic layers were washed with saturated aqueous NaCl (300 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The crude was recrystallized in 10 % Acetone in Hexanes (500 mL).
  • Step 4 6-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]pyridine- 2-carboxylic acid [00247] To a solution of methyl 6-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]pyridine-2-carboxylate (15.79 g, 36.477 mmol) in THF (180 mL) was added aqueous sodium hydroxide (182 mL of 1 M, 182.00 mmol). The reaction was stirred at RT for 1h.
  • the crude product was solubilized in EtOAc (150 mL) and the solution was washed with a 50% brine solution (100 mL) and brine (100 mL), then was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure.
  • the material was purified by trituration in MTBE/Heptane (2/1) (50 mL). The suspension was filtered, washed with MTBE/heptane (25 mL) and dried under high vacuum to give a yellow solid.
  • the product was then purified by normal phase chromatography eluting from 0-70% EtOAc in heptanes (no separation).
  • Step 2 3-Bromo-6-tert-butyl-5H-pyrrolo[2,3-b]pyrazine [00249] To a stirred solution of 6-bromo-3-(3,3-dimethylbut-1-ynyl)pyrazin-2-amine (7.50 g, 29.51 mmol) in t-BuOH (80 mL) was added sodium tert-butoxide (14.25 g, 148.3 mmol). The reaction mixture was stirred at 80 °C for 4 h, then allowed to cool down to ambient temperature. The volatiles were removed under reduced pressure and the residue was diluted with ice-cold water (250 mL).
  • the suspension was stirred at 0-4 °C (ice-water bath) for 15 min.
  • the solid was filtered and washed with water (3 x 30 ml) and air-dried.
  • the filter-cake was taken up in acetonitrile and concentrated under reduced pressure and the azeotropic water removal process repeated three times.
  • Step 3 3-Bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine
  • Sodium hydride (2.18 g, 54.505 mmol) was added portion-wise to a solution of 3-bromo-6-tert-butyl-5H-pyrrolo[2,3-b]pyrazine (6.71 g, 25.084 mmol) in anhydrous DMF (128 mL) at 0 ° C.
  • the suspension was stirred at this temperature for 30 min.
  • Dimethyl sulfate (4.7988 g, 3.6 mL, 38.046 mmol) was added and left stirring at 0 °C for 15 min.
  • the reaction was then stirred at room temperature for 1 h.
  • the reaction mixture was cooled down to 0 ° C and quenched with saturated aqueous ammonium chloride solution (250 mL).
  • the resulting precipitate was filtered and washed with water (2 x 50 mL).
  • the precipitate was dissolved in EtOAc (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.
  • the resulting precipitate was purified by reversed phase chromatography (Gradient: 5% MeOH in water, then 60-85% MeOH in water).
  • Example 7 Preparation of 3-bromo-6-tert-butyl-5-(trideuteriomethyl)pyrrolo[2,3- b]pyrazine
  • Step 1 3-Bromo-6-tert-butyl-5-(trideuteriomethyl)pyrrolo[2,3-b]pyrazine
  • DMF 25 mL
  • NaH in mineral oil
  • reaction mixture was stirred for 20 minutes then trideuterio(iodo)methane (1.75 g, 12.07 mmol) was added.
  • the reaction mixture was stirred for 1 h and ice was added carefully (caution! Rapid effervescence).
  • the suspension was diluted with water and was stirred at room temperature for 10 minutes.
  • Step 2 2-Bromo-6-tert-butyl-5H-pyrrolo[2,3-b]pyrazine [00253] To a stirred solution of 5-bromo-3-(3,3-dimethylbut-1-ynyl)pyrazin-2-amine (5.02 g, 19.75 mmol) in t-BuOH (50 mL) was added sodium tert-butoxide (9.49 g, 98.75 mmol). The reaction mixture was stirred at 80 °C for 4 h then was allowed to cool down to ambient temperature. The volatiles were removed under reduced pressure and the residue was taken up in water (50 mL). The resulting dark suspension was stirred at 0-4 °C for 15 min, then filtered off.
  • Step 3 2-Bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine
  • 2-bromo-6-tert-butyl-5H-pyrrolo[2,3-b]pyrazine 500 mg, 1.967 mmol
  • anhydrous DMF 8 mL
  • NaH in mineral oil
  • iodomethane 340 mg, 2.395 mmol
  • Example 9 Preparation of 2-bromo-7-isopropyl-quinoxaline Step 1: 1-Fluoro-4-isopropenyl-2-nitro-benzene [00255] To a solution of 4-bromo-1-fluoro-2-nitro-benzene (10.01 g, 45.501 mmol) and sodium carbonate (5.31 g, 50.100 mmol) in 1,4-dioxane (200 mL) and water (50 mL) was sparged with nitrogen. Bis(triphenylphosphine)palladium(II)chloride (659.3 mg, 0.9393 mmol) was added and the mixture was sparged with nitrogen.
  • Isopropenylboronic acid pinacol ester (8.3927 g, 9.43 mL, 49.944 mmol) was added and the mixture was stirred at 80 °C for 3 hours. The mixture was cooled to rt, filtered over Celite, washed with MTBE (3x 20 mL) and the volatiles of the filtrate were removed under reduced pressure. The mixture was diluted with water (200 mL) and extracted with MTBE (3x150 mL).
  • Step 2 Methyl 2-(4-isopropenyl-2-nitro-anilino)acetate
  • a solution of 1-fluoro-4-isopropenyl-2-nitro-benzene (8.2 g, 44.584 mmol) in anhydrous DMA (150 mL) was sparged with nitrogen.
  • Methyl 2-aminoacetate (hydrochloride salt) (7.59 g, 60.452 mmol) and DIPEA (21.518 g, 29 mL, 166.49 mmol) were added and the mixture was stirred at 80 °C for 1 hour.
  • the mixture was cooled to rt and poured into water (500 mL). The mixture was stirred at rt for 48 hours.
  • Step 4 7-Isopropyl-1H-quinoxalin-2-one [00258] To a solution of 7-isopropyl-3,4-dihydro-1H-quinoxalin-2-one (5.32 g, 27.545 mmol) in MeOH (200 mL) was added KotBu in THF (42 mL of 1 M, 42.000 mmol). Oxygen was flushed into the mixture for 5 min and the reaction was stirred under O 2 atmosphere for 16 hours at rt. The volatiles were removed under reduced pressure. The crude was diluted with water (100 mL) and TFA was added (5 mL) to neutralize the reaction. The aqueous layer was extracted with EtOAC (3 x 100 mL).
  • Step 5 2-Bromo-7-isopropyl-quinoxaline [00259] To a solution of 7-isopropyl-1H-quinoxalin-2-one (4.23 g, 22.428 mmol) in acetonitrile (150 mL) were added POBr 3 (9.05 g, 31.568 mmol) and potassium carbonate (4.52 g, 32.705 mmol) at room temperature. The mixture was stirred for 3 hours at 80 °C. The mixture was allowed to cool down and was placed into an ice bath at 0 °C. Water (150 mL) was added and the mixture was allowed to warm up to rt.
  • Example 10 Preparation of 5-bromo-2-tert-butyl-3-methyl-imidazo[4,5-b]pyrazine Step 1: 5-Bromo-N3-methyl-pyrazine-2,3-diamine [00260] Methyl amine in methanol (10 mL of 40 %w/v, 128.8 mmol) was added to 3,5- dibromopyrazin-2-amine (2.00 g, 7.908 mmol) and stirred for 10 min. Then water (2.0 mL) was added to the tan solution and the capped microwave tube was stirred in a microwave reactor at 130 °C (setting: very high absorption) for 1 h.
  • Step 2 5-Bromo-2-tert-butyl-3-methyl-imidazo[4,5-b]pyrazine [00261]
  • pivalic acid 34 g, 332.90 mmol
  • 2,2-dimethylpropanoyl 2,2- dimethylpropanoate (14.672 g, 16 mL, 78.777 mmol)
  • 5-bromo-N3-methyl-pyrazine-2,3- diamine 7.8 g, 38.186 mmol.
  • the flask was equipped with a Vigreux column and the reaction was stirred at 130 °C for 18 hours.
  • Step 2 3-Bromo-5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazine
  • 6-bromo-3-[2-[1-(trifluoromethyl)cyclopropyl]ethynyl]pyrazin-2- amine (19.66 g, 53.761 mmol) in anhydrous DMF (150 mL) at 0 ° C under nitrogen, was added NaH (4.5 g, 60 %w/w, 112.51 mmol). The mixture was stirred at 70 °C for 1 hour. The mixture was allowed to cool down to 0 °C.
  • Example 12 An alternative preparation of 3-bromo-5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine
  • Step 1 Triisopropyl-[3-(trifluoromethyl)but-3-en-1-ynyl]silane [00264]
  • CuI 41 mg, 0.2153 mmol
  • bis(triphenylphosphine)palladium(II) dichloride 77 mg, 0.1094 mmol
  • Step 2 Triisopropyl-[2-[1-(trifluoromethyl)cyclopropyl]ethynyl]silane [00265] LiHMDS (10.3 mL of 1 M, 10.300 mmol) was added to triisopropyl-[3- (trifluoromethyl)but-3-en-1-ynyl]silane (1 g, 3.4369 mmol) and diphenyl(methyl)sulfonium tetrafluoroborate (1.38 g, 4.7898 mmol) in solution in THF (2 mL) maintained at -78 °C.
  • the reaction mixture was left to slowly warm up to room temperature over 18 h.
  • the reaction mixture was diluted with pentane (15 mL) and filtered on a silica gel column and the desired product was eluted with pentane.
  • the filtrates were concentrated under reduced pressure.
  • the resulting residue was purified by reverse phase chromatography on a C18 column using a 5 to 100% gradient of acetonitrile in pure water.
  • the fractions containing the desired fraction were combined and extracted with pentane (3 x 50 mL).
  • Step 3 6-Bromo-3-[2-[1-(trifluoromethyl)cyclopropyl]ethynyl]pyrazin-2-amine
  • 3,6-dibromopyrazin-2- amine 8.5 g, 33.611 mmol
  • triisopropyl-[2-[1-(trifluoromethyl)cyclopropyl]ethynyl]silane 5 g, 16.354 mmol
  • CuI (625 mg, 3.2817 mmol
  • Pd(PPh3)2Cl2 574 mg, 0.8178 mmol
  • PPh3 430 mg, 1.6394 mmol
  • TEA 1.8150 g, 2.5 mL, 17.937 mmol
  • THF 35 mL
  • the vial was capped and the mixture was warmed-up to 60 °C, and TBAF (in THF) (17 mL of 1 M, 17.000 mmol) was introduced dropwise in the vial via a syringe.
  • the reaction mixture was left stirring at this temperature for 3 h and then left to cool to room temperature.
  • the mixture was filtered on a pad of silica gel, and the pad was eluted with ethyl acetate.
  • the combined filtrates were concentrated under reduced pressure.
  • the resulting residue was purified by flash chromatography on silica gel using a gradient of 0 to 100% ethyl acetate in heptanes. The collected fractions were combined and concentrated under reduced pressure.
  • the mixture was stirred at 70 °C for 1 hour. The mixture was allowed to cool down to 0 °C. A solution of dimethyl sulfate (13.566 g, 10.2 mL, 107.55 mmol) in anhydrous DMF (50 mL) was added drop-wise to the mixture at 0 °C over 15 minutes. The mixture was stirred at 0 °C for 1 h. Then, a saturated aqueous solution of ammonium chloride (250 mL) and water (250 mL) were added. The aqueous phase was extracted with EtOAc (3 x 250 mL).
  • Example 13 Preparation of 3-bromo-8,8-dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizine Step 1: tert-Butyl 2-(hydroxymethyl)-3,3-dimethyl-pyrrolidine-1-carboxylate [00268] To a solution of 1-tert-butoxycarbonyl-3,3-dimethyl-pyrrolidine-2-carboxylic acid (4.5 g, 18.496 mmol) in THF (80 mL) was added borane dimethyl sulfide (4.2400 g, 5.3 mL, 55.812 mmol) dropwise. The mixture was stirred at 50 °C for 10 min and cooled down to room temperature.
  • Step 2 tert-Butyl 2-formyl-3,3-dimethyl-pyrrolidine-1-carboxylate [00269] To a solution of tert-butyl 2-(hydroxymethyl)-3,3-dimethyl-pyrrolidine-1-carboxylate (4.2 g, 18.297 mmol) in water-saturated DCM (150 mL) was added DMP (15 g, 35.366 mmol). The mixture was stirred at room temperature for 16 h. Silica (20 g) was added, and the volatiles were removed under reduced pressure.
  • Step 4 (3,5-Dibromopyrazin-2-yl)-(3,3-dimethylpyrrolidin-2-yl)methanol
  • Concentrated HBr (in water) (6.4368 g, 9 mL, 38.186 mmol) was added to a solution of tert-butyl 2-[(3,5-dibromopyrazin-2-yl)-hydroxy-methyl]-3,3-dimethyl-pyrrolidine-1- carboxylate (1.4 g, 2.8772 mmol) in diethyl ether (18 mL) and water (9 mL).
  • the reaction mixture was stirred for 16 h at room temperature and then concentrated under reduced pressure.
  • reaction mixture was cooled to 0 °C and 1-ethynyl-1-(trifluoromethyl)-cyclobutane (910.5 mg, 6.147 mmol) was slowly added dropwise.
  • the reaction was stirred at 0 °C for 5 hours, then was diluted with EtOAc.
  • the suspension was filtered through Celite, and the pad was rinsed with EtOAc.
  • Example 15 Preparation of 3-bromo-5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl- ethyl)pyrrolo[2,3-b]pyrazine Step 1: 6-Bromo-3-(4,4,4-trifluoro-3,3-dimethyl-but-1-ynyl)pyrazin-2-amine [00276] In a flask under nitrogen atmosphere containing a solution of 4,4,4-trifluoro-3,3- dimethyl-but-1-yne (11.31 g, 83.092 mmol) in THF (250 mL) were added 3,6-dibromopyrazin- 2-amine (9.15 g, 36.181 mmol), triethylamine (12.342 g, 17 mL, 121.97 mmol), bis(triphenylphosphine)palladium(II) dichloride (3.52 g, 5.0150 mmol) and copper iodide (800 mg,
  • reaction mixture was filtered on Celite, and the cake was washed with acetonitrile (5 x 30 mL). Purification by reversed-phase chromatography (C 18 column. Gradient 5 - 80% acetonitrile in water with 0.1% formic acid) was performed. The fractions containing the product were combined and the volatiles were removed under reduced pressure. The aqueous phase was extracted with EtOAc (3 x 100 mL).
  • the mixture was stirred at 70 °C for 60 min. The mixture was allowed to cool down to 0 °C. A solution of dimethyl sulfate (8.7780 g, 6.6 mL, 69.594 mmol) in anhydrous DMF (100 mL) was added dropwise to the mixture at 0 °C over 45 min using a syringe driver. The mixture was stirred at 0 °C for an additional 10 min. Then, a saturated aqueous solution of NH 4 Cl (2 L) and water (2 L) were added. The mixture was stirred at rt for 10 min.
  • Example 16 Preparation of 3-bromo-5-methyl-6-spiro[2.3]hexan-5-yl-pyrrolo[2,3- b]pyrazine
  • Step 1 Methyl spiro[2.3]hexane-5-carboxylate
  • TFA 13.480 g, 9.2 mL, 117.04 mmol
  • Step 2 spiro[2.3]Hexane-5-carboxylic acid [00279] To a stirred solution of methyl spiro[2.3]hexane-5-carboxylate (37.5 g, 9.0954 mmol) in DCE (22 mL) was added NaOH (1.1 g, 27.502 mmol) in MeOH (10 mL). The mixture was stirred at room temperature for 16 h. Ether (50 mL) and water (80 mL) were added, and the phases were separated. The aqueous phase was washed with ether (2 x 80 mL) and then acidified with a 6 N HCl solution (5 mL).
  • Step 3 spiro[2.3]Hexan-5-ylmethanol [00280] To a solution of spiro[2.3]hexane-5-carboxylic acid (7.28 g, 46.974 mmol) in anhydrous THF (216 mL) at 0 ° C was added dropwise a borane-THF complex (60 mL of 1 M, 60.000 mmol) solution under nitrogen.
  • the reaction mixture was stirred at room temperature for 16 h, then was quenched with water (50 mL), a saturated aqueous solution of sodium bicarbonate (50 mL) and diethyl ether (400 mL) and water (350 mL) were added.
  • the aqueous phase was extracted with diethyl ether (2 x 100 mL) and the combined organics were washed with brine (600 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to afford spiro[2.3]hexan-5-ylmethanol (7.9 g, 100%) as a yellow liquid.
  • Step 4 spiro[2.3]Hexane-5-carbaldehyde [00281] To a stirring mixture of spiro[2.3]hexan-5-ylmethanol (7.467 g, 51.191 mmol) and TEMPO (925 mg, 5.9202 mmol) in DCM (80 mL) at 0 °C was added a solution of sodium bromide (3.1 g, 30.128 mmol) in water (40 mL).
  • Step 5 5-(2,2-Dibromovinyl)spiro[2.3]hexane [00282] To a solution of spiro[2.3]hexane-5-carbaldehyde (6.52 g, 59.190 mmol) in DCM (290 mL) was added triphenylphosphine (47 g, 177.40 mmol). The mixture was placed in a water bath and a solution of CBr4 (30 g, 89.558 mmol) in DCM (40 mL) was added dropwise over 1 hour. The mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure and the resulting precipitate was triturated in pentane (100 mL).
  • Step 6 5-Ethynylspiro[2.3]hexane [00283] To a solution of 5-(2,2-dibromovinyl)spiro[2.3]hexane (13.6 g, 31.651 mmol) in anhydrous THF (113 mL) was added n-BuLi (solution in hexanes) (28 mL of 2.5 M, 70.000 mmol) dropwise over 30 minutes at -78 °C. After addition, the reaction mixture was stirred at - 78 °C for 1 hour and 30 minutes.
  • n-BuLi solution in hexanes
  • Step 7 6-Bromo-3-(2-spiro[2.3]hexan-5-ylethynyl)pyrazin-2-amine
  • a solution of 5- ethynylspiro[2.3]hexane (3.36 g, 31.649 mmol) in THF (26 mL) were added 3,6- dibromopyrazin-2-amine (4.6 g, 17.826 mmol), TEA (7.3326 g, 10.1 mL, 72.464 mmol), PdCl 2 (PPh 3 ) 2 (1.3 g, 1.8151 mmol) and CuI (345 mg, 1.7934 mmol).
  • Step 8 3-Bromo-5-methyl-6-spiro[2.3]hexan-5-yl-pyrrolo[2,3-b]pyrazine
  • 6-bromo-3-(2-spiro[2.3]hexan-5-ylethynyl)pyrazin-2-amine 3.52 g, 12.642 mmol
  • NaH 1.1 g, 60 %w/w, 27.503 mmol
  • Example 17 Preparation of 3-bromo-5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl- ethyl)pyrrolo[2,3-b]pyrazine Step 1: 6-Bromo-3-(4,4,4-trifluoro-3,3-dimethyl-but-1-ynyl)pyrazin-2-amine [00286] In a flask under nitrogen atmosphere containing a solution of 4,4,4-trifluoro-3,3- dimethyl-but-1-yne (11.31 g, 83.092 mmol) in THF (250 mL) were added 3,6-dibromopyrazin- 2-amine (9.15 g, 36.181 mmol), triethylamine (12.342 g, 17 mL, 121.97 mmol), bis(triphenylphosphine)palladium(II) dichloride (3.52 g, 5.0150 mmol) and copper(I) iodide (
  • the mixture was stirred for 16 hours at rt under nitrogen atmosphere.
  • the reaction mixture was filtered on Celite, and the cake was washed with acetonitrile (5 x 30 mL).
  • Purification by reversed-phase chromatography (C18 column. Gradient 5 - 80% acetonitrile in water with 0.1% formic acid) was performed. The fractions containing the product were combined and the volatiles were removed under reduced pressure. The aqueous phase was extracted with EtOAc (3 x 100 mL).
  • the mixture was stirred at 70 °C for 60 minutes. The mixture was allowed to cool down to 0 °C. A solution of dimethyl sulfate (8.7780 g, 6.6 mL, 69.594 mmol) in anhydrous DMF (100 mL) was added dropwise to the mixture at 0 °C over 45 min using a syringe driver. The mixture was stirred at 0 °C for an additional 10 minutes. Then, a saturated aqueous solution of NH 4 Cl (2 L) and water (2 L) were added. The mixture was stirred at rt for 10 minutes.
  • Step 2 6-tert-Butyl-3-chloro-5-methyl-pyrrolo[2,3-b]pyrazine [00289] To a solution of 6-chloro-3-(3,3-dimethylbut-1-ynyl)pyrazin-2-amine (8.8 g, 41.466 mmol) in DMF (120 mL) was added potassium tert-butoxide (24 g, 213.88 mmol). The reaction mixture was heated at 80 °C for 30 minutes and was allowed to cool down to room temperature.
  • methyl iodide 11.400 g, 5 mL, 80.316 mmol
  • methyl iodide 11.400 g, 5 mL, 80.316 mmol
  • a saturated aqueous solution of sodium thiosulfate 50 mL was added, and the mixture was stirred for 1 hour.
  • the reaction mixture was diluted with water (100 mL), brine (100 mL) and ethyl acetate (100 mL) and the phases were separated.
  • Step 4 1-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2- dimethoxyphosphoryl-ethanone
  • LiHMDS in THF
  • THF THF
  • dimethyl methylphosphonate 2.5212 g, 2.2 mL, 20.320 mmol
  • THF 130 mL
  • Example 19 Preparation of methyl 5-methyl-6-(1-methylcyclopropyl)pyrrolo[2,3- b]pyrazine-3-carboxylate Step 1: Trimethyl-[2-(1-methylcyclopropyl)ethynyl]silane [00292] To a solution of 2-cyclopropylethynyl(trimethyl)silane (8.54 g, 10.047 mL, 61.758 mmol) in Et2O (70 mL) at room temperature was slowly added n-BuLi in hexanes (30 mL of 2.5 M, 75.000 mmol) over 20 minutes.
  • reaction mixture was stirred at room temperature for 18 hours, then the reaction was cooled down to -15 °C in a dry-ice/acetone bath.
  • To the reaction mixture was slowly added dropwise methyl iodide (9.5760 g, 4.2 mL, 67.466 mmol) over 10 minutes.
  • the resulting solution was stirred at -10 °C for 10 minutes, and then at 0 °C for 1 hour.
  • the reaction mixture was allowed to warm to room temperature for 1 hour.
  • the reaction mixture was diluted with diethyl ether (50 mL) and water (50 mL) and brine (50 mL) were added. The phases were separated, and the aqueous phase was extracted with diethyl ether (3 x 20 mL).
  • GC-MS method Column HP 5 ms Ultra Inert 30 m x 250 ⁇ m x 0.25 ⁇ m, control mode: flow: 3.3 mL/min, split ratio mode: 33:1, split flow: 165 mL/min., carrier gas: He.
  • Step 2 6-Chloro-3-[2-(1-methylcyclopropyl)ethynyl]pyrazin-2-amine [00293] To a sealed tube equipped with a stir bar were added copper(I) iodide (221 mg, 1.1604 mmol), bis(triphenylphosphine)palladium(II) chloride (330 mg, 0.4702 mmol), triphenylphosphine (234 mg, 0.8922 mmol), cesium fluoride (4.6 g, 30.282 mmol), 3-bromo-6- chloro-pyrazin-2-amine (3.15 g, 15.112 mmol), poly(ethylene glycol) 200 MW (3.0 mL), triethylamine (25 mL) and water (1.5 mL).
  • copper(I) iodide 221 mg, 1.1604 mmol
  • bis(triphenylphosphine)palladium(II) chloride 330 mg, 0.4702 m
  • Step 3 3-Chloro-5-methyl-6-(1-methylcyclopropyl)pyrrolo[2,3-b]pyrazine
  • 6-chloro-3-[2-(1-methylcyclopropyl)ethynyl]pyrazin-2-amine (2.23 g, 9.5682 mmol) in DMF (30 mL) was added potassium tert-butoxide (3.3 g, 29.409 mmol).
  • the reaction mixture was heated at 80 °C for 50 min then the reaction was allowed to cool down to room temperature before methyl iodide (2.7360 g, 1.2 mL, 19.276 mmol) was added dropwise.
  • Step 5 3-Chloro-5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazine
  • Sodium hydride (60% w/w in oil) (1.18 g, 29.503 mmol) was added portion-wise to a solution of 3-chloro-6-[1-(trifluoromethyl)cyclopropyl]-5H-pyrrolo[2,3-b]pyrazine (3.81 g, 14.242 mmol) in DMF (76 mL) at 0 ° C.
  • Step 6 Methyl 5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazine-3-carboxylate [00306] To a suspension of 3-chloro-5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazine (3.96 g, 14.351 mmol) in anhydrous methanol (72 mL) was added triethylamine (4.5012 g, 6.2 mL, 44.483 mmol).
  • Step 7 2-Dimethoxyphosphoryl-1-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]135xtracte
  • [methoxy(methyl)phosphoryl]oxymethane 190 ⁇ L, 1.753 mmol
  • THF 12.5 mL
  • Example 22 Preparation of 2-dimethoxyphosphoryl-1-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]135xtracte
  • Step 1 6-Chloro-3-(4,4,4-trifluoro-3,3-dimethyl-but-1-ynyl)pyrazin-2-amine
  • 3-bromo-6-chloro-pyrazin-2- amine 963 mg, 4.6199 mmol
  • CuI 93 mg, 0.4883 mmol
  • THF 20 mL
  • 4,4-Trifluoro- 3,3-dimethyl-but-1-yne (691 mg, 5.0766 mmol) and triethylamine (958.32 mg, 1.32 mL, 9.4705 mmol) were then added and the mixture was degassed with nitrogen for 10 min.
  • PdCl2(PPh3)2 (326 mg, 0.4645 mmol) was added and the mixture was re-degassed for another 5 min. The flask was sealed, and the reaction was stirred at room temperature for 2 hours. The mixture was filtered through Celite, washed with EtOAc (30 mL) and the filtrate concentrated in vacuo. Purification was done by reverse-phase chromatography (Gradient: 5-100 % MeOH in water with 0.1 % formic acid) to give 6-chloro-3-(4,4,4-trifluoro-3,3-dimethyl-but-1-ynyl)pyrazin-2- amine (654 mg, 54%) as an orange solid.
  • Step 2 3-Chloro-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)-5H-pyrrolo[2,3- b]pyrazine
  • 6-chloro-3-(4,4,4-trifluoro- 3,3-dimethyl-but-1-ynyl)pyrazin-2-amine (3.87 g, 14.459 mmol) in dry THF (60 mL).
  • the solution was cooled to 0 ° C and potassium tert-butoxide (1M solution in THF) (28 mL of 1 M, 28.000 mmol) was slowly added.
  • the reaction mixture was heated to reflux for 2 hours.
  • Step 4 Methyl 5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3- b]pyrazine-3-carboxylate [00311] To a suspension of 3-chloro-5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl- ethyl)pyrrolo[2,3-b]pyrazine (1.6 g, 5.0707 mmol) in anhydrous methanol (15 mL) was added triethylamine (1.0080 g, 1.4 mL, 9.9615 mmol).
  • Example 23 Preparation of 2-dimethoxyphosphoryl-1-(2-isopropyl-3-methyl-imidazo[4,5- b]pyrazin-5-yl)138xtracte Step 1: 5-Bromo-N3-methyl-pyrazine-2,3-diamine [00313] A suspension of 3,5-dibromopyrazin-2-amine (1 g, 3.9542 mmol) in MeNH2 in water (10 mL of 40 %w/v, 128.80 mmol) was heated in a microwave reactor at 130 °C for 1 h.
  • Step 2 5-Bromo-2-isopropyl-3-methyl-imidazo[4,5-b]pyrazine [00314] To a solution of 2-methylpropanoyl 2-methylpropanoate (248.04 mg, 0.260 mL, 1.5679 mmol) in isobutyric acid (2 mL) was added 5-bromo-N3-methyl-pyrazine-2,3-diamine (300 mg, 1.4775 mmol) and the reaction was stirred at room temperature for 10 minutes then heated in a microwave reactor at 130 °C for 5 h and 20 min.
  • Step 3 Methyl 2-isopropyl-3-methyl-imidazo[4,5-b]pyrazine-5-carboxylate [00315] To a solution of 5-bromo-2-isopropyl-1-methyl-imidazo[4,5-b]pyrazine (4.03 g, 15.797 mmol) in MeOH (80 mL) was added Pd(dppf)Cl2 – DCM complex (1.25 g, 1.5307 mmol) then TEA (32.670 g, 45 mL, 322.86 mmol) was added.
  • Step 4 2-Dimethoxyphosphoryl-1-(2-isopropyl-3-methyl-imidazo[4,5-b]pyrazin- 5-yl)140xtracte
  • [methoxy(methyl)phosphoryl]oxymethane 501 mg, 4.038 mmol
  • THF 20 mL
  • the solution was cooled to 0 °C.
  • a solution of [bis(trimethylsilyl)amino]lithium 8.1 mL of 1 M, 8.100 mmol was added dropwise.
  • Example 24 Preparation of 2-dimethoxyphosphoryl-1-[6-(1-methylcyclopropyl)furo[2,3- b]pyrazin-2-yl]141xtracte
  • Step 1 5-Chloro-1-[(4-methoxyphenyl)methyl]-3-[2-(1- methylcyclopropyl)ethynyl]pyrazin-2-one
  • the resulting material was purified by silica gel chromatography (40 g of silica) using a gradient eluent of 100% hexanes to 80% ethyl acetate in hexanes to afford as a yellow oil 5-chloro-1-[(4-methoxyphenyl)methyl]-3- [2-(1-methylcyclopropyl)ethynyl]pyrazin-2-one (287 mg, 71%), 1 H NMR (400 MHz, DMSO-d6) ⁇ 8.15 (s, 1H), 7.38 – 7.30 (m, 2H), 6.95 – 6.87 (m, 2H), 4.96 (s, 2H), 3.73 (s, 3H), 1.31 (s, 3H), 1.04 – 0.96 (m, 2H), 0.88 – 0.80 (m, 2H).
  • Step 3 Methyl 6-(1-methylcyclopropyl)furo[2,3-b]pyrazine-2-carboxylate
  • reaction mixture was heated to 100°C with 150 psi of carbon monoxide and stirred for 14h.
  • the reaction mixture was allowed to cool to rt.
  • the reaction mixture was filtered through a pad of celite, washed with ethyl acetate, filtrate was concentrated in vacuo.
  • Step 4 2-dimethoxyphosphoryl-1-[6-(1-methylcyclopropyl)furo[2,3-b]pyrazin-2- yl]142xtracte
  • a flask was flame-dried then cooled down to room temperature using a nitrogen stream and charged with dimethyl methylphosphonate (823.60 mg, 710 ⁇ L, 6.6379 mmol) and anhydrous THF (35 mL).
  • the layers were then separated, and the aqueous layers was extracted with ethyl acetate (3 x 40 mL) and the combined organic layers were washed with brine (100 mL), dried over magnesium sulfate, filtered and concentrated under reduced pressure.
  • the crude was purified by flash chromatography on silica-gel (gradient: 0 to 100% ethyl acetate in heptanes; 10 CV, then 100% ethyl acetate; 4 CV).
  • Example 25 Preparation of 1-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-2- dimethoxyphosphoryl-ethanone
  • Step 1 5-Chloro-3-(3,3-dimethylbut-1-ynyl)-1-[(4- methoxyphenyl)methyl]pyrazin-2-one
  • To a 20 mL microwave vial 3,5-dichloro-1-[(4- methoxyphenyl)methyl]pyrazin-2-one (457 mg, 1.603 mmol), bis(triphenylphosphine) palladium(II) dichloride (21 mg, 0.02992 mmol), CuI (14 mg, 0.07351 mmol) in DMF (3.5 mL) and TEA (1.3 mL, 9.327 mmol).
  • the resulting material was purified by silica gel chromatography (24 g of silica) using a gradient eluent of 100% hexanes to 80% ethyl acetate in hexanes to afford as a yellow oil 5-chloro-3-(3,3-dimethylbut-1-ynyl)-1-[(4-methoxyphenyl)methyl]pyrazin-2-one (457 mg, 86%), 1 H NMR (400 MHz, MeOH-d 4 ) ⁇ 7.77 (s, 1H), 7.38 – 7.29 (m, 2H), 6.96 – 6.88 (m, 2H), 5.05 (s, 2H), 3.78 (s, 3H), 1.34 (s, 9H).
  • Step 3 Methyl 6-tert-butylfuro[2,3-b]pyrazine-2-carboxylate [00323] A mixture of 6-tert-butyl-2-chloro-furo[2,3-b]pyrazine (295 mg, 1.400 mmol) and Pd(dppf)Cl 2 .DCM (115 mg, 0.1408 mmol) and TEA (900 ⁇ L, 6.457 mmol) in MeOH (10 mL) in a steel pressure vessel equipped with a stir bar was purged with carbon monoxide (4 g, 137.8 mmol) three times. The reaction mixture was heated to 100°C with 120 psi of carbon monoxide and stirred for 14 h.
  • reaction mixture was then stirred at 0 °C for 30 minutes and then a solution of methyl 6-tert-butylfuro[2,3-b]pyrazine-2-carboxylate (6.22 g, 26.553 mmol) in dry THF (62 mL) was added. The reaction was stirred at 0 °C for 90 minutes. The reaction was quenched by the addition of a saturated aqueous solution of ammonium chloride (20 mL) and the biphasic mixture was vigorously stirred at room temperature for 30 minutes. The layers were then separated, and the aqueous layer was extracted with EtOAc (3 x 25 mL).
  • Example 26 Preparation of –(7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-2- dimethoxyphosphoryl-ethanone
  • Step 1 2-Chloro-7-iodo-6-isopropyl-furo[2,3-b]pyrazine
  • N-chloro-1-[(4-methoxyphenyl)methyl]-3-(3-methylbut-1- ynyl)pyrazin-2-one (1.39 g, 4.3879 mmol) in dry DCM (28 mL)
  • Iodine 1. g, 5.5160 mmol
  • DCM 28 mL
  • reaction mixture was degassed for 10 minutes, then sealed and heated in an oil bath at 150 ° C for 90 minutes.
  • the reaction was cooled to RT, diluted with ethyl acetate, and washed with water, and brine. Then the organic layer was dried over sodium sulfate, filtered, and concentrated.
  • reaction mixture was heated to 100 ° C with 100 psi of carbon monoxide and stirred for 16 h.
  • the reaction mixture was allowed to cool to rt.
  • the reaction mixture was filtered through a pad of celite, washed with ethyl acetate, filtrate was concentrated in vacuo.
  • Step 4 1-(7-Cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-2- dimethoxyphosphoryl-ethanone
  • a flask was heat gun-dried then cooled down to room temperature using a nitrogen stream and charged with dimethyl methylphosphonate (92 ⁇ L, 0.8601 mmol) and anhydrous THF (5.0 mL). The solution was cooled down to 0 ° C and a solution of LiHMDS in THF (1.9 mL of 1 M, 1.900 mmol) was added dropwise.
  • Example 27 Preparation of 1-(5-cyclobutyl-6-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2- dimethoxyphosphoryl-ethanone Step 1: 6-Bromo-3-prop-1-ynyl-pyrazin-2-amine [00329] A solution of (E)-1-bromoprop-1-ene (2.3 g, 19.01 mmol) in dry THF (131 mL) was cooled -78 °C followed by the addition of butyllithium (14.8 mL of 2.5 M, 37.00 mmol) and the mixture was stirred for 20 min at -78 °C.
  • the biphasic mixture was vigorously stirred for 30 minutes, then the layers were separated, and the aqueous layer was extracted with ethyl acetate (3 x 75 mL). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure.
  • Example 28 Preparation of 1-(6-tert-butyl-7-methyl-furo[2,3-b]pyrazin-2-yl)-2- dimethoxyphosphoryl-ethanone
  • Step 1 Methyl 7-bromo-6-tert-butyl-furo[2,3-b]pyrazine-2-carboxylate
  • N-bromo-succinimide 8.25 g, 46.35 mmol.
  • reaction mixture was stirred then heated at 60 ° C for 4 h.
  • the reaction mixture was quenched with sodium bicarbonate (40 mL), diluted with EtOAc (200 mL), washed with sodium bisulfate solution (40 mL), water (3 x 40 mL), dried over sodium sulfate, filtered, concentrated in vacuo.
  • Step 2 Methyl 6-tert-butyl-7-methyl-furo[2,3-b]pyrazine-2-carboxylate [00335] Under an argon atmosphere, a vial with stir bar containing methyl 7-bromo-6-tert- butyl-furo[2,3-b]pyrazine-2-carboxylate (1.6 g, 5.109 mmol) was added tripotassium phosphate (3.25 g, 15.31 mmol), Pd(PPh 3 ) 4 (590 mg, 0.5106 mmol) and trimethylboroxine (1.5 mL, 10.25 mmol). Degassed DMF (48 mL) and degassed water (9.6 mL) were added to the mixture.
  • Step 3 1-(6-tert-Butyl-7-methyl-furo[2,3-b]pyrazin-2-yl)-2- dimethoxyphosphoryl-ethanone [00336] To a stirred solution of dimethyl methylphosphonate (500 ⁇ L, 4.675 mmol) in THF (22 mL) at -78 °C was added dropwise LiHMDS (in THF solution) (6.0 mL of 1 M, 6.000 mmol).
  • reaction mixture was stirred at -78 °C for 15 minutes and then a solution of methyl 6-tert-butyl-7-methyl-furo[2,3-b]pyrazine-2-carboxylate (753.3 mg, 3.034 mmol) in anhydrous THF (7.0 mL) was added dropwise over 5 minutes.
  • the reaction was stirred at -78 °C for 10 minutes.
  • the reaction was quenched by the addition of a saturated aqueous solution of ammonium chloride (5 mL) and the biphasic mixture was vigorously stirred at room temperature for 5 minutes. The layers were then separated, and the aqueous layer was extracted with EtOAc (2 x 5 mL).
  • Example 29 Preparation of 2-dimethoxyphosphoryl-1-[5-isopropyl-7-(2- methoxyethyl)pyrrolo[2,3-b]pyrazin-3-yl]153xtracte
  • Step 1 3-Chloro-5-isopropyl-pyrrolo[2,3-b]pyrazine
  • 3-Chloro-5H-pyrrolo[2,3-b]pyrazine (4.0 g, 26.05 mmol) was dissolved in anhydrous DMF (104.9 mL) then cooled to 0 °C.2-iodopropane (3.2 mL, 32.06 mmol) was added then sodium hydride (1.53 g of 60 %w/w, 38.25 mmol) was added in portions over 10 minutes.
  • Step 4 3-Chloro-5-isopropyl-7-(2-methoxyethyl)pyrrolo[2,3-b]pyrazine
  • 3-chloro-5-isopropyl-7-[(Z)-2-methoxyvinyl]pyrrolo[2,3-b]pyrazine ⁇ 1:1 mixture of isomers, 30 mg, 0.1192 mmol
  • EtOAc 0.1192 mmol
  • palladium on carbon 63.9 mg of 10 %w/w, 0.06005 mmol.
  • the reaction mixture was stirred at room temperature under a hydrogen atmosphere for 1 h.
  • the reaction mixture was filtered through Celite and concentrated under reduced pressure.
  • Step 6 2-Dimethoxyphosphoryl-1-[5-isopropyl-7-(2-methoxyethyl)pyrrolo[2,3- b]pyrazin-3-yl]156xtracte
  • a flask was flame-dried then cooled down to room temperature using a nitrogen stream, and charged with dimethyl methylphosphonate (0.22 mL, 2.057 mmol) and anhydrous THF (15.4 mL). The solution was cooled down to -78 °C and a solution of LiHMDS in THF (4.5 mL of 1 M, 4.500 mmol) was added dropwise.
  • Example 30 Preparation of 1-[6-tert-Butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl]-2-dimethoxyphosphoryl-ethanone
  • Step 1 3-Bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine-7-carbaldehyde
  • Step 3 methyl 6-tert-butyl-7-[(Z)-2-methoxyvinyl]-5-methyl-pyrrolo[2,3- b]pyrazine-3-carboxylate
  • methanol 2.0 mL
  • triethylamine 0.81 mL, 5.811 mmol
  • Nitrogen was bubbled into the mixture for 15 minutes and then Pd(dppf)Cl2.DCM (80.6 mg, 0.09870 mmol) was added.
  • Step 4 Methyl 6-tert-butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3-b]pyrazine- 3-carboxylate
  • Step 5 1-[6-tert-Butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]- 2-dimethoxyphosphoryl-ethanone [00347]
  • a flask was flame-dried then cooled down to room temperature using a nitrogen stream and charged with dimethyl methylphosphonate (0.27 mL, 2.524 mmol) and anhydrous THF (21 mL).
  • the solution was cooled down to -78 °C and a solution of LiHMDS in THF (5.3 mL of 1 M, 5.300 mmol) was added dropwise.
  • reaction was let stirred in the dry-ice bath for 20 hours allowing the temperature to slowly warm to room temperature.
  • a saturated aqueous solution of ammonium chloride 200 mL
  • EtOAc 200 mL
  • brine 200 mL
  • the aqueous layer was extracted with EtOAc (2 x 100 mL) and the combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo.
  • Step 2 tert-Butyl (4S)-4-[1-(3-tert-butoxy-3-oxo-propoxy)-3-methyl-butyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00349] To a solution of tert-butyl (4S)-4-(1-hydroxy-3-methyl-butyl)-2,2-dimethyl- oxazolidine-3-carboxylate (19.7 g, 65.119 mmol) in anhydrous THF (60 mL) was added portion- wise NaH (3.1 g, 60 %w/w, 77.507 mmol) at room temperature.
  • Step 3 tert-Butyl (4S)-4-[(1R)-1-(3-hydroxypropoxy)-3-methyl-butyl]-2,2- dimethyl-oxazolidine-3-carboxylate and tert-butyl (4S)-4-[(1S)-1-(3- hydroxypropoxy)-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00350]
  • LAH pellet form
  • THF 125 mL
  • Step 4 tert-Butyl (4S)-2,2-dimethyl-4-[(1S)-3-methyl-1-(3- oxopropoxy)butyl]oxazolidine-3-carboxylate [00351] To a solution of tert-butyl (4S)-4-[(1S)-1-(3-hydroxypropoxy)-3-methyl-butyl]-2,2- dimethyl-oxazolidine-3-carboxylate (3.87 g, 10.776 mmol) in dichloromethane (110 mL) cooled at 0 ° C were added water (40 mL), sodium bicarbonate (1.45 g, 17.261 mmol), sodium bromide (225 mg, 2.1867 mmol) and TEMPO (265 mg, 1.6961 mmol).
  • sodium hypochlorite 8 mL of 10 %w/v, 10.747 mmol
  • a saturated aqueous solution of sodium bicarbonate 100 mL was added.
  • the phases were separated, and the aqueous layer was extracted with DCM (2 x 50 mL).
  • Example 32 Preparation of tert-butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo- 2H-pyrrole-1-carboxylate Step 1: (5R)-5-[[tert-Butyl(diphenyl)silyl]oxymethyl]pyrrolidin-2-one [00352] To a stirred solution of (5R)-5-(hydroxymethyl)pyrrolidin-2-one (25.01 g, 212.89 mmol) in degassed dichloromethane (600 mL) were added imidazole (33.76 g, 490.95 mmol) and DMAP (2.8 g, 22.690 mmol) under nitrogen.
  • Step 2 tert-Butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo- pyrrolidine-1-carboxylate [00353] To a stirred solution of crude (5R)-5-[[tert-butyl(diphenyl)silyl]oxymethyl]pyrrolidin- 2-one (66.1 g, 171.45 mmol) in anhydrous acetonitrile (300 mL) were added DMAP (12.7 g, 103.96 mmol) and Boc2O (65 g, 297.83 mmol). The reaction was then stirred overnight at room temperature.
  • reaction mixture was maintained at this temperature for 1 h then phenyl selenohypochlorite (8.8 g, 45.949 mmol) in THF (50 mL) was added dropwise at a rate to avoid the reaction internal temperature warming up above -78 °C.
  • the reaction mixture was stirred at this temperature for 2 h then it was quenched with saturated aqueous ammonium chloride (100 mL) and left to warm up to room temperature. It was then left stirring at room temperature for 30 min. The phases were separated, and the aqueous phase was extracted with ethyl acetate (3 x 50 mL). The combined organics were dried over magnesium sulfate, filtered, and concentrated under reduced pressure.
  • Example 33 Preparation of tert-butyl (4R)-2,2-dimethyl-4-[(1R)-3-methyl-1-(2- oxoethyl)butyl]oxazolidine-3-carboxylate
  • Step 1 tert-Butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-isobutyl-5- oxo-pyrrolidine-1-carboxylate
  • the reaction was stirred at 50 o C for 1 hour and then more NaBH4 (12 g, 317.19 mmol) was added slowly. The resulting mixture was stirred at 50 °C for 30 minutes. To the reaction mixture was added water (500 mL) and the volatiles were concentrated under reduced pressure. To the resulting aqueous solution was added DCM (300 mL) and the phases were separated. The aqueous phase was extracted with DCM (3 x 150 mL).
  • the reaction was stirred in the ice-cold bath for 3 hours.
  • the reaction was quenched at 0 °C with a saturated aqueous sodium bicarbonate (500 mL) and EtOAc (250 mL).
  • the biphasic solution was stirred for 1 hour at room temperature and the layers were separated.
  • the aqueous layer was extracted with ethyl acetate (2 x 250 mL).
  • the combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.
  • the crude residue was purified in two different batches of 17g by reversed-phase chromatography (Column: C 18 . Gradient: 5-100 % methanol in water).
  • Step 6 tert-Butyl (4R)-2,2-dimethyl-4-[(1R)-3-methyl-1-(2- oxoethyl)butyl]oxazolidine-3-carboxylate
  • oxalyl chloride 1.550 g, 1.0 mL, 11.463 mmol
  • dichloromethane 15 mL
  • a solution of dimethylsulfoxide (1.5414 g, 1.4 mL, 19.728 mmol) in anhydrous dichloromethane (15 mL) was added dropwise.
  • Step 2 tert-Butyl (4R)-4-[(1S)-5-ethoxy-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate
  • Palladium (on carbon) (12.5 g, 5 %w/w, 5.8730 mmol) was added to a solution of tert-butyl (4R)-4-[(E,1R)-5-ethoxy-1-isobutyl-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3- carboxylate (25 g, 65.120 mmol) in methanol (0.65 L) and hydrogen gas was bubbled into the suspension for 5 min, then the mixture was stirred under hydrogen atmosphere for 1 h.
  • Step 3 tert-Butyl (4R)-4-[(1S)-5-hydroxy-1-isobutyl-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate
  • Lithium aluminum hydride (in THF) 34 mL of 2 M, 68.000 mmol
  • tert- butyl (4R)-4-[(1S)-5-ethoxy-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (24.5 g, 60.370 mmol) in THF (500 mL) at 0 °C and the reaction was stirred for 1 h.
  • Step 4 tert-Butyl (4R)-4-[(1S)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate
  • tert-butyl (4R)-4-[(1S)-5-hydroxy-1-isobutyl-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate 1.1 g, 3.1991 mmol
  • TEMPO 25 mg, 0.1600 mmol
  • Example 35 Preparation of racemic-tert-butyl (4R)-4-[(1S)-1-isobutyl-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate
  • Step 1 Racemic-tert-butyl (4R)-4-[(E,1R)-5-ethoxy-1-isobutyl-5-oxo-pent-3- enyl]-2,2-dimethyl-oxazolidine-3-carboxylate
  • Ethyl 2-diethoxyphosphorylacetate (29 g, 129.35 mmol) was added in portions at room temperature to a suspension of heptane-washed sodium hydride (in oil) (5.1 g, 60 %w/w, 127.51 mmol) in THF (200 mL).
  • reaction mixture was quenched with saturated aqueous sodium bicarbonate (500 mL) and extracted with ethyl acetate containing 20% heptane (200 mL) then with DCM (100 mL x 2). The combined organics were dried over magnesium sulfate, filtered, and concentrated under reduced pressure.
  • the resulting residue was purified by reverse phase chromatography on a C 18 column using a 5 to 100% gradient of acetonitrile in pure water to provide racemic-tert-tert-butyl (4R)-4-[(E,1R)-5-ethoxy-1-isobutyl- 5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (13.3 g, 79%) as a clear oil.
  • Step 2 Racemic-tert-butyl (4R)-4-[(1S)-5-ethoxy-1-isobutyl-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate
  • Palladium (on carbon) (12.5 g, 5 %w/w, 5.8730 mmol) was added to a solution of racemic-tert-butyl (4R)-4-[(E,1R)-5-ethoxy-1-isobutyl-5-oxo-pent-3-enyl]-2,2-dimethyl- oxazolidine-3-carboxylate (25 g, 65.120 mmol) in methanol (0.65 L) and hydrogen gas was injected into the suspension for 5 min then left stirring under hydrogen atmosphere for 1 h.
  • Step 3 Racemic-tert-butyl (4R)-4-[(1S)-5-hydroxy-1-isobutyl-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate
  • Lithium aluminum hydride (in THF) 34 mL of 2 M, 68.000 mmol) was added to racemic-tert-butyl (4R)-4-[(1S)-5-ethoxy-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (24.5 g, 60.370 mmol) in THF (500 mL) at 0 °C and the reaction was stirred for 1 h.
  • Step 4 Racemic-tert-butyl (4R)-4-[(1S)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate
  • oxalyl chloride 2.3280 g, 1.6 mL, 18.341 mmol
  • DCM 32 mL
  • a solution of dimethylsulfoxide 2.8626 g, 2.6 mL, 36.637 mmol
  • reaction was quenched by the addition of a saturated aqueous solution of ammonium chloride (30 mL). The mixture was vigorously stirred at room temperature for 30 minutes, then the layers were separated, and the aqueous layer was extracted with dichloromethane (2 x 75 mL).
  • Example 36 Preparation of tert-butyl (4R)-4-[(1R)-3,3-dimethyl-1-(2-oxoethyl)butyl]-2,2- dimethyl-oxazolidine-3-carboxylate
  • Step 1 tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-(2,2- dimethylpropyl)-5-oxo-pyrrolidine-1-carboxylate
  • a flask containing copper iodide (3.18 g, 16.697 mmol) was flame-dried until bright yellow under high vacuum then cooled down to room temperature using a nitrogen stream and charged with anhydrous THF (53 mL).
  • the heterogeneous mixture was cooled down to -78 °C and a solution of 2,2-dimethylpropylmagnesium chloride (in THF) (33 mL of 1 M, 33.000 mmol) was added dropwise over 15 min.
  • THF 2,2-dimethylpropylmagnesium chloride
  • the reaction was stirred at -78 °C for 30 min after which time a solution of tert-butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-2H- pyrrole-1-carboxylate (4.42 g, 8.3188 mmol) in anhydrous THF (53 mL) was added dropwise over 15 min.
  • the reaction was stirred for 1 hour at -78 °C then allowed to reach room temperature overnight.
  • the reaction was then quenched by the addition of a saturated aqueous solution of ammonium chloride (100 mL) and the mixture was vigorously stirred at room temperature for 15 minutes.
  • the biphasic mixture was filtered on a pad of Celite while washing with EtOAc (150 mL). The layers were separated, and the aqueous layer was extracted with ethyl acetate (5 x 50 mL) and the combined organic layers were washed with water (2 x 50 mL) and brine (3 x 50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure.
  • Step 2 tert-Butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-(2,2- dimethylpropyl)-5-hydroxy-pyrrolidine-1-carboxylate
  • a stirred solution of tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- (2,2-dimethylpropyl)-5-oxo-pyrrolidine-1-carboxylate (1.31 g, 2.4460 mmol) in anhydrous THF (16 mL) was cooled down to -78 °C then a solution of lithium triethylborohydride in THF (2.67 mL of 1 M, 2.6700 mmol) was added dropwise over 15 minutes.
  • reaction was then stirred at -78 °C for 1 hour then quenched by the addition of a saturated aqueous solution of sodium bicarbonate (50 mL) and slowly allowed to reach 0 °C after which time an aqueous solution of hydrogen peroxide (in water) (39.550 mg, 0.1 mL of 35 %w/w, 0.4070 mmol) was added dropwise.
  • the reaction was stirred for 30 minutes at 0 °C then diluted with EtOAc (50 mL) and stirred for another 30 minutes.
  • Step 3 tert-Butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2- hydroxyethyl)-4,4-dimethyl-pentyl]carbamate [00372] To a solution of tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-(2,2- dimethylpropyl)-5-hydroxy-pyrrolidine-1-carboxylate (1.27 g, 2.3453 mmol) in methanol (40 mL) was added sodium borohydride (3.1 g, 81.940 mmol).
  • the reaction mixture was stirred at room temperature for 1 hour then heated at 50 °C for 1 hour.
  • the reaction was then concentrated under reduced pressure and partitioned between water (100 mL) and dichloromethane (100 mL).
  • the biphasic mixture was vigorously stirred at room temperature for 30 minutes. The layers were separated, and the aqueous layer was extracted with dichloromethane (3 x 100 mL).
  • Step 4 tert-Butyl N-[(1R,2R)-2-(2-hydroxyethyl)-1-(hydroxymethyl)-4,4- dimethyl-pentyl]carbamate
  • TBAF in THF
  • the reaction was stirred at 0 °C for 1 hour then quenched by the addition of a saturated aqueous solution of ammonium chloride (50 mL) and diluted with EtOAc (50 mL). The mixture was stirred for 30 minutes at room temperature, then the layers were separated, and the aqueous layer was extracted with ethyl acetate (3 x 50 mL) and the combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude was purified by reverse phase chromatography (Column: C18. Gradient: 5-100% MeCN in water).
  • Step 5 tert-Butyl (4R)-4-[(1R)-1-(2-hydroxyethyl)-3,3-dimethyl-butyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00374] To a stirred solution of tert-butyl N-[(1R,2R)-2-(2-hydroxyethyl)-1-(hydroxymethyl)- 4,4-dimethyl-pentyl]carbamate (514 mg, 1.6872 mmol) in anhydrous DCM (10 mL) were added 2,2-dimethoxypropane (3.4850 g, 4.1 mL, 33.462 mmol) and paratoluenesulfonic acid hydrate (32 mg, 0.1682 mmol).
  • the reaction was stirred at room temperature for 90 minutes then concentrated under reduced pressure and partitioned between ethyl acetate (20 mL) and a saturated aqueous solution of sodium bicarbonate (20 mL). The mixture was vigorously stirred at room temperature for 20 minutes, then the layers were separated, and the aqueous layer was extracted with ethyl acetate (4 x 20 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude was purified by reverse phase chromatography on C 18 (gradient: 5 to 100% acetonitrile in water; 12 CV).
  • Step 6 tert-Butyl (4R)-4-[(1R)-3,3-dimethyl-1-(2-oxoethyl)butyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00375] To a solution of tert-butyl (4R)-4-[(1R)-1-(2-hydroxyethyl)-3,3-dimethyl-butyl]-2,2- dimethyl-oxazolidine-3-carboxylate (449 mg, 1.3614 mmol) in dichloromethane (14 mL) cooled at 0 ° C were added water (4.5 mL), sodium bicarbonate (181 mg, 2.1546 mmol), sodium bromide (24 mg, 0.2332 mmol) and TEMPO (32 mg, 0.2048 mmol).
  • Example 37 Preparation of tert-butyl (4R)-2,2-dimethyl-4-[(1S)-2-methyl-1-(2- oxoethyl)propyl]oxazolidine-3-carboxylate
  • Step 1 tert-Butyl (2R,3S)-2-[[tert-Butyl(diphenyl)silyl]oxymethyl]-3-isopropenyl- 5-oxo-pyrrolidine-1-carboxylate
  • a flask containing copper iodide (3.1 g, 16.277 mmol) was flame-dried under vacuum until bright yellow then cooled down to room temperature using a nitrogen stream and charged with anhydrous THF (80 mL).
  • the reaction was cooled down to 0 °C and quenched by the dropwise addition of a saturated aqueous solution of ammonium chloride (150 mL) over one hour. The layers were then separated, and the aqueous layer was extracted with ethyl acetate (4 x 100 mL), dried over magnesium sulfate, filtered on a pad of celite and concentrated under reduced pressure. The crude was purified by flash chromatography on silica-gel (gradient: 0 to 10% ethyl acetate in heptanes, 10 CV).
  • Step 2 tert-Butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-isopropyl-5- oxo-pyrrolidine-1-carboxylate
  • Nitrogen was bubbled for 15 minutes in a stirred solution of tert-butyl (2R,3S)-2- [[tert-butyl(diphenyl)silyl]oxymethyl]-3-isopropenyl-5-oxo-pyrrolidine-1-carboxylate (3.33 g, 6.7381 mmol) in anhydrous methanol (200 mL) after which time palladium on carbon (720 mg, 10 %w/w, 0.6766 mmol) was added.
  • Step 3 tert-Butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-hydroxy-3- isopropyl-pyrrolidine-1-carboxylate [00378] A stirred solution of crude tert-butyl (2R,3S)-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-3-isopropyl-5-oxo-pyrrolidine-1-carboxylate (3.107 g, 6.1986 mmol) in anhydrous THF (35 mL) was cooled down to -78 °C and a solution of lithium triethylborohydride (i.e.
  • Step 4 tert-Butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2- hydroxyethyl)-3-methyl-butyl]carbamate
  • Step 5 tert-Butyl N-[(1R,2S)-2-(2-hydroxyethyl)-1-(hydroxymethyl)-3-methyl- butyl]carbamate
  • a stirred solution of tert-butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2- (2-hydroxyethyl)-3-methyl-butyl]carbamate (2.392 g, 4.5470 mmol) in anhydrous THF (45 mL) was cooled down to 0 °C and a solution of tetrabutylammonium fluoride in THF (6.9 mL of 1 M, 6.9000 mmol) was added dropwise.
  • the reaction was stirred at 0 o C for 2 hours after which time the reaction was quenched by the addition of a saturated aqueous sodium bicarbonate (50 mL) and brine (50 mL). The mixture was vigorously stirred for 30 minutes, then the layers were separated, and the aqueous layer was extracted with ethyl acetate (4 x 30 mL). The combined organic layers were washed with brine (150 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude was purified by reverse-phase chromatography on C18 (gradient: 5 to 100% acetonitrile in water; 12 CV).
  • Step 6 tert-Butyl (4R)-4-[(1S)-1-(2-hydroxyethyl)-2-methyl-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00381] To a stirred solution of tert-butyl N-[(1R,2S)-2-(2-hydroxyethyl)-1-(hydroxymethyl)- 3-methyl-butyl]carbamate (1.02 g, 3.8988 mmol) in anhydrous dichloromethane (15 mL) were added 2,2-dimethoxypropane (8.5000 g, 10 mL, 81.615 mmol) and paratoluenesulfonic acid hydrate (75 mg, 0.3943 mmol).
  • Step 7 tert-Butyl (4R)-2,2-dimethyl-4-[(1S)-2-methyl-1-(2- oxoethyl)propyl]oxazolidine-3-carboxylate
  • oxalyl chloride 494.70 mg, 340 ⁇ L, 3.8975 mmol
  • dichloromethane 9 mL
  • the solution was cooled down to -78 °C and a solution of dimethylsulfoxide (605.55 mg, 550 ⁇ L, 7.7501 mmol) in anhydrous dichloromethane (9 mL) was added dropwise.
  • Example 38 Preparation of tert-butyl (4R)-4-[(1S)-2,2-dimethyl-1-(2-oxoethyl)propyl]-2,2- dimethyl-oxazolidine-3-carboxylate Step 1: (4S,5R)-4-tert-Butyl-5-[[tert-butyl(diphenyl)silyl]oxymethyl]pyrrolidin-2- one [00383] To a stirred solution of ZnCl 2 (in THF solution) (130 mL of 0.5 M, 65.000 mmol) at room temperature was added tert-butylmagnesium chloride (in THF solution) (66 mL of 1 M, 66.000 mmol) dropwise.
  • Step 2 tert-Butyl (2R,3S)-3-tert-butyl-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5- oxo-pyrrolidine-1-carboxylate
  • 4S,5R 4-tert-butyl-5-[[tert- butyl(diphenyl)silyl]oxymethyl]pyrrolidin-2-one (2.96 g, 6.6334 mmol) in MeCN (25 mL) at room temperature was added DMAP (433.8 mg, 3.5508 mmol) followed by Boc2O (4.69 g, 21.489 mmol). The mixture was stirred at room temperature for 15 minutes.
  • Step 4 tert-Butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2- hydroxyethyl)-3,3-dimethyl-butyl]carbamate [00386] To a stirred solution of crude tert-butyl (2R,3S)-3-tert-butyl-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-5-hydroxy-pyrrolidine-1-carboxylate (2.89 g, 5.1784 mmol) in methanol (50 mL) at room temperature was added sodium borohydride (2.3 g, 60.794 mmol) portion-wise over 15 minutes.
  • the reaction was stirred at room temperature for 1 hour and then at 75 °C for 1 hour. The mixture was then cooled down to room temperature and more sodium borohydride (2.3 g, 60.794 mmol) was added portion-wise over 15 minutes. The reaction was stirred at 75 °C for 2 hours. The mixture was then cooled down to room temperature and more sodium borohydride (2.15 g, 56.829 mmol) was added portion-wise over 10 minutes. The reaction was stirred at 75 °C for 2 hours and was then cooled down to room temperature and concentrated in vacuo. Water (250 mL) and EtOAc (250 mL) were added, and the mixture was extracted with EtOAc (3 x 250 mL).
  • Step 5 tert-Butyl N-[(1R,2S)-2-(2-hydroxyethyl)-1-(hydroxymethyl)-3,3-dimethyl- butyl]carbamate [00387] To a stirred solution of tert-butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]- 2-(2-hydroxyethyl)-3,3-dimethyl-butyl]carbamate (3 g, 4.3501 mmol) in dry THF (60 mL) at 0 °C was added a solution of tetrabutylammonium fluoride in THF (7 mL of 1 M, 7.0000 mmol) dropwise.
  • the reaction was stirred at 0 °C for 2 hours and was then quenched by the addition of a saturated solution of sodium bicarbonate (50 mL) and brine (50 mL). The mixture was vigorously stirred for 10 minutes, then the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (1 x 250 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C 18 cartridge, eluting with a gradient of 5 to 100% of MeCN in water. The fractions containing the desired product were combined and the organic solvent was evaporated.
  • Step 6 tert-Butyl (4R)-4-[(1S)-1-(2-hydroxyethyl)-2,2-dimethyl-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00388] To a stirred solution of tert-butyl N-[(1R,2S)-2-(2-hydroxyethyl)-1-(hydroxymethyl)- 3,3-dimethyl-butyl]carbamate (1.22 g, 4.2087 mmol) in dry DCM (20 mL) were added 2,2- dimethoxypropane (8.5000 g, 10 mL, 81.615 mmol) and paratoluenesulfonic acid hydrate (83.1 mg, 0.4369 mmol).
  • the reaction was stirred at room temperature for 90 minutes.
  • the reaction was quenched with a saturated aqueous solution of sodium bicarbonate (3mL), dried over sodium sulfate, filtered on a pad of celite, washed with DCM (2 x 25 mL) then concentrated in vacuo.
  • the crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeCN in water. The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (50 mL) was then added, and the mixture was extracted with EtOAc (3 x 50 mL).
  • Step 7 tert-Butyl (4R)-4-[(1S)-2,2-dimethyl-1-(2-oxoethyl)propyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00389] To a stirred solution of tert-butyl (4R)-4-[(1S)-1-(2-hydroxyethyl)-2,2-dimethyl- propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (373.4 mg, 1.1245 mmol) in DCM (11 mL) cooled at 0 °C were added a solution of sodium bicarbonate (160 mg, 1.9046 mmol) and sodium bromide (28.9 mg, 0.2809 mmol) in water (3 mL) followed by TEMPO (35.5 mg, 0.2272 mmol).
  • Example 39 Preparation of tert-butyl (4R)-4-[(1R)-1-(cyclopropylmethyl)-3-oxo-propyl]- 2,2-dimethyl-oxazolidine-3-carboxylate
  • Step 1 tert-Butyl (2R,3R)-3-allyl-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo- pyrrolidine-1-carboxylate [00390]
  • ZnCl2 in Me-THF
  • dry THF 80 mL
  • allyl(chloro)magnesium 22 mL of 2 M, 44.000 mmol
  • the reaction was then stirred at -78 °C for 90 minutes.
  • the mixture was then quenched by the addition of a saturated aqueous solution of sodium bicarbonate (50 mL) and slowly allowed to reach 0 °C after which time an aqueous solution of hydrogen peroxide (in water) (276.85 mg, 0.7 mL of 35 %w/w, 2.8487 mmol) was added dropwise.
  • the reaction was stirred for 30 minutes at 0 °C then diluted with EtOAc (50 mL) and stirred for another 30 minutes.
  • Step 4 tert-Butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2- (cyclopropylmethyl)-4-hydroxy-butyl]carbamate [00393] To a stirred solution of tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- (cyclopropylmethyl)-5-hydroxy-pyrrolidine-1-carboxylate (2.77 g, 5.1949 mmol) in methanol (80 mL) at room temperature was added portion-wise NaBH 4 (4.6 g, 121.59 mmol).
  • Step 6 tert-Butyl (4R)-4-[(1R)-1-(cyclopropylmethyl)-3-hydroxy-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00395] To a solution of tert-butyl N-[(1R,2R)-2-(cyclopropylmethyl)-4-hydroxy-1- (hydroxymethyl)butyl]carbamate (7.65 g, 26.585 mmol) in dichloromethane (75 mL) at room temperature was added 2,2-dimethoxypropane (13.600 g, 16 mL, 130.58 mmol) followed by p- toluenesulfonic acid (507.8 mg, 2.6696 mmol).
  • the reaction mixture was stirred at room temperature for 2 hours.
  • the reaction was quenched with a saturated aqueous solution of sodium bicarbonate (50 mL) and the mixture was stirred at room temperature for 5 minutes.
  • the layers were separated, and the aqueous layer was extracted with DCM (3 x 50 mL).
  • the combined organic layers were washed with brine (1 x 100 mL), dried over sodium sulfate, filtered, and concentrated in vacuo.
  • the crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeCN in water. The fractions containing the desired product were combined and the organic solvent was evaporated.
  • Step 7 tert-Butyl (4R)-4-[(1R)-1-(cyclopropylmethyl)-3-oxo-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00396] To a stirred solution of tert-butyl (4R)-4-[(1R)-1-(cyclopropylmethyl)-3-hydroxy- propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (6.4 g, 19.398 mmol) in DCM (200 mL) cooled at 0 °C were added a solution of sodium bicarbonate (2.8 g, 33.331 mmol) and sodium bromide (400 mg, 3.8875 mmol) in water (60 mL) followed by TEMPO (606 mg, 3.8785 mmol).
  • Step 2 tert-Butyl (4S)-4-(1-hydroxypent-4-enyl)-2,2-dimethyl-oxazolidine-3- carboxylate [00398] To a solution of tert-butyl (4S)-4-formyl-2,2-dimethyl-oxazolidine-3-carboxylate (15 g, 65.424 mmol) in THF (120 mL) was slowly added at -78 °C bromo(but-3-enyl)magnesium in THF (65 mL of 1 M, 65.000 mmol).
  • the solution was stirred at this temperature for 2 hours, then the reaction mixture was allowed to warm up to room temperature and stirred for 16 h.
  • the reaction was quenched using a saturated aqueous solution of ammonium chloride (150 mL).
  • the product was extracted three times using ethyl acetate (75 mL x 3) and combined organic layers were washed using brine (200 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure.
  • Step 3 tert-Butyl (4S)-2,2-dimethyl-4-pent-4-enoyl-oxazolidine-3-carboxylate [00399] To a solution of tert-butyl (4S)-4-(1-hydroxypent-4-enyl)-2,2-dimethyl-oxazolidine-3- carboxylate (17.4 g, 60.911 mmol) in dichloromethane (150 mL) at 0 ° C was added Dess-Martin periodinane (31 g, 73.089 mmol) and the reaction was stirred for 1 hour at 0 ° C and then at room temperature for 1 hour.
  • Step 5 tert-Butyl (4S)-4-(1-isopropoxypent-4-enyl)-2,2-dimethyl-oxazolidine-3- carboxylate [00401] To a solution of tert-butyl (4S)-4-(1-hydroxypent-4-enyl)-2,2-dimethyl-oxazolidine-3- carboxylate (19.2 g, 53.218 mmol) in 2-iodopropane (365.50 g, 215 mL, 2.1501 mol) was added silver (I) oxide (68 g, 293.44 mmol) at room temperature. The solution was stirred at 45 °C for 24 h.
  • reaction mixture was filtered through a pad of Celite and washed with EtOAc (3 x 25 mL).
  • the crude mixture was put on a silica pad (50 g) and was washed with a mixture of heptanes/EtOAc (2 x 475 mL / 25 mL) to afford, after evaporation, tert-butyl (4S)-4-(1- isopropoxypent-4-enyl)-2,2-dimethyl-oxazolidine-3-carboxylate (13.6 g, 66%) as a yellow oil.
  • Step 6 tert-Butyl (4S)-4-(5-hydroxy-1-isopropoxy-pentyl)-2,2-dimethyl- oxazolidine-3-carboxylate ⁇ 85:15 mixture of diastereomers, the major i somer having the 4S,1R configuration
  • tert-butyl (4S)-4-(1-isopropoxypent-4-enyl)-2,2-dimethyl- oxazolidine-3-carboxylate (13.6 g, 35.344 mmol) in tetrahydrofuran (150 mL) was added borane tetrahydrofuran complex (141 mL of 1 M, 141.00 mmol) and the solution was stirred for 1 h at that temperature.
  • Example 41 Preparation of tert-butyl (4S)-2,2-dimethyl-4-[(1R)-5-oxo-1-[1,2,2,2- tetradeuterio-1-(trideuteriomethyl)ethoxy]pentyl]oxazolidine-3-carboxylate
  • Step 1 tert-Butyl (4S)-4-hex-5-enoyl-2,2-dimethyl-oxazolidine-3-carboxylate
  • 5-bromopent-1-ene 8.580 g, 57.57 mmol
  • THF 67.78 mL
  • magnesium turnings 1.515 g, 57.56 mmol
  • Step 2 (4S)-4-[(1R)-1-Hydroxyhex-5-enyl]-2,2-dimethyl-oxazolidine-3- carboxylate [00405] To a solution of tert-butyl (4S)-4-hex-5-enoyl-2,2-dimethyl-oxazolidine-3-carboxylate (14.73 g, 49.53 mmol) in THF (147.3 mL) was added magnesium chloride (4.716 g, 49.53 mmol). The resulting white suspension was stirred at room temperature for 10 min. Then borane (t-butylamine salt) (5.169 g, 59.44 mmol) was added in one portion.
  • borane t-butylamine salt
  • the reaction mixture was stirred at room temperature for 1 h.
  • the reaction was quenched at room temperature with aqueous saturated NH 4 Cl (100 mL), the organics were then diluted with EtOAc (100 mL) and washed once with a 100 mL half brine solution.
  • the organics were dried filtered and evaporated then purified using silica gel chromatography with a shallow gradient from 100% hexanes to 100% EtOAc to afford tert-butyl (4S)-4-[(1R)-1-hydroxyhex-5-enyl]-2,2-dimethyl-oxazolidine- 3-carboxylate (14.83 g, 100%) as a clear oil.
  • Step 3 tert-butyl (4S)-2,2-dimethyl-4-[(1R)-1-[1,2,2,2-tetradeuterio-1- (trideuteriomethyl)ethoxy]hex-5-enyl]oxazolidine-3-carboxylate [00406] To a solution of tert-butyl (4S)-4-[(1R)-1-hydroxyhex-5-enyl]-2,2-dimethyl- oxazolidine-3-carboxylate (700 mg, 2.338 mmol) in 1,1,1,2,3,3,3-heptadeuterio-2-iodo-propane (2.8 mL, 28.06 mmol) was added silver(I) oxide (2.709 g, 11.69 mmol).
  • Example 42 Preparation of tert-butyl (4S)-4-[(1R)-1-isopropoxy-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate
  • Step 1 tert-Butyl (4S)-4-[(1R)-1-isopropoxyhex-5-enyl]-2,2-dimethyl- oxazolidine-3-carboxylate
  • reaction mixture was partitioned between ethyl acetate and water. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate once more. The combined organic layers were dried over sodium sulfate, filtered and concentrated, then purified by silica gel chromatography using a shallow gradient from 100% hexanes to 100% EtOAc giving as a clear oil, tert-butyl (4S)-4- [(1R)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (4.61 g, 82%) ESI- MS m/z calc.343.23587, found 344.4 (M+1) + ; Retention time: 1.78 minutes.
  • Step 2 tert-Butyl (4S)-4-[(1S)-1-[3-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin- 3-yl)-3-oxo-propoxy]-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00411] To a solution of tert-butyl (4S)-4-[(1S)-1-[3-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-hydroxy-propoxy]-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate (820 mg, 1.5377 mmol) in anhydrous DCE (20 mL) was added MnO2 (activated) (2.98 g, 29.136 mmol).
  • the suspension was heated to 70 °C for 45 minutes then the reaction was allowed to cool down to room temperature.
  • the suspension was filtered through Celite, and the pad was rinsed with DCM (3 x 20 mL).
  • the combined filtrates were concentrated in vacuo to afford 850 mg of the desired product (85 % purity 1 H NMR, 89% yield) as a mixture of diastereomers.
  • Step 3 3-[(1S)-1-[(1S)-1-Amino-2-hydroxy-ethyl]-3-methyl-butoxy]-1-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)propan-1-one [00412] To a solution of tert-butyl (4S)-4-[(1S)-1-[3-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-oxo-propoxy]-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate (380 mg, 0.6752 mmol) in Et 2 O (8 mL) and water (2 mL) at room temperature was added a solution of HCl (in diethyl ether) (5 mL of 2 M, 10.000 mmol).
  • Step 4 [(2S,3S,5S)-5-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2- isobutyl-1,4-oxazepan-3-yl]methanol and [(2S,3S,5R)-5-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-2-isobutyl-1,4-oxazepan-3-yl]methanol [00413] To a solution of 3-[(1S)-1-[(1S)-1-amino-2-hydroxy-ethyl]-3-methyl-butoxy]-1-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)propan-1-one (264 mg, 0.6760 mmol) in anhydrous DCM (20 mL) at room temperature was added NaBH(Oac) 3
  • the reaction was stirred at room temperature for 18 hours and a saturated aqueous solution of sodium bicarbonate (20 mL) was added. The phases were separated, and the aqueous solution was extracted with DCM (3 x 10 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.
  • N-methylmorpholine (15.7 ⁇ L, 0.1428 mmol) was added by syringe and the reaction was stirred at 0 °C for 30 minutes. The ice bath was then removed and stirring was continued for an additional 48 hours at room temperature. The reaction mixture was then partitioned between 25 mL 1M HCl and 25 mL ethyl acetate. The layers were separated, and the aqueous layer was extracted with an additional 25 mL ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated.
  • N-methylmorpholine (16.4 ⁇ L, 0.1492 mmol) was added by syringe and the reaction was stirred at 0 °C for 30 minutes. The ice bath was then removed and stirring was continued for an additional 16 hours at room temperature. The reaction mixture was then partitioned between 25 mL 1M HCl and 25 mL ethyl acetate. The layers were separated, and the aqueous layer was extracted with an additional 25 mL ethyl acetate. The combined organic layers were washed 2 x 25 mL with brine, dried over magnesium sulfate, filtered, and concentrated.
  • Step 2 tert-Butyl (4R)-4-[(1S)-5-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-1-isobutyl- 5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate
  • Nitrogen was bubbled through a stirred solution of tert-butyl (4R)-4-[(1R)-5-(6-tert- butylfuro[2,3-b]pyrazin-2-yl)-1-isobutyl-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3- carboxylate (201 mg, 0.3612 mmol) in methanol (40 mL) for 10 minutes after which time palladium on carbon (38 mg, 10 %w/w, 0.0357 mmol) was added.
  • Step 4 [(2R,3S)-7-(6-tert-Butylfuro[2,3-b]pyrazin-2-yl)-3-isobutyl-azepan-2- yl]methanol [00421] To a solution of (5S)-5-[(1R)-1-amino-2-hydroxy-ethyl]-1-(6-tert-butylfuro[2,3- b]pyrazin-2-yl)-7-methyl-octan-1-one (hydrochloride salt) (143 mg, 0.3468 mmol) in anhydrous DCM (7.5 mL) at room temperature was added NaBH(Oac) 3 (245 mg, 1.156 mmol).
  • Step 7 (4S,8S,9R)-4-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-14-(2,6- dimethylphenyl)-8-isobutyl-18,18-dioxo-11-oxa-18 ⁇ 6 -thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one [00424] In a 4 mL vial, 3-[[4-[[(2R,3S,7S)-7-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-3-isobutyl- azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (
  • Step 2 (5S)-5-[(1R)-1-Amino-2-hydroxy-ethyl]-1-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-7-methyl-octan-1-one
  • HCl (in dioxane) 14 mL of 4 M, 56.00 mmol
  • tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-isobutyl-5-oxo- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate 328 mg, 0.5707 mmol
  • dioxane (12 mL
  • water 4.8 mL
  • reaction mixture was stirred at this temperature for 2 h.
  • the reaction mixture was quenched with a saturated sodium bicarbonate aqueous solution (25 mL) and this mixture was left stirring for 30 min.
  • the aqueous phase was separated and extracted with DCM (3x 50 mL). The combined organics were dried over magnesium sulfate, filtered, and concentrated under reduced pressure.
  • reaction mixture was stirred for 30 minutes at room temperature then warmed to 40 °C for 2 hours.
  • the reaction mixture was cooled to room temperature and diluted with EtOAc and 1 M HCl.
  • the organic phase was separated, and the aqueous layer was extracted with EtOAc (5 x).
  • Step 2 tert-Butyl (4S)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate, tert- butyl (4S)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1- isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate, and tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-isopropoxy- 5-oxo-pentyl]-2,2-dimethyl-ox
  • the resulting suspension was stirred at 70 °C for 1.5 h.
  • the suspension was filtered on a pad of Celite and washed with DCM (2 x 50 mL).
  • the crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C 18 cartridge, eluting with a gradient of 5 to 100% of MeOH in acidic water (0.1% v/v of formic acid in water).
  • the fractions containing the desired product were combined and the volatiles were removed under reduced pressure.
  • To the remaining aqueous phase was added DCM (50 mL) and water (200 mL).
  • the aqueous phase was extracted with DCM (2 x 50 mL).
  • Step 3 [(2S,3R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol and (5R,6S)-6-amino-1- (6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-7-hydroxy-5-isopropoxy- heptan-1-one [00432] To a solution of tert-butyl (4S)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.45 g, 2.7049 mmol) in 1,4-dioxane (30 mL
  • Step 4 [(2S,3R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methanol [00433] To a solution of (5R,6S)-6-amino-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-7-hydroxy-5-isopropoxy-heptan-1-one (892.2 mg, 2.1681 mmol) in dichloromethane (35 mL) was added sodium triacetoxyborohydride (1.08 g, 5.0958 mmol). The reaction was stirred at rt for 0.5 h.
  • Step 5 [(2S,3R,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methanol and [(2S,3R,7R)-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methanol [00434] A solution of a first batch of [(2S,3R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin- 3-yl)-3-isopropoxy-azepan-2-yl]methanol (690 mg, 1.7116 mmol) in acetonitrile (50 mL) and a solution of a second batch of [(2S,3R)-7-(6-tert-butyl-5-methyl-pyrrolo
  • the product was purified by preparative HPLC (Flow: 42,0 mL/min, Run time: 19 min, mobile phase conditions: initial 100% H 2 O (10 mM NH4CO3H) and 0% MeCN for 3 min, reach 30% MeCN over 1 min, reach 50% MeCN over 11 min and finally reach 95% MeCN over 2 min with a 2 min hold at 95%). Three fractions were obtained. The first fraction afforded, after evaporation and lyophilization, [(2S,3R,7S)-7-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methanol (342.6 mg, 42%) as a white solid.
  • reaction mixture was stirred for 30 minutes at room temperature then warmed to 40 °C for 2 hours.
  • the reaction mixture was cooled to room temperature and diluted with EtOAc and 1 M HCl.
  • the organic phase was separated, and the aqueous layer was extracted with EtOAc (5 x).
  • Step 7 (4S,8R,9S)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-8-isopropoxy-18,18-dioxo-11-oxa-18 ⁇ 6 -thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one [00436] 3-[[4-[[(2S,3R,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (101.8 mg,
  • Example 48 Preparation of Compound 17 Step 1: 3-[[4-[[(2S,3R,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin- 2-yl]sulfamoyl]benzoic acid [00437] Into a reaction vial was added [(2S,3R,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methanol (55 mg, 0.1469 mmol) and 3-[[4-chloro-6- (2,6-dimethylphenyl)-5-methyl-pyrimidin-2-yl]sulfamoyl]benzoic acid (65 mg, 0.
  • Step 2 (4S,8R,9S)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-8-isopropoxy-13-methyl-18,18-dioxo-11-oxa-18 ⁇ 6 -thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one [00438] To a scintillation vial with stir bar was added 3-[[4-[[(2S,3R,7S)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)-5-methyl-pyrimidin
  • NMM 35 ⁇ L, 0.3183 mmol
  • NMM 7.1 ⁇ L, 0.06458 mmol
  • NMM 49.0 ⁇ L, 0.4457 mmol
  • NMM 49.0 ⁇ L, 0.4457 mmol
  • additional NMM 12.2 ⁇ L, 0.1110 mmol
  • stirring an additional hour more NMM (12.2 ⁇ L, 0.111 mmol) was added and the reaction was stirred overnight.
  • Step 2 tert-Butyl (4R)-4-[(1S)-5-(7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin- 2-yl)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate
  • the mixture was evacuated and then stirred under a hydrogen atmosphere using a hydrogen-filled balloon at room temperature for 5 hours.
  • the mixture was filtered over a plug of celite, washed with methanol and concentrated.
  • the crude was then purified by silica chromatography (40 g of silica) using a gradient eluent of 100% hexanes to 100% ethyl acetate over 20 minutes (product elutes at 32% EtOAc).
  • Step 3 [(2R,3S)-7-(7-Cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-3- isobutyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00443] To a stirred solution of tert-butyl (4R)-4-[(1S)-5-(7-cyclopropyl-6-isopropyl-furo[2,3- b]pyrazin-2-yl)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (54 mg, 0.09968 mmol) in a mixture of dioxane (1.5 mL) and water (750 ⁇ L) was added a solution of hydrogen chloride (4M in dioxane) (2.5 mL of 4 M, 10.0 mmol) dropwise.
  • Step 4 [(2R,3S)-7-(7-Cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-3- isobutyl-azepan-2-yl]methanol [00444] To a solution of [(2R,3S)-7-(7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-3- isobutyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (38 mg, 0.09908 mmol) in anhydrous dichloromethane (2 mL) at room temperature was added sodium triacetoxyborohydride (117 mg, 0.5520 mmol).
  • the reaction was stirred at room temperature for 1 hour and a saturated aqueous solution of sodium bicarbonate (10 mL) was added. The phases were separated, and the aqueous solution was extracted with DCM (2 x). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude was then purified by silica chromatography (12 g of silica) using a gradient eluent of 100% dichloromethane to 20% methanol in DCM over 20 minutes (product eluted at 12% MeOH).
  • Step 5 3-[[4-[[(2R,3S)-7-(7-Cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-3- isobutyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00445] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (31 mg, 0.07419 mmol) and [(2R,3S)-7-(7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-3-isobutyl- azepan-2-yl]methanol (28 mg, 0.07262 mmol) were combined in a flask and dissolved in THF (500
  • the reaction mixture was stirred for 60 minutes at room temperature then warmed to 40 ° C for 30 minutes then 50 ° C for 1 hour.
  • the reaction mixture was cooled to room temperature and diluted with EtOAc and 1 M HCl.
  • the organic phase was separated, and the aqueous layer was extracted with EtOAc (5x).
  • the organic layers were combined, washed with brine, and dried over sodium sulfate.
  • the mixture was filtered, and then concentrated under reduced pressure.
  • the crude was then purified by silica chromatography (12 g of silica) using a gradient eluent of 100% hexanes to 100% ethyl acetate over 20 minutes followed by a gradient of 100% dichloromethane to 20% methanol in dichloromethane.
  • Step 6 (8S,9R)-4-(7-Cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-14-(2,6- dimethylphenyl)-8-isobutyl-18,18-dioxo-11-oxa-18 ⁇ 6 -thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one, diastereomer 1 (Compound 20), and (8S,9R)-4-(7-cyclopropyl-6- isopropyl-furo[2,3-b]pyrazin-2-yl)-14-(2,6-dimethylphenyl)-8-isobutyl-18,18- dioxo-11-oxa-18 ⁇ 6 -thia-3,15,17,24- tetrazatetracycl
  • the crude material was purified by silica chromatography (40 g of silica) using a gradient of 100% dichloromethane to 20% methanol in dichloromethane.
  • the desired fractions were concentrated under reduced pressure, and dried under high vacuum to give a light yellow solid 3-[[4- [[(2S,3R,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2- yl]methoxy]-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (134.6 mg, 90%).
  • Step 2 [(2R,3S)-7-[6-tert-Butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl]-3-isobutyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00450] To a stirred solution of tert-butyl (4R)-4-[(1S)-5-[6-tert-butyl-7-(2-methoxyethyl)-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl]-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (452.6 mg, 0.7713 mmol) in a mixture of dioxane (81 mL) and water (41.0 mL) was added a solution of hydrogen chloride (19.3 mL of 4 M, 77.20
  • the reaction was stirred at room temperature for 1.5 hours and a saturated aqueous solution of sodium bicarbonate (100 mL) was added. The phases were separated, and the aqueous solution was extracted with DCM (2 x). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.
  • Step 4 3-[[4-[[(2R,3S)-7-[6-tert-Butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl]-3-isobutyl-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid
  • reaction mixture was stirred for 30 minutes at room temperature then warmed to 40 °C for 1 hour.
  • the reaction mixture was cooled to room temperature then diluted with EtOAc and 1 M HCl.
  • the organic phase was separated, and the aqueous layer was extracted with EtOAc (5X).
  • NMM (39.5 ⁇ L, 0.3593 mmol) was added and the reaction was stirred at room temperature overnight.
  • the reaction mixture was diluted with 1M HCl and extracted with ethyl acetate. The layers were separated, and the aqueous layer was further extracted with ethyl acetate (2 x).
  • Step 2 tert-Butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-1-isopropyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate
  • Nitrogen was bubbled through a stirred solution of tert-butyl (4R)-4-[(E,1S)-5-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-isopropyl-5-oxo-pent-3-enyl]-2,2-dimethyl- oxazolidine-3-carboxylate (300 mg, 0.5559 mmol) in anhydrous methanol (10 mL) for 10 minutes at which time palladium on carbon (60 mg, 10 %w/w, 0.0564 mmol) was then added.
  • Step 3 [(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00456] To a stirred solution of tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-isopropyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (127 mg, 0.2344 mmol) in diethyl ether (2.5 mL) was added a solution of hydrogen chloride in diethyl ether (3 mL of 2 M, 6.0000 mmol) and water (0.650 mL).
  • the reaction was stirred at room temperature overnight then neutralized with a saturated aqueous solution of sodium bicarbonate (5 mL) then concentrated under reduced pressure to remove the volatiles.
  • the aqueous layer was extracted with dichloromethane (5 x 5 mL) and the combined organic layers were washed with brine (20 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure then dried under high vacuum.
  • the crude was purified by reverse-phase chromatography on C 18 (gradient: 5 to 100% methanol in water; 12CV). The desired fractions were concentrated under reduced pressure and the remaining water was co-evaporated with methanol (6 x 5 mL).
  • Step 4 [(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropyl-azepan-2-yl]methanol, diastereomer 1, and [(2R,3S)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropyl-azepan-2-yl]methanol, diastereomer 2 D iastereomer 1 Diastereomer 2 [00457] To a stirred solution of [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-isopropyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (41 mg, 0.1093 mmol) in anhydrous DCM (5 mL) was
  • the reaction was stirred overnight at room temperature then quenched by the addition of a saturated aqueous solution of sodium bicarbonate (10 mL).
  • the biphasic mixture was vigorously stirred at room temperature for 30 minutes.
  • the layers were then separated, and the aqueous layer was extracted with DCM (5 x 5 mL).
  • the combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure.
  • the desired fractions were concentrated under reduced pressure and the remaining water was co-evaporated with acetonitrile (5 x 5 mL) then freeze- dried to afford a white fluffy powder.
  • the 1:1 diastereomeric mixture was subjected to SFC separation using the following conditions: a Cellulose 4 column (250 x 30 mm), 5 ⁇ M column at 40 °C, eluent: 40% methanol and 60% CO 2 (0.1% diethylamine), flow rate: 150 mL/min, concentration : 4.6 mg/mL in methanol (no modifier), injection volume : 100 ⁇ L, pressure : 100 bars, wavelength: 210 nm.
  • Step 5 3-[[4-[[(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid D iastereomer 1 [00458] [(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropyl-azepan-2- yl]methanol (diastereomer 1, 6.7 mg, 0.01869 mmol) was combined with 3-[[4-chloro-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (7.9 mg, 0.01891 mmol) in THF (
  • Sodium tert-butoxide (12 mg, 0.1249 mmol) was added and the reaction was placed in an aluminum heating block at 35 °C for 3 hours. The reaction mixture was then partitioned between 15 mL 1M HCl and 15 mL ethyl acetate. The layers were separated and the aqueous was extracted with an additional 10 ml ethyl acetate. The aqueous was then diluted with an equal volume of brine and extracted with an additional 2 x 10 mL ethyl acetate.
  • Step 6 (8S,9R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-8-isopropyl-18,18-dioxo-11-oxa-18 ⁇ 6 -thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one, diastereomer 1, compound 35 Diastereomer 1 Diastereomer 1 [00459] 3-[[4-[[(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropyl- azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-y
  • N-methylmorpholine (approximately 11.34 mg, 12.33 ⁇ L, 0.1121 mmol) was added and the reaction was stirred for 18 hours at room temperature. The reaction mixture was then partitioned between 1M HCl and ethyl acetate. The layers were separated, and the aqueous layer was extracted 3x with ethyl acetate. The combined organics were washed with water and brine, dried over sodium sulfate and concentrated.
  • Example 54 Preparation of Compound 36 Step 1: 3-[[4-[[(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid D iastereomer 2 [00460] [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropyl-azepan-2- yl]methanol (diastereomer 2, 4.5 mg, 0.01255 mmol) was combined with 3-[[4-chloro-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (5.3 mg, 0.01268
  • Step 2 (8S,9R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-8-isopropyl-18,18-dioxo-11-oxa-18 ⁇ 6 -thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one, diastereomer 2, compound 36 Diastereomer 2 Diastereomer 2 [00461] 3-[[4-[[(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropyl- azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-y
  • N-methylmorpholine (approximately 7.428 mg, 8.074 ⁇ L, 0.07344 mmol) was added and the reaction was stirred for 18 hours at room temperature. The reaction mixture was then partitioned between 1M HCl and ethyl acetate. The layers were separated, and the aqueous phase was extracted with 3x ethyl acetate. The combined organics were washed with water and brine, dried over sodium sulfate and concentrated.

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Abstract

This disclosure provides modulators of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR), pharmaceutical compositions containing at least one such modulator, methods of treatment of CFTR mediated diseases, including cystic fibrosis, using such modulators and pharmaceutical compositions, combination pharmaceutical compositions and combination therapies employing those modulators, and processes and intermediates for making such modulators.

Description

MODULATORS OF CYSTIC FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR [0001] This application claims the benefit of priority of U.S. Provisional Application No. 63/587,955, filed October 4, 2023, the contents of which are incorporated herein by reference in their entirety. [0002] The disclosure relates to modulators of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR), pharmaceutical compositions containing the modulators, methods of treatment of CFTR mediated diseases, including cystic fibrosis, using such modulators, combination therapies and combination pharmaceutical compositions employing such modulators, and processes and intermediates for making such modulators. [0003] Cystic fibrosis (CF) is a recessive genetic disease that affects approximately 92,000 children and adults worldwide. Despite progress in the treatment of CF, there is no cure. [0004] In patients with CF, mutations in CFTR endogenously expressed in respiratory epithelia lead to reduced apical anion secretion causing an imbalance in ion and fluid transport. The resulting decrease in anion transport contributes to increased mucus accumulation in the lung and accompanying microbial infections that ultimately cause death in CF patients. In addition to respiratory disease, CF patients typically suffer from gastrointestinal problems and pancreatic insufficiency that, if left untreated, result in death. In addition, the majority of males with cystic fibrosis are infertile, and fertility is reduced among females with cystic fibrosis. [0005] Sequence analysis of the CFTR gene has revealed a variety of disease-causing mutations (Cutting, G. R. et al. (1990) Nature 346:366-369; Dean, M. et al. (1990) Cell 61:863:870; and Kerem, B-S. et al. (1989) Science 245:1073-1080; Kerem, B-S et al. (1990) Proc. Natl. Acad. Sci. USA 87:8447-8451). To date, greater than 2000 mutations in the CFTR gene have been identified; currently, the CFTR2 database contains information on only 432 of these identified mutations, with sufficient evidence to define 352 mutations as disease-causing. The most prevalent disease-causing mutation is a deletion of phenylalanine at position 508 of the CFTR amino acid sequence and is commonly referred to as the F508del mutation. This mutation occurs in many of the cases of cystic fibrosis and is associated with severe disease. [0006] The deletion of residue 508 in CFTR prevents the nascent protein from folding correctly. This results in the inability of the mutant protein to exit the endoplasmic reticulum (ER) and traffic to the plasma membrane. As a result, the number of CFTR channels for anion transport present in the membrane is far less than observed in cells expressing wild-type CFTR, i.e., CFTR having no mutations. In addition to impaired trafficking, the mutation results in defective channel gating. Together, the reduced number of channels in the membrane and the defective gating lead to reduced anion and fluid transport across epithelia. (Quinton, P. M. (1990), FASEB J.4: 2709-2727). The channels that are defective because of the F508del mutation are still functional, albeit less functional than wild-type CFTR channels. (Dalemans et al. (1991), Nature Lond.354: 526-528; Pasyk and Foskett (1995), J. Cell. Biochem.270: 12347- 50). In addition to F508del, other disease-causing mutations in CFTR that result in defective processing, trafficking, synthesis, and/or channel gating could modify anion secretion and therefore modify disease progression and/or severity. [0007] CFTR is a cAMP/ATP-mediated anion channel that is expressed in a variety of cell types, including absorptive and secretory epithelia cells, where it regulates anion flux across the membrane, as well as the activity of other ion channels and proteins. In epithelial cells, normal functioning of CFTR is critical for the maintenance of electrolyte transport throughout the body, including respiratory and digestive tissue. CFTR is composed of 1480 amino acids that encode a protein which is made up of a tandem repeat of transmembrane domains, each containing six transmembrane helices and a nucleotide binding domain. The two transmembrane domains are linked by a large, polar, regulatory (R)-domain with multiple phosphorylation sites that regulate channel activity and cellular trafficking. [0008] Chloride transport takes place by the coordinated activity of ENaC and CFTR present on the apical membrane and the Na+-K+-ATPase pump and Cl- channels expressed on the basolateral surface of the cell. Secondary active transport of chloride from the luminal side leads to the accumulation of intracellular chloride, which can then passively leave the cell via Cl- channels, resulting in a vectorial transport. Arrangement of Na+/2Cl-/K+ co-transporter, Na+- K+-ATPase pump and the basolateral membrane K+ channels on the basolateral surface and CFTR on the luminal side coordinate the secretion of chloride via CFTR on the luminal side. Because water is probably never actively transported itself, its flow across epithelia depends on tiny transepithelial osmotic gradients generated by the bulk flow of sodium and chloride. [0009] A number of CFTR modulating compounds have recently been identified. However, compounds that can treat or reduce the severity of cystic fibrosis and other CFTR mediated diseases, and particularly the more severe forms of these diseases, are still needed. [0010] One aspect of the disclosure provides novel compounds, including compounds of Formula I, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing. Formula I encompasses compounds falling within the following structure: tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing (i.e., pharmaceutically acceptable salts of compounds, tautomers, and deuterated derivatives), wherein: Ring A is selected from 7- and 8-membered heterocycles containing 0-2 additional heteroatoms selected from O and N, wherein Ring A is substituted with 0-4 groups selected from: • C3-C6 cycloalkyl (substituted with 0-4 groups selected from C1-C4 alkyl), • C1-C5 alkyl (substituted with 0-4 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, oxo, and 4- to 6-membered heterocycle); • C1-C4 alkoxy (substituted with 0-4 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, and oxo); • 5- and 6-membered heterocycle; • OH; • NH2; and • oxo; wherein two substituents on Ring A taken together with the atoms to which they are bonded may form a ring selected from C3-C6 cycloalkyl (further substituted with 0-2 groups selected from C1-C4 alkyl and C1-C4 alkoxy); Ring B is a 9-12 membered heteroaryl group containing 1-4 heteroatoms selected from N, O, and S, wherein Ring B is substituted with 1-4 groups selected from: • C1-C6 alkyl (substituted with 0-3 groups selected from halogen, C1-C3 alkoxy, and OH); • C3-C6 cycloalkyl, substituted with 0-3 groups selected from: o C1-C6 alkyl (substituted with 0-3 groups selected from C1-C3 alkoxy and halogen); o OH; and o halogen; • halogen; and • 5- and 6-membered heterocycle; each X is independently selected from CH and N; Z is selected from CH and N; R1 is selected from hydrogen, halogen, C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy (substituted with 0-5 halogens); each R2 is independently selected from C1-C5 alkyl, C1-C3 alkoxy, OH, and halogen; each R3 is independently selected from C1-C3 alkyl, C1-C3 alkoxy, OH, and halogen; n is an integer selected from 2 to 5; and p is an integer selected from zero to 3. [0011] Formula I also includes compounds of Formula I′: tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing (i.e., pharmaceutically acceptable salts of compounds, tautomers, and deuterated derivatives), wherein: Ring A is selected from 7- and 8- membered heterocycles containing 0-2 additional heteroatoms selected from O and N, wherein Ring A is substituted with 0-2 groups selected from: • C3-C6 cycloalkyl (substituted with 0-2 groups selected from C1-C4 alkyl), • C1-C5 alkyl (substituted with 0-2 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, oxo, and C4-C6 heterocycle); • C1-C4 alkoxy (substituted with 0-2 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, and oxo); • 5- and 6-membered heterocycle; • OH; • NH2; and • oxo; Ring B is a 9-12 membered heteroaryl group containing 1-4 heteroatoms selected from N, O, and S, wherein Ring B is substituted with 1-4 groups selected from: • C1-C6 alkyl (substituted with 0-3 groups selected from halogen, C1-C3 alkoxy, and OH); • C3-C6 cycloalkyl, substituted with 0-3 groups selected from: o C1-C6 alkyl (substituted with 0-3 groups selected from C1-C3 alkoxy and halogen); o OH; and o halogen; • halogen; and • 5- and 6-membered heterocycle; each X is independently selected from CH and N; Z is selected from CH and N; R1 is selected from hydrogen, halogen, C1-C5 alkyl, C1-C5 haloalkyl, and C1-C5 alkoxy (substituted with 0-5 halogens); each R2 is independently selected from C1-C5 alkyl, C1-C3 alkoxy, OH, and halogen; each R3 is independently selected from C1-C3 alkyl, C1-C3 alkoxy, OH, and halogen; n is an integer selected from 2 to 5; and p is an integer selected from zero to 3. [0012] Formula I also includes compounds of Formula Ia: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein all variables are as defined for Formula I′. [0013] Formula I also includes compounds of Formula Ib: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein all variables are as defined for Formula I′. [0014] Formula I also includes compounds of Formula Ic: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein all variables are as defined for Formula I′. [0015] One aspect of the disclosure provides novel compounds, including compounds of Formula II, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing. Formula II encompasses compounds falling within the following structure: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein R4 is selected from: • C3-C6 cycloalkyl (substituted with 0-2 groups selected from C1-C4 alkyl), • C1-C5 alkyl (substituted with 0-2 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, oxo, and C3-C6 heterocycle); • C1-C4 alkoxy (substituted with 0-2 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, and oxo); • 5- to 6-membered heterocycle; and • OH; • NH2; and • oxo; and wherein all other variables are as defined for Formula I′. [0016] Formula II also includes compounds of Formula IIa: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein R4 is as defined for Formula II, and wherein all other variables are as defined for Formula I′. [0017] Formula II also includes compounds of Formula IIb: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein R4 is as defined for Formula II, and wherein all other variables are as defined for Formula I′. [0018] Formula II also includes compounds of Formula IIc: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein R4 is as defined for Formula II, and wherein all other variables are as defined for Formula I′. [0019] Formula I also includes compounds of Formula IIIa: (IIIa), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein: Y1 is selected from O and -CH2-; Y2 is selected from NR9 and CH; Y3 is selected from O and CH; R4 is selected from: C1-C4 alkyl (substituted with 0-3 groups selected from halogen and cyclopropyl); C1-C4 alkoxy; and C3-C4 cycloalkyl; and R8 is selected from C1-C4 alkyl and C3-C4 cycloalkyl, each of which is substituted with 0-1 trifluoromethyl groups and, if R9 is present, R9 is methyl; or R8 and R9 are taken together to form a five-membered heterocyclic ring, substituted with 0-2 methyl groups. [0020] Formula I also includes compounds of Formula IIIb: (IIIb), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein: R4 is selected from: C1-C4 alkyl (substituted with 0-3 groups selected from halogen and cyclopropyl); C1-C4 alkoxy; and C3-C4 cycloalkyl; and R8 is selected from C3 alkyl and C3-C4 cycloalkyl, each of which is substituted with 0-1 trifluoromethyl groups and R9 is methyl; or R8 and R9 are taken together to form a five-membered heterocyclic ring, substituted with 0-2 methyl groups. [0021] In some embodiments, the compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb are chosen from Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing. [0022] Another aspect of the disclosure provides pharmaceutical compositions comprising at least one compound chosen from the novel compounds disclosed herein, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and at least one pharmaceutically acceptable carrier, which compositions may further include at least one additional active pharmaceutical ingredient. In some embodiments of the pharmaceutical compositions disclosed herein, the at least one additional active pharmaceutical ingredient is at least one other CFTR modulator. In some embodiments, the at least one other CFTR modulator is selected from CFTR potentiators and CFTR modulators. [0023] Thus, another aspect of the disclosure provides methods of treating the CFTR- mediated disease cystic fibrosis comprising administering at least one compound chosen from the novel compounds disclosed herein, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and at least one pharmaceutically acceptable carrier, optionally as part of a pharmaceutical composition comprising at least one additional component, to a subject in need thereof. In some embodiments, the at least one additional active pharmaceutical ingredient in the methods of treating disclosed herein is at least one other CFTR modulator. In some embodiments, the at least one other CFTR modulator is selected from CFTR potentiators and CFTR correctors. [0024] In certain embodiments, the pharmaceutical compositions of the disclosure comprise at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing. In certain embodiments, the pharmaceutical compositions of the disclosure comprise at least one compound chosen from Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing. [0025] In some embodiments, compositions comprising at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing may optionally further comprise (a) at least one (i.e., one or more) compound chosen from (R)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-N-(1-(2,3- dihydroxypropyl)-6-fluoro-2-(1-hydroxy-2-methylpropan-2-yl)-1H-indol-5- yl)cyclopropanecarboxamide (tezacaftor), 3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5- yl)cyclopropane carboxamido)-3-methylpyridin-2-yl)benzoic acid (lumacaftor), deuterated derivatives of tezacaftor and lumacaftor, and pharmaceutically acceptable salts of any of the foregoing; and/or (b) at least one (i.e., one or more) compound chosen from N-[2,4-bis(1,1- dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide (ivacaftor), N-(2- (tert-butyl)-5-hydroxy-4-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenyl)-4-oxo-1,4- dihydroquinoline-3-carboxamide (deutivacaftor), (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, and (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol; deuterated derivatives of ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa- 3,4,18-triazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol, and (6R)-17-amino- 12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol; and pharmaceutically acceptable salts of any of the foregoing. [0026] In some embodiments, compositions comprising at least one compound chosen from Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing may optionally further comprise (a) at least one (i.e., one or more) compound chosen from (R)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-N-(1-(2,3- dihydroxypropyl)-6-fluoro-2-(1-hydroxy-2-methylpropan-2-yl)-1H-indol-5- yl)cyclopropanecarboxamide (tezacaftor), 3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5- yl)cyclopropane carboxamido)-3-methylpyridin-2-yl)benzoic acid (lumacaftor), deuterated derivatives of tezacaftor and lumacaftor, and pharmaceutically acceptable salts of any of the foregoing, and/or (b) at least one (i.e., one or more) compound chosen from N-[2,4-bis(1,1- dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide (ivacaftor), N-(2- (tert-butyl)-5-hydroxy-4-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenyl)-4-oxo-1,4- dihydroquinoline-3-carboxamide (deutivacaftor), (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, and (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol; deuterated derivatives of ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa- 3,4,18-triazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol, and (6R)-17-amino- 12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo [12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol; and pharmaceutically acceptable salts of any of the foregoing. [0027] Another aspect of the disclosure provides methods of treating the CFTR-mediated disease cystic fibrosis comprising administering to a patient in need thereof at least one compound chosen from the novel compounds disclosed herein, pharmaceutically acceptable salts thereof, and deuterated derivatives of any of the foregoing, and optionally further administering one or more additional CFTR modulating agents selected from tezacaftor, ivacaftor, and lumacaftor. [0028] In a further aspect, compounds of the disclosure (e.g., compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), and pharmaceutical compositions comprising those compounds, and optionally further comprising one or more CFTR modulating agents, are used in therapy or in the manufacture of a medicament. In some embodiments, the one or more additional CFTR modulating agents are selected from CFTR potentiators. In some embodiments, the one or more additional CFTR modulating agents are selected from CFTR correctors. In some embodiments, the one or more additional CFTR modulating agents are selected from tezacaftor, lumacaftor, ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and (6R)-17-amino-12,12- dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5] nonadeca- 1(18),2,4,14,16-pentaen-6-ol; and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing. [0029] A further aspect of the disclosure provides intermediates and methods for making the compounds and compositions disclosed herein. Definitions [0030] “Tezacaftor” as used herein, refers to (R)-1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-N- (1-(2,3-dihydroxypropyl)-6-fluoro-2-(1-hydroxy-2-methylpropan-2-yl)-1H-indol-5- yl)cyclopropanecarboxamide, which can be depicted with the following structure: . Tezacaftor may be in the form of a deuterated derivative, a pharmaceutically acceptable salt, or a pharmaceutically acceptable salt of a deuterated derivative. Tezacaftor and methods of making and using tezacaftor are disclosed in WO 2010/053471, WO 2011/119984, WO 2011/133751, WO 2011/133951, WO 2015/160787, and US 2009/0131492, each of which is incorporated herein by reference. [0031] “Ivacaftor” as used throughout this disclosure refers to N-[2,4-bis(1,1-dimethylethyl)- 5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide, which is depicted by the structure: . Ivacaftor may also be in the form of a deuterated derivative, a pharmaceutically acceptable salt, or a pharmaceutically acceptable salt of a deuterated derivative. Ivacaftor and methods of making and using ivacaftor are disclosed in WO 2006/002421, WO 2007/079139, WO 2010/108162, and WO 2010/019239, each of which is incorporated herein by reference. [0032] In some embodiments, a deuterated derivative of ivacaftor (deutivacaftor) is employed in the compositions and methods disclosed herein. A chemical name for deutivacaftor is N-(2-(tert-butyl)-5-hydroxy-4-(2-(methyl-d3)propan-2-yl-1,1,1,3,3,3-d6)phenyl)-4-oxo-1,4- dihydroquinoline-3-carboxamide, as depicted by the structure: . Deutivacaftor may be in the form of a further deuterated derivative, a pharmaceutically acceptable salt, or a pharmaceutically acceptable salt of a deuterated derivative. Deutivacaftor and methods of making and using deutivacaftor are disclosed in WO 2012/158885, WO 2014/078842, and US Patent No.8,865,902, each of which is incorporated herein by reference. [0033] “Lumacaftor” as used herein, refers to 3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5- yl)cyclopropanecarboxamido)-3-methylpyridin-2-yl)benzoic acid, which is depicted by the chemical structure: . [0034] Lumacaftor may be in the form of a deuterated derivative, a pharmaceutically acceptable salt, or a pharmaceutically acceptable salt of a deuterated derivative. Lumacaftor and methods of making and using lumacaftor are disclosed in WO 2007/056341, WO 2009/073757, and WO 2009/076142, each of which is incorporated herein by reference. [0035] (6R,12R)-17amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, deuterated derivatives, and pharmaceutically acceptable salts and methods of making and using (6R,12R)-17-amino-12- methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca- 1(18),2,4,14,16-pentaen-6-ol are described in WO 2022/032068, WO 2023/150236, and WO 2023/012230, each of which is incorporated herein by reference. [0036] (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo [12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, deuterated derivatives, and pharmaceutically acceptable salts and methods of making and using (6R)-17-amino-12,12- dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo [12.3.1.12,5]nonadeca- 1(18),2,4,14,16-pentaen-6-ol are described in WO 2022/109573, WO 2023/224924, and WO 2023/224931, each of which is incorporated herein by reference. [0037] As used herein, the term “alkyl” refers to a saturated or partially saturated, branched, or unbranched (linear) aliphatic hydrocarbon containing carbon atoms (such as, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms), which may contain a double (alkenyl) or triple (alkynyl) bond between one or more sets of adjacent carbon atoms. Alkyl groups may be substituted or unsubstituted. [0038] The term “aliphatic” or “aliphatic group,” as used herein, means a straight-chain (i.e., unbranched) or branched, substituted, or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic (also referred to herein as “cycloaliphatic,” “carbocycle,” or “cycloalkyl”), that has a single point of attachment to the rest of the molecule. Unless otherwise specified, aliphatic groups contain 1-20 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-10 aliphatic carbon atoms. In other embodiments, unless otherwise specified, aliphatic groups contain 1-8 aliphatic carbon atoms. In still other embodiments, unless otherwise specified, aliphatic groups contain 1-6 aliphatic carbon atoms, and in yet other embodiments, unless otherwise specified, aliphatic groups contain 1-4 aliphatic carbon atoms. In some embodiments, “cycloaliphatic” (or “carbocycle” or “cycloalkyl”) refers to a monocyclic C3-8 hydrocarbon or bicyclic or tricyclic C8-14 hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic, that has a single point of attachment to the rest of the molecule wherein any individual ring in said bicyclic ring system has 3-7 members. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and hybrids thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, and (cycloalkyl)alkenyl. Suitable cycloaliphatic groups include cycloalkyl, bicyclic cycloalkyl (e.g., decalin), bridged bicycloalkyl such as norbornyl or [2.2.2]bicyclo-octyl, and bridged tricyclic such as adamantyl. [0039] As used herein, the term “unsaturated” means that a moiety has one or more units of unsaturation. [0040] As used herein, the term “pi bond” means a covalent bond formed by the p orbitals of adjacent atoms. Pi bonds exist where there is a multiple bond, i.e., a double or triple bond, between two atoms. For example, a carbon-carbon double bond consists of one pi bond, and a carbon-carbon triple bond consists of two pi bonds. [0041] As used herein, the term “haloalkyl group” refers to an alkyl group substituted with one or more halogen atoms, e.g., fluoroalkyl, which refers to an alkyl group substituted with one or more fluorine atoms. In some embodiments, one carbon atom of the alkyl group is substituted with one or more halogen atoms. In some embodiments, each carbon atom of the alkyl group is substituted with one or more halogen atoms. In some embodiments, one or more carbon atoms of the alkyl group is a perhalo carbon atom (i.e., all hydrogen atoms of the alkyl group are substituted by halogen atoms). In some embodiments, each carbon atom of the alkyl group is a perhalo carbon atom. Non-limiting examples of fluoroalkyl include —CHF2, —CH2F, —CF3, —CF2—, and perhaloalkyl, such as —CF2CF3. [0042] As used herein, the term “halogen” or “halo” means F, Cl, Br, or I. [0043] As used herein, the terms “oxo” and “=O” refer to a substituent oxygen atom connected to another atom by a double bond. [0044] The term “alkoxy,” as used herein, refers to an alkyl or cycloalkyl covalently bonded to an oxygen atom. Alkoxy groups may be linear or branched, and substituted or unsubstituted. [0045] As used herein, “cycloalkyl” refers to a cyclic, bicyclic, tricyclic, or polycyclic non- aromatic hydrocarbon groups having 3 to 12 carbons (such as, for example 3-10 carbons) and may include one or more unsaturated bonds. “Cycloalkyl” groups encompass monocyclic, bicyclic, tricyclic, bridged, fused, and spiro rings, including mono spiro and dispiro rings. Non- limiting examples of cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantyl, norbornyl, dispiro[2.0.2.1]heptane, bicyclo[1.1.1]pentane, and spiro[2,3]hexane. Cycloalkyl groups may be substituted or unsubstituted. [0046] The term “aryl,” as used herein, is a functional group or substituent derived from an aromatic ring and encompasses monocyclic aromatic rings and bicyclic, tricyclic, and fused ring systems wherein at least one ring in the system is aromatic. An aryl group may be optionally substituted with one or more substituents. Non-limiting examples of aryl groups include phenyl, naphthyl, and 1,2,3,4-tetrahydronaphthalenyl. [0047] The term “heteroatom” means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the quaternized form of any basic nitrogen; and a substitutable nitrogen of a heterocyclic ring, for example, N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR+ (as in N-substituted pyrrolidinyl)). In some embodiments, “heteroatom” refers to oxygen, sulfur, and nitrogen. [0048] The term “heteroaliphatic,” as used herein, means aliphatic groups wherein one or more carbon atoms are independently replaced with one or more heteroatoms, for example, oxygen, sulfur, nitrogen, phosphorus, or silicon. In some embodiments, the heteroatoms are selected from oxygen, sulfur, and nitrogen. Heteroaliphatic groups may be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and include “heterocycle,” “heterocyclyl,” “heterocycloaliphatic,” and “heterocyclic” groups. [0049] The term “heteroaryl ring,” as used herein, refers to an aromatic ring comprising at least one ring atom that is a heteroatom, such as O, N, S, P, and Si. In some embodiments, the heteroaryl ring contains 1-5 heteroatoms selected from oxygen, sulfur, and nitrogen. In some embodiments, the heteroaryl ring contains 1-3 heteroatoms selected from oxygen, sulfur, and nitrogen. Heteroaryl groups encompass monocyclic rings and bicyclic, tricyclic, bridged, fused, and spiro ring systems (including monospiro and dispiro rings) having a total of five to fourteen ring members, wherein at least one ring in the system is aromatic, at least one ring in the system contains one or more heteroatoms, and wherein each ring in the system contains three to seven ring members. Heteroaryl groups also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloalkyl, or heterocyclyl rings, where the point of attachment is on the heteroaromatic ring. Non-limiting examples of heteroaryl rings include pyridine, quinoline, indole, indoline, azaindole, pyrrolopyrazine, furopyridine, and furopyrazine. A heteroaryl group may be optionally substituted with one or more substituents. In certain embodiments, the term “heteroaryl ring” encompasses heteroaryl rings with various oxidation states, such as heteroaryl rings containing N-oxides and sulfoxides. Non-limiting examples of such heteroaryl rings include pyrimidine N-oxides, quinoline N-oxides, thiophene S-oxides, and pyrimidine N-oxides. [0050] As used herein, the term “heterocycle” is used interchangeably with “heterocyclyl ring,” and refers to a non-aromatic hydrocarbon containing 3 to 12 atoms in a ring (such as, for example 3-10 atoms) comprising at least one ring atom that is a heteroatom, such as O, N, S, P, or Si and may include one or more unsaturated bonds. In some embodiments, the heterocyclyl ring contains 1-5 heteroatoms selected from oxygen, sulfur, and nitrogen. In some embodiments, the heterocyclyl ring contains 1-3 heteroatoms selected from oxygen, sulfur, and nitrogen. Heterocyclyl rings encompass monocyclic, bicyclic, tricyclic, polycyclic, bridged, fused, and spiro rings, including monospiro and dispiro rings. [0051] In some embodiments, a C3-C6 heterocycle is a heterocycle selected from, for example: [0052] In some embodiments, a C4-C6 heterocycle is a heterocycle selected from, for example: [0053] The bond designation “ ” is intended to reflect the presence of an aromatic (i.e., conjugated) ring system. [0054] It will be appreciated that certain compounds of this disclosure may exist as separate stereoisomers or enantiomers and/or mixtures of those stereoisomers or enantiomers. As used in the chemical structures disclosed herein, a “wedge” ( ) or “hash” ( ) bond to a stereogenic atom indicates a chiral center of known absolute stereochemistry (i.e., one stereoisomer). As used in the chemical structures disclosed herein, a “wavy” bond ( ) to a stereogenic atom indicates a chiral center of unknown absolute stereochemistry (i.e., one stereoisomer). As used in the chemical structures disclosed herein, a “wavy” bond ( ) to a double-bonded carbon indicates a mixture of E/Z isomers. As used in the chemical structures disclosed herein, a (“straight”) bond to a stereogenic atom indicates where there is a mixture (e.g., racemic or enriched). As used herein, two (“straight”) bonds to a double-bonded carbon indicates that the double bond possesses the E/Z stereochemistry as drawn. As used in the chemical structures A disclosed herein, a (a “wavy” line perpendicular to a “straight” bond to group “A”) indicates that group “A” is a substituent whose point of attachment is at the end of the bond that terminates at the “wavy” line. As used herein, a stereogenic atom that is notated with an (R) or (S) indicates the stereochemical designation of the stereogenic atom under the Cahn-Ingold- Prelog convention. [0055] Certain compounds can exist as atropisomers. It will be appreciated that certain compounds of this disclosure may exist as separated atropisomers and/or mixtures of those atropisomers, i.e., a subclass of stereoisomers resulting from hindered rotation about single bonds or chirality axis and that can be isolated as separate chemical species. As used herein, a stereogenic unit that is notated with a (P) or (M) indicates the stereochemical designation of the stereogenic unit under the Cahn-Ingold-Prelog convention (basic terminology of stereochemistry, IUPAC Recommendations 1996, Pure & Appl. Chem., Vol 68, No.12, pp. 2193-2222, 1996). [0056] Certain compounds disclosed herein may exist as tautomers and both tautomeric forms are intended, even though only a single tautomeric structure is depicted. For example, a description of Compound X is understood to include its tautomer Compound Y and vice versa, as well as mixtures thereof: Compound X Compound Y . Unless otherwise stated, all tautomeric forms of the compounds of the disclosure are within the scope of the disclosure. [0057] “Tert” and “t-” are used interchangeably and mean tertiary. [0058] Compounds described herein may optionally be substituted with one or more substituents, such as are illustrated generally above, or as exemplified by particular classes, subclasses, and species of the disclosure. It will be appreciated that the phrase “optionally substituted” is used interchangeably with the phrase “substituted or unsubstituted.” “Substituted,” whether preceded by the term “optionally” or not, indicates that at least one hydrogen of the “substituted” group is replaced by a substituent. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent chosen from a specified group, the substituent may be either the same or different at each position. Combinations of substituents envisioned by this disclosure are preferably those that result in the formation of stable or chemically feasible compounds. [0059] The term “stable,” as used herein, refers to compounds that are not substantially altered when subjected to conditions to allow for their production, detection, and preferably their recovery, purification, and use for one or more of the purposes disclosed herein. [0060] The term “stable compounds,” as used herein, refers to compounds which possess sufficient stability to allow for their manufacture and which maintain the integrity of the compounds for a sufficient period of time to be useful for the purposes detailed herein (e.g., formulation into therapeutic products, intermediates for use in production of therapeutic compounds, isolatable or storable intermediates, and/or treating a disease or condition responsive to therapeutic agents). [0061] In the compounds of this disclosure, any atom not specifically designated as a particular isotope is meant to represent any stable isotope of that atom. Unless otherwise stated, when a position is designated specifically as “H” or “hydrogen,” the position is understood to have hydrogen at its natural abundance isotopic composition. [0062] As used herein, the term “derivative” refers to a collection of molecules having a chemical structure identical to a compound of this disclosure, except that one or more atoms of the molecule may have been substituted with another atom. Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of hydrogen by deuterium or tritium, or the replacement of a carbon by a 13C or 14C, are within the scope of this disclosure. Such compounds are useful as, for example, analytical tools, probes in biological assays, or compounds with improved therapeutic profiles. [0063] As used herein, “deuterated derivative(s)” refers to a compound having the same chemical structure as a reference compound, with one or more hydrogen atoms replaced by a deuterium atom. In some embodiments, the one or more hydrogens replaced by deuterium are part of an alkyl group. In some embodiments, the one or more hydrogens replaced by deuterium are part of a methyl group. In chemical structures, deuterium may be represented as “D.” In compounds identified as a deuterated derivative, or containing a “D” substitution, the deuterium atom is present in amounts far exceeding its natural abundance; for example, the compound may have an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom). [0064] As used herein, “CFTR” means cystic fibrosis transmembrane conductance regulator. [0065] As used herein, the term “modulator” refers to a compound that increases the activity of a biological compound or molecule such as a protein. [0066] As used herein, the term “CFTR modulator” refers to a compound that increases the activity of CFTR. The increase in activity resulting from a CFTR modulator includes, but is not limited to, compounds that correct, potentiate, stabilize, and/or amplify CFTR. [0067] As used herein, the terms “corrector” and “CFTR corrector” are used interchangeably and refer to a compound that facilitates the processing and trafficking of CFTR to increase the amount of CFTR at the cell surface. The novel compounds disclosed herein are CFTR correctors. Tezacaftor and lumacaftor, and deuterated derivatives and pharmaceutically acceptable salts thereof as referenced herein, are correctors. Other CFTR correctors include those disclosed in WO 2023/034946, WO 2023/034992, WO 2021/097054, WO 2021/097057, WO 2021/113806, WO 2021/113808, and WO 2021113809, such as SION-109, SION-638, SION-719, and SION-451. Additional CFTR correctors include ABBV-2851, disclosed in WO 2017/009804, ABBV-3748; ABBV-3903; and ABBV-119. [0068] As used herein, the terms “potentiator” and “CFTR potentiator” refer to a compound that increases the channel activity of CFTR protein located at the cell surface, resulting in enhanced ion transport. Ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, and (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol; as referenced herein, are CFTR potentiators. It will be appreciated that a description of a combination of compounds that includes a compound of the disclosure (e.g., compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing) will typically but not necessarily include a CFTR potentiator, such as, e.g., ivacaftor, deutivacaftor, (6R,12R)-17-amino-12- methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5] nonadeca- 1(18),2,4,14,16-pentaen-6-ol, and (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19- oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and (6R)-17- amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol; or a deuterated derivative or pharmaceutically acceptable salt of any of the foregoing. In addition, the combination will typically, but not necessarily, include only a single potentiator, but may include more than one corrector. Thus, in some embodiments, a combination of at least one compound of the disclosure (e.g., a compound selected from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), will include a potentiator selected from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19- dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and (6R)-17- amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol; or deuterated derivatives or pharmaceutically acceptable salts thereof and may also include another CFTR corrector, such as, e.g., a corrector compound selected from tezacaftor, lumacaftor, and deuterated derivatives and pharmaceutically acceptable salts thereof. In some embodiments, such a combination may also include a CFTR potentiator enhancer. [0069] As used herein, the term “CFTR potentiator enhancer,” “CFTR potentiation enhancer,” and “CFTR co-potentiator” are used interchangeably and refer to a compound that enhances CFTR potentiation. [0070] The term “compound,” when referring to a compound of this disclosure, refers to a collection of molecules having an identical chemical structure, except that there may be isotopic variation among the constituent atoms of the molecules. [0071] The phrase “a novel compound of the disclosure” refers to a compound chosen from compounds of any one of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing. Specifically excluded from the phrase “a novel compound of the disclosure is any compound disclosed in PCT/US2021/053858 (incorporated herein by reference). [0072] The term “at least one compound selected from,” as used herein, refers to the selection of one or more of the compounds from a specified group. “Selected from” and “chosen from” may be used interchangeably herein. [0073] A reference to “Compounds 1 to 139” herein is intended to represent a reference to each of Compounds 1 through 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139) encompassed by Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc IIIa, and/or IIIb, and disclosed in Tables 2 and 4 herein, individually or as a group. [0074] As used herein, the term “active pharmaceutical ingredient” or “therapeutic agent” (“API”) refers to a biologically active compound. [0075] The terms “patient” and “subject” are used interchangeably and refer to an animal, including a human. [0076] The terms "effective dose" and "effective amount" are used interchangeably herein and refer to that amount of a compound that produces the desired effect for which it is administered (e.g., improvement in CF or a symptom of CF, or lessening the severity of CF or a symptom of CF). The exact amount of an effective dose will depend on the purpose of the treatment and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lloyd (1999) The Art, Science and Technology of Pharmaceutical Compounding). [0077] As used herein, the terms "treatment," "treating," and the like generally mean the improvement in one or more symptoms of CF or lessening the severity of CF or one or more symptoms of CF in a subject. “Treatment,” as used herein, includes, but is not limited to, the following: increased growth of the subject, increased weight gain, reduction of mucus in the lungs, improved pancreatic and/or liver function, reduction of chest infections, and/or reductions in coughing or shortness of breath. Improvements in or lessening the severity of any of these symptoms can be readily assessed according to standard methods and techniques known in the art. [0078] It should be understood that references herein to methods of treatment (e.g., methods of treating a CFTR mediated disease or a method of treating cystic fibrosis) using one or more compounds of the disclosure optionally in combination with one or more additional CFTR modulating agents (e.g., a compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, optionally in combination with one or more additional CFTR modulating agents) should also be interpreted as references to: - one or more compounds of the disclosure (e.g., a compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, optionally in combination with one or more additional CFTR modulating agents) for use in methods of treating, e.g., cystic fibrosis, optionally in combination with one or more additional CFTR modulating agents; and/or - the use of one or more compounds of the disclosure (e.g., a compound chosen compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, optionally in combination with one or more additional CFTR modulating agents) in the manufacture of a medicament for treating, e.g., cystic fibrosis. [0079] It should be also understood that references herein to methods of treatment (e.g., methods of treating a CFTR mediated disease or a method of treating cystic fibrosis) using a pharmaceutical composition of the disclosure (e.g., a pharmaceutical composition comprising at least one compound chosen from compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and optionally further comprising one or more additional CFTR modulating agents) should also be interpreted as references to: - a pharmaceutical composition (e.g., a pharmaceutical composition comprising at least one compound chosen from compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and optionally further comprising one or more additional CFTR modulating agents) for use in methods of treating, e.g., cystic fibrosis; and/or - the use of a pharmaceutical composition (e.g., a pharmaceutical composition comprising at least one compound chosen from compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and optionally further comprising one or more additional CFTR modulating agents) in the manufacture of a medicament for treating, e.g., cystic fibrosis. [0080] As used herein, the term “in combination with,” when referring to two or more compounds, agents, or additional active pharmaceutical ingredients, means the administration of two or more compounds, agents, or active pharmaceutical ingredients to the patient prior to, concurrent with, or subsequent to each other. [0081] As used herein, the terms “about” and “approximately,” when used in connection with amounts, volumes, reaction times, reaction temperatures, etc. mean an acceptable error for a particular value as determined by one of ordinary skill in the art, which depends in part on how the value is measured or determined. In some embodiments, the terms “about” and “approximately” mean within 1, 2, 3, or 4 standard deviations. In certain embodiments, the terms “about” and “approximately” mean within 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, or 0.05% of a given value or range. In some embodiments, “about” and “approximately” mean within 15% of a given value or range. . In some embodiments, “about” and “approximately” mean within 10% of a given value or range. . In some embodiments, “about” and “approximately” mean within 5% of a given value or range. As used herein, the symbol appearing immediately before a numerical value has the same meaning as the terms “about” and “approximately.” [0082] The term “at least one” refers to one or more. [0083] As used herein, the term “solvent” refers to any liquid in which the product is at least partially soluble (solubility of product >1 g/L). [0084] Non-limiting examples of suitable solvents that may be used in this disclosure include, for example, water (H2O), methanol (MeOH), methylene chloride or dichloromethane (DCM; CH2Cl2), acetonitrile (MeCN; CH3CN; ACN), N,N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), methyl acetate (MeOAc), ethyl acetate (EtOAc), isopropyl acetate (IPAc), tert-butyl acetate (t-BuOAc), isopropyl alcohol (IPA), tetrahydrofuran (THF), 2-methyl tetrahydrofuran (2-MeTHF), methyl ethyl ketone (MEK), tert-butanol, diethyl ether (Et2O), methyl tert-butyl ether (MTBE), 1,4-dioxane, and N-methylpyrrolidone (NMP). [0085] As used herein, the term “ambient conditions” means room temperature, open air condition and uncontrolled humidity condition. As used herein, the term “room temperature” or “ambient temperature” means 15 °C to 30 °C. [0086] As used herein, “mutations” can refer to mutations in the CFTR gene or the CFTR protein. A “CFTR gene mutation” refers to a mutation in the CFTR gene, and a “CFTR protein mutation” refers to a mutation in the CFTR protein. In general, a genetic defect or mutation, or a change in the nucleotides in a gene, results in a mutation in the CFTR protein translated from that gene, or a frame shift(s). [0087] As used herein, “minimal function (MF) mutations” refer to CFTR gene mutations associated with minimal CFTR function (little-to-no functioning CFTR protein) and include, for example, mutations associated with severe defects in ability of the CFTR channel to open and close, known as defective channel gating or “gating mutations”; mutations associated with severe defects in the cellular processing of CFTR and its delivery to the cell surface; mutations associated with no (or minimal) CFTR synthesis; and mutations associated with severe defects in channel conductance. [0088] As used herein, the term “F508del” refers to a mutant CFTR protein which is lacking the amino acid phenylalanine at position 508, or to a mutant CFTR gene which encodes for a CFTR protein lacking the amino acid phenylalanine at position 508. [0089] The disclosure also provides processes for preparing salts of the compounds of the disclosure. A salt of a compound of this disclosure is formed between an acid and a basic group of the compound, such as an amino functional group, or a base and an acidic group of the compound, such as a carboxyl functional group. In some embodiments, the salt is a pharmaceutically acceptable salt. [0090] The term “pharmaceutically acceptable,” as used herein, refers to a component that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and other mammals without undue toxicity, irritation, allergic response, and the like, and is commensurate with a reasonable benefit/risk ratio. [0091] As used herein, the term “pharmaceutically acceptable salt” means any non-toxic salt that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this disclosure. Pharmaceutically acceptable salts of the compounds of this disclosure include those derived from suitable inorganic and organic acids and bases. A “pharmaceutically acceptable counterion” is an ionic portion of a salt that is not toxic when released from the salt upon administration to a recipient. One of ordinary skill in the art would recognize that, when an amount of “a compound or a pharmaceutically acceptable salt thereof” is disclosed, the amount of the pharmaceutically acceptable salt form of the compound is the amount equivalent to the concentration of the free base of the compound. [0092] A “free base” form of a compound does not contain an ionically bonded salt. It is noted that the disclosed amounts of the compounds or their pharmaceutically acceptable salts thereof herein are based upon their free base form. For example, “10 mg of at least one compound chosen from Compound I and pharmaceutically acceptable salts thereof” includes 10 mg of Compound I and a concentration of a pharmaceutically acceptable salt of Compound I equivalent to 10 mg of Compound I. [0093] Suitable pharmaceutically acceptable salts are, for example, those disclosed in S. M. Berge, et al. J. Pharmaceutical Sciences, 1977, 66, 1-19. For example, Table 1 of that article provides the following pharmaceutically acceptable salts: Table 1 of S. M. Berge, et al.: [0094] Non-limiting examples of pharmaceutically acceptable acid addition salts include: salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, or perchloric acid; salts formed with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid; and salts formed by using other methods used in the art, such as ion exchange. Non-limiting examples of pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy- ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, and valerate salts. Pharmaceutically acceptable salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium, and N+(C1-4alkyl)4 salts. This disclosure also envisions the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. Suitable non-limiting examples of alkali and alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium. Further non-limiting examples of pharmaceutically acceptable salts include ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate. Other suitable, non-limiting examples of pharmaceutically acceptable salts include besylate and glucosamine salts. CFTR Modulator Compounds [0095] One aspect of the disclosure provides novel compounds, including compounds of Formula I, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing. Formula I encompasses compounds falling within the following structure: tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing (i.e., salts of compounds, tautomers, and deuterated derivatives), wherein: Ring A is selected from 7- and 8-membered heterocycles containing 0-2 additional heteroatoms selected from O and N, wherein Ring A is substituted with 0-4 groups selected from: • C3-C6 cycloalkyl (substituted with 0-4 groups selected from C1-C4 alkyl); • C1-C5 alkyl (substituted with 0-4 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, oxo, and 4- to 6-membered heterocycle); • C1-C4 alkoxy (substituted with 0-4 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, and oxo); • 5- and 6-membered heterocycle; • OH; • NH2; and • oxo; wherein two substituents on Ring A taken together with the atoms to which they are bonded may form a ring selected from C3-C6 cycloalkyl (further substituted with 0-2 groups selected from C1-C4 alkyl and C1-C4 alkoxy); Ring B is a 9-12 membered heteroaryl group containing 1-4 heteroatoms selected from N, O, and S, wherein Ring B is substituted with 1-4 groups selected from: • C1-C6 alkyl (substituted with 0-3 groups selected from halogen, C1-C3 alkoxy, and OH); • C3-C6 cycloalkyl, substituted with 0-3 groups selected from: o C1-C6 alkyl (substituted with 0-3 groups selected from C1-C3 alkoxy and halogen), o OH, and o halogen; • halogen; and • 5- and 6-membered heterocycle; each X is independently selected from CH and N; Z is selected from CH and N; R1 is selected from hydrogen, halogen, C1-C5 alkyl, C1-C5 haloalkyl, and C1-C5 alkoxy (optionally substituted with 0-5 halogens); each R2 is independently selected from C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy (substituted with 0-5 halogens), OH, and halogens; each R3 is independently selected from C1-C3 alkyl, C1-C3 alkoxy, OH, and halogen; n is an integer selected from 2 to 5; and p is an integer selected from 0 to 3. [0096] Formula I also includes compounds of Formula I′: tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing (i.e., salts of compounds, tautomers, and deuterated derivatives), wherein: Ring A is selected from 7- and 8- membered heterocycles containing 0-2 additional heteroatoms selected from O and N, wherein Ring A is substituted with 0-2 groups selected from: • C3-C6 cycloalkyl (substituted with 0-2 groups selected from C1-C4 alkyl), • C1-C5 alkyl (substituted with 0-2 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, oxo, and C4-C6 heterocycle); • C1-C4 alkoxy (substituted with 0-2 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, and oxo); • 5- and 6-membered heterocycle; • OH; • NH2; and • oxo; Ring B is a 9-12 membered heteroaryl group containing 1-4 heteroatoms selected from N, O, and S, wherein Ring B is substituted with 1-4 groups selected from: • C1-C6 alkyl (substituted with 0-3 groups selected from halogen, C1-C3 alkoxy, and OH); • C3-C6 cycloalkyl, substituted with 0-3 groups selected from: o C1-C6 alkyl (substituted with 0-3 groups selected from: C1-C3 alkoxy and halogen); o OH; and o halogen; • halogen; and • 5- and 6-membered heterocycle; each X is independently selected from CH and N; Z is selected from CH and N; R1 is selected from hydrogen, halogen, C1-C5 alkyl, C1-C5 haloalkyl, and C1-C5 alkoxy (substituted with 0-5 halogens); each R2 is independently selected from C1-C5 alkyl, C1-C3 alkoxy, OH, and halogen; each R3 is independently selected from C1-C3 alkyl, C1-C3 alkoxy, OH, and halogen; n is an integer selected from 2 to 5; and p is an integer selected from zero to 3. [0097] Formula I also includes compounds of Formula Ia: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein all variables are as defined for Formula I′. [0098] Formula I also includes compounds of Formula Ib: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein all variables are as defined for Formula I′. [0099] Formula I also includes compounds of Formula Ic: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein all variables are as defined for Formula I′. [00100] One aspect of the disclosure provides novel compounds, including compounds of Formula II, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing. Formula II encompasses compounds falling within the following structure: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein R4 is selected from: • C3-C6 cycloalkyl (substituted with 0-2 groups selected from C1-C4 alkyl), • C1-C5 alkyl (substituted with 0-2 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, oxo, and C3-C6 heterocycle); • C1-C4 alkoxy (substituted with 0-2 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, and oxo); • 5- and 6-membered heterocycle; • OH; • NH2; and • oxo; and wherein all other variables are as defined for Formula I′. [00101] Formula I also includes compounds of Formula II: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein R4 is selected from: • C3-C6 cycloalkyl (substituted with 0-2 groups selected from C1-C4 alkyl), • C1-C5 alkyl (substituted with 0-2 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, oxo, and C4-C6 heterocycle); • C1-C4 alkoxy (substituted with 0-2 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, and oxo); • 5- and 6-membered heterocycle; • OH; • NH2; and • oxo; and wherein all other variables are as defined for Formula I′. [00102] Formulae I and II also include compounds of Formula IIa: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein R4 is as defined for Formula II, and wherein all other variables are as defined for Formula I′. [00103] Formulae I and II also include compounds of Formula IIb: (IIb), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein R4 is as defined for Formula II, and wherein all other variables are as defined for Formula I′. [00104] Formulae I and II also include compounds of Formula IIc: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein R4 is as defined for Formula II, and wherein all other variables are as defined for Formula I′. [00105] Formulae I and II also include compounds of Formula IIIa: (IIIa), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein: Y1 is selected from O and -CH2-; Y2 is selected from NR9 and CH; Y3 is selected from O and CH; R4 is selected from: C1-C4 alkyl (substituted with 0-3 groups selected from halogen and cyclopropyl); C1-C4 alkoxy; and C3-C4 cycloalkyl; and R8 is selected from C1-C4 alkyl and C3-C4 cycloalkyl, each of which is substituted with 0-1 trifluoromethyl groups and, if R9 is present, R9 is methyl; or R8 and R9 are taken together to form a five-membered heterocyclic ring, substituted with 0-2 methyl groups. [00106] Formulae I and II also include compounds of Formula IIIb: tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein: R4 is selected from: C1-C4 alkyl (substituted with 0-3 groups selected from halogen and cyclopropyl); C1-C4 alkoxy; and C3-C4 cycloalkyl; and R8 is selected from C3 alkyl and C3-C4 cycloalkyl, each of which is substituted with 0-1 trifluoromethyl groups and R9 is methyl; or R8 and R9 are taken together to form a five-membered heterocyclic ring, substituted with 0-2 methyl groups. [00107] In some embodiments of Formula I, Ring A is selected from azepane or azocane, wherein one CH2 group is optionally replaced by an O atom, and wherein Ring A is substituted with 0-2 groups selected from: • C3 or C4 cycloalkyl (substituted with 0-1 methyl groups), • C1-C5 alkyl (substituted with 0-1 groups selected from C3-C4 cycloalkyl); and • C1-C3 alkoxy (substituted with 0-1 groups selected from C3-C4 cycloalkyl); Ring B is a 9-12 membered heteroaryl group containing 2-4 heteroatoms independently selected from N and O, wherein Ring B is substituted with 1-3 groups selected from: • C1-C4 alkyl, substituted with 0-3 groups selected from: o OMe, o halogen, and o OH; • C3-C4 cycloalkyl, substituted with 0-1 groups selected from: o C1 and C2 alkyl (substituted with 0-3 halogen atoms) and o CH2OCH3; • halogen; and • 6-membered heterocycle; X is N; Z is selected from CH and N; R1 is selected from H, CH3, OCH3, and F; each R2 is CH3; n is 2; and p is zero and R3 is absent. [00108] In some embodiments, Ring A in the compound, tautomer, deuterated derivative, or salt of Formulae I, I′, Ia, Ib, Ic, is selected from: [00109] In some embodiments, Ring A in the compound, tautomer, deuterated derivative, or salt of Formulae I, I′, Ia, Ib, Ic, is selected from: [00110] In some embodiments, Ring B in the compound, tautomer, deuterated derivative, or salt of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb is selected from: Q is selected from C and N; each W is independently selected from CH and N, W1 is selected from N, CH, and CF; Y is selected from N, NR, O, S, and CR; wherein Ring B contains 1 to 4 nitrogen atoms; R is selected from H and C1-C3 alkyl (substituted with 0-1 groups selected from C1-C3 alkoxy and OH); R5 is selected from H, C1-C4 alkyl, and C3 and C4 cycloalkyl; R6 is selected from: • H, • C1-C5 alkyl, • C1-C5 haloalkyl, • C3 and C4 cycloalkyl, substituted with 0-1 groups selected from: o C1-C4 alkyl, o CH2OCH3, and o C1-C3 haloalkyl; • 5- and 6-membered heterocycle; R7 is selected from C1-C3 alkyl; and R8 is selected from C1-C4 alkyl; and wherein R5 and R6 are not both hydrogen. [00111] In some embodiments, Ring B in the compound, tautomer, deuterated derivative, or salt of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb is selected from:
R is selected from H and C1-C3 (alkyl substituted with 0-1 groups selected from C1-C3 alkoxy and OH); R5 is selected from H, C1-C4 alkyl, and C3- and C4-cycloalkyl; R6 is selected from: • H, • C1-C5 alkyl, • C1-C5 haloalkyl, • C3 and C4 cycloalkyl, substituted with 0-1 groups selected from: o C1-C4 alkyl, o CH2OCH3, and o C1-C3 haloalkyl; • 5- and 6-membered heterocycle; R7 is selected from C1-C3 alkyl; R8 is selected from C1-C4 alkyl; and G is selected from halogens; and wherein R, R5 and R6 are not all simultaneously hydrogen, R5 and R6 are not simultaneously hydrogen when R is not present, and R6 is not hydrogen when R and R5 are not present. [00112] In some embodiments, Ring B in the compound, tautomers, deuterated derivatives, or salts of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb is selected from:
[00113] In some embodiments, Ring B in the compounds, tautomers, deuterated derivatives, or salts of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb is selected from: [00114] In some embodiments, the compounds of Formulae I or I′ are chosen from Compounds 1 to 107, tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
[00115] In some embodiments, the compounds of Formula I or I′ are chosen from Table 3. Table 3.
[00116] In some embodiments, the compounds of Formula I or I′ are chosen from Compounds 108 to 139, tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing. Table 4. Structures of Compounds 108 to 139
[00117] Because the general properties of silicon and germanium are similar to those of carbon, replacement of carbon by silicon or germanium can result in compounds with similar biological activity to a carbon-containing original compound. [00118] In some embodiments, the compounds of the invention include silicon derivatives of the compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein at least one carbon atom has been replaced with silicon. In some embodiments, the at least one carbon atom replaced with silicon may be a non-aromatic carbon. In some embodiments, the at least one carbon atom replaced with silicon may be an aromatic carbon. In certain embodiments, the silicon derivatives of the invention may also have one or more hydrogen atoms replaced with deuterium. In certain embodiments, the silicon derivatives of the invention may also have one or more carbon atoms replaced with germanium. [00119] In some embodiments, the compounds of the invention include germanium derivatives of the compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein at least one carbon atom has been replaced with germanium. In some embodiments, the at least one carbon atom replaced with germanium may be a non-aromatic carbon. In some embodiments, the at least one carbon atom replaced with germanium may be an aromatic carbon. In certain embodiments, the germanium derivatives of the invention may also have one or more hydrogen atoms replaced with deuterium. In certain embodiments, the germanium derivatives of the invention may also have one or more carbon atoms replaced with silicon. [00120] Exemplary embodiments of the invention include, but are not limited to, the following silicon and germanium derivatives:
Methods of Treatment [00121] Any of the novel compounds disclosed herein, such as, for example, a compound chosen from compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, can act as a CFTR modulator, i.e., it modulates CFTR activity in the body. Individuals suffering from a mutation in the gene encoding CFTR may benefit from receiving a CFTR modulator. A CFTR mutation may affect the CFTR quantity, i.e., the number of CFTR channels at the cell surface, or it may impact CFTR function, i.e., the functional ability of each channel to open and transport ions. Mutations affecting CFTR quantity include mutations that cause defective synthesis (Class I defect), mutations that cause defective processing and trafficking (Class II defect), mutations that cause reduced synthesis of CFTR (Class V defect), and mutations that reduce the surface stability of CFTR (Class VI defect). Mutations that affect CFTR function include mutations that cause defective gating (Class III defect) and mutations that cause defective conductance (Class IV defect). Some CFTR mutations exhibit characteristics of multiple classes. Certain mutations in the CFTR gene result in cystic fibrosis. [00122] Thus, in some embodiments, the disclosure provides methods of treating, lessening the severity of, or symptomatically treating cystic fibrosis in a patient comprising administering to the patient an effective amount of any of the novel compounds disclosed herein, such as, for example, a compound chosen from compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, alone or in combination with another active ingredient, such as one or more CFTR modulating agents. In some embodiments, the one or more CFTR modulating agents are selected from ivacaftor, deutivacaftor, lumacaftor, and tezacaftor. In some embodiments, the patient has an F508del/minimal function (MF) genotype, F508del/F508del genotype (homozygous for the F508del mutation), F508del/gating genotype, or F508del/residual function (RF) genotype. In some embodiments, the patient is heterozygous and has one F508del mutation. In some embodiments, the patient is homozygous for the N1303K mutation. [00123] In some embodiments, 1 mg to 500 mg of a compound disclosed herein, a tautomer thereof, deuterated derivatives of the compound and tautomer, or a pharmaceutically acceptable salt of any of the foregoing are administered daily. [00124] In some embodiments, the patient has at least one F508del mutation in the CFTR gene. In some embodiments, the patient has a CFTR gene mutation that is responsive to a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt of the invention based on in vitro data. In some embodiments, the disclosure also is directed to methods of treatment using isotope-labelled compounds of the afore-mentioned compounds, or pharmaceutically acceptable salts thereof, wherein the formula and variables of such compounds and salts are each and independently as described above or any other embodiments described above, provided that one or more atoms therein have been replaced by an atom or atoms having an atomic mass or mass number which differs from the atomic mass or mass number of the atom which usually occurs naturally (isotope labelled). Examples of isotopes which are commercially available and suitable for the disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, for example 2H, 3H, 13C, 14C, 15N, 18O, 17O, 31P, 32P, 35S, 18F, and 36Cl, respectively. [00125] The isotope-labelled compounds and salts can be used in a number of beneficial ways. They can be suitable for medicaments and/or various types of assays, such as substrate tissue distribution assays. For example, tritium (3H)- and/or carbon-14 (14C)-labelled compounds are particularly useful for various types of assays, such as substrate tissue distribution assays, due to relatively simple preparation and excellent detectability. For example, deuterium (2H)-labelled ones are therapeutically useful with potential therapeutic advantages over the non-2H-labelled compounds. In general, deuterium (2H)-labelled compounds and salts can have higher metabolic stability as compared to those that are not isotope-labelled owing to the kinetic isotope effect described below. Higher metabolic stability translates directly into an increased in vivo half-life or lower dosages, which could be desired. The isotope-labelled compounds and salts can usually be prepared by carrying out the procedures disclosed in the synthesis schemes and the related description, in the example part and in the preparation part in the present text, replacing a non-isotope-labelled reactant by a readily available isotope-labelled reactant. [00126] In some embodiments, the isotope-labelled compounds and salts are deuterium (2H)- labelled ones. In some specific embodiments, the isotope-labelled compounds and salts are deuterium (2H)-labelled, wherein one or more hydrogen atoms therein have been replaced by deuterium. In chemical structures, deuterium may be represented as “D” or “2H.” [00127] The concentration of the isotope(s) (e.g., deuterium) incorporated into the isotope- labelled compounds and salt of the disclosure may be defined by the isotopic enrichment factor. The term “isotopic enrichment factor” as used herein means the ratio between the isotopic abundance and the natural abundance of a specified isotope. In some embodiments, if a substituent in a compound of the disclosure is denoted as deuterium, such compound has an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation). Combination Therapies [00128] One aspect disclosed herein provides methods of treating cystic fibrosis and other CFTR mediated diseases using any of the novel compounds disclosed herein, such as, for example, compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, in combination with at least one additional active pharmaceutical ingredient. [00129] In some embodiments, at least one additional active pharmaceutical ingredient is selected from mucolytic agents, bronchodilators, antibiotics, anti-infective agents, and anti- inflammatory agents. [00130] In some embodiments, the additional therapeutic agent is an antibiotic. Exemplary antibiotics useful herein include tobramycin, including tobramycin inhaled powder (TIP), azithromycin, aztreonam, including the aerosolized form of aztreonam, amikacin, including liposomal formulations thereof, ciprofloxacin, including formulations thereof suitable for administration by inhalation, levofloxacin, including aerosolized formulations thereof, and combinations of two antibiotics, e.g., fosfomycin and tobramycin. [00131] In some embodiments, the additional agent is a mucolyte. Exemplary mucolytes useful herein include Pulmozyme®. [00132] In some embodiments, the additional agent is a bronchodilator. Exemplary bronchodilators include albuterol, metaproterenol sulfate, pirbuterol acetate, salmeterol, or terbutaline sulfate. [00133] In some embodiments, the additional agent is an anti-inflammatory agent, i.e., an agent that can reduce the inflammation in the lungs. Exemplary such agents useful herein include ibuprofen, docosahexaenoic acid (DHA), sildenafil, inhaled glutathione, pioglitazone, hydroxychloroquine, or simvastatin. [00134] In some embodiments, the additional agent is a nutritional agent. Exemplary nutritional agents include pancrelipase (pancreatic enzyme replacement), including Pancrease®, Pancreacarb®, Ultrase®, or Creon®, Liprotomase® (formerly Trizytek®), Aquadeks®, or glutathione inhalation. In some embodiments, the additional nutritional agent is pancrelipase. [00135] In some embodiments, at least one additional active pharmaceutical ingredient is selected from CFTR modulating agents. In some embodiments, the at least one additional active pharmaceutical ingredient is selected from CFTR potentiators. In some embodiments, the potentiator is selected from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing. In some embodiments, the at least one additional active pharmaceutical ingredient is chosen from CFTR correctors. In some embodiments, the correctors are selected from lumacaftor, tezacaftor, and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing. [00136] In some embodiments, the at least one additional active pharmaceutical ingredient is chosen from (a) tezacaftor, lumacaftor, and deuterated derivatives and pharmaceutically acceptable salts thereof; and/or (b) ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing. [00137] In some embodiments, the at least one additional active pharmaceutical ingredient is (a) a compound chosen from (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19- dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol and (6R)-17- amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, deuterated derivatives of the compound, and pharmaceutically acceptable salts of any of the foregoing, and may optionally include tezacaftor, deuterated derivatives, or pharmaceutically acceptable salts of any of the foregoing. [00138] Thus, in some embodiments, the combination therapies provided herein comprise (a) a compound selected from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing; and (b) at least one compound selected from tezacaftor, lumacaftor, and deuterated derivatives and pharmaceutically acceptable salts thereof; or (c) at least one compound selected from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing. In other embodiments, the combination therapies provided herein comprise (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing; (b) at least one compound selected from tezacaftor and pharmaceutically acceptable salts thereof; and (c) at least one compound selected from ivacaftor, deutivacaftor, and pharmaceutically acceptable salts thereof. In still other embodiments, the combination therapies provided herein comprise (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing; (b) at least one compound selected from tezacaftor, lumacaftor, and deuterated derivatives and pharmaceutically acceptable salts thereof; and/or (c) at least one compound selected from (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino-12,12- dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5]nonadeca- 1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof. In still other embodiments, the combination therapies provided herein comprise (a) a compound chosen from (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa- 3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol and (6R)-17-amino- 12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol, deuterated derivatives of the compound, and pharmaceutically acceptable salts of any of the foregoing, and (b) optionally tezacaftor, deuterated derivatives, or pharmaceutically acceptable salts of any of the foregoing. [00139] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in combination with at least one compound chosen from tezacaftor and pharmaceutically acceptable salts thereof. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in combination with at least one compound chosen from lumacaftor and pharmaceutically acceptable salts thereof. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in combination with at least one compound chosen from ivacaftor and pharmaceutically acceptable salts thereof. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in combination with at least one compound chosen from deutivacaftor and pharmaceutically acceptable salts thereof. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in combination with at least one compound chosen from (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof. [00140] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in combination with at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof and at least one compound chosen from ivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in combination with at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof and at least one compound chosen from deutivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in combination with at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof and at least one compound chosen from (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof. [00141] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in combination with at least one compound chosen from lumacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof and at least one compound chosen from ivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in combination with at least one compound chosen from lumacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof and at least one compound chosen from deutivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in combination with at least one compound chosen from lumacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof and at least one compound chosen from (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof. [00142] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in combination with at (a) a compound chosen from (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol and (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, deuterated derivatives of the compound, and pharmaceutically acceptable salts of any of the foregoing, and (b) optionally tezacaftor, deuterated derivatives, or pharmaceutically acceptable salts of any of the foregoing. [00143] Each of the compounds of the disclosure, (e.g., Compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), independently can be administered once daily, twice daily, or three times daily. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered once daily. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered twice daily. [00144] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof are administered once daily. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof are administered twice daily. [00145] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and at least one compound chosen from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing are administered once daily. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and at least one compound chosen from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof are administered twice daily. [00146] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, and at least one compound chosen from ivacaftor, deutivacaftor, (6R,12R)-17- amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino-12,12- dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5]nonadeca- 1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof are administered once daily. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, at least one compound chosen from tezacaftor and pharmaceutically acceptable salts thereof, and at least one compound chosen from ivacaftor, deutivacaftor, (6R,12R)-17- amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino-12,12- dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5]nonadeca- 1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof are administered twice daily. [00147] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, at least one compound chosen from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof, and at least one compound chosen from lumacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, are administered once daily. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, at least one compound chosen from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12- methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca- 1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa- 3,4,13,18-tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof, and at least one compound chosen from lumacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, are administered twice daily. [00148] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, are administered once daily and at least one compound chosen from ivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, are administered twice daily. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and at least one compound chosen from lumacaftor and pharmaceutically acceptable salts thereof, are administered once daily and at least one compound chosen from ivacaftor and pharmaceutically acceptable salts thereof, are administered twice daily. [00149] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, are administered once daily and at least one compound chosen from (6R,12R)-17- amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo [12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15- bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof, are administered once or twice daily. In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and at least one compound chosen from lumacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, are administered once daily and at least one compound chosen from (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa- 3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino-12,12- dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5]nonadeca- 1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof, are administered once or twice daily. [00150] Compounds of the disclosure, (e.g., compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), as well as additional CFTR modulator compounds, such as, e.g., tezacaftor, lumacaftor, ivacaftor, deutivacaftor, (6R,12R)- 17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino-12,12- dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5]nonadeca- 1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof, can be administered in a single pharmaceutical composition or separate pharmaceutical compositions. Such pharmaceutical compositions can be administered once daily or multiple times daily, such as twice daily. As used herein, the phrase that a given amount of API (e.g., tezacaftor, lumacaftor, ivacaftor, deutivacaftor (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof) is administered once or twice daily or per day means that said given amount is administered per dosing once or twice daily. [00151] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in a first pharmaceutical composition; at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a second pharmaceutical composition; and at least one compound chosen from ivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a third pharmaceutical composition. [00152] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in a first pharmaceutical composition; at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a second pharmaceutical composition; at least one compound chosen from deutivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a third pharmaceutical composition. [00153] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in a first pharmaceutical composition; at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a second pharmaceutical composition; at least one compound chosen from (6R,12R)-17-amino-12- methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca- 1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa- 3,4,13,18-tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a third pharmaceutical composition. [00154] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in a first pharmaceutical composition; at least one compound chosen from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino-12,12- dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5]nonadeca- 1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a second pharmaceutical composition; at least one compound chosen from lumacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a third pharmaceutical composition. [00155] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, is administered in a first pharmaceutical composition; and at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof and at least one compound chosen from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)- 13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17- amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts thereof are administered in a second pharmaceutical composition. In some embodiments, the second pharmaceutical composition comprises a half of a daily dose of said at least one compound chosen from ivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, and the other half of the daily dose of said at least one compound chosen from ivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof is administered in a third pharmaceutical composition. [00156] In some embodiments, at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, at least one compound chosen from tezacaftor and pharmaceutically acceptable salts thereof, and at least one compound chosen from ivacaftor, deutivacaftor, and pharmaceutically acceptable salts thereof are administered in a first pharmaceutical composition. In some embodiments, the first pharmaceutical composition is administered to the patient twice daily. In some embodiments, the first pharmaceutical composition is administered once daily. In some embodiments, the first pharmaceutical composition is administered once daily and, when the first composition comprises ivacaftor, a second composition comprising only ivacaftor is administered once daily. [00157] Any suitable pharmaceutical compositions can be used for compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, tezacaftor, ivacaftor, deutivacaftor, lumacaftor and tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing. Some exemplary pharmaceutical compositions for tezacaftor and its pharmaceutically acceptable salts can be found in WO 2011/119984 and WO 2014/014841, all of which is incorporated herein by reference. Some exemplary pharmaceutical compositions for ivacaftor and its pharmaceutically acceptable salts can be found in WO 2007/134279, WO 2010/019239, WO 2011/019413, WO 2012/027731, and WO 2013/130669, and some exemplary pharmaceutical compositions for deutivacaftor and its pharmaceutically acceptable salts can be found in US 8,865,902, US 9,181,192, US 9,512,079, WO 2017/053455, and WO 2018/080591, all of which are incorporated herein by reference. Some exemplary pharmaceutical compositions for lumacaftor and its pharmaceutically acceptable salts can be found in WO 2010/037066, WO 2011/127421, and WO 2014/071122, all of which are incorporated herein by reference. [00158] In some embodiments, this disclosure provides a method of treating cystic fibrosis comprising the step of administering a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt disclosed herein (e.g., compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), or a pharmaceutical composition disclosed herein (e.g., pharmaceutical compositions comprising compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing). [00159] In some embodiments, the method of treating cystic fibrosis further comprises administering at least one additional active therapeutic agent. [00160] In some embodiments, this disclosure provides use of a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt disclosed herein (e.g., compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), or a pharmaceutical composition disclosed herein (e.g., pharmaceutical compositions comprising compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), in the manufacture of a medicament for treating cystic fibrosis. [00161] In some embodiments, the medicament is formulated to be administered with at least one additional active therapeutic agent. In some embodiments, the medicament comprises at least one additional active therapeutic agent. [00162] In some embodiments, this disclosure provides a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt disclosed herein (e.g., compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), or a pharmaceutical composition disclosed herein (e.g., pharmaceutical compositions comprising compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), for use in the treatment of cystic fibrosis. In some embodiments, this disclosure provides a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt disclosed herein (e.g., compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), or a pharmaceutical composition disclosed herein (e.g., pharmaceutical compositions comprising compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing) for use in a method of treating cystic fibrosis. [00163] In some embodiments, the treatment further comprises administering at least one additional active therapeutic agent. [00164] In some embodiments, this disclosure provides the use of a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt disclosed herein (e.g., compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), or a pharmaceutical composition disclosed herein (e.g., pharmaceutical compositions comprising compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing) in the treatment of cystic fibrosis. In some embodiments, this disclosure provides the use of a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt disclosed herein (e.g., compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), or a pharmaceutical composition disclosed herein (e.g., pharmaceutical compositions comprising compounds of any of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing) in a method of treating cystic fibrosis. [00165] In some embodiments, the use further comprises use of at least one additional active therapeutic agent. [00166] In some embodiments, the at least one additional active therapeutic agent to be administered is selected from mucolytic agents, bronchodilators, antibiotics, anti-infective agents, anti-inflammatory agents, nutritional agents, and CFTR modulators. [00167] In some embodiments, the at least one additional active therapeutic agent to be administered is selected from CFTR potentiators. [00168] In some embodiments, the at least one CFTR potentiator to be administered is selected from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)- 13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17- amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing. [00169] In some embodiments, the at least one additional active therapeutic agent to be administered is selected from CFTR correctors. [00170] In some embodiments, the at least one CFTR corrector to be administered is selected from lumacaftor, tezacaftor, and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing. [00171] In some embodiments, the at least one additional active therapeutic agent to be administered is (a) at least one compound selected from tezacaftor, lumacaftor, and deuterated derivatives and pharmaceutically acceptable salts thereof; and (b) at least one compound selected from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)- 13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17- amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing. [00172] In some embodiments, the at least one additional active therapeutic agent to be administered is (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol, and optionally tezacaftor. [00173] In some embodiments, the at least one additional active therapeutic agent to be administered is (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and optionally tezacaftor. Pharmaceutical Compositions [00174] Another aspect of the disclosure provides a pharmaceutical composition comprising at least one novel compound of the disclosure (e.g., a compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing), and at least one pharmaceutically acceptable carrier. [00175] In some embodiments, the disclosure provides pharmaceutical compositions comprising at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, in combination with at least one additional active pharmaceutical ingredient. In some embodiments, the at least one additional active pharmaceutical ingredient is a CFTR modulator. In some embodiments, the at least one additional active pharmaceutical ingredient is a CFTR corrector. In some embodiments, the at least one additional active pharmaceutical ingredient is a CFTR potentiator. In some embodiments, the pharmaceutical composition comprises at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, and at least two additional active pharmaceutical ingredients, one of which is a CFTR corrector and one of which is a CFTR potentiator. [00176] In some embodiments, the disclosure provides a pharmaceutical composition comprising (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, (b) at least one compound chosen from tezacaftor and pharmaceutically acceptable salts thereof, and (c) at least one pharmaceutically acceptable carrier. [00177] In some embodiments, the disclosure provides a pharmaceutical composition comprising (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, (b) at least one compound chosen from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19- dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino- 12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing, and (c) at least one pharmaceutically acceptable carrier. [00178] In some embodiments, the disclosure provides a pharmaceutical composition comprising (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, (b) at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, (c) at least one compound chosen from ivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, and (d) at least one pharmaceutically acceptable carrier. [00179] In some embodiments, the disclosure provides a pharmaceutical composition comprising (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, (b) at least one compound chosen from tezacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, (c) at least one compound chosen from deutivacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, and (d) at least one pharmaceutically acceptable carrier. [00180] In some embodiments, the disclosure provides a pharmaceutical composition comprising (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, (a) at least one compound chosen from (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino-12,12- dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18-tetrazatricyclo[12.3.1.12,5]nonadeca- 1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing, (b) optionally tezacaftor, deuterated derivatives, or pharmaceutically acceptable salts of any of the foregoing, and (c) at least one pharmaceutically acceptable carrier. [00181] In some embodiments, the disclosure provides a pharmaceutical composition comprising (a) at least one compound chosen from compounds of Formulae I, I′, Ia, Ib, Ic, II, IIa, IIb, IIc, IIIa, and IIIb, Compounds 1 to 139 (i.e., Compounds 1 to 107 and/or Compounds 108 to 139), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, (b) at least one compound chosen from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19- dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, (6R)-17-amino- 12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing, (c) at least one compound chosen from lumacaftor and deuterated derivatives and pharmaceutically acceptable salts thereof, and (d) at least one pharmaceutically acceptable carrier. [00182] Any pharmaceutical composition disclosed herein may comprise at least one pharmaceutically acceptable carrier. In some embodiments, the at least one pharmaceutically acceptable carrier is chosen from pharmaceutically acceptable vehicles and pharmaceutically acceptable adjuvants. In some embodiments, the at least one pharmaceutically acceptable is chosen from pharmaceutically acceptable fillers, disintegrants, surfactants, binders, and lubricants. [00183] The pharmaceutical compositions described herein are useful for treating cystic fibrosis and other CFTR mediated diseases. [00184] As described above, pharmaceutical compositions disclosed herein may optionally further comprise at least one pharmaceutically acceptable carrier. The at least one pharmaceutically acceptable carrier may be chosen from adjuvants and vehicles. The at least one pharmaceutically acceptable carrier, as used herein, includes any and all solvents, diluents, other liquid vehicles, dispersion aids, suspension aids, surface active agents, isotonic agents, thickening agents, emulsifying agents, preservatives, solid binders, and lubricants, as suited to the particular dosage form desired. Remington: The Science and Practice of Pharmacy, 21st edition, 2005, ed. D.B. Troy, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York discloses various carriers used in formulating pharmaceutical compositions and known techniques for the preparation thereof. Except insofar as any conventional carrier is incompatible with the compounds of this disclosure, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutical composition, its use is contemplated to be within the scope of this disclosure. Non-limiting examples of suitable pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffer substances (such as phosphates, glycine, sorbic acid, and potassium sorbate), partial glyceride mixtures of saturated vegetable fatty acids, water, salts, and electrolytes (such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, and zinc salts), colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, wool fat, sugars (such as lactose, glucose and sucrose), starches (such as corn starch and potato starch), cellulose and its derivatives (such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate), powdered tragacanth, malt, gelatin, talc, excipients (such as cocoa butter and suppository waxes), oils (such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil), glycols (such as propylene glycol and polyethylene glycol), esters (such as ethyl oleate and ethyl laurate), agar, buffering agents (such as magnesium hydroxide and aluminum hydroxide), alginic acid, pyrogen-free water, isotonic saline, Ringer’s solution, ethyl alcohol, phosphate buffer solutions, non-toxic compatible lubricants (such as sodium lauryl sulfate and magnesium stearate), coloring agents, releasing agents, coating agents, sweetening agents, flavoring agents, perfuming agents, preservatives, and antioxidants. Examples I. Abbreviation List ACN: acetonitrile aq: aqueous Boc2O: di-tert-butyl dicarbonate CDMT: 2-chloro-4,6-dimethoxy-1,3,5-triazine Cmpd: compound CV: column volume(s) DCM: dichloromethane de: diastereomeric excess DI: deionized DIBAL: diisobutylaluminum hydride DIEA, DIPEA, DiPEA: N,N-diisopropylethylamine DMA: N,N-dimethylacetamide DMAP: 4-dimethylaminopyridine DMF: N,N-dimethylformamide DMSO: dimethyl sulfoxide de: diastereomeric excess ee: enantiomeric excess ESI-MS: electrospray ionization-mass spectrometry FA: formic acid FID: free induction decay GC: gas chromatography GCMS: gas chromatography mass spectrometry HMDS: hexamethyldisilazide HPLC: high-performance liquid chromatography IPA: isopropanol LAH: lithium aluminum hydride LC: liquid chromatography LCMS: liquid chromatography mass spectrometry LCMS Rt: LCMS retention time MTBE: methyl tert-butyl ether MeTHF or 2-MeTHF: 2-methyltetrahydrofuran MS: mass spectrometry MSD: mass-selective detector NMP: N-methyl-2-pyrrolidone NMM: N-methylmorpholine NMR: nuclear magnetic resonance Pd(dppf)Cl2: [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) rac: racemic rt, RT: room temperature SFC: supercritical fluid chromatography TBAF: tetra-n-butylammonium fluoride TBDPS: tert-butyldiphenylsilyl TBS: tert-butyldimethylsilyl TEA: triethylamine TEMPO: 2,2,6,6-tetramethylpiperidine 1-oxyl TFA: trifluoroacetic acid THF: tetrahydrofuran TMEDA, TEMED: tetramethylethylenediamine TMS: trimethylsilyl T3P: propanephosphonic acid anhydride TsCl: Tosyl chloride or p-toluenesulfonyl chloride UPLC: Ultra Performance Liquid Chromatography II. General Methods [00185] Reagents and starting materials were obtained by commercial sources unless otherwise stated and were used without purification. [00186] Proton and carbon NMR spectra were acquired on either a Bruker Biospin DRX 400 MHz FTNMR spectrometer operating at a 1H and 13C resonant frequency of 400 and 100 MHz respectively, or on a 300 MHz NMR spectrometer. One dimensional proton and carbon spectra were acquired using a broadband observe (BBFO) probe with 20 Hz sample rotation at 0.1834 and 0.9083 Hz/Pt digital resolution respectively. All proton and carbon spectra were acquired with temperature control at 30 °C using standard, previously published pulse sequences and routine processing parameters. [00187] NMR (1D & 2D) spectra were also recorded on a Bruker AVNEO 400 MHz spectrometer operating at 400 MHz and 100 MHz respectively equipped with a 5 mm multinuclear Iprobe. [00188] NMR spectra were also recorded on a Varian Mercury NMR instrument at 300 MHz for 1H using a 45 degree pulse angle, a spectral width of 4800 Hz and 28860 points of acquisition. FID were zero-filled to 32k points and a line broadening of 0.3Hz was applied before Fourier transform. 19F NMR spectra were recorded at 282 MHz using a 30 degree pulse angle, a spectral width of 100 kHz and 59202 points were acquired. FID were zero-filled to 64k points and a line broadening of 0.5 Hz was applied before Fourier transform. [00189] NMR spectra were also recorded on a Bruker Avance III HD NMR instrument at 400 MHz for 1H using a 30 degree pulse angle, a spectral width of 8000 Hz and 128k points of acquisition. FID were zero-filled to 256k points and a line broadening of 0.3 Hz was applied before Fourier transform.19F NMR spectra were recorded at 377 MHz using a 30 deg pulse angle, a spectral width of 89286 Hz and 128k points were acquired. FID were zero-filled to 256k points and a line broadening of 0.3 Hz was applied before Fourier transform. [00190] NMR spectra were also recorded on a Bruker AC 250 MHz instrument equipped with a: 5 mm QNP(H1/C13/F19/P31) probe (type: 250-SB, s#23055/0020) or on a Varian 500 MHz instrument equipped with an ID PFG, 5 mm, 50-202/500 MHz probe (model/part# 99337300). III. General UPLC/HPLC Analytical Methods [00191] LC method A: Analytical reverse phase UPLC using an Acquity UPLC BEH C18 column (50 × 2.1 mm, 1.7 μm particle) made by Waters (pn: 186002350), and a dual gradient run from 1-99% mobile phase B over 2.9 minutes. Mobile phase A = H2O (0.05 % CF3CO2H). Mobile phase B = CH3CN (0.035 % CF3CO2H). Flow rate = 1.2 mL/min, injection volume = 1.5 μL, and column temperature = 60 °C. [00192] LC method B: Acquity UPLC BEH C18 column (30 × 2.1 mm, 1.7 μm particle) made by Waters (pn: 186002349), and a dual gradient run from 1-99% mobile phase B over 1.0 minute. Mobile phase A = H2O (0.05 % CF3CO2H). Mobile phase B = CH3CN (0.035 % CF3CO2H). Flow rate = 1.5 mL/min, injection volume = 1.5 μL, and column temperature = 60 °C. [00193] LC method C: Reversed phase UPLC using an Acquity UPLC BEH C18 column (50 × 2.1 mm, 1.7 μm particle) made by Waters (pn: 186002350), and a dual gradient run from 30- 99% mobile phase B over 2.9 minutes. Mobile phase A = H2O (0.05 % CF3CO2H). Mobile phase B = CH3CN (0.035 % CF3CO2H). Flow rate = 1.2 mL/min, injection volume = 1.5 μL, and column temperature = 60 °C. [00194] LC method D: Merckmillipore Chromolith SpeedROD C18 column (50 x 4.6 mm) and a dual gradient run from 5 – 100% mobile phase B over 12 minutes. Mobile phase A = water (0.1 % CF3CO2H). Mobile phase B = acetonitrile (0.1 % CF3CO2H). [00195] LC method E: Merckmillipore Chromolith SpeedROD C18 column (50 x 4.6 mm) and a dual gradient run from 5 – 100% mobile phase B over 6 minutes. Mobile phase A = water (0.1 % CF3CO2H). Mobile phase B = acetonitrile (0.1 % CF3CO2H). [00196] LC method F: Kinetex Polar C183.0 x 50 mm 2.6 μm, 6 min, 5-95% I in H2O (0.1% Formic Acid) 1.2 mL/min. [00197] LC method G: Acquity UPLC BEH C18 column (50 × 2.1 mm, 1.7 μm particle) made by Waters (pn: 186002350), and a dual gradient run from 1-30% mobile phase B over 2.9 minutes. Mobile phase A = H2O (0.05 % CF3CO2H). Mobile phase B = CH3CN (0.035 % CF3CO2H). Flow rate = 1.2 mL/min, injection volume = 1.5 μL, and column temperature = 60 [00198] LC method H: water Cortex 2.7 μ C18 (3.0 mm x 50 mm), Temp: 55 oC; Flow: 1.2 mL/min; mobile phase: 100% water with 0.1% trifluoroacetic (TFA) acid then 100% acetonitrile with 0.1% TFA, gradient run from 5%-100% mobile phase B over 4 min, with stay at 100% B for 0.5 min, equilibration to 5% B over 1.5 min. [00199] LC method I: UPLC Luna C18(2) 50 x 3 mm 3 μm. Run: 2.5 min. Mobile phase: Initial 95% H2O 0.1% FA / 5%MeCN 0.1% FA, linear gradient to 95% MeCN 0.1% FA over 1.3 min, hold 1.2 min 95% CH3CN 0.1% FA,T: 45 oC, Flow: 1.5 mL/min. [00200] LC method J: UPLC SunFire C1875 x 4.6 mm 3.5 μm, run: 6 min. Mobile phase conditions: Initial 95% H2O + 0.1% FA/5% CH3CN + 0.1% FA, linear gradient to 95% CH3CN for 4 min, hold for 2 min at 95% CH3CN. T:45 oC, Flow:1.5 mL/min. [00201] LC method K: Xbridge C184.6 x 75 mm, 5μm, Initial Gradient at 95% NH4HCO3/5% MeCN 6 min run with 1 min equilibration gradient 0 to 3 min at 95% MeCN and hold for 3 minutes. Flow 1.5 mL/min. [00202] LC method L: Luna C183.0 x 50 mm 3.0 μM, Temp: 45 oC, Flow: 2.0 mL/min, Run Time: 3 minutes. Mobile Phase: Initial 95% H2O (0.1% Formic Acid) and 5% CH3CN (0.1% FA) linear gradient to 95% CH3CN (0.1% FA) for 2.0 min then hold at 95% CH3CN (0.1% FA) for 1.0 min. [00203] LC method M: Poroshell 120 EC-C183.0 x 50 mm 2.7 μM, Temp:45 oC, Flow: 1,5 mL/min, RunTime: 3 min. Mobile phase conditions: Initial.95% H2O (0.1% Formic Acid) and 5% CH3CN (0.1% FA) linear gradient to 95% CH3CN (0.1% FA) for 1.5 min then hold at 95% CH3CN (0.1% FA) for 1.5 min. [00204] LC method N: Reversed phase UPLC using an Acquity UPLC BEH C18 column (50 × 2.1 mm, 1.7 μm particle) made by Waters (pn: 186002350), and a dual gradient run from 1- 99% mobile phase B over 4.5 minutes. Mobile phase A = H2O (0.05 % TFA). Mobile phase B = acetonitrile (0.035 % TFA). Flow rate = 1.2 mL/min, injection volume = 1.5 μL, and column temperature = 60 °C. [00205] LC method O: Kinetex Polar C182.6 µm, 50 x 3.0 mm. Temperature: 45 oC, Flow: 1.2 mL/min, Run time: 3 min. Mobile phase conditions: Initial 95% water + 0.1% formic acid / 5% acetonitrile + 0.1% formic acid then linear gradient to 95% acetonitrile for 1.5 min then hold for 1.5 min at 95% acetonitrile. [00206] LC method P: Kinetex Polar C182.6 µm, 50 x 3.0 mm. Temperature: 45 oC, Flow: 1.2mL/min, Run time: 6 min. Mobile phase conditions: Initial 95% water + 0.1% formic acid / 5% acetonitrile + 0.1% formic acid then linear gradient to 95% acetonitrile for 3.5 min then hold for 2.5 min at 95% acetonitrile. [00207] LC method Q: Luna C18(2) 50 x 3 mm, 3 µm. Temperature: 45 oC, Flow: 1.5 mL/min, Run time: 3.5 min. Mobile phase conditions: Initial 50% H2O 0.1% FA / 50% MeCN 0.1% FA, linear gradient to 100% MeCN 0.1% FA over 1.5 min then hold for 2 minute at 100% CH3CN 0.1% FA. ESI Positive. [00208] LC method R: Luna C182.0 x 50mm 3 µm Gradient from 50%H2O + 0.1% FA / 50% MeCN + 0.1% FA to 95% MeCN + 0.1% in 1.5 min and hold for 1.5 minutes. Column temperature: 45 oC Runtime: 3 min. ESI Positive. [00209] LC method S: Luna C18(2) 50 X 3 mm, 3 µm. Temperature: 45 oC, Flow: 1.5 mL/min, Run time: 3.5 min. Mobile phase conditions: Initial 95% H2O 0.1% FA / 5% MeCN 0.1% FA, linear gradient to 95% MeCN 0.1% FA over 1.3 min then hold for 2.2 minutes at 95% CH3CN 0.1% FA. ESI Positive. [00210] LC method T: Luna C18(2) 50 X 3 mm, 3 µm. Temperature: 45 oC, Flow: 1.5 mL/min, Run time: 3 min. Mobile phase conditions: Initial 50% water 0.1% FA / 50% MeCN 0.1% FA, linear gradient to 95% MeCN 0.1% FA over 1.5min then hold for 1.5 min at 95% acetonitrile 0.1% FA. [00211] LC method U: Luna C18(2) 50 X 3 mm, 3 µm. Temperature: 45 oC, Flow: 1.5 mL/min, Run time: 3 min. Mobile phase conditions: Initial 95% water 0.1% FA / 5% MeCN 0.1% FA, linear gradient to 95% MeCN 0.1% FA over 1.5 min then hold for 1.5 min at 95% acetonitrile 0.1% FA. [00212] LC method V: Xbridge C184.6 x 75 mm 5 µm. Temperature: 45 oC, Flow: 1.5mL/min, Run time: 6 min. Mobile phase conditions: Initial 50% water 10 mM NH4HCO3 / 50% acetonitrile then linear gradient to 95% acetonitrile for 3 min then hold for 3 min at 95% acetonitrile. [00213] LC method W: Xbridge C184.6 x 75 mm 5 µm. Temperature: 45 oC, Flow: 1.5 mL/min, Run time: 6 min. Mobile phase conditions: Initial 95% water 10 mM NH4HCO3 / 5% acetonitrile then linear gradient to 95% acetonitrile for 3 min then hold for 3 min at 95% acetonitrile. [00214] LC method X: Xbridge C184.6 x 50 mm 3.5 µm. Temperature: 45 oC, Flow: 2 mL/min, Run time: 3 min. Mobile phase conditions: Initial 95% water 10 mM NH4HCO3 / 5% acetonitrile then linear gradient to 95% acetonitrile for 1.5 min then hold for 1.5 min at 95% acetonitrile. [00215] LC method Y: Xbridge C184.6 x 50 mm 3.5 µm. Temperature: 45 oC, Flow: 2 mL/min, Run time: 4 min. Mobile phase conditions: Initial 95% water 10 mM NH4HCO3 / 5% acetonitrile then linear gradient to 95% acetonitrile for 1.5 min then hold for 2.5 min at 95% acetonitrile. [00216] LC method Z: Luna C18(2) 50 x 3 mm, 3 µm. Temperature: 45 oC, Flow: 1.5 mL/min, Run time: 6 min. Mobile phase conditions: Initial 50% water 0.1% FA / 50% MeCN 0.1% FA, linear gradient to 95% MeCN 0.1% FA over 4 min then hold for 2 min at 95% acetonitrile 0.1% FA. [00217] LC method 1A: Xbridge C184.6 x 75 mm 5 µm. Temperature: 45 oC, Flow: 1.5 mL/min, run time: 10 min. Mobile phase conditions: Initial 95% water 10 mM NH4HCO3 / 5% acetonitrile then linear gradient to 95% acetonitrile for 6.5 min then hold for 3.5 min at 95% acetonitrile. [00218] LC method 1B: Luna C18(2) 50 x 3 mm, 3 µm.Temperature: 45 oC, Flow: 1.5 mL/min, Run time: 6 min. Mobile phase conditions: Initial 95% H2O 0.1% FA / 5% MeCN 0.1% FA, linear gradient to 95% MeCN 0.1% FA over 4 min then hold for 2 minute at 95% CH3CN 0.1% FA. ESI Positive. [00219] LC method 1C: Waters Acquity BEH C181.7 µm 2.1 x 100 mm. Column temp: 40 oC, flow rate: 0.5 mL/min. Eluent A: water 0.1 % FA, eluent B: acetonitrile. Conditions: 95% A 5% B, linear gradient to 5% A 95% B for 3 min, hold at 5% A 95% B for 2 min. Total run time: 5 min. [00220] LC method 1D: Luna C18(2) 50 x 3 mm, 3 µm. Temperature: 45 oC, Flow: 1.5 mL/min, Run time: 4 min. Mobile phase conditions: Initial 95% water 0.1% FA / 5% MeCN 0.1% FA, linear gradient to 95% MeCN 0.1% FA over 1.5 min then hold for 2.5 min at 95% acetonitrile 0.1% FA. [00221] LC method 1E: Waters Acquity BEH C181.7 µm 2.1 x 50 mm. Column temp: 40 oC, flow rate: 0.5 mL/min. Eluent A: water 0.1 % FA, eluent B: acetonitrile. Conditions: 50% A 50% B, linear gradient to 5% A 95% B for 3 min, hold at 5% A 95% B for 2 min. Total run time: 5 min. Synthesis of Intermediates IV. Synthesis of substituted chloropyrimidine intermediates Example 1: Preparation of 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid Step 1: tert-Butyl N-tert-butoxycarbonyl-N-(4,6-dichloropyrimidin-2-yl)carbamate [00222] To a solution of 4,6-dichloropyrimidin-2-amine (300 g, 1.829 mol) in DCM (2.1 L) was added (Boc)2O (838 g, 3.840 mol) followed by DMAP (5.6 g, 45.84 mmol). The mixture was stirred at ambient temperature for 6 h. Additional DMAP (5.6 g, 45.84 mmol) was added and the reaction was continued to stir at ambient temperature for 24 h. The mixture was diluted with water (2.1 L) and the organic phase separated. The organic phase was washed with water (2.1 L), 2.1L of brine, dried over magnesium sulfate, filtered over Celite and concentrated in vacuo affording a light orange oil which had a silt in the slurry. The mixture was diluted with ~500 mL of heptane and filtered using an M filter. The precipitate (starting material) was washed with 250 mL of heptane. The filtrate was concentrated in vacuo affording a thick orange oil which was seeded with solid from a previous experiment and crystallized on standing, affording a light orange hard solid. Tert-butyl N-tert-butoxycarbonyl-N-(4,6-dichloropyrimidin- 2-yl)carbamate (645 g, 97%).1H NMR (400 MHz, DMSO-d6) δ 8.07 (s, 1H), 1.44 (s, 18H). ESI- MS m/z calc.363.07526, found 364.1 (M+1)+; Retention time: 2.12 minutes (LC method A). Step 2: tert-Butyl N-tert-butoxycarbonyl-N-[4-chloro-6-(2,6-dimethylphenyl)pyrimidin- 2-yl]carbamate. [00223] All solvents were degassed prior to use. To a slurry of tert-butyl N-tert- butoxycarbonyl-N-(4,6-dichloropyrimidin-2-yl)carbamate (88 g, 241.6 mmol), (2,6- dimethylphenyl)boronic acid (approximately 36.24 g, 241.6 mmol) and Cs2CO3 (approximately 196.8 g, 604.0 mmol) in DME (704 mL) and water (176 mL) were added. Pd(dppf)Cl2 (approximately 8.839 g, 12.08 mmol) was added and the mixture was vigorously stirred under nitrogen at 80 ºC (reflux) for 1 h (no SM remained). The reaction was cooled to ambient temperature and diluted with water (704 mL). The aqueous phase was separated and extracted with EtOAc (704 mL). The organic phase was washed with 700 mL of brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The crude product was chromatographed on a 1500 g silica gel column eluting with 0-30% EtOAc/hexanes. The product fractions (eluted at 15% EtOAc) were combined, and concentrated in vacuo affording the product as a clear oil which crystallized on standing. Tert-butyl N-tert-butoxycarbonyl-N-[4-chloro-6-(2,6- dimethylphenyl)pyrimidin-2-yl]carbamate (81.3 g, 78%).1H NMR (400 MHz, DMSO-d6) δ 7.88 (s, 1H), 7.30 (dd, J = 8.2, 7.0 Hz, 1H), 7.21 – 7.16 (m, 2H), 2.03 (s, 6H), 1.38 (s, 18H). ESI-MS m/z calc.433.17682, found 434.1 (M+1)+; Retention time: 2.32 minutes (LC method A). Step 3: 4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-amine (hydrochloride salt) [00224] Tert-Butyl N-tert-butoxycarbonyl-N-[4-chloro-6-(2,6-dimethylphenyl) pyrimidin-2- yl]carbamate (514.8 g, 915.9 mmol) was dissolved in dichloromethane (4 L). Hydrogen chloride in p-dioxane (1 L, 4 mol) was added and the mixture was stirred overnight at room temperature. The resulting precipitate was collected by vacuum filtration and dried in vacuo to obtain 4- chloro-6-(2,6-dimethylphenyl)pyrimidin-2-amine hydrochloride (213.5 g, 64%) as a white solid (213.5 g, 82%).1H NMR (250 MHz, DMSO-d6) δ 7.45-6.91 (m, 3H), 6.73 (s, 1H), 2.08 (s, 6H). ESI-MS m/z calc.233.072, found 234.1 (M+1)+; Retention time: 2.1 minutes (LC Method C). Step 4: 4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-amine [00225] 4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-amine (hydrochloride salt) (166 g, 614.5 mmol) and 4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-amine (hydrochloride salt) (30 g, 111.0 mmol) were suspended in DCM (2.5 L), treated with NaOH (725 mL of 1 M, 725.0 mmol) and stirred at room temperature for 1 hour.The mixture was transferred into a separatory funnel and left standing over night. The DCM phase was separated and the aqueous phase with insoluble material was extracted twice more with DCM (2 x 500mL). The combined brown DCM phases were stirred over magnesium sulfate and charcoal for 1 hour, filtered and the yellow solution concentrated to a volume of ~ 500 mL. The solution was diluted with heptane (750 mL) and DCM was removed under reduced pressure at 60 °C to give a cream suspension. It was stirred at room temperature for 1 hour, filtered, washed with cold heptane and dried to give 4-chloro-6- (2,6-dimethylphenyl)pyrimidin-2-amine (157 g, 91%) as a cream solid.1H NMR (400 MHz, DMSO-d6) δ 7.28 – 7.14 (m, 3H), 7.10 (d, J = 7.5 Hz, 2H), 6.63 (s, 1H), 2.06 (s, 6H). ESI-MS m/z calc.233.07198, found 234.0 (M+1)+; Retention time: 1.45 minutes (LC method A). Step 5: 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid [00226] 4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-amine (235 g, 985.5 mmol) was dissolved in MeTHF (2.3 L) and cooled in an ice bath under stirring and nitrogen. To the cold solution methyl 3-chlorosulfonylbenzoate (347 g, 1.479 mol) was added in one portion (seems slightly endothermic) and to the cold pale-yellow solution a solution of 2-methyl-butan-2-ol (Lithium salt) (875 mL of 3.1 M, 2.712 mol) (in heptane) was added dropwise over 1.25 hour (exothermic, internal temperature from 0 to 10 °C). The ice bath was removed, and the greenish solution was stirred for 4 hours at room temperature. To the greenish solution cold HCl (2 L of 1.5 M, 3.000 mol) was added, the phases separated and the organic phase was washed once with water (1L) and once with brine (500 mL). The aqueous phases were back extracted once with MeTHF (350 mL) and the organic phases were combined. This yellow MeTHF solution of methyl 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoate (ESI-MS m/z calc.431.07065, found 432.0 (M+1)+; Retention time: 1.81 minutes) was treated with NaOH (2.3 L of 2 M, 4.600 mol) and stirred at room temperature for 1 hour. The phases were separated and the NaOH phase was washed twice with MeTHF (2 x 500 mL) and the combined organic phases were extracted once with 2 M NaOH (1 x 250 mL). The combined NaOH phases were combined, stirred in an ice bath and slowly acidified by addition of HCl (416 mL of 36 %w/w, 4.929 mol) while keeping the internal temperature between 10 and 20 °C. At the end of the addition (pH ~5-6) the final pH was adjusted to 2-3 by addition of solid citric acid. The formed yellow tacky suspension was stirred at room temperature overnight to give a cream crisp suspension. The solid was collected by filtration, washed with plenty of water and sucked dry for 3 hours. The solid was dried under reduced pressure with a nitrogen leak at 45-50 °C for 120 hours 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (395 g, 96%) was isolated as an off-white solid.1H NMR (400 MHz, DMSO-d6) δ 13.44 (s, 1H), 12.46 (s, 1H), 8.48 – 8.39 (m, 1H), 8.25 – 8.15 (m, 1H), 8.15 – 8.08 (m, 1H), 7.68 (t, J = 7.8 Hz, 1H), 7.31 (s, 1H), 7.28 – 7.18 (m, 1H), 7.10 (d, J = 7.6 Hz, 2H), 1.84 (s, 6H). ESI-MS m/z calc. 417.055, found 418.0 (M+1)+; Retention time: 1.56 minutes. (LC method A).
Example 2: Preparation of 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin-2- yl]sulfamoyl]benzoic acid Step 1: tert-Butyl N-tert-butoxycarbonyl-N-(4,6-dichloro-5-methyl-pyrimidin-2- yl)carbamate [00227] To a solution of 4,6-dichloro-5-methyl-pyrimidin-2-amine (57.85 g, 318.47 mmol) in DCM (580 mL) was added tert-butoxycarbonyl tert-butyl carbonate (159.92 g, 168.34 mL, 710.77 mmol) and DMAP (3.96 g, 32.090 mmol) at RT. The reaction was stirred for 3 h. The reaction mixture was quenched with DI H2O (250 mL). DCM (100 mL) was added. The layers were separated, and the aqueous layer was extracted with DCM (2 x 250 mL). The combined organic layers were washed with aqueous saturated NaCl (250 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. Crude Mass = 125.71 g (Yellow Solid). The yellow solid was triturated with hexanes (300 mL, 3 h), filtered through a Type “M” glass filter by vacuum, and the solids were rinsed with hexanes (2 x 200 mL). Final Product (101.00 g) was obtained as a yellow solid. Tert-butyl N-tert-butoxycarbonyl-N-(4,6-dichloro-5- methyl-pyrimidin-2-yl)carbamate (101.00 g, 80%).1H NMR (500 MHz, Chloroform-d) δ 2.48 (s, 3H), 1.47 (s, 18H). ESI-MS m/z calc.377.0909, found 378.0 (M+1)+; Retention time: 3.39 minutes; LC method E. Step 2: tert-Butyl N-tert-butoxycarbonyl-N-[4-chloro-6-(2,6-dimethylphenyl)-5- methyl-pyrimidin-2-yl]carbamate [00228] To a solution of tert-butyl N-tert-butoxycarbonyl-N-(4,6-dichloro-5-methyl- pyrimidin-2-yl)carbamate (120.85 g, 319.50 mmol) dissolved in DME (850 mL) and water (120 mL) was added (2,6-dimethylphenyl)boronic acid (57.5 g, 383.38 mmol) and cesium carbonate (271 g, 831.75 mmol) at room temperature. The solution was stirred for 10 min while being bubbled with a nitrogen stream. Then Pd(dppf)Cl2 (11.7 g, 15.990 mmol) was added to the solution and heated to 80 °C overnight. The solution was cooled to room temperature before being diluted with water (500 mL) and extracted with ethyl acetate (2 x 1 L). The combined organic layer was washed with brine (1 L) and dried over sodium sulfate before being concentrated in vacuum. The organic residue was filtered through a pad of silica gel and washed with a solution of 1:3 ethyl acetate-hexanes (3 x 1 L) to give tert-butyl N-tert-butoxycarbonyl-N- [4-chloro-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin-2-yl]carbamate (100.71 g, 58%). ESI-MS m/z calc.447.19247, found 448.1 (M+1)+; Retention time: 4.24 minutes; LC method E. Step 3: 4-Chloro-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin-2-amine [00229] To a solution of tert-butyl N-tert-butoxycarbonyl-N-[4-chloro-6-(2,6- dimethylphenyl)-5-methyl-pyrimidin-2-yl]carbamate (100.71 g, 224.82 mmol) in DCM (500 mL) was added HCl (200 mL of 4 M, 800.00 mmol) in dioxane. The solution was stirred at room temperature overnight before being concentrated in vacuum. The residue was then basified with sodium bicarbonate (500 mL) and extracted with ethyl acetate (1 L). The organic layer was washed with brine (400 mL) and dried over sodium sulfate. The organic phase was concentrated then the residue was triturated with hexanes (2 x 200 mL) to give 4-chloro-6-(2,6- dimethylphenyl)-5-methyl-pyrimidin-2-amine (54.88 g, 99%) as an off-white solid.ESI-MS m/z calc.247.08763, found 248.2 (M+1)+; Retention time: 2.94 minutes; LC method E. Step 4: Methyl 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin-2- yl]sulfamoyl]benzoate [00230] To a solution of 4-chloro-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin-2-amine (35 g, 141.29 mmol) in THF (400 mL) at 0 °C was added methyl 3-chlorosulfonylbenzoate (50 g, 213.08 mmol). Then Lithium tert-amoxide (46.428 g, 159 mL of 40 %w/w, 197.40 mmol) was added to the solution dropwise keeping the temperature below 5 °C. The solution was allowed to warm to room temperature while it stirred for 3 hours. The solution was acidified with 1 M HCl (200 mL) and extracted with ethyl acetate (3 x 200 mL). The organic layer was washed with brine (300 mL) and dried over sodium sulfate. The organic layer was then concentrated under vacuum to give methyl 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methyl- pyrimidin-2-yl]sulfamoyl]benzoate (63.01 g, 100%) as a yellow solid. ESI-MS m/z calc. 445.0863, found 446.2 (M+1)+; Retention time: 3.63 minutes; LC method E. Step 5: 3-[[4-Chloro-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin-2- yl]sulfamoyl]benzoic acid [00231] To a solution of methyl 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin- 2-yl]sulfamoyl]benzoate (59.51 g, 133.45 mmol) in THF (500 mL) was added an aqueous solution of NaOH (300 mL of 2 M, 600.00 mmol) and the mixture was stirred for 2 hours at room temperature. The solution was acidified using 3 M HCl (500 mL) and extracted with ethyl acetate (2 x 500 mL) before being washed with brine (500 mL). The organic layer was dried over sodium sulfate and concentrated under vacuum. The organic residue was then recrystallized with ethanol and filtered to give 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin-2- yl]sulfamoyl]benzoic acid (34.44 g, 56%) as a white solid.1H NMR (500 MHz, DMSO-d6) δ 8.43 (t, J = 1.8 Hz, 1H), 8.18 (dt, J = 7.8, 1.4 Hz, 1H), 8.10 (ddd, J = 7.9, 2.0, 1.2 Hz, 1H), 7.66 (t, J = 7.8 Hz, 1H), 7.25 (t, J = 7.6 Hz, 1H), 7.12 (d, J = 7.6 Hz, 2H), 1.85 (s, 3H), 1.74 (s, 6H). ESI-MS m/z calc.431.07065, found 432.4 (M+1)+; Retention time: 2.43 minutes; LC method E. Example 3: Preparation of 3-[[4-Chloro-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2- yl]sulfamoyl]benzoic acid Step 1: 2-Amino-5-fluoro-pyrimidine-4,6-diol [00232] To a 2 L flask charged with ethanol (750 mL), solid chunks of sodium metal (21.5 g, 935.20 mmol) were added gradually and carefully, and the mixture was stirred until completely dissolved. Once cooled back to room temperature guanidine (hydrochloride salt) (34.5 g, 361.14 mmol) and diethyl 2-fluoropropanedioate (50 g, 280.65 mmol) were successively added and the reaction was heated at 80 °C for 17 hours. Once cooled to room temperature, the crude mixture was concentrated under reduced pressure to remove most of the ethanol. Water was added to complete dissolution, the resulting solution was cooled in an ice bath and acidified to pH of 1-2 using concentrated HCl. The solids were filtered and washed with water (2 x 200 mL) then with acetone (2 x 50 mL) and dried under high vacuum to provide 2-amino-5-fluoro-pyrimidine-4,6-diol hydrate (41.8 g, 91%)) as a pink solid.1H NMR (300 MHz, DMSO-d6) ppm 6.53 (br. S., 1H), 10.97 (br. S., 1H).19F NMR (282 MHz, DMSO-d6) ppm -196.1 (br. S., 1F). ESI-MS m/z calc.145.0288, found 146.1 (M+1)+; Retention time: 0.2 minutes (LC method M). Step 2: 4,6-Dichloro-5-fluoro-pyrimidin-2-amine [00233] A solution of 2-amino-5-fluoro-pyrimidine-4,6-diol hydrate (21.09 g, 126.46 mmol) in phosphorus oxychloride (101.99 g, 62 mL, 665.16 mmol) was heated to 90 °C and N,N-diethylaniline (25.326 g, 27 mL, 169.71 mmol) was added slowly. The reaction was then heated for 3 hours at 105 °C. The solution was poured in water and neutralized to pH ~5 with 50% aqueous sodium hydroxide solution and refluxed for 1 h. The solution was cooled on ice and the precipitate was filtered and dried. The solid was triturated in dichloromethane (~150 mL), filtered and dried. The solid obtained was then triturated in a mix of acetone/heptanes (1:1, 40 mL), filtered and dried provide the desired 4,6-dichloro-5-fluoro-pyrimidin-2-amine (13.75 g, 59%) as a yellow solid.1H NMR (300 MHz, DMSO-d6) δ 7.44 (br. S., 2H).19F NMR (282 MHz, DMSO-d6) δ -149.01 (s, 1F). ESI-MS m/z calc.180.961, found 182.0 (M+1)+; Retention time: 2.17 minutes. LC method F. Step 3: tert-Butyl N-(4,6-dichloro-5-fluoro-pyrimidin-2-yl)carbamate [00234] To a stirring solution of 4,6-dichloro-5-fluoro-pyrimidin-2-amine (5.153 g, 27.495 mmol) and Boc anhydride (8.43 g, 38.626 mmol) in anhydrous THF (100 mL) at -78 °C under nitrogen was dropwise added a solution of LiHMDS (50 mL of 1.3 M, 65.000 mmol) in THF. After the addition was complete, the reaction mixture was stirred at this temperature for 2 h. The reaction was quenched cold with saturated aqueous ammonium chloride (20 mL) and allowed to warm up to room temperature. The product was extracted with ethyl acetate (3 x 30 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate and concentrated. The crude was purified by silica gel chromatography using 0 – 15% ethyl acetate in hexane to afford tert-butyl N-(4,6-dichloro-5-fluoro-pyrimidin-2- yl)carbamate (6.632 g, 86%) as a white solid.1H NMR (500 MHz, DMSO-d6) δ 10.74 (s, 1H), 1.45 (s, 9H). Step 4: tert-Butyl N-[4-chloro-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2- yl]carbamate [00235] A stirring solution of tert-butyl N-(4,6-dichloro-5-fluoro-pyrimidin-2- yl)carbamate (6.815 g, 22.467 mmol) and (2,6-dimethylphenyl)boronic acid (3.06 g, 20.402 mmol) in a mixture of 1,2-dimethoxyethane (55 mL) and water (15 mL) at room temperature was degassed with nitrogen for 30 minutes. Under nitrogen, cesium carbonate (18.3 g, 56.166 mmol) and [1,1’-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (1.65 g, 2.2550 mmol) were added. The reaction mixture was heated to 80 °C for 4 h. After cooling to room temperature, water (150 mL) was added, and the product was extracted with ethyl acetate (3 x 150 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, and concentrated. The crude was purified by silica gel chromatography using 0 – 5% hexanes – ethyl acetate to afford tert-butyl N-[4-chloro-6-(2,6-dimethylphenyl)-5-fluoro- pyrimidin-2-yl]carbamate (2.43 g, 29%) as white solid. ESI-MS m/z calc.351.115, found 352.4 (M+1)+; Retention time: 6.51 minutes, LC method D. Step 5: 4-Chloro-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2-amine [00236] To a stirring solution of tert-butyl N-[4-chloro-6-(2,6-dimethylphenyl)-5-fluoro- pyrimidin-2-yl]carbamate (2.43 g, 6.4238 mmol) in DCM (23 mL) at room temperature was added a solution of HCl (6.5 mL of 4 M, 26.000 mmol) in 1,4-dioxane. The reaction mixture was stirred at this temperature for 2 h. The reaction mixture was evaporated to dryness. The obtained white solid was resuspended in saturated aqueous sodium bicarbonate (100 mL) and stirred at room temperature for 15 minutes. The product was extracted with ethyl acetate (3 x 100 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate and concentrated. The crude was purified by silica gel chromatography using 0 – 15% hexanes – ethyl acetate to afford 4-chloro-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2- amine (1.42 g, 83%) as white solid. ESI-MS m/z calc.251.0626, found 252.3 (M+1)+; Retention time: 5.06 minutes, LC method D. Step 6: Methyl 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2- yl]sulfamoyl]benzoate [00237] To a stirring solution of 4-chloro-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2- amine (2.54 g, 10.092 mmol) and methyl 3-chlorosulfonylbenzoate (4.25 g, 18.112 mmol) in anhydrous THF (70 mL) at 0 °C under nitrogen was added dropwise a solution of lithium tert- amoxide (5.8400 g, 20 mL of 40 %w/w, 24.830 mmol) in heptanes. After the addition was complete, the reaction mixture was stirred at this temperature for 1 h. The reaction was quenched cold with 1 M aqueous HCl (180 mL) and then allowed to warm up to room temperature. Volatiles were removed under vacuum and the product was extracted with ethyl acetate (3 x 100 mL). The combined organic layers were washed with brine (60 mL), dried over anhydrous sodium sulfate, and concentrated. The crude was purified by silica gel chromatography using 0 – 20% hexanes – ethyl acetate. The obtained product was triturated with hexanes (100 mL), collected by filtration and dried under vacuum to afford methyl 3-[[4- chloro-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2-yl]sulfamoyl]benzoate (4.381 g, 93%) as white solid. ESI-MS m/z calc.449.0612, found 450.1 (M+1)+; Retention time: 2.72 minutes, LC method E.1H NMR (500 MHz, DMSO-d6) δ 12.49 (s, 1H), 8.43 (t, J = 1.8 Hz, 1H), 8.21 (dt, J = 7.8, 1.4 Hz, 1H), 8.14 (dt, J = 8.0, 1.5 Hz, 1H), 7.73 (t, J = 7.8 Hz, 1H), 7.32 (t, J = 7.6 Hz, 1H), 7.14 (d, J = 7.6 Hz, 2H), 3.82 (s, 3H), 1.85 (s, 6H). Step 7: 3-[[4-Chloro-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2- yl]sulfamoyl]benzoic acid [00238] To a stirring solution of methyl 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-fluoro- pyrimidin-2-yl]sulfamoyl]benzoate (490 mg, 1.0892 mmol) in THF (9 mL) at room temperature was added an aqueous solution of NaOH (4.5 mL of 1 M, 4.5000 mmol). The reaction mixture was stirred at this temperature for 2 h. Water was added (20 mL), and the reaction mixture was acidified with 2 M aqueous HCl to pH ~1. The product was extracted with ethyl acetate (3 x 40 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate and concentrated to afford 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-fluoro- pyrimidin-2-yl]sulfamoyl]benzoic acid (463 mg, 93%) as white solid. ESI-MS m/z calc. 435.0456, found 436.4 (M+1)+; Retention time: 5.21 minutes; LC method D. Example 4: Preparation of 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2- yl]sulfamoyl]benzoic acid Step 1: tert-Butyl N-tert-butoxycarbonyl-N-(4,6-dichloro-5-methoxy-pyrimidin-2- yl)carbamate [00239] To a solution of 4,6-dichloro-5-methoxy-pyrimidin-2-amine (11.73 g, 60.458 mmol) in DCM (200 mL) was added DMAP (591 mg, 4.8376 mmol) and Boc2O (27.7 g, 29.158 mL, 126.92 mmol). The reaction was stirred for 3 hours at room temperature before being washed with brine (200 mL) and water (200 mL). The organic layer was concentrated in vacuum and dried over sodium sulfate to give tert-butyl N-tert-butoxycarbonyl-N-(4,6-dichloro-5-methoxy- pyrimidin-2-yl)carbamate (21.55 g, 90%). ESI-MS m/z calc.393.08582, found 394.0 (M+1)+; Retention time: 3.44 minutes; LC method E. Step 2: tert-Butyl N-tert-butoxycarbonyl-N-[4-chloro-6-(2,6-dimethylphenyl)-5- methoxy-pyrimidin-2-yl]carbamate [00240] To a solution of tert-butyl N-tert-butoxycarbonyl-N-(4,6-dichloro-5-methoxy- pyrimidin-2-yl)carbamate (31.58 g, 80.101 mmol) dissolved in DME (225 mL) and water (31 mL) was added (2,6-dimethylphenyl)boronic acid (16.5 g, 110.01 mmol) and cesium carbonate (68 g, 208.71 mmol) at room temperature. The solution was stirred for 10 min while being bubbled with a nitrogen stream. Then Pd(dppf)Cl2 (5.86 g, 8.0087 mmol) was added to the solution and heated to 80 °C overnight. The solution was cooled to room temperature before being diluted with water (250 mL) and extracted with ethyl acetate (2 x 300mL). The combined organic layers were washed with brine (400 mL) and dried over sodium sulfate before being concentrated under vacuum. The organic residue was purified by silica gel chromatography eluting 0-60% ethyl acetate-hexanes to give tert-butyl N-tert-butoxycarbonyl-N-[4-chloro-6- (2,6-dimethylphenyl)-5-methoxy-pyrimidin-2-yl]carbamate (50.35 g, 135%). ESI-MS m/z calc. 463.1874, found 464.2 (M+1)+; Retention time: 3.68 minutes; LC method E. Step 3: 4-Chloro-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2-amine [00241] To a solution of tert-butyl N-tert-butoxycarbonyl-N-[4-chloro-6-(2,6- dimethylphenyl)-5-methoxy-pyrimidin-2-yl]carbamate (50.35 g, 108.52 mmol) in DCM (500 mL) was added HCl (100 mL of 4 M, 400.00 mmol) in dioxane. The solution was stirred at room temperature overnight before being concentrated under vacuum. The residue was then basified with sodium bicarbonate (400 mL) and extracted with ethyl acetate (500 mL). The organic layer was washed with brine (500 mL) and dried over sodium sulfate. The organic phase was concentrated then triturated with hexanes (2 x 50 mL) to give 4-chloro-6-(2,6- dimethylphenyl)-5-methoxy-pyrimidin-2-amine (10.16 g, 36%). ESI-MS m/z calc.263.08255, found 264.1 (M+1)+; Retention time: 2.73 minutes; LC method E. Step 4: Methyl 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2- yl]sulfamoyl]benzoate [00242] To a solution of 4-chloro-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2-amine (223 mg, 0.8456 mmol) in THF (6 mL) at 0°C was added methyl 3-chlorosulfonylbenzoate (496 mg, 2.1137 mmol). Then Lithium tert-amoxide (584.00 mg, 2 mL of 40 %w/w, 2.4830 mmol) was added to the solution dropwise keeping the temperature below 5 °C. The solution was allowed to warm to room temperature while it stirred for 3 hours. The solution was acidified with 1 M HCl (5 mL) and extracted with ethyl acetate (20 mL). The organic layer was washed with brine (20 mL) and dried over sodium sulfate. The organic layer was then concentrated under vacuum to give methyl 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2-yl]sulfamoyl]benzoate (386 mg, 99%) as a yellow solid. ESI-MS m/z calc.461.0812, found 462.1 (M+1)+; Retention time: 3.18 minutes; LC method E. Step 5: 3-[[4-Chloro-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2- yl]sulfamoyl]benzoic acid [00243] To a solution of methyl 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2- yl]sulfamoyl]benzoate (386 mg, 0.8356 mmol) in THF (10 mL) was added an aqueous solution of NaOH (5 mL of 1 M, 5.0000 mmol) and stirred for 1 hour at room temperature. The solution was acidified using 1 M HCl (5 mL) and extracted with ethyl acetate (2 x 20 mL) before being washed with brine (20 mL). The organic layer was dried over sodium sulfate and concentrated under vacuum to give 3-[[4-chloro-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2- yl]sulfamoyl]benzoic acid (314 mg, 84%) as a white solid. ESI-MS m/z calc.447.06558, found 448.1 (M+1)+; Retention time: 2.9 minutes; LC method E. Example 5: Preparation of 6-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]pyridine-2-carboxylic acid Step 1: Methyl 6-benzylsulfanylpyridine-2-carboxylate [00244] To a solution of phenylmethanethiol (28.408 g, 26.800 mL, 228.72 mmol) in THF (600 mL) was added NaH (11.200 g, 60 %w/w, 280.03 mmol) in a few portions at 0°C. The slurry was warmed to room temperature and stirred for 30 min, then methyl 6-bromopyridine-2- carboxylate (50 g, 231.45 mmol) was added as a single portion. After 3 h, the reaction was diluted with ether (800 mL) and quenched with water (400 mL) and saturated sodium bicarbonate (50 mL). The layers were separated, and the organic layer was washed with brine, dried over sodium sulfate, and concentrated under reduced pressure to yield methyl 6- benzylsulfanylpyridine-2-carboxylate (56.35 g, 89%) as a yellow oil.1H NMR (500 MHz, DMSO-d6) δ 7.84 – 7.77 (m, 1H), 7.77 – 7.73 (m, 1H), 7.52 (m, 1H), 7.48 (d, J = 7.8 Hz, 2H), 7.28(t, J = 7.2, 7.2 Hz, 2H), 7.24 – 7.18 (m, 1H), 4.44 (s, 2H), 3.90 (d, J = 1.2 Hz, 3H). ESI-MS m/z calc.259.0667, found 260.1 (M+1)+; Retention time: 3.2 minutes; LC method E. Step 2: Methyl 6-chlorosulfonylpyridine-2-carboxylate [00245] A solution of methyl 6-benzylsulfanylpyridine-2-carboxylate (121.62 g, 431.47 mmol) in DCM (950 mL) and DI water (300 mL) was cooled in a -1 – 0 °C ice bath and, with vigorous stirring, sulfuryl chloride (228.14 g, 140 mL, 1.6396 mol) was added dropwise while the temperature was maintained below 5 °C. After the addition, the organic phase was separated, washed with DI water (2 x 500 mL), dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue was dissolved in DCM (500 mL). Hexanes (1000 mL) was added, and the DCM was slowly evaporated off. The white precipitate was filtered by vacuum and the solids were washed with Hexanes (2 x 500 mL). The filtered solids were collected. The residue solids in the filtrate were filtered and dissolved in DCM (500 mL). The DCM solution was transferred to a 1 L round-bottom flask and concentrated under vacuum. The residue was dissolved in DCM (200 mL). Hexanes (600 mL) was added and the DCM was slowly evaporated off. The white precipitation was filtered by vacuum and the solids were washed with hexanes (2 x 500 mL) After drying, methyl 6-chlorosulfonylpyridine-2-carboxylate (56.898 g, 55%) was isolated.1H NMR (500 MHz, Chloroform-d) δ 8.48 (dd, J = 7.8, 1.1 Hz, 1H), 8.31 (dd, J = 7.9, 1.1 Hz, 1H), 8.25 (t, J = 7.8 Hz, 1H), 4.08 (s, 3H). ESI-MS m/z calc. 234.97061, found 236.1 (M+1)+; Retention time: 1.74 minutes; LC method E. Step 3: Methyl 6-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]pyridine-2-carboxylate [00246] A solution of 4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-amine (16.63 g, 71.161 mmol) and methyl 6-chlorosulfonylpyridine-2-carboxylate (16.8 g, 71.294 mmol) dissolved in anhydrous THF (680 mL) was cooled to – 78 °C. Then Lithium bis(trimethylsilyl)amide (143 mL of 1 M, 143.00 mmol) in solution in THF was added dropwise. The mixture was allowed to warm up to 0 °C slowly and then 1M aqueous HCl (146 mL) was added, followed by DI water (680 mL). The THF was evaporated, and the aqueous phase was extracted with chloroform (3 x 250 mL). The combined organic layers were washed with saturated aqueous NaCl (300 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The crude was recrystallized in 10 % Acetone in Hexanes (500 mL). The white precipitate was filtered and rinsed with acetone (2 x 100 mL) to give methyl 6-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin- 2-yl]sulfamoyl]pyridine-2-carboxylate (15.79 g, 50%). ESI-MS m/z calc.432.06592, found 433.3 (M+1)+; Retention time: 5.5 minutes; LC method D. Step 4: 6-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]pyridine- 2-carboxylic acid [00247] To a solution of methyl 6-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]pyridine-2-carboxylate (15.79 g, 36.477 mmol) in THF (180 mL) was added aqueous sodium hydroxide (182 mL of 1 M, 182.00 mmol). The reaction was stirred at RT for 1h. The THF was evaporated, and the aqueous layer was washed with diethyl ether (2 x 200 mL). The aqueous layer was acidified to pH 2 with 1 M Aqueous HCl (250 mL). The precipitate was filtered and the white solid was rinsed with DI water (2 x 250 mL). The solids were dried under vacuum to give 6-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]pyridine-2- carboxylic acid (14.3444 g, 93%).1H NMR (250 MHz, DMSO-d6) δ 8.14 – 7.99 (m, 3H), 7.21 – 7.11 (m, 1H), 7.03 (d, J = 7.7 Hz, 2H), 6.92 (s, 1H), 1.78 (s, 6H). ESI-MS m/z calc.418.05026, found 419.1 (M+1)+; Retention time: 2.61 minutes; LC method E. V. Synthesis of substituted bicyclic bromopyrazine and bromoindole intermediates Example 6: Preparation of 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine Step 1: 6-Bromo-3-(3,3-dimethylbut-1-ynyl)pyrazin-2-amine [00248] A solution of 3,6-dibromopyrazin-2-amine (10.085 g, 39.878 mmol) in anhydrous THF (100 mL) was bubbled with nitrogen for 5 minutes, then triethylamine (5.8080 g, 8 mL, 57.397 mmol), CuI (764 mg, 4.0116 mmol) and PdCl2(PPh3)2 (1.388 g, 1.9775 mmol) were added. The reaction was stirred at room temperature for 30 minutes. To the reaction mixture was slowly added dropwise 3,3-dimethylbut-1-yne (3.3350 g, 5 mL, 40.600 mmol) at room temperature. The reaction was stirred at room temperature for 1 h then was diluted with EtOAc (100 mL). The suspension was filtered through Celite, and the pad was rinsed with EtOAc (3 x 75 mL). The combined organic filtrates evaporated under reduced pressure (to remove THF). The crude product was solubilized in EtOAc (150 mL) and the solution was washed with a 50% brine solution (100 mL) and brine (100 mL), then was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The material was purified by trituration in MTBE/Heptane (2/1) (50 mL). The suspension was filtered, washed with MTBE/heptane (25 mL) and dried under high vacuum to give a yellow solid. The product was then purified by normal phase chromatography eluting from 0-70% EtOAc in heptanes (no separation). The product was then purified by reverse phase chromatography eluting from 5-60% I in water (0,1% FA) (the product came out in the I flush). The product was then repurified by reverse phase chromatography eluting from 55-85% I in water (0.1% FA) to give 2 fractions. Fraction 1 contained the pure desired compound while fraction 2 contained the desired product with a dimeric impurity. Fraction 2 was repurified by reverse phase chromatography eluting from 40- 80% I in water (0.1 % FA) to give the pure desired compound. Both lots of pure compound were mixed together to give 6-bromo-3-(3,3-dimethylbut-1-ynyl)pyrazin-2-amine (5.8 g, 57%) as a light yellow solid. ESI-MS m/z calc.253.0215, found 254.0 (M+1)+; Retention time: 1.97 minutes.1H NMR (400 MHz, DMSO-d6) δ 7.79 (s, 1H), 6.87 (br. S., 2H), 1.31 (s, 9H). LC method O. Step 2: 3-Bromo-6-tert-butyl-5H-pyrrolo[2,3-b]pyrazine [00249] To a stirred solution of 6-bromo-3-(3,3-dimethylbut-1-ynyl)pyrazin-2-amine (7.50 g, 29.51 mmol) in t-BuOH (80 mL) was added sodium tert-butoxide (14.25 g, 148.3 mmol). The reaction mixture was stirred at 80 °C for 4 h, then allowed to cool down to ambient temperature. The volatiles were removed under reduced pressure and the residue was diluted with ice-cold water (250 mL). The suspension was stirred at 0-4 °C (ice-water bath) for 15 min. The solid was filtered and washed with water (3 x 30 ml) and air-dried. The filter-cake was taken up in acetonitrile and concentrated under reduced pressure and the azeotropic water removal process repeated three times. Then the solid was dried under reduced pressure to furnish 3-bromo-6-tert- butyl-5H-pyrrolo[2,3-b]pyrazine as a tan solid (7.23 g, 96%), 1H NMR (400 MHz, DMSO) δ 12.25 (s, 1H), 8.37 (s, 1H), 6.37 (s, 1H), 1.37 (s, 9H)., ESI-MS m/z calc.253.02145, found 255.9 (M+3)+; Retention time: 1.47 minutes. LC method A. Step 3: 3-Bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine [00250] Sodium hydride (60% w/w in oil) (2.18 g, 54.505 mmol) was added portion-wise to a solution of 3-bromo-6-tert-butyl-5H-pyrrolo[2,3-b]pyrazine (6.71 g, 25.084 mmol) in anhydrous DMF (128 mL) at 0 °C. The suspension was stirred at this temperature for 30 min. Dimethyl sulfate (4.7988 g, 3.6 mL, 38.046 mmol) was added and left stirring at 0 °C for 15 min. The reaction was then stirred at room temperature for 1 h. The reaction mixture was cooled down to 0 °C and quenched with saturated aqueous ammonium chloride solution (250 mL). The resulting precipitate was filtered and washed with water (2 x 50 mL). The precipitate was dissolved in EtOAc (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting precipitate was purified by reversed phase chromatography (Gradient: 5% MeOH in water, then 60-85% MeOH in water). The desired fractions were combined and concentrated under reduced pressure to afford 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazine (5.96 g, 84%) as a light yellow solid.1H NMR (400 MHz, CDCl3) δ 8.42 (s, 1H), 6.52 (s, 1H), 4.01 (s, 3H), 1.51 (s, 9H). ESI-MS m/z calc.267.0371, found 268.2 (M+1)+; Retention time: 1.95 minutes. LC method I. Example 7: Preparation of 3-bromo-6-tert-butyl-5-(trideuteriomethyl)pyrrolo[2,3- b]pyrazine Step 1: 3-Bromo-6-tert-butyl-5-(trideuteriomethyl)pyrrolo[2,3-b]pyrazine [00251] To a solution of 3-bromo-6-tert-butyl-5H-pyrrolo[2,3-b]pyrazine (2.50 g, 9.837 mmol) in DMF (25 mL) at 0 °C was added NaH (in mineral oil) (650 mg of 60 %w/w, 16.25 mmol). The reaction mixture was stirred for 20 minutes then trideuterio(iodo)methane (1.75 g, 12.07 mmol) was added. The reaction mixture was stirred for 1 h and ice was added carefully (caution! Rapid effervescence). The suspension was diluted with water and was stirred at room temperature for 10 minutes. The mixture was extracted with ethyl acetate (3 x 25 mL) and the combined organics were washed with water (20 mL), brine (20 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure to furnish 3-bromo-6-tert-butyl-5- (trideuteriomethyl)pyrrolo[2,3-b]pyrazine as an orange-brown oil (1.88 g, 70%), 1H NMR (400 MHz, CDCl3) δ 8.41 (s, 1H), 6.48 (s, 1H), 1.50 (s, 9H)., ESI-MS m/z calc.270.05594, found 271.08 (M+1)+; Retention time: 1.71 minutes. LC method A. Example 8: Preparation of 2-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine Step 1: 5-Bromo-3-(3,3-dimethylbut-1-ynyl)pyrazin-2-amine [00252] A stirred solution of 3,5-dibromopyrazin-2-amine (5.00 g, 19.77 mmol) in anhydrous THF (80 mL) was sparged with nitrogen for 10 min. Then triethylamine (3.5 mL, 25.11 mmol), CuI (305 mg, 1.601 mmol) and PdCl2(PPh3)2 (556 mg, 0.7921 mmol) were added, in that order, at ambient temperature. The heterogeneous yellow mixture was cooled to 0 °C and 3,3- dimethylbut-1-yne (1.62 g, 19.72 mmol) was added dropwise over 5 min. After stirring the reaction for 3 h at that temperature, it was diluted with ethyl acetate (100 mL). The suspension was filtered over a pad of Celite, and the filter cake was rinsed with ethyl acetate (3 x 50 mL). The filtrate was washed with water (2 x 50 mL). The aqueous layer was extracted with ethyl acetate (50 mL). The combined organics were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to furnish 5-bromo-3-(3,3- dimethylbut-1-ynyl)pyrazin-2-amine (5.02 g, 100%) as an orange-yellow solid. ESI-MS m/z calc.253.02145, found 254.0 (M+1)+; Retention time: 1.57 minutes. LC method A. Step 2: 2-Bromo-6-tert-butyl-5H-pyrrolo[2,3-b]pyrazine [00253] To a stirred solution of 5-bromo-3-(3,3-dimethylbut-1-ynyl)pyrazin-2-amine (5.02 g, 19.75 mmol) in t-BuOH (50 mL) was added sodium tert-butoxide (9.49 g, 98.75 mmol). The reaction mixture was stirred at 80 °C for 4 h then was allowed to cool down to ambient temperature. The volatiles were removed under reduced pressure and the residue was taken up in water (50 mL). The resulting dark suspension was stirred at 0-4 °C for 15 min, then filtered off. The solid was washed with water (3 x 20 mL) and dried under reduced pressure to furnish 2- bromo-6-tert-butyl-5H-pyrrolo[2,3-b]pyrazine (4.65 g, 93%) as a tan solid.1H NMR (400 MHz, DMSO) δ 12.20 (s, 1H), 8.25 (s, 1H), 6.36 (d, J = 2.0 Hz, 1H), 1.38 (s, 9H). ESI-MS m/z calc. 253.02145, found 254.0 (M+1)+; Retention time: 1.39 minutes. LC method A. Step 3: 2-Bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine [00254] To a stirred solution of 2-bromo-6-tert-butyl-5H-pyrrolo[2,3-b]pyrazine (500 mg, 1.967 mmol) in anhydrous DMF (8 mL) was added NaH (in mineral oil) (115 mg of 60 %w/w, 2.875 mmol) at 0 °C under nitrogen. After stirring the reaction mixture for 20 min, iodomethane (340 mg, 2.395 mmol) was added. After another 1 h, at that temperature, ice was added very carefully (caution! Rapid effervescence). The suspension was diluted with cold water (100 mL) and stirring was continued at that temperature for 10 min. The solid was filtered and washed with water (3 x 25 mL) and dried in vacuo to furnish 2-bromo-6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazine as a dark brown solid (492 mg, 93%).1H NMR (400 MHz, DMSO-d6) δ 8.32 (s, 1H), 6.49 (s, 1H), 3.96 (s, 3H), 1.46 (s, 9H). ESI-MS m/z calc.267.0371, found 267.9 (M+1)+; Retention time: 1.58 minutes. LC method A. Example 9: Preparation of 2-bromo-7-isopropyl-quinoxaline Step 1: 1-Fluoro-4-isopropenyl-2-nitro-benzene [00255] To a solution of 4-bromo-1-fluoro-2-nitro-benzene (10.01 g, 45.501 mmol) and sodium carbonate (5.31 g, 50.100 mmol) in 1,4-dioxane (200 mL) and water (50 mL) was sparged with nitrogen. Bis(triphenylphosphine)palladium(II)chloride (659.3 mg, 0.9393 mmol) was added and the mixture was sparged with nitrogen. Isopropenylboronic acid pinacol ester (8.3927 g, 9.43 mL, 49.944 mmol) was added and the mixture was stirred at 80 °C for 3 hours. The mixture was cooled to rt, filtered over Celite, washed with MTBE (3x 20 mL) and the volatiles of the filtrate were removed under reduced pressure. The mixture was diluted with water (200 mL) and extracted with MTBE (3x150 mL). The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure to afford the crude 1-fluoro-4-isopropenyl-2-nitro-benzene (10.5 g, 125%) without further purification as a brown semi-solid. ESI-MS m/z calc.181.16, no ionization; Retention time: 2.12 minutes.1H NMR (400 MHz, Chloroform-d) δ 8.12 (dd, J = 7.1, 2.4 Hz, 1H), 7.75 – 7.69 (m, 1H), 7.29 – 7.21 (overlapped with chloroform, m, 1H), 5.43 (s, 1H), 5.24 (s, 1H), 2.17 (s, 3H).19F NMR (377 MHz, Chloroform-d) δ -119.85 (s, 1F). LC method O. Step 2: Methyl 2-(4-isopropenyl-2-nitro-anilino)acetate [00256] A solution of 1-fluoro-4-isopropenyl-2-nitro-benzene (8.2 g, 44.584 mmol) in anhydrous DMA (150 mL) was sparged with nitrogen. Methyl 2-aminoacetate (hydrochloride salt) (7.59 g, 60.452 mmol) and DIPEA (21.518 g, 29 mL, 166.49 mmol) were added and the mixture was stirred at 80 °C for 1 hour. The mixture was cooled to rt and poured into water (500 mL). The mixture was stirred at rt for 48 hours. The resulting precipitate was filtered and washed with water (2 x 20 mL). Purification by silica gel chromatography (Gradient 0 – 80% EtOAc in heptane) afforded after evaporation of the volatiles methyl 2-(4-isopropenyl-2-nitro- anilino)acetate (7.20 g, 64%) as a first crop as an orange solid. ESI-MS m/z calc.250.0954, found 251.1 (M+1)+; Retention time: 2.22 minutes 1H NMR (400 MHz, Chloroform-d) δ 8.47 – 8.37 (m, 1H), 8.29 (d, J = 2.4 Hz, 1H), 7.67 (dd, J = 8.8, 2.2 Hz, 1H), 6.68 (d, J = 8.8 Hz, 1H), 5.35 (s, 1H), 5.10 – 5.05 (m, 1H), 4.14 (d, J = 5.4 Hz, 2H), 3.84 (s, 3H), 2.15 (br s, 3H). The fractions containing a second crop were combined and evaporated under reduced pressure to afford methyl 2-(4-isopropenyl-2-nitro-anilino)acetate (1.3 g, 12%) as an orange solid. ESI-MS m/z calc.250.0954, found 251.1 (M+1)+; Retention time: 2.28 minutes 1H NMR (400 MHz, Chloroform-d) δ 8.47 – 8.36 (m, 1H), 8.29 (d, J = 2.2 Hz, 1H), 7.67 (dd, J = 8.8, 2.2 Hz, 1H), 6.68 (d, J = 8.8 Hz, 1H), 5.38 – 5.34 (m, 1H), 5.10 – 5.05 (m, 1H), 4.14 (d, J = 5.4 Hz, 2H), 3.84 (s, 3H), 2.15 (s, 3H). LC method O. Step 3: 7-Isopropyl-3,4-dihydro-1H-quinoxalin-2-one [00257] A solution of methyl 2-(4-isopropenyl-2-nitro-anilino)acetate (7.2 g, 28.742 mmol) in MeOH (200 mL) was sparged with nitrogen. Palladium on carbon (10% wt, 50% wet) (696.3 mg, 5 %w/w, 0.3271 mmol) was added and the mixture was sparged with nitrogen and then with hydrogen. The mixture was stirred under hydrogen atmosphere (balloon) at rt for 48 hours. Palladium on carbon (10% wt, 50% wet) (621.8 mg, 5 %w/w, 0.2921 mmol) was added and the mixture was flushed with nitrogen and then with hydrogen. The mixture was stirred under hydrogen atmosphere (balloon) at rt for 16 hours. The mixture was filtered over Celite, and the volatiles of the filtrate were removed under reduced pressure to afford 7-isopropyl-3,4-dihydro- 1H-quinoxalin-2-one (5.24 g, 88%) as a yellow oil. ESI-MS m/z calc.190.1106, found 191.2 (M+1)+; Retention time: 1.85 minutes 1H NMR (400 MHz, DMSO-d6) δ 10.11 (s, 1H), 6.66 – 6.55 (m, 3H), 5.75 (s, 1H), 3.65 (d, J = 1.7 Hz, 2H), 2.70 (dt, J = 13.6, 6.8 Hz, 1H), 1.12 (d, J = 6.8 Hz, 6H). LC method O. Step 4: 7-Isopropyl-1H-quinoxalin-2-one [00258] To a solution of 7-isopropyl-3,4-dihydro-1H-quinoxalin-2-one (5.32 g, 27.545 mmol) in MeOH (200 mL) was added KotBu in THF (42 mL of 1 M, 42.000 mmol). Oxygen was flushed into the mixture for 5 min and the reaction was stirred under O2 atmosphere for 16 hours at rt. The volatiles were removed under reduced pressure. The crude was diluted with water (100 mL) and TFA was added (5 mL) to neutralize the reaction. The aqueous layer was extracted with EtOAC (3 x 100 mL). The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to afford 7-isopropyl- 1H-quinoxalin-2-one (4.23 g, 81%) as a yellow solid. ESI-MS m/z calc.188.095, found 189.2 (M+1)+; Retention time: 1.84 minutes 1H NMR (400 MHz, DMSO-d6) δ 12.33 (br s, 1H), 8.09 (s, 1H), 7.70 (d, J = 8.3 Hz, 1H), 7.22 (dd, J = 8.3, 2.0 Hz, 1H), 7.14 (d, J = 2.0 Hz, 1H), 2.98 (spt, J = 6.8 Hz, 1H), 1.23 (d, J = 6.8 Hz, 6H). LC method O. Step 5: 2-Bromo-7-isopropyl-quinoxaline [00259] To a solution of 7-isopropyl-1H-quinoxalin-2-one (4.23 g, 22.428 mmol) in acetonitrile (150 mL) were added POBr3 (9.05 g, 31.568 mmol) and potassium carbonate (4.52 g, 32.705 mmol) at room temperature. The mixture was stirred for 3 hours at 80 °C. The mixture was allowed to cool down and was placed into an ice bath at 0 °C. Water (150 mL) was added and the mixture was allowed to warm up to rt. Acetonitrile was removed under reduced pressure and the remaining aqueous layer was extracted with EtOAc (3 x 100 mL). The organic phases were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. Purification by silica gel chromatography (Gradient 0 – 20% EtOAc in heptane) afforded after evaporation of the volatiles 2-bromo-7-isopropyl-quinoxaline (4.73 g, 83%) as an orange oil. ESI-MS m/z calc.250.0106, found 251.1 (M+1)+; Retention time: 3.34 minutes 1H NMR (400 MHz, DMSO-d6) δ 8.96 (s, 1H), 8.09 – 8.01 (m, 1H), 7.89 – 7.82 (m, 2H), 3.16 (spt, J = 6.9 Hz, 1H), 1.30 (d, J = 6.8 Hz, 6H). LC method P. Example 10: Preparation of 5-bromo-2-tert-butyl-3-methyl-imidazo[4,5-b]pyrazine Step 1: 5-Bromo-N3-methyl-pyrazine-2,3-diamine [00260] Methyl amine in methanol (10 mL of 40 %w/v, 128.8 mmol) was added to 3,5- dibromopyrazin-2-amine (2.00 g, 7.908 mmol) and stirred for 10 min. Then water (2.0 mL) was added to the tan solution and the capped microwave tube was stirred in a microwave reactor at 130 °C (setting: very high absorption) for 1 h. The reaction was allowed to cool to ambient temperature and extracted with dichloromethane (3 x 20 mL), washed with brine (20 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to furnish 5-bromo-N3- methyl-pyrazine-2,3-diamine as a tan solid (1.580 g, 98%) 1H NMR (400 MHz, CDCl3) δ 7.46 (s, 1H), 4.28 (s, 1H), 4.09 (s, 2H), 3.02 (d, J = 5.0 Hz, 3H). ESI-MS m/z calc.201.98541, found 203.0 (M+1)+; Retention time: 0.61 minutes. LC method A. Step 2: 5-Bromo-2-tert-butyl-3-methyl-imidazo[4,5-b]pyrazine [00261] In flask was added pivalic acid (34 g, 332.90 mmol), 2,2-dimethylpropanoyl 2,2- dimethylpropanoate (14.672 g, 16 mL, 78.777 mmol) and 5-bromo-N3-methyl-pyrazine-2,3- diamine (7.8 g, 38.186 mmol). The flask was equipped with a Vigreux column and the reaction was stirred at 130 °C for 18 hours. The reaction was allowed to cool down to room temperature and was diluted with ethyl acetate (100 mL), washed with a 1M aqueous solution of NaOH (2 x 100 mL), brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude 5-bromo-2-tert-butyl-3-methyl-imidazo[4,5-b]pyrazine (21.6 g, 95%) as a dark red oil.1H NMR (400 MHz, CDCl3) δ 8.53 (s, 1H), 4.02 (s, 3H), 1.59 (s, 9H). ESI-MS m/z calc.268.0324, found 269.2 (M+1)+; Retention time: 1.69 minutes. LC method I. Example 11: Preparation of 3-bromo-5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine Step 1: 6-Bromo-3-[2-[1-(trifluoromethyl)cyclopropyl]ethynyl]pyrazin-2-amine [00262] In a flask under nitrogen was added THF (200 mL). nitrogen was flushed into the mixture for 10 min.1-Ethynyl-1-(trifluoromethyl)cyclopropane (10 g, 70.843 mmol), 3,6- dibromopyrazin-2-amine (35.83 g, 141.68 mmol), triethylamine (21.780 g, 30 mL, 215.24 mmol), bis(triphenylphosphine)palladium(II) dichloride (987.7 mg, 1.4072 mmol) and copper iodide (272.2 mg, 1.4292 mmol) were added. The resulting mixture was stirred at room temperature under nitrogen atmosphere for 4 hours. The reaction mixture was filtered on Celite and the cake was washed with acetonitrile (5 x 30 mL). Water with 0.1% formic acid (200 mL) was added in the filtrate and the precipitate was collected by filtration. The solid was dissolved in I (50 mL) and water + 0.1% formic acid was added until a precipitate formed. The solid was collected by filtration. This step was done 4 times until the starting material was removed from the crude. The desired product 6-bromo-3-[2-[1-(trifluoromethyl)cyclopropyl]ethynyl]pyrazin-2- amine (19.66 g, 76%) was obtained as a yellow solid. ESI-MS m/z calc.304.9775, found 306.0 (M+1)+; Retention time: 2.05 minutes.1H NMR (400 MHz, Chloroform-d) δ 7.97 (s, 1H), 5.15 (br s, 2H), 1.50 – 1.44 (m, 2H), 1.41 – 1.34 (m, 2H).19F NMR (377 MHz, Chloroform-d) δ - 70.07 (s, 3F). LC method O. Step 2: 3-Bromo-5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazine [00263] To a solution of 6-bromo-3-[2-[1-(trifluoromethyl)cyclopropyl]ethynyl]pyrazin-2- amine (19.66 g, 53.761 mmol) in anhydrous DMF (150 mL) at 0 °C under nitrogen, was added NaH (4.5 g, 60 %w/w, 112.51 mmol). The mixture was stirred at 70 °C for 1 hour. The mixture was allowed to cool down to 0 °C. A solution of dimethyl sulfate (13.566 g, 10.2 mL, 107.55 mmol) in anhydrous DMF (50 mL) was added dropwise to the mixture at 0 °C over 15 minutes. The mixture was stirred at 0 °C for 1 hour. Then, a saturated aqueous solution of ammonium chloride (250 mL) and water (250 mL) were added. The aqueous phase was extracted with EtOAc (3 x 250 mL). The organic phases were combined, washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. Purification by silica gel chromatography (Gradient 0 – 10% EtOAc in heptane) afforded after evaporation of the volatiles 3-bromo-5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine (14.03 g, 81%) as a light yellow solid. ESI-MS m/z calc.318.9932, found 320.0 (M+1)+; Retention time: 2.17 minutes 1H NMR (400 MHz, Chloroform-d) δ 8.51 (s, 1H), 6.77 (s, 1H), 3.92 (s, 3H), 1.64 – 1.57 (overlapped with water, m, 2H), 1.28 – 1.22 (m, 2H).19F NMR (377 MHz, Chloroform-d) δ -68.75 (s, 3F). LC method O. Example 12: An alternative preparation of 3-bromo-5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine Step 1: Triisopropyl-[3-(trifluoromethyl)but-3-en-1-ynyl]silane [00264] To a pressure tube were added CuI (41 mg, 0.2153 mmol) and bis(triphenylphosphine)palladium(II) dichloride (77 mg, 0.1094 mmol). The mixture was dried under vacuum for 15 min. Then, THF (2 mL) and TEA (2.5410 g, 3.5 mL, 25.111 mmol) were added, followed by ethynyl(triisopropyl)silane (2 g, 10.966 mmol) and cold 2-bromo-3,3,3- trifluoro-prop-1-ene (2.5 g, 14.290 mmol). The tube was closed and heated for 20 min at 50 °C in an oil bath. The reaction mixture was cooled to room temperature, partitioned between pentane (100 mL) and saturated aqueous ammonium chloride (50 mL) containing HCl (30 mL of 1 M, 30.000 mmol) and water (50 mL). The aqueous phase was separated and extracted with pentane (2 x 50 mL). The combined organics were dried on magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified on silica gel by elution in 100% pentane to provide triisopropyl-[3-(trifluoromethyl)but-3-en-1-ynyl]silane (2.76 g, 87%) as a clear oil 1H NMR (400 MHz, CDCl3) δ 6.05 (q, J = 1.3 Hz, 1H), 5.89 (q, J = 1.2 Hz, 1H), 1.12 - 1.08 (m, 21H).19F NMR (377 MHz, CDCl3) δ -68.20 (s, 3F). Step 2: Triisopropyl-[2-[1-(trifluoromethyl)cyclopropyl]ethynyl]silane [00265] LiHMDS (10.3 mL of 1 M, 10.300 mmol) was added to triisopropyl-[3- (trifluoromethyl)but-3-en-1-ynyl]silane (1 g, 3.4369 mmol) and diphenyl(methyl)sulfonium tetrafluoroborate (1.38 g, 4.7898 mmol) in solution in THF (2 mL) maintained at -78 °C. The reaction mixture was left to slowly warm up to room temperature over 18 h. The reaction mixture was diluted with pentane (15 mL) and filtered on a silica gel column and the desired product was eluted with pentane. The filtrates were concentrated under reduced pressure. The resulting residue was purified by reverse phase chromatography on a C18 column using a 5 to 100% gradient of acetonitrile in pure water. The fractions containing the desired fraction were combined and extracted with pentane (3 x 50 mL). The pentane phases were combined, washed with water (20 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to provide triisopropyl-[2-[1-(trifluoromethyl)cyclopropyl]ethynyl]silane (550 mg, 52%) as a clear oil.1H NMR (400 MHz, CDCl3) δ 1.30 - 1.26 (m, 2H), 1.17 - 1.12 (m, 2H), 1.08 - 1.04 (m, 21H).19F NMR (377 MHz, CDCl3) δ -70.80 (s, 3F). Step 3: 6-Bromo-3-[2-[1-(trifluoromethyl)cyclopropyl]ethynyl]pyrazin-2-amine [00266] Under a nitrogen atmosphere in a microwave vial were added 3,6-dibromopyrazin-2- amine (8.5 g, 33.611 mmol), triisopropyl-[2-[1-(trifluoromethyl)cyclopropyl]ethynyl]silane (5 g, 16.354 mmol), CuI (625 mg, 3.2817 mmol), Pd(PPh3)2Cl2 (574 mg, 0.8178 mmol), PPh3 (430 mg, 1.6394 mmol), TEA (1.8150 g, 2.5 mL, 17.937 mmol), and THF (35 mL). The vial was capped and the mixture was warmed-up to 60 °C, and TBAF (in THF) (17 mL of 1 M, 17.000 mmol) was introduced dropwise in the vial via a syringe. The reaction mixture was left stirring at this temperature for 3 h and then left to cool to room temperature. The mixture was filtered on a pad of silica gel, and the pad was eluted with ethyl acetate. The combined filtrates were concentrated under reduced pressure. The resulting residue was purified by flash chromatography on silica gel using a gradient of 0 to 100% ethyl acetate in heptanes. The collected fractions were combined and concentrated under reduced pressure. The resulting residue was purified using multiple reverse phase chromatography passes (3 x) on a C18 column using a 5 to 100% gradient of acetonitrile in pure water to provide 6-bromo-3-[2-[1- (trifluoromethyl)cyclopropyl]ethynyl]pyrazin-2-amine (3.31 g, 63%) as a yellow solid ESI-MS m/z calc.304.9775, found 305.9 (M+1)+; Retention time: 2.27 minutes Step 4: 3-Bromo-5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazine [00267] To a solution of 6-bromo-3-[2-[1-(trifluoromethyl)cyclopropyl]ethynyl]pyrazin-2- amine (19.66 g, 53.761 mmol) in anhydrous DMF (150 mL) at 0 °C under nitrogen, was added NaH (4.5 g, 60 %w/w, 112.51 mmol). The mixture was stirred at 70 °C for 1 hour. The mixture was allowed to cool down to 0 °C. A solution of dimethyl sulfate (13.566 g, 10.2 mL, 107.55 mmol) in anhydrous DMF (50 mL) was added drop-wise to the mixture at 0 °C over 15 minutes. The mixture was stirred at 0 °C for 1 h. Then, a saturated aqueous solution of ammonium chloride (250 mL) and water (250 mL) were added. The aqueous phase was extracted with EtOAc (3 x 250 mL). The organic phases were combined, washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. Purification by silica gel chromatography (Gradient 0 - 10% EtOAc in heptane) afforded after evaporation of the volatiles 3-bromo-5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine (14.03 g, 81%) as a light yellow solid. ESI-MS m/z calc.318.9932, found 320.0 (M+1)+; Retention time: 2.17 minutes 1H NMR (400 MHz, Chloroform-d) δ 8.51 (s, 1H), 6.77 (s, 1H), 3.92 (s, 3H), 1.64 - 1.57 (overlapped with water, m, 2H), 1.28 - 1.22 (m, 2H).19F NMR (377 MHz, Chloroform-d) δ -68.75 (s, 3F). Example 13: Preparation of 3-bromo-8,8-dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizine Step 1: tert-Butyl 2-(hydroxymethyl)-3,3-dimethyl-pyrrolidine-1-carboxylate [00268] To a solution of 1-tert-butoxycarbonyl-3,3-dimethyl-pyrrolidine-2-carboxylic acid (4.5 g, 18.496 mmol) in THF (80 mL) was added borane dimethyl sulfide (4.2400 g, 5.3 mL, 55.812 mmol) dropwise. The mixture was stirred at 50 °C for 10 min and cooled down to room temperature. MeOH (10 mL) was added, and the mixture was stirred at room temperature for 1 h. Concentration of the reaction mixture under reduced pressure afforded tert-butyl 2- (hydroxymethyl)-3,3-dimethyl-pyrrolidine-1-carboxylate (4.2 g, 99%) as a colorless oil.1H NMR (400 MHz, Chloroform-d) δ 3.80 – 3.70 (m, 1H), 3.60 – 3.30 (m, 5H), 1.74 – 1.55 (m, 2H), 1.48 (s, 9H), 1.08 (s, 3H), 0.95 (s, 3H). ESI-MS m/z calc.229.1678, found 174.2 (M-55)+; Retention time: 1.97 minutes. LC method O. Step 2: tert-Butyl 2-formyl-3,3-dimethyl-pyrrolidine-1-carboxylate [00269] To a solution of tert-butyl 2-(hydroxymethyl)-3,3-dimethyl-pyrrolidine-1-carboxylate (4.2 g, 18.297 mmol) in water-saturated DCM (150 mL) was added DMP (15 g, 35.366 mmol). The mixture was stirred at room temperature for 16 h. Silica (20 g) was added, and the volatiles were removed under reduced pressure. Purification by silica gel chromatography (Gradient: 0-10 % EtOAc in heptanes) yielded after evaporation of the volatiles tert-butyl 2-formyl-3,3- dimethyl-pyrrolidine-1-carboxylate (4.2 g, 96%) as a colorless oil ESI-MS m/z calc.227.1521, found 172.2 (M-55)+; Retention time: 2.03 minutes.1H NMR (400 MHz, Chloroform-d) δ 9.54 – 9.41 (m, 1H), 3.74 – 3.46 (m, 3H), 1.77 – 1.65 (m, 2H), 1.48 – 1.40 (m, 9H), 1.17 (s, 3H), 1.09 – 1.02 (m, 3H). LC method O. Step 3: tert-Butyl 2-[(3,5-dibromopyrazin-2-yl)-hydroxy-methyl]-3,3-dimethyl- pyrrolidine-1-carboxylate [00270] A dried 100-mL flask under nitrogen was charged with 2,2,6,6-tetramethylpiperidine (510 mg, 3.6105 mmol) and THF (20 mL). The mixture was cooled to -40 °C and a n-BuLi solution in hexanes (1.5 mL of 2.5 M, 3.7500 mmol) was added dropwise. The mixture was stirred at 0 °C for 20 minutes and cooled to -100 °C. A solution of 2,6-dibromopyrazine (710 mg, 2.9847 mmol) in THF (7 mL) was added dropwise over 40 minutes. After addition, the mixture was slowly warmed to -90 °C over 23 minutes and kept at -90 °C - -86 °C (bath temperature) for 8 minutes. The mixture was re-cooled to -100 °C and a solution of tert-butyl 2- formyl-3,3-dimethyl-pyrrolidine-1-carboxylate (617 mg, 2.5788 mmol) in THF (5 mL) was added dropwise over 15 minutes. After addition, the mixture was stirred at -100 °C for 10 minutes. A solution of HOAc (180 mg, 3 mmol) in THF (3 mL) was added dropwise to quench the reaction. The cooling bath was removed and heptanes (75 mL) was added, followed by saturated aqueous sodium bicarbonate (10 mL). The two layers were separated, and the aqueous phase was extracted with heptanes-EtOAc (3:1, 28 mL) and EtOAc (30 mL). The combined organic layers were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (Gradient: 5-80% MeCN in water with 0.1% formic acid). The desired fractions were collected, neutralized with saturated aqueous sodium bicarbonate (2 mL) and concentrated under reduced pressure to remove most of MeCN. The residue was extracted with DCM (3 x 20 mL). The combined organic layers were dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give tert-butyl 2-[(3,5-dibromopyrazin-2-yl)-hydroxy-methyl]-3,3-dimethyl- pyrrolidine-1-carboxylate (1.087 g, 91%) as a light-brown oil. ESI-MS m/z calc.463.0106, found 364.0 (M-99)+; Retention time: 2.21 minutes. LC method O. Step 4: (3,5-Dibromopyrazin-2-yl)-(3,3-dimethylpyrrolidin-2-yl)methanol [00271] Concentrated HBr (in water) (6.4368 g, 9 mL, 38.186 mmol) was added to a solution of tert-butyl 2-[(3,5-dibromopyrazin-2-yl)-hydroxy-methyl]-3,3-dimethyl-pyrrolidine-1- carboxylate (1.4 g, 2.8772 mmol) in diethyl ether (18 mL) and water (9 mL). The reaction mixture was stirred for 16 h at room temperature and then concentrated under reduced pressure. The resulting residue was quenched with aqueous sodium bicarbonate (25 mL) until pH = 4-5. The resulting aqueous solution was purified by reversed-phase chromatography (C18 column. Gradient: 0-20 % MeCN in water with 0.1 % formic acid). The volatiles were removed under reduced pressure from the fractions containing the product to afford (3,5-dibromopyrazin-2-yl)- (3,3-dimethylpyrrolidin-2-yl)methanol (Hydrobromic Acid (1)) (1.1 g, 83%) as a white solid. ESI-MS m/z calc.362.9582, found 363.9 (M+1)+; Retention time: 1.54 minutes.1H NMR (400 MHz, DMSO-d6) δ 9.25 (br s, 1H), 9.01 (s, 1H), 8.86 (br s, 1H), 6.63 (d, J = 4.2 Hz, 1H), 5.21 (dd, J = 7.1, 4.2 Hz, 1H), 3.72 – 3.57 (m, 1H), 3.20 (br s, 2H), 1.90 – 1.72 (m, 2H), 1.01 (s, 3H), 0.74 (s, 3H). LC method O. Step 5: 3-bromo-8,8-dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizine [00272] To a solution of (3,5-dibromopyrazin-2-yl)-(3,3-dimethylpyrrolidin-2-yl)methanol (trifluoroacetate salt) (4.7 g, 9.8005 mmol) in MeCN (85 mL) was added TEA (10.164 g, 14 mL, 100.44 mmol). The reaction mixture was stirred at reflux for 1.5 h. The reaction mixture was cooled down to 0 °C and Ms2O (5 g, 28.703 mmol) was added portion wise. The reaction mixture was stirred at 0 °C for 1.5 h. Water (30 mL) and brine (40 mL) were added and the mixture was extracted with EtOAc (3 x 75 mL). The combined organic layers were washed with saturated aqueous solution of sodium bicarbonate (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by silica gel chromatography (Gradient: 0-20 % EtOAc in heptanes) yielded after evaporation of the volatiles and dried under high vacuum 3-bromo-8,8-dimethyl-6H,7H,8H-pyrazino[2,3- b]pyrrolizine (2.4 g, 91%) as a tan solid.1H NMR (400 MHz, Chloroform-d) δ 8.40 (s, 1H), 6.30 (s, 1H), 4.26 (br. T, J = 6.7 Hz, 2H), 2.42 (br. T, J = 6.8 Hz, 2H), 1.46 (s, 6H). ESI-MS m/z calc.265.0215, found 266.0 (M+1)+; Retention time: 2.87 minutes. LC method P. Example 14: Preparation of 3-bromo-5-methyl-6-[1- (trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazine Step 1: 6-Bromo-3-[2-[1-(trifluoromethyl)cyclobutyl]ethynyl]pyrazin-2-amine [00273] A solution of 3,6-dibromopyrazin-2-amine (1.56 g, 6.169 mmol) in anhydrous THF (31 mL) was bubbled with nitrogen for 5 minutes, then triethylamine (1.05 mL, 7.533 mmol), CuI (94 mg, 0.4936 mmol) and PdCl2(PPh3)2 (175 mg, 0.2493 mmol) were added. The reaction mixture was cooled to 0 °C and 1-ethynyl-1-(trifluoromethyl)-cyclobutane (910.5 mg, 6.147 mmol) was slowly added dropwise. The reaction was stirred at 0 °C for 5 hours, then was diluted with EtOAc. The suspension was filtered through Celite, and the pad was rinsed with EtOAc. The product was purified by silica gel chromatography (120 g column, Gradient: 0-50 % EtOAc in Hexane, 30 min) and reverse phase chromatography (C18, water/ I (0-100% over 25 min) to afford 6-bromo-3-[2-[1-(trifluoromethyl)cyclobutyl]ethynyl]pyrazin-2-amine as a yellow solid (1.1408 g, 57%), 1H NMR (400 MHz, DMSO) δ 7.86 (s, 1H), 7.12 (s, 2H), 2.70 – 2.53 (m, 4H), 2.21 – 2.12 (m, 1H), 2.07 – 1.95 (m, 1H). ESI-MS m/z calc.318.9932, found 320.022 (M+1)+; Retention time: 0.672 minutes. LC method B. Step 2: 3-Bromo-6-[1-(trifluoromethyl)cyclobutyl]-5H-pyrrolo[2,3-b]pyrazine [00274] To a suspension of 6-bromo-3-[2-[1-(trifluoromethyl)cyclobutyl]ethynyl]pyrazin-2- amine (1.24 g, 3.874 mmol) in t-BuOH (13 mL) was added sodium tert-butoxide (2.18 g, 19.43 mmol). The reaction mixture was stirred at 90 °C for 4 hours then was allowed to cool down to room temperature. The volatiles were removed under reduced pressure and the residue was suspended in water (10 ml). The suspension was stirred for 10 minutes in an ice-cold bath and extracted with EtOAc. The organic layers were concentrated under reduced pressure. The orange solid was dried under reduced pressure to afford 3-bromo-6-[1-(trifluoromethyl)cyclobutyl]-5H- pyrrolo[2,3-b]pyrazine (1.24 g, 99%).1H NMR (400 MHz, DMSO) δ 8.43 (s, 1H), 6.74 (s, 1H), 2.74 – 2.59 (m, 4H), 2.12 – 1.91 (m, 2H). ESI-MS m/z calc.318.9932, found 319.9 (M+1)+; Retention time: 0.62 minutes. LC method B. Step 3: 3-Bromo-5-methyl-6-[1-(trifluoromethyl)cyclobutyl]pyrrolo[2,3- b]pyrazine [00275] To a solution of 3-bromo-6-[1-(trifluoromethyl)cyclobutyl]-5H-pyrrolo[2,3- b]pyrazine (909.4 mg, 2.812 mmol) in DMF (20 mL) at 0 °C was added NaH (in mineral oil) (136 mg of 60 %w/w, 3.400 mmol). The reaction mixture was stirred for 25 minutes then methyl iodide (196 µL, 3.148 mmol) was added. The reaction mixture was stirred for 1 hour in an ice bath and then ice was added to the reaction. The suspension was diluted with water and was stirred at room temperature for 10 minutes. The solid was filtered and washed with water and dried in vacuo to afford 3-bromo-5-methyl-6-[1-(trifluoromethyl)cyclobutyl]pyrrolo[2,3- b]pyrazine (889.6 mg, 94%) as a beige solid. 1H NMR (400 MHz, CDCl3) δ 8.42 (s, 1H), 6.54 (s, 1H), 3.71 (s, 3H), 2.87 – 2.78 (m, 2H), 2.74 – 2.62 (m, 2H), 2.37 – 2.22 (m, 1H), 2.07 – 1.94 (m, 1H). ESI-MS m/z calc.333.00885, found 333.97 (M+1)+; Retention time: 0.71 minutes. LC method B. Example 15: Preparation of 3-bromo-5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl- ethyl)pyrrolo[2,3-b]pyrazine Step 1: 6-Bromo-3-(4,4,4-trifluoro-3,3-dimethyl-but-1-ynyl)pyrazin-2-amine [00276] In a flask under nitrogen atmosphere containing a solution of 4,4,4-trifluoro-3,3- dimethyl-but-1-yne (11.31 g, 83.092 mmol) in THF (250 mL) were added 3,6-dibromopyrazin- 2-amine (9.15 g, 36.181 mmol), triethylamine (12.342 g, 17 mL, 121.97 mmol), bis(triphenylphosphine)palladium(II) dichloride (3.52 g, 5.0150 mmol) and copper iodide (800 mg, 4.2006 mmol). The resulting mixture was stirred at room temperature under nitrogen atmosphere for 2 hours. Bis(triphenylphosphine)palladium(II) dichloride (1.57 g, 2.2368 mmol) and copper iodide (450 mg, 2.3628 mmol) were added to the mixture and the mixture was flushed with nitrogen for 5 min. The mixture was stirred for 2 hours at rt under nitrogen atmosphere. Additional Bis(triphenylphosphine)palladium(II) dichloride (1.583 g, 2.2553 mmol) and copper iodide (472 mg, 2.4783 mmol) were added to the mixture and the mixture was flushed with nitrogen for 5 min. The mixture was stirred for 16 hours at rt under nitrogen atmosphere. The reaction mixture was filtered on Celite, and the cake was washed with acetonitrile (5 x 30 mL). Purification by reversed-phase chromatography (C18 column. Gradient 5 - 80% acetonitrile in water with 0.1% formic acid) was performed. The fractions containing the product were combined and the volatiles were removed under reduced pressure. The aqueous phase was extracted with EtOAc (3 x 100 mL). The organic phases were combined, washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure to afford 6-bromo-3-(4,4,4-trifluoro-3,3-dimethyl-but-1-ynyl)pyrazin-2-amine (6.82 g, 60%) as a yellow solid. ESI-MS m/z calc.306.9932, found 308.1 (M+1)+; Retention time: 2.13 minutes 1H NMR (400 MHz, Chloroform-d) δ 7.98 (s, 1H), 5.11 (br s, 2H), 1.56 (overlapped with water, s, 6H).19F NMR (377 MHz, Chloroform-d) δ -77.85 (s, 3F). LC method O. Step 2: 3-Bromo-5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3- b]pyrazine [00277] To a solution of 6-bromo-3-(4,4,4-trifluoro-3,3-dimethyl-but-1-ynyl)pyrazin-2-amine (10.8 g, 35.019 mmol) in anhydrous DMF (280 mL), in a flame dried flask, at 0 °C under nitrogen, was added NaH (3.15 g, 60 % w/w, 78.758 mmol). The mixture was stirred at 70 °C for 60 min. The mixture was allowed to cool down to 0 °C. A solution of dimethyl sulfate (8.7780 g, 6.6 mL, 69.594 mmol) in anhydrous DMF (100 mL) was added dropwise to the mixture at 0 °C over 45 min using a syringe driver. The mixture was stirred at 0 °C for an additional 10 min. Then, a saturated aqueous solution of NH4Cl (2 L) and water (2 L) were added. The mixture was stirred at rt for 10 min. The precipitate was collected by filtration, washed with cold water (3 x 50 mL) and dried under high vacuum to afford 3-bromo-5-methyl- 6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3-b]pyrazine (10 g, 87%) as a brown solid. ESI- MS m/z calc.321.0088, found 322.0 (M+1)+; Retention time: 2.18 minutes. LC method O. Example 16: Preparation of 3-bromo-5-methyl-6-spiro[2.3]hexan-5-yl-pyrrolo[2,3- b]pyrazine Step 1: Methyl spiro[2.3]hexane-5-carboxylate [00278] To a solution of diethylzinc in hexanes (119 mL of 1 M, 119.00 mmol) in DCM (27.5 mL) was added a solution of TFA (13.480 g, 9.2 mL, 117.04 mmol) in DCM (27.5 mL) dropwise at 0 °C over 30 min. A solution of diiodomethane (30.239 g, 9.2 mL, 111.77 mmol) in DCM (27.5 mL) was added dropwise at 0 °C over 45 min. The reaction mixture was stirred at 0 °C for 1 h. A solution of methyl 3-methylenecyclobutanecarboxylate (5 g, 38.445 mmol) in DCM (27.5 mL) was added to the reaction mixture. The reaction mixture was allowed to warm to room temperature for 48 h. A saturated ammonium chloride solution (100 mL) was added, and the mixture was extracted with DCM (3 x 50 mL). The organic phases were combined, washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and carefully evaporated under reduced pressure to afford the crude methyl spiro[2.3]hexane-5-carboxylate (12.5 g, 100%) as a yellow liquid.1H NMR (400 MHz, Chloroform-d) δ 3.71 (s, 3H), 3.34 - 3.24 (m, 1H), 2.55 - 2.46 (m, 2H), 2.29 - 2.19 (m, 2H), 0.53 - 0.38 (m, 4H). ESI-MS m/z calc. 140.0837, found 141.1 (M+1)+; Retention time: 1.91 minutes. LC method O. Step 2: spiro[2.3]Hexane-5-carboxylic acid [00279] To a stirred solution of methyl spiro[2.3]hexane-5-carboxylate (37.5 g, 9.0954 mmol) in DCE (22 mL) was added NaOH (1.1 g, 27.502 mmol) in MeOH (10 mL). The mixture was stirred at room temperature for 16 h. Ether (50 mL) and water (80 mL) were added, and the phases were separated. The aqueous phase was washed with ether (2 x 80 mL) and then acidified with a 6 N HCl solution (5 mL). The aqueous phase was extracted with DCM (3 x 100 mL) and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford spiro[2.3]hexane-5-carboxylic acid (890 mg, 76%) as a clear oil.1H NMR (400 MHz, DMSO-d6) δ 12.06 (br s, 1H), 3.17 (tt, J = 9.4, 7.3 Hz, 1H), 2.38 - 2.29 (m, 2H), 2.23 - 2.13 (m, 2H), 0.50 - 0.32 (m, 4H). ESI-MS m/z calc.126.0681, found 127.2 (M+1)+; Retention time: 1.59 minutes. LC method O. Step 3: spiro[2.3]Hexan-5-ylmethanol [00280] To a solution of spiro[2.3]hexane-5-carboxylic acid (7.28 g, 46.974 mmol) in anhydrous THF (216 mL) at 0 °C was added dropwise a borane-THF complex (60 mL of 1 M, 60.000 mmol) solution under nitrogen. The reaction mixture was stirred at room temperature for 16 h, then was quenched with water (50 mL), a saturated aqueous solution of sodium bicarbonate (50 mL) and diethyl ether (400 mL) and water (350 mL) were added. The aqueous phase was extracted with diethyl ether (2 x 100 mL) and the combined organics were washed with brine (600 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to afford spiro[2.3]hexan-5-ylmethanol (7.9 g, 100%) as a yellow liquid.1H NMR (400 MHz, Chloroform-d) δ 3.73 - 3.69 (m, 2H), 2.67 - 2.54 (m, 1H), 2.21 - 2.12 (m, 2H), 1.90 - 1.82 (m, 2H), 0.46 - 0.34 (m, 4H), (1H missing, labile proton). Step 4: spiro[2.3]Hexane-5-carbaldehyde [00281] To a stirring mixture of spiro[2.3]hexan-5-ylmethanol (7.467 g, 51.191 mmol) and TEMPO (925 mg, 5.9202 mmol) in DCM (80 mL) at 0 °C was added a solution of sodium bromide (3.1 g, 30.128 mmol) in water (40 mL). This suspension was stirred at that temperature for 15 min then a solution of sodium hypochlorite (in water) (53 mL of 10 % w/v, 71.196 mmol) with sodium bicarbonate (8.9 g, 105.94 mmol) was added dropwise over 1 h, while avoiding using the excess of sodium bicarbonate at the bottom of the flask. The mixture was stirred at 0 °C for 30 minutes then it was diluted with distilled water (40 mL). The aqueous phase was separated then extracted with DCM (3 x 70 mL). The combined organics were washed with brine (250 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford spiro[2.3]hexane-5-carbaldehyde (5.6389 g, 100%) as a light yellow liquid.1H NMR (400 MHz, Chloroform-d) δ 9.80 (d, J = 2.3 Hz, 1H), 3.32 - 3.19 (m, 1H), 2.45 - 2.21 (m, 4H), 0.54 - 0.35 (m, 4H). Step 5: 5-(2,2-Dibromovinyl)spiro[2.3]hexane [00282] To a solution of spiro[2.3]hexane-5-carbaldehyde (6.52 g, 59.190 mmol) in DCM (290 mL) was added triphenylphosphine (47 g, 177.40 mmol). The mixture was placed in a water bath and a solution of CBr4 (30 g, 89.558 mmol) in DCM (40 mL) was added dropwise over 1 hour. The mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure and the resulting precipitate was triturated in pentane (100 mL). The solvent was filtered on a silica pad and the silica cake was washed with pentane (3 x 100 mL). The solvent was removed under reduced pressure. Purification by silica gel chromatography (isocratic: 100% heptanes) yielded after evaporation of the volatiles 5-(2,2- dibromovinyl)spiro[2.3]hexane (16.7 g, 66%) as a colorless liquid.1H NMR (400 MHz, Chloroform-d) δ 6.67 (d, J = 8.6 Hz, 1H), 3.38 - 3.23 (m, 1H), 2.37 - 2.27 (m, 2H), 2.16 - 2.07 (m, 2H), 0.52 - 0.36 (m, 4H). ESI-MS m/z calc.305.8867, found no ionization; Retention time: 2.39 minutes. LC method O. Step 6: 5-Ethynylspiro[2.3]hexane [00283] To a solution of 5-(2,2-dibromovinyl)spiro[2.3]hexane (13.6 g, 31.651 mmol) in anhydrous THF (113 mL) was added n-BuLi (solution in hexanes) (28 mL of 2.5 M, 70.000 mmol) dropwise over 30 minutes at -78 °C. After addition, the reaction mixture was stirred at - 78 °C for 1 hour and 30 minutes. Then it was quenched with an aqueous saturated ammonium chloride (2.2 mL) solution, dried over anhydrous sodium sulfate, and filtered over cotton to give 5-ethynylspiro[2.3]hexane (3.3602 g, 100%) as a solution in THF (113 mL). The product was used as a solution for the next step. Step 7: 6-Bromo-3-(2-spiro[2.3]hexan-5-ylethynyl)pyrazin-2-amine [00284] In a flask under nitrogen atmosphere containing a solution of 5- ethynylspiro[2.3]hexane (3.36 g, 31.649 mmol) in THF (26 mL) were added 3,6- dibromopyrazin-2-amine (4.6 g, 17.826 mmol), TEA (7.3326 g, 10.1 mL, 72.464 mmol), PdCl2(PPh3)2 (1.3 g, 1.8151 mmol) and CuI (345 mg, 1.7934 mmol). The resulting mixture was stirred at room temperature under nitrogen atmosphere for 16 hours. The reaction mixture was filtered on celite, and the cake was washed with acetonitrile (5 x 50 mL). The organics were concentrated under reduced pressure and the resulting residue was purified by reversed-phase chromatography (C18 column. Gradient: 5-100 % MeCN in water with 0.1 % formic acid) to afford after evaporation of the MeCN and extraction with DCM (3 x 150 mL) 6-bromo-3-(2- spiro[2.3]hexan-5-ylethynyl)pyrazin-2-amine (3.52 g, 71%) as a yellow solid.1H NMR (400 MHz, DMSO-d6) δ 7.79 (s, 1H), 6.99 (br s, 2H), 3.50 (tt, J = 8.9, 7.7 Hz, 1H), 2.49 - 2.34 (m, 4H), 0.52 - 0.40 (m, 4H). ESI-MS m/z calc.277.0215, found 278.1 (M+1)+; Retention time: 2.17 minutes. LC method O. Step 8: 3-Bromo-5-methyl-6-spiro[2.3]hexan-5-yl-pyrrolo[2,3-b]pyrazine [00285] To a solution of 6-bromo-3-(2-spiro[2.3]hexan-5-ylethynyl)pyrazin-2-amine (3.52 g, 12.642 mmol) in anhydrous DMF (22 mL), in a flame-dried flask at 0 °C under nitrogen, was added NaH (1.1 g, 60 %w/w, 27.503 mmol) portion-wise. The mixture was allowed to warm up to room temperature and was stirred for 16 hours. Dimethyl sulfate (3.5838 g, 2.7 mL, 28.356 mmol) in DMF (7 mL) was added dropwise at 0 °C over 15 minutes, and the reaction was stirred for 30 minutes. Water (150 mL), a saturated aqueous solution of ammonium chloride (50 mL) and EtOAc (200 mL) were added, and the phases were separated. The organic phase was washed with water (2 x 200 mL) and brine (200 mL), dried over anhydrous sodium sulfate, filtered, and evaporated under reduced pressure. Purification by silica gel chromatography (Gradient 0 - 10% EtOAc in heptanes) afforded after evaporation of the volatiles and freeze-drying 3-bromo-5- methyl-6-spiro[2.3]hexan-5-yl-pyrrolo[2,3-b]pyrazine (1.65 g, 44%) as a white solid.1H NMR (400 MHz, Chloroform-d) δ 8.42 (s, 1H), 6.56 (d, J = 0.9 Hz, 1H), 3.94 - 3.83 (m, 1H), 3.72 (s, 3H), 2.60 - 2.47 (m, 4H), 0.64 - 0.56 (m, 2H), 0.54 - 0.46 (m, 2H). ESI-MS m/z calc.291.0371, found 292.1 (M+1)+; Retention time: 3.31 minutes. LC method P.
Example 17: Preparation of 3-bromo-5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl- ethyl)pyrrolo[2,3-b]pyrazine Step 1: 6-Bromo-3-(4,4,4-trifluoro-3,3-dimethyl-but-1-ynyl)pyrazin-2-amine [00286] In a flask under nitrogen atmosphere containing a solution of 4,4,4-trifluoro-3,3- dimethyl-but-1-yne (11.31 g, 83.092 mmol) in THF (250 mL) were added 3,6-dibromopyrazin- 2-amine (9.15 g, 36.181 mmol), triethylamine (12.342 g, 17 mL, 121.97 mmol), bis(triphenylphosphine)palladium(II) dichloride (3.52 g, 5.0150 mmol) and copper(I) iodide (800 mg, 4.2006 mmol). The resulting mixture was stirred at room temperature under nitrogen atmosphere for 2 hours. Bis(triphenylphosphine)palladium(II) dichloride (1.57 g, 2.2368 mmol) and copper(I) iodide (450 mg, 2.3628 mmol) were added to the mixture and the mixture was flushed with nitrogen for 5 min. The mixture was stirred for 2 hours at rt under nitrogen atmosphere. Additional Bis(triphenylphosphine)palladium(II) dichloride (1.583 g, 2.2553 mmol) and copper iodide (472 mg, 2.4783 mmol) were added to the mixture and the mixture was flushed with nitrogen for 5 min. The mixture was stirred for 16 hours at rt under nitrogen atmosphere. The reaction mixture was filtered on Celite, and the cake was washed with acetonitrile (5 x 30 mL). Purification by reversed-phase chromatography (C18 column. Gradient 5 - 80% acetonitrile in water with 0.1% formic acid) was performed. The fractions containing the product were combined and the volatiles were removed under reduced pressure. The aqueous phase was extracted with EtOAc (3 x 100 mL). The organic phases were combined, washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure to afford 6-bromo-3-(4,4,4-trifluoro-3,3-dimethyl-but-1-ynyl)pyrazin-2-amine (6.82 g, 60%) as a yellow solid. ESI-MS m/z calc.306.9932, found 308.1 (M+1)+; Retention time: 2.13 minutes 1H NMR (400 MHz, Chloroform-d) δ 7.98 (s, 1H), 5.11 (br s, 2H), 1.56 (overlapped with water, s, 6H).19F NMR (377 MHz, Chloroform-d) δ -77.85 (s, 3F). LC method O. Step 2: 3-Bromo-5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3- b]pyrazine [00287] To a solution of 6-bromo-3-(4,4,4-trifluoro-3,3-dimethyl-but-1-ynyl)pyrazin-2-amine (10.8 g, 35.019 mmol) in anhydrous DMF (280 mL), in a flame dried flask, at 0 °C under nitrogen, was added NaH (3.15 g, 60 %w/w, 78.758 mmol). The mixture was stirred at 70 °C for 60 minutes. The mixture was allowed to cool down to 0 °C. A solution of dimethyl sulfate (8.7780 g, 6.6 mL, 69.594 mmol) in anhydrous DMF (100 mL) was added dropwise to the mixture at 0 °C over 45 min using a syringe driver. The mixture was stirred at 0 °C for an additional 10 minutes. Then, a saturated aqueous solution of NH4Cl (2 L) and water (2 L) were added. The mixture was stirred at rt for 10 minutes. The precipitate was collected by filtration, washed with cold water (3 x 50 mL) and dried under high vacuum to afford 3-bromo-5-methyl- 6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3-b]pyrazine (10 g, 87%) as a brown solid. ESI- MS m/z calc.321.0088, found 322.0 (M+1)+; Retention time: 2.18 minutes. LC method O. VI. Synthesis of substituted bicyclic esters and alkyl-phosphonate intermediates Example 18: Preparation of 1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2- dimethoxyphosphoryl-ethanone Step 1: 6-Chloro-3-(3,3-dimethylbut-1-ynyl)pyrazin-2-amine [00288] In a stirred solution of 3-bromo-6-chloro-pyrazin-2-amine (10 g, 47.975 mmol) in triethylamine (100 mL) nitrogen was bubbled for 10 minutes.3,3-Dimethylbut-1-yne (3.9200 g, 5.6 mL, 47.721 mmol) was added, followed by bis(triphenylphosphine)palladium(II) dichloride (1.67 g, 2.3793 mmol) and copper iodide (450 mg, 2.3628 mmol). Nitrogen was bubbled into the mixture for 5 minutes. The mixture was stirred at room temperature for 3 hours. The mixture was diluted by EtOAc (200 mL), was filtered on a pad of Celite and the pad was rinsed with EtOAc (250 mL). Water (250 mL) was then added and the filtrate was washed with water (3 x 250 mL) and brine (1 x 200 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford crude of 6-chloro-3-(3,3-dimethylbut-1-ynyl)pyrazin-2-amine (10 g, 94%) as an brownish solid; 1H NMR (400 MHz, DMSO-d6) δ 7.72 (s, 1H), 6.85 (br. S., 2H), 1.31 (s, 9H). ESI-MS m/z calc.209.072, found 210.2 (M+1)+; Retention time: 1.76 minutes. The crude was used for the next step without further purification. ESI-MS m/z calc.209.07198, found 210.2 (M+1)+; Retention time: 1.76 minutes; LC method I. Step 2: 6-tert-Butyl-3-chloro-5-methyl-pyrrolo[2,3-b]pyrazine [00289] To a solution of 6-chloro-3-(3,3-dimethylbut-1-ynyl)pyrazin-2-amine (8.8 g, 41.466 mmol) in DMF (120 mL) was added potassium tert-butoxide (24 g, 213.88 mmol). The reaction mixture was heated at 80 °C for 30 minutes and was allowed to cool down to room temperature. To the reaction mixture at room temperature was added methyl iodide (11.400 g, 5 mL, 80.316 mmol) dropwise over 20 minutes and the reaction was stirred for 45 minutes. To the reaction mixture at room temperature was again added methyl iodide (11.400 g, 5 mL, 80.316 mmol) dropwise over 20 minutes and the reaction was stirred for 15 minutes. A saturated aqueous solution of sodium thiosulfate (50 mL) was added, and the mixture was stirred for 1 hour. The reaction mixture was diluted with water (100 mL), brine (100 mL) and ethyl acetate (100 mL) and the phases were separated. The aqueous layer was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (3 x 150 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude 6-tert-butyl-3- chloro-5-methyl-pyrrolo[2,3-b]pyrazine (9.6 g, 76%) as dark red oil.1H NMR (400 MHz, CDCl3) δ 8.34 (s, 1H), 6.49 (s, 1H), 3.99 (s, 3H), 1.51 (s, 9H). ESI-MS m/z calc.223.0876, found 224.2 (M+1)+; Retention time: 1.93 minutes. LC method I. Step 3: Methyl 6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine-3-carboxylate [00290] To a suspension of 6-tert-butyl-3-chloro-5-methyl-pyrrolo[2,3-b]pyrazine (8.65 g, 36.850 mmol) in MeOH (50 mL) was added triethylamine (6.5340 g, 9 mL, 64.572 mmol). Nitrogen was bubbled into the mixture for 15 minutes and then Pd(dppf)Cl2.DCM (906 mg, 1.1094 mmol) was added. Nitrogen bubbling was then continued for 5 minutes. The mixture was then stirred at 80 °C under 60 psi carbon monoxide pressure for 18 hours. The mixture was then allowed to cool down to room temperature, was filtered on a Celite pad followed by a silica gel pad and concentrated in vacuo. Water (100 mL) and EtOAc (100 mL) were then added, and the mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (100 mL) and dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford crude methyl 6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine-3-carboxylate (9.53 g, 90%) as an orange oil. It was engaged in the following step without any further purification.1H NMR (400 MHz, CDCl3) δ 9.17 (s, 1H), 6.60 (s, 1H), 4.10 (s, 3H), 4.04 (s, 3H), 1.55 (s, 9H). ESI-MS m/z calc.247.1321, found 248.2 (M+1)+; Retention time: 1.7 minutes. LC method I. Step 4: 1-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2- dimethoxyphosphoryl-ethanone [00291] LiHMDS (in THF) (41 mL of 1 M, 41.000 mmol) was added to a solution of dimethyl methylphosphonate (2.5212 g, 2.2 mL, 20.320 mmol) in THF (130 mL) maintained at 0 °C. This solution was left stirring for 30 min at this temperature. A solution of methyl 6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazine-3-carboxylate (4.53 g, 18.300 mmol) in THF (50 mL) was added at 0 °C. The reaction was stirred at 0 °C for 90 minutes. The reaction was quenched by the addition of a saturated aqueous solution of ammonium chloride (50 mL).1 N aqueous HCl (100 mL) was added dropwise to the suspension until the solid mostly dissolved. The layers were then separated, and the aqueous layer was extracted with EtOAc (3 x 100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by silica gel chromatography (Gradient: 0-5 % MeOH in DCM) to afford 2.53 g of pure product as a yellow solid. The mixed fractions were combined, evaporated under reduced pressure, and purified by reversed-phase chromatography (5%-100% MeCN in water) to afford 3.04 g of pure product as a yellow solid. The two products were combined to afford 1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2- dimethoxyphosphoryl-ethanone (5.45 g, 88%) as a yellow solid.1H NMR (400 MHz, CDCl3) δ 9.18 (s, 1H), 6.66 (s, 1H), 4.11 (s, 3H), 4.07 (d, J = 22.3 Hz, 2H), 3.82 (s, 3H), 3.80 (s, 3H), 1.56 (s, 9H). ESI-MS m/z calc.339.1348, found 340.2 (M+1)+; Retention time: 3.62 minutes. LC method J. Example 19: Preparation of methyl 5-methyl-6-(1-methylcyclopropyl)pyrrolo[2,3- b]pyrazine-3-carboxylate Step 1: Trimethyl-[2-(1-methylcyclopropyl)ethynyl]silane [00292] To a solution of 2-cyclopropylethynyl(trimethyl)silane (8.54 g, 10.047 mL, 61.758 mmol) in Et2O (70 mL) at room temperature was slowly added n-BuLi in hexanes (30 mL of 2.5 M, 75.000 mmol) over 20 minutes. The reaction mixture was stirred at room temperature for 18 hours, then the reaction was cooled down to -15 °C in a dry-ice/acetone bath. To the reaction mixture was slowly added dropwise methyl iodide (9.5760 g, 4.2 mL, 67.466 mmol) over 10 minutes. The resulting solution was stirred at -10 °C for 10 minutes, and then at 0 °C for 1 hour. The reaction mixture was allowed to warm to room temperature for 1 hour. The reaction mixture was diluted with diethyl ether (50 mL) and water (50 mL) and brine (50 mL) were added. The phases were separated, and the aqueous phase was extracted with diethyl ether (3 x 20 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by silica gel chromatography (Eluent: 100% Heptane) afforded trimethyl-[2-(1-methylcyclopropyl)ethynyl]silane (2.63 g, 19%) as a light yellow liquid. GC-MS m/z calc.152.1021, found 152.00 (M), retention time 5.041 minutes. GC-MS method: Column HP 5 ms Ultra Inert 30 m x 250 μm x 0.25 μm, control mode: flow: 3.3 mL/min, split ratio mode: 33:1, split flow: 165 mL/min., carrier gas: He. Injector temperature 250 oC, FID temperature 300 oC, MSD transfer line 250 oC, solvent delay: 3 min. Oven temperature: isotherm at 40 oC for 1 min, then linear heating at 10 oC/min until 100 oC, then 20 oC/min until 220 oC then isotherm 220 oC for 4 min. Run time 17 min. Step 2: 6-Chloro-3-[2-(1-methylcyclopropyl)ethynyl]pyrazin-2-amine [00293] To a sealed tube equipped with a stir bar were added copper(I) iodide (221 mg, 1.1604 mmol), bis(triphenylphosphine)palladium(II) chloride (330 mg, 0.4702 mmol), triphenylphosphine (234 mg, 0.8922 mmol), cesium fluoride (4.6 g, 30.282 mmol), 3-bromo-6- chloro-pyrazin-2-amine (3.15 g, 15.112 mmol), poly(ethylene glycol) 200 MW (3.0 mL), triethylamine (25 mL) and water (1.5 mL). The vial was capped and the mixture sparged with nitrogen for 15 minutes while being stirred at room temperature. Trimethyl-[2-(1- methylcyclopropyl)ethynyl]silane (2.63 g, 11.621 mmol) was then added and the tube was sealed. The reaction mixture was allowed to stir at 80 °C for 17 hours. The mixture was allowed to cool to room temperature then was diluted with ethyl acetate (50 mL), water (50 mL) and brine (50 mL). The phases were separated, and the organic layer was washed with a 50% aqueous saturated sodium chloride solution (2 x 50 mL). The combined aqueous layers were extracted with ethyl acetate (2 x 50 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by silica gel chromatography (Gradient: 0-30% EtOAc in heptane) afforded 6-chloro-3-[2-(1-methylcyclopropyl)ethynyl]pyrazin-2-amine (2.23 g, 82%) as a yellow solid. ESI-MS m/z calc.207.05632, found 208.2 (M+1)+; Retention time: 1.76 minutes; LC method I. Step 3: 3-Chloro-5-methyl-6-(1-methylcyclopropyl)pyrrolo[2,3-b]pyrazine [00294] To a solution of 6-chloro-3-[2-(1-methylcyclopropyl)ethynyl]pyrazin-2-amine (2.23 g, 9.5682 mmol) in DMF (30 mL) was added potassium tert-butoxide (3.3 g, 29.409 mmol). The reaction mixture was heated at 80 °C for 50 min then the reaction was allowed to cool down to room temperature before methyl iodide (2.7360 g, 1.2 mL, 19.276 mmol) was added dropwise. The reaction was stirred at room temperature for 1 hour then potassium tert-butoxide (1.02 g, 1.1308 mL, 9.0899 mmol) and methyl iodide (2.2800 g, 1 mL, 16.063 mmol) were again added. The reaction mixture was stirred at room temperature for 18 hours then was diluted with H2O (100 mL), ethyl acetate (50 mL) and brine (100 mL). The phases were separated, and the aqueous layer was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with a 50% solution of brine (4 x 100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was diluted with EtOAc (30 mL) and activated carbon (1.5 g was added). The suspension was stirred for 10 minutes then the solution was filtered on a silica pad and washed with EtOAc (200 mL). The filtrate was concentrated under reduced pressure to afford 3-chloro-5-methyl-6-(1- methylcyclopropyl)pyrrolo[2,3-b]pyrazine (1.88 g, 82%) as a brown solid. ESI-MS m/z calc. 221.07198, found 222.2 (M+1)+; Retention time: 1.88 minutes; LC method I. Step 4: Methyl 5-methyl-6-(1-methylcyclopropyl)pyrrolo[2,3-b]pyrazine-3- carboxylate [00295] To a solution of 3-chloro-5-methyl-6-(1-methylcyclopropyl)pyrrolo[2,3-b]pyrazine (4.56 g, 19.788 mmol) in methanol (40 mL) was added triethylamine (5.8080 g, 8 mL, 57.397 mmol). The solution was sparged with nitrogen for 15 minutes then Pd(dppf)Cl2.DCM (872 mg, 1.0678 mmol) was added. The suspension was sparged with nitrogen for 5 minutes then was stirred at 100 °C under 50 psi of carbon monoxide for 18 hours. The mixture was allowed to cool down to room temperature and was filtered through Celite, washed with EtOAc (2 x 25 mL) and the filtrate was concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-50% EtOAc in heptane) afforded methyl 5-methyl-6-(1- methylcyclopropyl)pyrrolo[2,3-b]pyrazine-3-carboxylate (4.24 g, 85%) as a light yellow solid. 1H NMR (400 MHz, CDCl3) δ 9.17 (s, 1H), 6.52 (s, 1H), 4.04 (s, 3H), 4.02 (s, 3H), 1.49 (s, 3H), 1.07 – 1.00 (m, 2H), 0.95 – 0.89 (m, 2H). ESI-MS m/z calc.245.11642, found 246.2 (M+1)+; Retention time: 1.66 minutes; LC method I. Example 20: Preparation of 1-(6-tert-butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-2- dimethoxyphosphoryl-ethanone Step 1: 5-Bromo-2-chloro-N-methyl-pyrimidin-4-amine [00296] To a solution of 5-bromo-2,4-dichloro-pyrimidine (16 g, 70.214 mmol) in methanol (130 mL) was added dropwise methylamine in ethanol (461.54 mg, 1.85 mL of 33 %w/w, 4.9041 mmol) at 0 °C. The mixture was stirred for 20 minutes at 0 °C and for 3 hours at room temperature. The solvent was evaporated, and the residue was solubilized with ethyl acetate (100 mL). The organic layer was washed with a saturated aqueous solution of sodium bicarbonate (2 X 100 mL) and a saturated aqueous solution of NaCl (100 mL). The organic layer was dried over sodium sulfate, filtered, concentrated under reduced pressure and recrystallized in heptane and ethyl acetate. The solid was filtered on Buchner funnel, rinsed with heptane and dried under high vacuum to give 5-bromo-2-chloro-N-methyl-pyrimidin-4-amine (13.7 g, 86%) as a white crystalline solid 1H NMR (400 MHz, CDCl3) δ 8.10 (s, 1H), 5.62 (br. S, 1H), 3.09 (d, J = 4.9 Hz, 3H) ESI-MS m/z calc.220.9355, found 222.0 (M+1)+; Retention time: 1.51 minutes. LC method I. Step 2: 2-Chloro-5-(3,3-dimethylbut-1-ynyl)-N-methyl-pyrimidin-4-amine [00297] 5-Bromo-2-chloro-N-methyl-pyrimidin-4-amine (8.459 g, 36.122 mmol) was solubilized in THF (80 mL) and triethylamine (80 mL) was added dropwise to the solution. CuI (340 mg, 1.7852 mmol) and bis(triphenylphosphine)palladium(II)chloride (1.27 g, 1.8094 mmol) were added. The mixture was purged with nitrogen for 5 minutes and 3,3-dimethyl-1- butyne (3.6018 g, 5.4 mL, 43.848 mmol) was added. The mixture was stirred for 1.5 hours at 50 °C. The reaction was diluted in ethyl acetate (200 mL) and filtered over a pad of celite. Then, the celite was rinsed with ethyl acetate (100 mL). The filtrate was concentrated under reduced pressure and purified by reverse flash chromatography (gradient : acetonitrile in aqueous formic acid solution 0.1 %, 5 to 80 %, 12 CV) to afford 2-chloro-5-(3,3-dimethylbut-1-ynyl)-N-methyl- pyrimidin-4-amine (5.725 g, 57%) 1H NMR (400 MHz, CDCl3) δ 8.00 (s, 1H), 5.54 (br. S, 1H), 3.08 (d, J = 4.9 Hz, 3H), 1.33 (s, 9H) as a yellow solid ESI-MS m/z calc.223.0876, found 224.2 (M+1)+; Retention time: 1.88 minutes. LC method I. Step 3: 6-tert-Butyl-2-chloro-7-methyl-pyrrolo[2,3-d]pyrimidine [00298] To a solution of 2-chloro-5-(3,3-dimethylbut-1-ynyl)-N-methyl-pyrimidin-4-amine (5.725 g, 20.422 mmol) in THF (140 mL) was added TBAF in THF (61 mL of 1 M, 61.000 mmol) and the mixture was stirred overnight at 65 °C. The solvent was removed under reduced pressure and the residue was dissolved in EtOAc (400 mL) and the mixture was washed with water (2 x 200 mL) and brine (2 x 300 mL). The organic phase was dried over sodium sulfate, filtered, and concentrated under reduced pressure. Then, the crude product was purified by reverse flash chromatography on silica gel (gradient: 0 to 80 % acetonitrile in aqueous formic acid solution, 12 CV). The pure fractions were concentrated under reduce pressure. The aqueous phase was extracted with EtOAc (50 mL). Organic layer was washed with sodium bicarbonate (3 x 100 mL) and brine (100 mL). Then, the organic layer was dried under sodium sulfate and evaporated under reduced pressure to afford 6-tert-butyl-2-chloro-7-methyl-pyrrolo[2,3- d]pyrimidine (4.166 g, 91%) as a white solid 1H NMR (400 MHz, CDCl3) δ 8.64 (s, 1H), 6.32 (s, 1H), 3.97 (s, 3H), 1.48 (s, 9H) ESI-MS m/z calc.223.0876, found 220.2 (M+1)+; Retention time: 1.25 minutes. LC method I. Step 4: Methyl 6-tert-butyl-7-methyl-pyrrolo[2,3-d]pyrimidine-2-carboxylate [00299] To a stirred solution of 6-tert-butyl-2-chloro-7-methyl-pyrrolo[2,3-d]pyrimidine (4.166 g, 18.604 mmol) and triethylamine (5.8080 g, 8 mL, 57.397 mmol) in methanol (45 mL) under nitrogen atmosphere was added Pd(dppf)Cl2.DCM (3.34 g, 4.0899 mmol). Nitrogen was bubbled for 15 minutes to the mixture and the reaction was stirred overnight at 100 °C under 60 psi of carbon monoxide. The reaction was then cooled down to room temperature and concentrated under reduced pressure. The crude was purified by flash chromatography on silica- gel (gradient : 5 to 60 % ethyl acetate in heptanes; 15 CV) and the volatiles were concentrated under reduced pressure to afford methyl 6-tert-butyl-7-methyl-pyrrolo[2,3-d]pyrimidine-2- carboxylate (3.05 g, 66%) as a white solid 1H NMR (400 MHz, CDCl3) δ 8.95 (s, 1H), 6.45 (s, 1H), 4.10 (s, 3H), 4.07 (s, 3H),1.51 (s, 9H) ESI-MS m/z calc.247.1321, found 248.2 (M+1)+; Retention time: 1.6 minutes. LC method I. Step 5: 1-(6-tert-Butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-2- dimethoxyphosphoryl-ethanone [00300] To a stirred solution of dimethyl methylphosphonate (400 µL, 3.740 mmol) in THF (17.6 mL) at -78 °C was added dropwise LiHMDS (in THF solution) (4.85 mL of 1 M, 4.850 mmol). The reaction mixture was stirred at -78 °C for 15 minutes and then a solution of methyl 6-tert-butyl-7-methyl-pyrrolo[2,3-d]pyrimidine-2-carboxylate (602 mg, 2.434 mmol) in anhydrous THF (5.6 mL) was added dropwise over 5 minutes. The reaction was stirred at -78 °C for 10 minutes. The reaction was quenched by the addition of a saturated aqueous solution of ammonium chloride (5 mL) and the biphasic mixture was vigorously stirred at room temperature for 5 minutes. The layers were then separated, and the aqueous layer was extracted with EtOAc (2 x 5 mL). The combined organic layers were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by normal phase chromatography (40 g of silica gel, 0-100% of EtOAc-Hex, 15 minute run) to give a light yellow oil of 1-(6-tert-butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-2-dimethoxyphosphoryl-ethanone (583 mg, 71%) ESI-MS m/z calc.339.1348, found 340.606 (M+1)+; Retention time: 1.001 minutes. LC method A. Example 21: Preparation of 2-dimethoxyphosphoryl-1-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]131xtracte Step 1: 1-(2,2-Dibromovinyl)-1-(trifluoromethyl)cyclopropane [00301] To a solution of 1-(trifluoromethyl)cyclopropanecarbaldehyde (13.886 g, 100.56 mmol) in DCM was added CBr4 (50 g, 150.77 mmol). The mixture was placed in a water bath and triphenylphosphine (80 g, 305.01 mmol) was added portion-wise over 1 hour. The mixture was stirred at room temperature for 16 hours. The precipitate was filtered on a Celite pad and the Celite cake was washed with DCM (2 x 20 mL). The filtrate was carefully evaporated under reduced pressure and the product was triturated in pentanes (300 mL) for 4 hours. The precipitate was filtered on a silica pad and the silica cake was washed with pentane (3 x 50 mL). The solvent was carefully removed under reduced pressure to afford 1-(2,2-dibromovinyl)-1- (trifluoromethyl)cyclopropane (27.5 g, 70%) as a colorless liquid. Step 2: 1-Ethynyl-1-(trifluoromethyl)cyclopropane [00302] To a solution of 1-(2,2-dibromovinyl)-1-(trifluoromethyl)cyclopropane (27.5 g, 70.175 mmol) in anhydrous THF (200 mL) was added n-BuLi (solution in hexanes) (62 mL of 2.5 M, 155.00 mmol) dropwise over 30 minutes at -78°C. After addition, the reaction mixture was stirred at -78°C for 1.5 h. Then it was quenched with ammonium chloride (7 mL), dried over magnesium sulfate and filtered on a pad of celite to give 1-ethynyl-1- (trifluoromethyl)cyclopropane as a solution in THF. The product was used as a solution for the next step. Step 3: 6-Chloro-3-[2-[1-(trifluoromethyl)cyclopropyl]ethynyl]pyrazin-2-amine [00303] In a flask under nitrogen atmosphere containing a solution of 1-ethynyl-1- (trifluoromethyl)cyclopropane (9.4104 g, 70.175 mmol) in THF were added 3-bromo-6-chloro- pyrazin-2-amine (7.29 g, 34.973 mmol), triethylamine (11.616 g, 16 mL, 114.79 mmol), bis(triphenylphosphine)palladium(II) dichloride (2.53 g, 3.6045 mmol) and copper iodide (725 mg, 3.8068 mmol). The resulting mixture was stirred at room temperature under nitrogen atmosphere (sealed tube) for 16 hours. The reaction mixture was filtered on Celite and the cake was washed with acetonitrile (5 x 30 mL). The organics were concentrated under reduced pressure and the resulting residue was purified by silica gel chromatography (0-100% EtOAc in Heptanes) to afford 6-chloro-3-[2-[1-(trifluoromethyl)cyclopropyl]ethynyl]pyrazin-2-amine (8.80 g, 96%) as an orange solid. ESI-MS m/z calc.261.0281, found 262.2 (M+1)+; Retention time: 1.8 minutes. LC method I. Step 4: 3-Chloro-6-[1-(trifluoromethyl)cyclopropyl]-5H-pyrrolo[2,3-b]pyrazine [00304] Potassium tert-butoxide (12 g, 106.94 mmol) was added to a solution of 6-chloro-3- [2-[1-(trifluoromethyl)cyclopropyl]ethynyl]pyrazin-2-amine (8.80 g, 33.602 mmol) in tert- butanol (170 mL) and heated at 70 °C for 16 h. The mixture was allowed to cool down to room temperature, then quenched with water (20 mL). The volatiles were removed under reduced pressure and saturated aqueous ammonium chloride solution (300 mL) was added. The mixture was cooled at 0 °C and stirred for 20 minutes. The resulting precipitate was filtered and washed with distilled water (2 x 50 mL). The precipitate was dissolved in EtOAc (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to provide 3-chloro- 6-[1-(trifluoromethyl)cyclopropyl]-5H-pyrrolo[2,3-b]pyrazine (8.74 g, 99%) as a brown solid. ESI-MS m/z calc.261.0281, found 262.2 (M+1)+; Retention time: 1.74 minutes. This product was used in the next step without further purification. LC method I. Step 5: 3-Chloro-5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazine [00305] Sodium hydride (60% w/w in oil) (1.18 g, 29.503 mmol) was added portion-wise to a solution of 3-chloro-6-[1-(trifluoromethyl)cyclopropyl]-5H-pyrrolo[2,3-b]pyrazine (3.81 g, 14.242 mmol) in DMF (76 mL) at 0 °C. The suspension was stirred at this temperature for 15 min. Dimethyl sulfate (2.7993 g, 2.1 mL, 22.193 mmol) was added and left stirring at 0 °C for 15 min. The reaction was then stirred at room temperature for 1 h. The reaction mixture was cooled back to 0 °C and quenched with saturated aqueous ammonium chloride solution (100 mL). The resulting precipitate was filtered, washed with pure water (2 x 25mL) and a small volume of heptanes (5 mL) then dried under high vacuum to provide 3.65 g of the desired product as a brown solid. The filtrate was extracted with EtOAc (3 x 50 mL) and the combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The resulting precipitate was purified by reversed phase chromatography (Gradient : 5-100 % MeCN in water) to provide 318 mg of the desired product as a dark yellow solid. The two product obtained were combined to afford 3-chloro-5-methyl-6- [1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine (3.96 g, 101%) as a brown solid. ESI- MS m/z calc.275.0437, found 276.2 (M+1)+; Retention time: 1.89 minutes. LC method I. Step 6: Methyl 5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazine-3-carboxylate [00306] To a suspension of 3-chloro-5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazine (3.96 g, 14.351 mmol) in anhydrous methanol (72 mL) was added triethylamine (4.5012 g, 6.2 mL, 44.483 mmol). Nitrogen was bubbled into the mixture for 15 minutes and then Pd(dppf)Cl2.DCM (592 mg, 0.7249 mmol) was added. Nitrogen bubbling was then continued for 5 minutes. The mixture was then stirred at 100 °C under 50 psi of carbon monoxide pressure for 18 hours. The mixture was then allowed to cool down to room temperature and was filtered on a Celite pad. The Celite cake was washed with EtOAc (2 x 30 mL) and concentrated under reduced pressure. Water (100 mL) and EtOAc (100 mL) were then added, and the mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (100 mL) and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting crude was purified by silica gel chromatography (Gradient: 0-60% EtOAc in heptane) to afford methyl 5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine-3-carboxylate (3.72 g, 85%) as a light yellow solid.1H NMR (400 MHz, CDCl3) δ 9.24 (s, 1H), 6.86 (s, 1H), 4.06 (s, 3H), 4.03 (s, 3H), 1.69 – 1.64 (m, 2H), 1.32 – 1.27 (m, 2H).19F NMR (377 MHz, CDCl3) δ -68.55 (s, 3F). ESI-MS m/z calc.299.0882, found 300.0 (M+1)+; Retention time: 3.96 minutes. LC method J. Step 7: 2-Dimethoxyphosphoryl-1-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]135xtracte [00307] In an oven-dried round bottom flask, [methoxy(methyl)phosphoryl]oxymethane (190 µL, 1.753 mmol) was dissolved in THF (12.5 mL). It was cooled to 0 °C before the dropwise addition of a solution of [bis(trimethylsilyl)amino]lithium (3.4 mL of 1 M, 3.400 mmol) in THF. After stirring for 30 minutes at 0 °C, a solution of methyl 5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine-3-carboxylate (500 mg, 1.671 mmol) in THF (5 mL) was then added. The reaction mixture was stirred at 0 °C for 3 hours and at room temperature overnight. The reaction was quenched with the addition of saturated aqueous ammonium chloride solution (50 mL). After stirring for 1 hour, it was extracted with EtOAc (2× 50 mL). Organic layers were combined, dried over sodium sulfate, filtered, and concentrated under reduced pressure. Purification by flash chromatography on silica gel (0-100% EtOAc/hexanes over 15 minutes) provided 2-dimethoxyphosphoryl-1-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]135xtracte (340 mg, 52%) as a yellow oil that crystallized upon standing. ESI-MS m/z calc.391.09088, found 392.1 (M+1)+; Retention time: 0.57 minutes. LC method B. Example 22: Preparation of 2-dimethoxyphosphoryl-1-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]135xtracte Step 1: 6-Chloro-3-(4,4,4-trifluoro-3,3-dimethyl-but-1-ynyl)pyrazin-2-amine [00308] In a flask under nitrogen atmosphere were dissolved 3-bromo-6-chloro-pyrazin-2- amine (963 mg, 4.6199 mmol) and CuI (93 mg, 0.4883 mmol) in THF (20 mL).4,4,4-Trifluoro- 3,3-dimethyl-but-1-yne (691 mg, 5.0766 mmol) and triethylamine (958.32 mg, 1.32 mL, 9.4705 mmol) were then added and the mixture was degassed with nitrogen for 10 min. PdCl2(PPh3)2 (326 mg, 0.4645 mmol) was added and the mixture was re-degassed for another 5 min. The flask was sealed, and the reaction was stirred at room temperature for 2 hours. The mixture was filtered through Celite, washed with EtOAc (30 mL) and the filtrate concentrated in vacuo. Purification was done by reverse-phase chromatography (Gradient: 5-100 % MeOH in water with 0.1 % formic acid) to give 6-chloro-3-(4,4,4-trifluoro-3,3-dimethyl-but-1-ynyl)pyrazin-2- amine (654 mg, 54%) as an orange solid.1H NMR (400 MHz, Chloroform-d) δ 7.89 (s, 1H), 5.10 (br. S., 2H), 1.57 (s, 6H).19F NMR (377 MHz, Chloroform-d) δ -77.87 (s, 3F). ESI-MS m/z calc.263.0437, found 264.2 (M+1)+; Retention time: 1.84 minutes. LC method I. Step 2: 3-Chloro-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)-5H-pyrrolo[2,3- b]pyrazine [00309] In a dried-flask under nitrogen atmosphere was dissolved 6-chloro-3-(4,4,4-trifluoro- 3,3-dimethyl-but-1-ynyl)pyrazin-2-amine (3.87 g, 14.459 mmol) in dry THF (60 mL). The solution was cooled to 0 °C and potassium tert-butoxide (1M solution in THF) (28 mL of 1 M, 28.000 mmol) was slowly added. The reaction mixture was heated to reflux for 2 hours. The crude was cooled down to room temperature, diluted with EtOAc (20 mL) and quenched with water (15 mL). The aqueous layer was extracted with EtOAc (2 x 15 mL). The combined organic layers were washed with brine (2 x 15 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The crude was purified by reverse-phase chromatography (Gradient: 5- 100 % MeOH in water with 0.1 % formic acid) to afford 3-chloro-6-(2,2,2-trifluoro-1,1- dimethyl-ethyl)-5H-pyrrolo[2,3-b]pyrazine (2.4 g, 32%) 1H NMR (400 MHz, Chloroform-d) δ 8.78 (br. S., 1H), 8.46 (s, 1H), 6.71 (d, J = 2.0 Hz, 1H), 1.66 (s, 6H).19F NMR (377 MHz, Chloroform-d) δ -75.83 (s, 3F). ESI-MS m/z calc.263.0437, found 264.2 (M+1)+; Retention time: 1.77 minutes. LC method I. Step 3: 3-Chloro-5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3- b]pyrazine [00310] To a solution of 3-chloro-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)-5H-pyrrolo[2,3- b]pyrazine (3.55 g, 13.465 mmol) in dry DMF (80 mL) at 0 °C was added NaH (60% dispersion in mineral oil) (1.7 g, 42.504 mmol) portion-wise. The mixture was stirred at 0 °C for 10 minutes then dimethyl sulfate (3.4580 g, 2.6 mL, 27.416 mmol) was added dropwise over 20 min. The reaction was stirred at room temperature for 2 hours and cooled to 0 °C, water (20 mL) was slowly added followed by MeTHF (20 mL). The aqueous phase was extracted with MeTHF (4 x 15 mL). The combined organic layers were then washed with brine (2 x 15 mL), dried over anhydrous sodium sulfate, and filtered. The crude mixture was concentrated in vacuo and was then purified by reverse-phase chromatography (Gradient: 5-100 % MeCN in water with 0.1 % formic acid) to give 3-chloro-5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3- b]pyrazine (3 g, 80%) as a tan solid.1H NMR (400 MHz, DMSO-d6) δ 8.53 (s, 1H), 6.99 (s, 1H), 3.95 (s, 3H), 1.76 (s, 6H).19F NMR (377 MHz, DMSO-d6) δ -73.79 (s, 3F). ESI-MS m/z calc.277.0594, found 278.1 (M+1)+; Retention time: 4.54 minutes. LC method I. Step 4: Methyl 5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3- b]pyrazine-3-carboxylate [00311] To a suspension of 3-chloro-5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl- ethyl)pyrrolo[2,3-b]pyrazine (1.6 g, 5.0707 mmol) in anhydrous methanol (15 mL) was added triethylamine (1.0080 g, 1.4 mL, 9.9615 mmol). Nitrogen was bubbled into the mixture for 15 minutes and then Pd(dppf)Cl2.DCM (410 mg, 0.5021 mmol) was added. Nitrogen was bubbled for 5 minutes. The mixture was then stirred at 90 °C under 50 psi of carbon monoxide pressure for 3 hours. The crude was filtered on a Celite pad, washed with EtOAc (15 mL) and concentrated under reduced pressure. Water (15 mL) was added, and the mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (10 mL) and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting crude was purified by silica gel chromatography (Gradient: 0-70% EtOAc in heptane) to afford methyl 5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3-b]pyrazine-3- carboxylate (960 mg, 62%) as a light-yellow solid. ESI-MS m/z calc.301.1038, found 302.2 (M+1)+; Retention time: 4.05 minutes.1H NMR (400 MHz, DMSO-d6) δ 9.09 (s, 1H), 7.05 (s, 1H), 4.04 (s, 3H), 3.94 (s, 3H), 1.79 (s, 6H).19F NMR (377 MHz, DMSO-d6) δ -73.62 (s, 3F). ESI-MS m/z calc.301.10382, found 302.2 (M+1)+; Retention time: 4.05 minutes; LC method J. Step 5: 2-Dimethoxyphosphoryl-1-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]138xtracte [00312] In an oven-dried round bottom flask, [methoxy(methyl)phosphoryl]oxymethane (190 µL, 1.753 mmol) was dissolved in THF (12.5 mL). It was cooled to 0 °C before the dropwise addition of a solution of [bis(trimethylsilyl)amino]lithium (3.4 mL of 1 M, 3.400 mmol) in THF. After stirring for 30 minutes at 0 °C, a solution of methyl 5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine-3-carboxylate (500 mg, 1.671 mmol) in THF (5 mL) was then added. The reaction mixture was stirred at 0 °C for 3 hours and at room temperature overnight. The reaction was quenched with the addition of saturated aqueous ammonium chloride solution (50 mL). After stirring for 1 hour, it was extracted with EtOAc (2× 50 mL). Organic layers were combined, dried over sodium sulfate, filtered, and concentrated under reduced pressure. Purification by flash chromatography on silica gel (0- 100% EtOAc/hexanes over 15 minutes) provided 2-dimethoxyphosphoryl-1-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]138xtracte (340 mg, 52%) as a yellow oil that crystallized upon standing. ESI-MS m/z calc.391.09088, found 392.1 (M+1)+; Retention time: 0.57 minutes. LC method B. Example 23: Preparation of 2-dimethoxyphosphoryl-1-(2-isopropyl-3-methyl-imidazo[4,5- b]pyrazin-5-yl)138xtracte Step 1: 5-Bromo-N3-methyl-pyrazine-2,3-diamine [00313] A suspension of 3,5-dibromopyrazin-2-amine (1 g, 3.9542 mmol) in MeNH2 in water (10 mL of 40 %w/v, 128.80 mmol) was heated in a microwave reactor at 130 °C for 1 h. The reaction was cooled to room temperature and extracted with DCM (3 x 15 mL), dried over MgSO4, filtered and concentrated in vacuo to provide 5-bromo-N3-methyl-pyrazine-2,3-diamine (802 mg, 100%) as a tan solid.1H NMR (400 MHz, Chloroform-d) δ 7.43 (s, 1H), 4.48 (s, 1H), 4.24 (s, 2H), 2.99 (d, J = 5.0 Hz, 3H). ESI-MS m/z calc.201.98541, found 203.1 (M+1)+; Retention time: 2.4 minutes; LC method D. Step 2: 5-Bromo-2-isopropyl-3-methyl-imidazo[4,5-b]pyrazine [00314] To a solution of 2-methylpropanoyl 2-methylpropanoate (248.04 mg, 0.260 mL, 1.5679 mmol) in isobutyric acid (2 mL) was added 5-bromo-N3-methyl-pyrazine-2,3-diamine (300 mg, 1.4775 mmol) and the reaction was stirred at room temperature for 10 minutes then heated in a microwave reactor at 130 °C for 5 h and 20 min. To the reaction was added 2- methylpropanoyl 2-methylpropanoate (477.00 mg, 0.500 mL, 3.0153 mmol) and it was heated at 130° C for 2 h. The reaction was cooled to room temperature and diluted with DCM (30 mL) and the organic phase was washed with 1 M NaOH (30 mL). The aqueous phase was back extracted with DCM (3 x 25 mL) and filtered through filter paper to remove emulsions. The organic phase was washed with brine, dried over MgSO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography (loaded onto a 12 g silica cartridge with benzene, and eluted with 0-40% EtOAc in hexanes over a 30 min gradient) to provide 5-bromo- 2-isopropyl-3-methyl-imidazo[4,5-b]pyrazine (270 mg, 72%) as an off white solid.1H NMR (500 MHz, Chloroform-d) δ 8.50 (s, 1H), 3.83 (s, 3H), 3.25 (hept, J = 6.8 Hz, 1H), 1.47 (d, J = 6.8 Hz, 6H).13C NMR (126 MHz, CDCl3) δ 165.38, 147.56, 141.09, 132.60, 28.70, 27.58, 20.77. ESI-MS m/z calc.254.01671, found 255.4 (M+1)+; Retention time: 2.87 minutes; LC method D. Step 3: Methyl 2-isopropyl-3-methyl-imidazo[4,5-b]pyrazine-5-carboxylate [00315] To a solution of 5-bromo-2-isopropyl-1-methyl-imidazo[4,5-b]pyrazine (4.03 g, 15.797 mmol) in MeOH (80 mL) was added Pd(dppf)Cl2 – DCM complex (1.25 g, 1.5307 mmol) then TEA (32.670 g, 45 mL, 322.86 mmol) was added. The vessel was sealed and pressurized with CO gas (100 psi) and vented 3 times, then re-pressurized at 100 psi and heated at 80° C for 30 minutes. The reaction was cooled to room temperature and stirred at 100 psi for 12 h. Then the pressure was released, and the mixture was concentrated in vacuo. The reaction was diluted with toluene and the TEA salts were filtered off through a pad of celite. The supernatant was concentrated in vacuo and the residue was purified by flash chromatography (loaded with toluene onto a 120 g silica cartridge pre-equilibrated with 1% TEA in hexanes, and eluted with 0-50% EtOAc in hexanes over a 60 min gradient and held at 47% EtOAc for 15 min) to provide methyl 2-isopropyl-1-methyl-imidazo[4,5-b]pyrazine-5-carboxylate (2.8 g, 76%) as a colorless solid. ESI-MS m/z calc.234.11168, found 235.3 (M+1)+; Retention time: 2.18 minutes; LC method D. Step 4: 2-Dimethoxyphosphoryl-1-(2-isopropyl-3-methyl-imidazo[4,5-b]pyrazin- 5-yl)140xtracte [00316] In an oven-dried round bottom flask, [methoxy(methyl)phosphoryl]oxymethane (501 mg, 4.038 mmol) was dissolved in THF (20 mL). The solution was cooled to 0 °C. A solution of [bis(trimethylsilyl)amino]lithium (8.1 mL of 1 M, 8.100 mmol) was added dropwise. After stirring at 0 °C for 30 minutes, a solution of methyl 2-isopropyl-3-methyl-imidazo[4,5- b]pyrazine-5-carboxylate (946 mg, 4.038 mmol) in THF (5 mL) was added dropwise. The reaction mixture was stirred at 0 °C for 30 minutes and at room temperature for 1 hour. The reaction was quenched with the addition of saturated aqueous ammonium chloride solution (50 mL). After stirring for 1 hour, it was extracted with EtOAc (2× 50 mL). Organic layers were combined, dried over sodium sulfate, filtered, and concentrated under reduced pressure. Purification by flash chromatography on silica gel (0-100% EtOAc/hexanes over 30 minutes) provided dimethyl (2-(2-isopropyl-1-methyl-1H-imidazo[4,5-b]pyrazin-6-yl)-2- oxoethyl)phosphonate (382 mg, 29%) as a yellow oil. ESI-MS m/z calc.326.11438, found 327.2 (M+1)+; Retention time: 0.79 minutes. LC method A. Example 24: Preparation of 2-dimethoxyphosphoryl-1-[6-(1-methylcyclopropyl)furo[2,3- b]pyrazin-2-yl]141xtracte Step 1: 5-Chloro-1-[(4-methoxyphenyl)methyl]-3-[2-(1- methylcyclopropyl)ethynyl]pyrazin-2-one [00317] To a 20 mL microwave vial was charged 3,5-dichloro-1-[(4- methoxyphenyl)methyl]pyrazin-2-one (350 mg, 1.228 mmol), bis(triphenylphosphine)palladium(II) dichloride (16 mg, 0.02280 mmol), CuI (10 mg, 0.05251 mmol) in DMF (2.5 mL) and TEA (1.0 mL, 7.175 mmol). Then, 1-ethynyl-1-methyl- cyclopropane (100 mg, 1.248 mmol) was added, and the vessel was sealed. The mixture was irradiated at 80°C µW for 10 min. The reaction mixture was allowed to cool, diluted with DCM (20 mL) and washed with water (2x10 mL). The organic layer was washed with brine (10 mL), dried with sodium sulfate, filtered, and concentrated under vacuo. The resulting material was purified by silica gel chromatography (40 g of silica) using a gradient eluent of 100% hexanes to 80% ethyl acetate in hexanes to afford as a yellow oil 5-chloro-1-[(4-methoxyphenyl)methyl]-3- [2-(1-methylcyclopropyl)ethynyl]pyrazin-2-one (287 mg, 71%), 1H NMR (400 MHz, DMSO-d6) δ 8.15 (s, 1H), 7.38 – 7.30 (m, 2H), 6.95 – 6.87 (m, 2H), 4.96 (s, 2H), 3.73 (s, 3H), 1.31 (s, 3H), 1.04 – 0.96 (m, 2H), 0.88 – 0.80 (m, 2H). ESI-MS m/z calc.328.09787, found 329.2 (M+1)+; Retention time: 1.83 minutes; LC method A. Step 2: 2-Chloro-6-(1-methylcyclopropyl)furo[2,3-b]pyrazine [00318] To a solution of 5-chloro-1-[(4-methoxyphenyl)methyl]-3-[2-(1- methylcyclopropyl)ethynyl]pyrazin-2-one (283 mg, 0.8607 mmol) in DCM (5.0 mL), trifluoromethanesulfonate (6 mg, 0.02335 mmol) (silver salt) and TFA (600 µL, 7.788 mmol) were added and stirred at room temperature for 90 minutes. The residue was concentrated in vacuo and purified by silica gel chromatography (40 g of silica) using a gradient eluent of 100% hexanes to 30% ethyl acetate in hexanes to give as a pale yellow solid 2-chloro-6-(1- methylcyclopropyl)furo[2,3-b]pyrazine (148.0 mg, 82%), 1H NMR (400 MHz, MeOH-d4) δ 8.14 (s, 1H), 6.73 (s, 1H), 1.55 (s, 3H), 1.41 – 1.34 (m, 2H), 1.07 – 0.99 (m, 2H). ESI-MS m/z calc. 208.04034, found 209.2 (M+1)+; Retention time: 1.76 minutes; LC method A. Step 3: Methyl 6-(1-methylcyclopropyl)furo[2,3-b]pyrazine-2-carboxylate [00319] A mixture of 2-chloro-6-(1-methylcyclopropyl)furo[2,3-b]pyrazine (144 mg, 0.6902 mmol) and Pd(dppf)Cl2.DCM (58 mg, 0.07102 mmol) and TEA (450 µL, 3.229 mmol) in MeOH (6 mL) in a steel bomb equipped with a stir bar was purged with carbon monoxide (2 g, 68.92 mmol) three times. The reaction mixture was heated to 100°C with 150 psi of carbon monoxide and stirred for 14h. The reaction mixture was allowed to cool to rt. The reaction mixture was filtered through a pad of celite, washed with ethyl acetate, filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography (12 g of silica) using a gradient eluent of 100% hexanes to 50% ethyl acetate in hexanes to give as a white solid methyl 6-(1- methylcyclopropyl)furo[2,3-b]pyrazine-2-carboxylate (77 mg, 48%), 1H NMR (400 MHz, Chloroform-d) δ 8.94 (s, 1H), 6.71 (s, 1H), 4.05 (s, 3H), 1.56 (s, 3H), 1.47 – 1.42 (m, 2H), 1.03 – 0.99 (m, 2H). ESI-MS m/z calc.232.0848, found 233.3 (M+1)+; Retention time: 1.44 minutes; LC method A. Step 4: 2-dimethoxyphosphoryl-1-[6-(1-methylcyclopropyl)furo[2,3-b]pyrazin-2- yl]142xtracte [00320] A flask was flame-dried then cooled down to room temperature using a nitrogen stream and charged with dimethyl methylphosphonate (823.60 mg, 710 μL, 6.6379 mmol) and anhydrous THF (35 mL). The solution was cooled down to 0 °C and a solution of LiHMDS in THF (13.0 mL of 1 M, 13.000 mmol) was added dropwise. The reaction was stirred at 0 °C for 30 minutes after which time a solution of methyl 6-(1-methylcyclopropyl)furo[2,3-b]pyrazine-2- carboxylate (1.5 g, 6.4525 mmol) in anhydrous THF (15 mL) was added dropwise. The reaction was stirred at 0 °C for 1 hour then quenched by the addition of a saturated aqueous solution of ammonium chloride (100 ml). The mixture was allowed to reach room temperature and vigorously stirred for 30 minutes. The layers were then separated, and the aqueous layers was extracted with ethyl acetate (3 x 40 mL) and the combined organic layers were washed with brine (100 mL), dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude was purified by flash chromatography on silica-gel (gradient: 0 to 100% ethyl acetate in heptanes; 10 CV, then 100% ethyl acetate; 4 CV). The desired fractions were concentrated under reduced pressure, dried under high vacuum to afford 2-dimethoxyphosphoryl-1-[6-(1- methylcyclopropyl)furo[2,3-b]pyrazin-2-yl]-acetaldehyde (1.58 g, 75%) as a light yellow powder. ESI-MS m/z calc.324.0875, found 325.2 (M+1)+; Retention time: 1.64 minutes. LC method I. Example 25: Preparation of 1-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-2- dimethoxyphosphoryl-ethanone Step 1: 5-Chloro-3-(3,3-dimethylbut-1-ynyl)-1-[(4- methoxyphenyl)methyl]pyrazin-2-one [00321] To a 20 mL microwave vial was charged 3,5-dichloro-1-[(4- methoxyphenyl)methyl]pyrazin-2-one (457 mg, 1.603 mmol), bis(triphenylphosphine) palladium(II) dichloride (21 mg, 0.02992 mmol), CuI (14 mg, 0.07351 mmol) in DMF (3.5 mL) and TEA (1.3 mL, 9.327 mmol). Then, 3,3-dimethylbut-1-yne (290 µL, 2.429 mmol) was added and the vessel was sealed. The mixture was irradiated at 80°C µW for 10 min. The reaction mixture was allowed to cool, diluted with DCM (20 mL) and washed with water (2x10 mL). The organic layer was washed with brine (10 mL), dried with sodium sulfate, filtered and concentrated under vacuo. The resulting material was purified by silica gel chromatography (24 g of silica) using a gradient eluent of 100% hexanes to 80% ethyl acetate in hexanes to afford as a yellow oil 5-chloro-3-(3,3-dimethylbut-1-ynyl)-1-[(4-methoxyphenyl)methyl]pyrazin-2-one (457 mg, 86%), 1H NMR (400 MHz, MeOH-d4) δ 7.77 (s, 1H), 7.38 – 7.29 (m, 2H), 6.96 – 6.88 (m, 2H), 5.05 (s, 2H), 3.78 (s, 3H), 1.34 (s, 9H). ESI-MS m/z calc.330.1135, found 331.2 (M+1)+; Retention time: 1.92 minutes; LC method A. Step 2: 6-tert-Butyl-2-chloro-furo[2,3-b]pyrazine [00322] To a solution of 5-chloro-3-(3,3-dimethylbut-1-ynyl)-1-[(4- methoxyphenyl)methyl]pyrazin-2-one (453 mg, 1.369 mmol) in DCM (9.0 mL), trifluoromethanesulfonate (18 mg, 0.07006 mmol) (silver salt) and TFA (950 µL, 12.33 mmol) were added and stirred at room temperature for 90 minutes. The residue was concentrated in vacuo and purified by silica gel chromatography (40 g of silica) using a gradient eluent of 100% hexanes to 30% ethyl acetate in hexanes to give as an off-white solid 6-tert-butyl-2-chloro- furo[2,3-b]pyrazine (258.0 mg, 89%), 1H NMR (400 MHz, MeOH-d4) δ 8.21 (s, 1H), 6.73 (s, 1H), 1.43 (s, 9H). ESI-MS m/z calc.210.05598, found 211.2 (M+1)+; Retention time: 1.9 minutes. LC method A. Step 3: Methyl 6-tert-butylfuro[2,3-b]pyrazine-2-carboxylate [00323] A mixture of 6-tert-butyl-2-chloro-furo[2,3-b]pyrazine (295 mg, 1.400 mmol) and Pd(dppf)Cl2.DCM (115 mg, 0.1408 mmol) and TEA (900 µL, 6.457 mmol) in MeOH (10 mL) in a steel pressure vessel equipped with a stir bar was purged with carbon monoxide (4 g, 137.8 mmol) three times. The reaction mixture was heated to 100°C with 120 psi of carbon monoxide and stirred for 14 h. The reaction mixture was allowed to cool to rt. The reaction mixture was filtered through a pad of celite, washed with ethyl acetate, filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography (40 g of silica) using a gradient eluent of 100% hexanes to 50% ethyl acetate in hexanes to give as a white solid methyl 6-tert- butylfuro[2,3-b]pyrazine-2-carboxylate (238 mg, 73%) 1H NMR (400 MHz, Chloroform-d) δ 9.01 (s, 1H), 6.69 (s, 1H), 4.06 (s, 3H), 1.45 (s, 9H). ESI-MS m/z calc.234.10045, found 235.3 (M+1)+; Retention time: 1.53 minutes. LC method A. Step 4: 1-(6-tert-Butylfuro[2,3-b]pyrazin-2-yl)-2-dimethoxyphosphoryl-ethanone [00324] To a stirred solution of dimethyl methylphosphonate (3.48 g, 3.0000 mL, 28.047 mmol) in dry THF (155 mL) at 0 °C was added dropwise LiHMDS (in THF solution) (53 mL of 1 M, 53.000 mmol). The reaction mixture was then stirred at 0 °C for 30 minutes and then a solution of methyl 6-tert-butylfuro[2,3-b]pyrazine-2-carboxylate (6.22 g, 26.553 mmol) in dry THF (62 mL) was added. The reaction was stirred at 0 °C for 90 minutes. The reaction was quenched by the addition of a saturated aqueous solution of ammonium chloride (20 mL) and the biphasic mixture was vigorously stirred at room temperature for 30 minutes. The layers were then separated, and the aqueous layer was extracted with EtOAc (3 x 25 mL). The combined organic layers were washed with brine (50 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The crude mixture was purified by silica gel chromatography, eluting with a gradient of 0 to 40% of acetone in heptane. The fractions containing the desired product were combined and the organic solvent was evaporated in vacuo to afford 1-(6-tert- butylfuro[2,3-b]pyrazin-2-yl)-2-dimethoxyphosphoryl-ethanone (8.29 g, 96%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 8.96 (s, 1H), 6.68 (s, 1H), 4.02 (d, J = 22.7 Hz, 2H), 3.80 (s, 3H), 3.78 (s, 3H), 1.45 (s, 9H). ESI-MS m/z calc.326.1032, found 327.2 (M+1)+; Retention time: 3.92 minutes. LC method J. Example 26: Preparation of –(7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-2- dimethoxyphosphoryl-ethanone Step 1: 2-Chloro-7-iodo-6-isopropyl-furo[2,3-b]pyrazine [00325] To a solution of 5-chloro-1-[(4-methoxyphenyl)methyl]-3-(3-methylbut-1- ynyl)pyrazin-2-one (1.39 g, 4.3879 mmol) in dry DCM (28 mL), Iodine (1.4 g, 5.5160 mmol) in DCM (28 mL) was added slowly for 30 minutes and the resulting mixture was stirred at room temperature for 30 min. The solvent was evaporated, and the residue was subjected to silica gel column chromatography in 0% to 10% ethyl acetate in hexanes. Collected fractions were combined and concentrated. The residue was triturated with hexane (10 mL) for 30 minutes. The precipitate was filtered and washed with hexane to afford 2-chloro-7-iodo-6-isopropyl-furo[2,3- b]pyrazine (1.1 g, 74%) as a pale yellow solid.1H NMR (500 MHz, CDCl3) δ 8.15 (s, 1H), 3.43 (hept, J = 6.9 Hz, 1H), 1.40 (d, J = 6.9 Hz, 6H). ESI-MS m/z calc.321.93698, found 323.1 (M+1)+; Retention time: 3.14 minutes; LC method E. Step 2: 2-Chloro-7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazine [00326] To a 250 mL screw top vessel was added 2-chloro-7-iodo-6-isopropyl-furo[2,3- b]pyrazine (0.953 g, 2.955 mmol), potassium cyclopropyltrifluoroborate (434 mg, 2.933 mmol), cesium carbonate (3.104 g, 9.527 mmol), Xantphos (122 mg, 0.2108 mmol), and PdOAc2 (36 mg, 0.1603 mmol) in THF (20 mL). The reaction mixture was degassed for 10 minutes, then sealed and heated in an oil bath at 150 °C for 90 minutes. The reaction was cooled to RT, diluted with ethyl acetate, and washed with water, and brine. Then the organic layer was dried over sodium sulfate, filtered, and concentrated. The crude material was then purified by silica chromatography (80 g of silica, collect by ELSD) using a gradient eluent of 100% hexane to 40% ethyl acetate in hexanes over 25 min (compound elutes at 25% ethyl acetate) followed by a second purification using reverse phase HPLC chromatography (1-99% acetonitrile/5 mM aqueous HCl over 20 min) which afforded the desired white solid as 2-chloro-7-cyclopropyl-6- isopropyl-furo[2,3-b]pyrazine (135 mg, 19%), 1H NMR (400 MHz, CDCl3) δ 8.08 (s, 1H), 3.50 – 3.35 (m, 1H), 1.82 – 1.71 (m, 1H), 1.40 (s, 3H), 1.38 (s, 3H), 1.19 – 1.11 (m, 2H), 1.03 – 0.91 (m, 2H). ESI-MS m/z calc.236.07164, found 237.1 (M+1)+; Retention time: 2.09 minutes. LC method A. Step 3: Methyl 7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazine-2-carboxylate [00327] A mixture of 2-chloro-7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazine (135 mg, 0.5703 mmol) and Pd(dppf)Cl2.DCM (47 mg, 0.05755 mmol) and TEA (400 µL, 2.870 mmol) in MeOH (2.0 mL) in a steel pressure vessel equipped with a stir bar was purged with excess carbon monoxide (1.6 g, 55.14 mmol). The reaction mixture was heated to 100 °C with 100 psi of carbon monoxide and stirred for 16 h. The reaction mixture was allowed to cool to rt. The reaction mixture was filtered through a pad of celite, washed with ethyl acetate, filtrate was concentrated in vacuo. The residue was purified by silica gel chromatography (40 g of silica) using a gradient eluent of 100% hexanes to 55% ethyl acetate in hexanes (compound elutes at 25% ethyl acetate) to afford a white solid methyl 7-cyclopropyl-6-isopropyl-furo[2,3- b]pyrazine-2-carboxylate (118 mg, 80%). ESI-MS m/z calc.260.1161, found 261.1 (M+1)+; Retention time: 1.75 minutes. LC method A. Step 4: 1-(7-Cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-2- dimethoxyphosphoryl-ethanone [00328] A flask was heat gun-dried then cooled down to room temperature using a nitrogen stream and charged with dimethyl methylphosphonate (92 µL, 0.8601 mmol) and anhydrous THF (5.0 mL). The solution was cooled down to 0 °C and a solution of LiHMDS in THF (1.9 mL of 1 M, 1.900 mmol) was added dropwise. The reaction was stirred at 0 °C for 30 minutes after which time a solution of methyl 7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazine-2- carboxylate (118 mg, 0.4533 mmol) and methyl 7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazine- 2-carboxylate (100 mg, 0.3842 mmol) in anhydrous THF (2.2 mL) was added dropwise. The reaction was stirred at 0 °C for 90 minutes then allowed to warm to rt and stirred an additional 30 minutes. The mixture was then quenched by the addition of a saturated aqueous solution of ammonium chloride (10 mL). The mixture was allowed to stir at room temperature vigorously stirred for 30 minutes. The layers were then separated, and the aqueous layers was extracted with ethyl acetate (2 x 10 mL) and the combined organic layers were washed with brine (10 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude was then purified by silica chromatography (40 g of silica) using a gradient eluent of 100% hexanes to 100% ethyl acetate over 20 minutes followed by a gradient of 100% dichloromethane to 20% methanol in dichloromethane over 25 minutes. The desired fractions were concentrated under reduced pressure, and dried under high vacuum to afford 1-(7-cyclopropyl-6-isopropyl-furo[2,3- b]pyrazin-2-yl)-2-dimethoxyphosphoryl-ethanone (170 mg, 56%), ESI-MS m/z calc.352.1188, found 353.3 (M+1)+; Retention time: 1.6 minutes. LC method A. Example 27: Preparation of 1-(5-cyclobutyl-6-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2- dimethoxyphosphoryl-ethanone Step 1: 6-Bromo-3-prop-1-ynyl-pyrazin-2-amine [00329] A solution of (E)-1-bromoprop-1-ene (2.3 g, 19.01 mmol) in dry THF (131 mL) was cooled -78 °C followed by the addition of butyllithium (14.8 mL of 2.5 M, 37.00 mmol) and the mixture was stirred for 20 min at -78 °C. Then the mixture was warmed to 0 °C and water (992 µL, 55.07 mmol) was added and the mixture was stirred for 5 min at this temperature. Then, 3,6- dibromopyrazin-2-amine (4,350 mg, 17.20 mmol), trans-dichloro-bis(triphenylphosphine) palladium(II) (1.8 g, 2.564 mmol), copper iodide (491 mg, 2.578 mmol) and triethylamine (7.6 mL, 54.53 mmol) were added and the mixture was slowly warmed to room temperature and stirred for 22 hours. The dark mixture was diluted with 50 ml of ethyl acetate and filtered through thin layer of celite and dried under reduced pressure. This was then purified by silica chromatography (40 g, 0% to 20% EtOAc in Hexane (over 7 min), 10% MeOH in DCM (over 23 Min). This gave 2 aliquots of the desired product 6-bromo-3-prop-1-ynyl-pyrazin-2-amine (1.11 g, 30%) ESI-MS m/z calc.210.9745, found 213.944 (M+1)+; Retention time: 0.903 minutes and 6-bromo-3-prop-1-ynyl-pyrazin-2-amine (2.34 g, 49%) ESI-MS m/z calc.210.9745, found 213.944 (M+1)+; Retention time: 0.895 minutes. LC method A. Step 2: 3-Bromo-6-methyl-5H-pyrrolo[2,3-b]pyrazine [00330] To a solution of 6-bromo-3-prop-1-ynyl-pyrazin-2-amine (1.21 g, 4.964 mmol) in t- BuOH (10.4 mL) was added potassium tert-butoxide (2.79 g, 24.86 mmol). The reaction mixture was stirred at 100 °C for 17 hours then was allowed to cool down to room temperature. The volatiles were removed under reduced pressure and the residue was diluted with water (250 mL) and stirred at 0 °C for 5 minutes and was filtered. The solid was washed with water (3 x 100 mL) and was diluted in acetonitrile (500 mL). The solution was concentrated under reduced pressure and the solid was triturated in acetonitrile (3 x 100 mL) and filtered. The solid was dried under reduced pressure to afford 3-bromo-6-methyl-5H-pyrrolo[2,3-b]pyrazine (1.07 g, 91%).1H NMR (400 MHz, DMSO) δ 12.14 (s, 1H), 8.38 (s, 1H), 6.39 (d, J = 1.1 Hz, 1H), 2.48 – 2.45 (m, 3H). ESI-MS m/z calc.210.9745, found 213.999 (M+1)+; Retention time: 1.088 minutes. LC method N. Step 3: 3-Bromo-5-cyclobutyl-6-methyl-pyrrolo[2,3-b]pyrazine [00331] 3-Bromo-6-methyl-5H-pyrrolo[2,3-b]pyrazine (3.65 g, 14.29 mmol) was dissolved in anhydrous DMF (60.6 mL) then cooled to 0 °C. Cyclobutane iodide (6.76 g, 37.14 mmol) was added then sodium hydride (1.6 g of 60 %w/w, 40.00 mmol) was added in 4 portions over 10 minutes. Once addition was complete the reaction was stirred at 0 °C for 30 minutes then heated to 75 °C and stirred for 2.5 hours. The reaction was quenched by cautious, dropwise addition of ammonium chloride, then evaporated to dryness under reduced pressure. The residue was partitioned between EtOAc and water, and the layers were separated. The aqueous layer was further extracted with EtOAc (2 times). The combined extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. This was then purified by silica chromatography (80 g, 0% to 25% EtOAc in hexanes over 25 Min) to give 3-bromo-5- cyclobutyl-6-methyl-pyrrolo[2,3-b]pyrazine (1.20 g, 29%) 1H NMR (400 MHz, DMSO) δ 8.43 (s, 1H), 6.49 – 6.45 (m, 1H), 5.03 – 4.90 (m, 1H), 3.12 – 2.96 (m, 2H), 2.52 (d, J = 1.0 Hz, 3H), 2.47 – 2.36 (m, 2H), 1.98 – 1.74 (m, 2H). ESI-MS m/z calc.265.02145, found 267.977 (M+1)+; Retention time: 2.288 minutes. LC method N. Step 4: Methyl 5-cyclobutyl-6-methyl-pyrrolo[2,3-b]pyrazine-3-carboxylate [00332] To a suspension of 3-bromo-5-cyclobutyl-6-methyl-pyrrolo[2,3-b]pyrazine (1.20 g, 4.329 mmol) in MeOH (27.6 mL) was added Triethylamine (1.37 g, 13.54 mmol). Nitrogen was bubbled into the mixture for 15 minutes and then Pd(dppf)Cl2.DCM (3.7 g, 4.531 mmol) was added. Nitrogen bubbling was then continued for 5 minutes. The mixture was then stirred at 100 °C under 125 psi of carbon monoxide pressure for 1 hour. The reaction mixture was dried under reduced pressure and taken up in EtOAc and passed through celite and the liquid was dried under reduced pressure. This was then purified by silica chromatography (40 g, 0% to 25% EtOAc in Hexane (over 25 Min)). This material was then triturated in EtOAc and hexanes and filtered which gave the desired product methyl 5-cyclobutyl-6-methyl-pyrrolo[2,3-b]pyrazine-3- carboxylate (469 mg, 44%) 1H NMR (400 MHz, DMSO) δ 8.98 (s, 1H), 6.57 (s, 1H), 5.07 (ttd, J = 9.4, 8.0, 1.0 Hz, 1H), 3.93 (s, 3H), 3.22 – 3.06 (m, 2H), 2.61 (s, 3H), 2.51 – 2.38 (m, 2H), 2.02 – 1.76 (m, 2H). ESI-MS m/z calc.245.11642, found 246.078 (M+1)+; Retention time: 1.63 minutes. LC method N. Step 5: 1-(5-Cyclobutyl-6-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2- dimethoxyphosphoryl-ethanone [00333] A flask was flame-dried then cooled down to room temperature using a nitrogen stream and charged with [methoxy(methyl)phosphoryl]oxymethane (587 µL, 5.417 mmol) and anhydrous THF (25.5 mL). The solution was cooled down to -78 °C and a solution of (diisopropylamino)lithium (2.6 mL of 2 M, 5.200 mmol) was added dropwise. The reaction was stirred for 1 hour at -78 °C and a solution of methyl 5-cyclobutyl-6-methyl-pyrrolo[2,3- b]pyrazine-3-carboxylate (664 mg, 2.707 mmol) in anhydrous THF (6.8 mL) was added dropwise. The reaction was then stirred at -78 °C for 30 minutes then slowly taken to 0 °C for 30 minutes. The reaction was then quenched by the addition of a saturated aqueous solution of ammonium chloride (50 mL). The biphasic mixture was vigorously stirred for 30 minutes, then the layers were separated, and the aqueous layer was extracted with ethyl acetate (3 x 75 mL). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. This was then purified by silica chromatography (24 g, 0% to 25% EtOAc in Hexane (over 25 Min), 25% to 100% EtOAc in hexanes (over 5 Min), 0% to 15% MeOH in DCM (over 10 Min)) This gave the desired product 1-(5-cyclobutyl-6-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-2-dimethoxyphosphoryl-ethanone (416 mg, 45%) ESI-MS m/z calc. 337.11914, found 338.24 (M+1)+; Retention time: 1.417 minutes. LC method N. Example 28: Preparation of 1-(6-tert-butyl-7-methyl-furo[2,3-b]pyrazin-2-yl)-2- dimethoxyphosphoryl-ethanone Step 1: Methyl 7-bromo-6-tert-butyl-furo[2,3-b]pyrazine-2-carboxylate [00334] To a solution of methyl 6-tert-butylfuro[2,3-b]pyrazine-2-carboxylate (3.1 g, 13.23 mmol) in DMF (62 mL) at 0 °C was added N-bromo-succinimide (8.25 g, 46.35 mmol). The reaction mixture was stirred then heated at 60 °C for 4 h. The reaction mixture was quenched with sodium bicarbonate (40 mL), diluted with EtOAc (200 mL), washed with sodium bisulfate solution (40 mL), water (3 x 40 mL), dried over sodium sulfate, filtered, concentrated in vacuo. The residue was purified by normal phase flash chromatography (120 g of silica gel, 0-40% EtOAc to Hex, 15 minute run) to give a clear oil of methyl 7-bromo-6-tert-butyl-furo[2,3- b]pyrazine-2-carboxylate (3.0 g, 72%) ESI-MS m/z calc.312.01096, found 312.985 (M+1)+; Retention time: 1.583 minutes. LC method A. Step 2: Methyl 6-tert-butyl-7-methyl-furo[2,3-b]pyrazine-2-carboxylate [00335] Under an argon atmosphere, a vial with stir bar containing methyl 7-bromo-6-tert- butyl-furo[2,3-b]pyrazine-2-carboxylate (1.6 g, 5.109 mmol) was added tripotassium phosphate (3.25 g, 15.31 mmol), Pd(PPh3)4 (590 mg, 0.5106 mmol) and trimethylboroxine (1.5 mL, 10.25 mmol). Degassed DMF (48 mL) and degassed water (9.6 mL) were added to the mixture. The mixture was sealed under argon and heated at 170 °C in a microwave reactor for 30 min, then cooled to room temperature. To the reaction mixture was added 1 M NaOH (until pH=10) and washed with diethyl ether (50 mL) and EtOAc (50 mL). The aqueous was acidified with 1 M HCl (until pH = 3), then extracted with EtOAc (3 x 25 mL). washed with 1 M HCl (until pH = 3), dried over sodium sulfate, filtered, and concentrated in vacuo. The mixture was purified via normal phase flash chromatography (80g of silica gel, 0-50% EtOAc-hexanes, 15 minute run) to give a crude clear oil of 6-tert-butyl-7-methyl-furo[2,3-b]pyrazine-2-carboxylic acid (958 mg, 70%) ESI-MS m/z calc.234.10045, found 235.018 (M+1)+; Retention time: 1.261 minutes. To a solution of the crude intermediate 6-tert-butyl-7-methyl-furo[2,3-b]pyrazine-2-carboxylic acid (958 mg, 70%) in anhydrous DMF (21 mL) was added potassium carbonate (2.09 g, 15.12 mmol) and methyl iodide (965 µL, 15.50 mmol) and stirred 90 minutes at room temperature. The reaction was quenched with saturated aqueous and extracted with EtOAc (3 x 25 mL). The combined organics were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by normal phase flash chromatography (80g of silica gel, 0- 20 EtOAc-hexanes, 15 minute run) to give a clear oil of methyl 6-tert-butyl-7-methyl-furo[2,3- b]pyrazine-2-carboxylate (761 mg, 59%) ESI-MS m/z calc.248.11609, found 249.489 (M+1)+; Retention time: 1.488 minutes. LC method A.
Step 3: 1-(6-tert-Butyl-7-methyl-furo[2,3-b]pyrazin-2-yl)-2- dimethoxyphosphoryl-ethanone [00336] To a stirred solution of dimethyl methylphosphonate (500 µL, 4.675 mmol) in THF (22 mL) at -78 °C was added dropwise LiHMDS (in THF solution) (6.0 mL of 1 M, 6.000 mmol). The reaction mixture was stirred at -78 °C for 15 minutes and then a solution of methyl 6-tert-butyl-7-methyl-furo[2,3-b]pyrazine-2-carboxylate (753.3 mg, 3.034 mmol) in anhydrous THF (7.0 mL) was added dropwise over 5 minutes. The reaction was stirred at -78 °C for 10 minutes. The reaction was quenched by the addition of a saturated aqueous solution of ammonium chloride (5 mL) and the biphasic mixture was vigorously stirred at room temperature for 5 minutes. The layers were then separated, and the aqueous layer was extracted with EtOAc (2 x 5 mL). The combined organic layers were washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by normal phase chromatography (80 g of silica gel, 20-99% of EtOAc-Hex, 15 minutes run) to give a light- yellow oil of 1-(6-tert-butyl-7-methyl-furo[2,3-b]pyrazin-2-yl)-2-dimethoxyphosphoryl- ethanone (766 mg, 74%) ESI-MS m/z calc.340.1188, found 341.432 (M+1)+; Retention time: 1.417 minutes. LC method A. Example 29: Preparation of 2-dimethoxyphosphoryl-1-[5-isopropyl-7-(2- methoxyethyl)pyrrolo[2,3-b]pyrazin-3-yl]153xtracte Step 1: 3-Chloro-5-isopropyl-pyrrolo[2,3-b]pyrazine [00337] 3-Chloro-5H-pyrrolo[2,3-b]pyrazine (4.0 g, 26.05 mmol) was dissolved in anhydrous DMF (104.9 mL) then cooled to 0 °C.2-iodopropane (3.2 mL, 32.06 mmol) was added then sodium hydride (1.53 g of 60 %w/w, 38.25 mmol) was added in portions over 10 minutes. Once addition was complete the reaction was stirred at 0 °C for 30 minutes then warmed up to room temperature and stirred for 90 minutes. The reaction was quenched by cautious, dropwise addition of ammonium chloride, then evaporated to dryness under reduced pressure. The residue was partitioned between EtOAc and water, and the layers were separated. The aqueous layer was further extracted with EtOAc (2 x) The combined extracts were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by flash chromatography on silica gel using 0-100% EtOAc/hexanes to give 3- chloro-5-isopropyl-pyrrolo[2,3-b]pyrazine (3.690 g, 72%) as a yellow solid.1H NMR (400 MHz, CDCl3) δ 8.41 (s, 1H), 7.58 (d, J = 3.7 Hz, 1H), 6.70 (d, J = 3.7 Hz, 1H), 5.08 (hept, J = 6.8 Hz, 1H), 1.55 (s, 3H), 1.53 (s, 3H). ESI-MS m/z calc.195.05632, found 196.9 (M+1)+; Retention time: 1.39 minutes. LC method A. Step 2: 3-Chloro-5-isopropyl-pyrrolo[2,3-b]pyrazine-7-carbaldehyde [00338] POCl3 (1.8 mL, 19.31 mmol) was added dropwise to a solution of 3-chloro-5- isopropyl-pyrrolo[2,3-b]pyrazine (3.69 g, 18.86 mmol) in DMF (18.63 mL) at 0 °C under argon. The reaction mixture was stirred for 20 minutes at room temperature then the reaction mixture was heated at 60 °C for 1 hour. After 1 hour, the mixture was carefully added to an ice/water bath while stirring. Afterwards, NaOAc was added to the ice/water solution. The aqueous solution was extracted with DCM (3X). The organic layers were combined, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by flash chromatography on silica gel using 0-100% EtOAc/hexanes to give 3-chloro-5- isopropyl-pyrrolo[2,3-b]pyrazine-7-carbaldehyde (1.800 g, 43%) as a white solid.1H NMR (400 MHz, CDCl3) δ 10.26 (s, 1H), 8.62 (s, 1H), 8.24 (s, 1H), 5.12 (hept, J = 6.8 Hz, 1H), 1.61 (d, J = 6.8 Hz, 6H). ESI-MS m/z calc.223.05124, found 224.1 (M+1)+; Retention time: 1.23 minutes. LC method A. Step 3: 3-Chloro-5-isopropyl-7-[(Z)-2-methoxyvinyl]pyrrolo[2,3-b]pyrazine [00339] To a suspension of methoxymethyl(triphenyl)phosphonium chloride (4.553 g, 12.88 mmol) in THF (39 mL) at 0 °C was added NaHMDS in THF (16.1 mL of 1 M, 16.10 mmol) dropwise. The mixture was stirred at 0 °C for 45 min. A solution of 3-chloro-5-isopropyl- pyrrolo[2,3-b]pyrazine-7-carbaldehyde (1.8 g, 8.048 mmol) in THF (21.13 mL) was added dropwise. The mixture was stirred at 0 °C for 1 hour. The mixture was partitioned between ethyl acetate (100 mL) and water (100 mL). The layers were separated, and the aqueous phase was extracted with ethyl acetate (2 x 100 mL). The combined organic layers were washed with brine (50 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel using 0-100% EtOAc/hexanes over 25 minutes to give 3-chloro-5-isopropyl-7-[(Z)-2-methoxyvinyl]pyrrolo[2,3-b]pyrazine (1.400 g, 69%) as a yellow oil (mixture of Z and E isomer, ~1:1 ratio). ESI-MS m/z calc.251.08253, found 252.1 (M+1)+; Retention time: 1.82 minutes and 1.76 minutes. LC method A. Step 4: 3-Chloro-5-isopropyl-7-(2-methoxyethyl)pyrrolo[2,3-b]pyrazine [00340] To a solution of 3-chloro-5-isopropyl-7-[(Z)-2-methoxyvinyl]pyrrolo[2,3-b]pyrazine (~1:1 mixture of isomers, 30 mg, 0.1192 mmol) in EtOAc (12.0 mL) under nitrogen was added palladium on carbon (63.9 mg of 10 %w/w, 0.06005 mmol). The reaction mixture was stirred at room temperature under a hydrogen atmosphere for 1 h. The reaction mixture was filtered through Celite and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel using 0-100% EtOAc/hexanes over 20 minutes to give 3-chloro-5-isopropyl-7-(2-methoxyethyl)pyrrolo[2,3-b]pyrazine (19.8 mg, 65%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 8.36 (s, 1H), 7.44 (s, 1H), 5.03 (hept, J = 6.8 Hz, 1H), 3.74 (t, J = 6.7 Hz, 2H), 3.39 (s, 3H), 3.09 (td, J = 6.6, 1.0 Hz, 2H), 1.52 (d, J = 6.8 Hz, 6H). ESI-MS m/z calc.253.09819, found 254.1 (M+1)+; Retention time: 1.57 minutes. LC method A. Step 5: Methyl 5-isopropyl-7-(2-methoxyethyl)pyrrolo[2,3-b]pyrazine-3- carboxylate [00341] To a suspension of 3-chloro-5-isopropyl-7-(2-methoxyethyl)pyrrolo[2,3-b]pyrazine (422.3 mg, 1.664 mmol) in MeOH (9.7 mL) was added triethylamine (0.7 mL, 5.022 mmol). Nitrogen was bubbled into the mixture for 15 minutes and then Pd(dppf)Cl2.DCM (137.4 mg, 0.1683 mmol) was added. Nitrogen bubbling was then continued for 5 minutes. The mixture was then stirred at 100 °C under 80-100 psi of carbon monoxide pressure for 16 hours. The reaction mixture was filtered and concentrated under reduced pressure. The crude product was purified by flash chromatography on silica gel using 0-100% EtOAc/hexanes over 20 minutes to give methyl 5-isopropyl-7-(2-methoxyethyl)pyrrolo[2,3-b]pyrazine-3-carboxylate (401.7 mg, 87%) as a yellow-orange oil.1H NMR (400 MHz, CDCl3) δ 9.18 (s, 1H), 7.72 – 7.71 (m, 1H), 5.27 (hept, J = 6.8 Hz, 1H), 4.04 (s, 3H), 3.76 (t, J = 6.6 Hz, 2H), 3.39 (s, 3H), 3.14 (t, J = 0.9 Hz, 2H), 1.53 (d, J = 6.8 Hz, 6H). ESI-MS m/z calc.277.14264, found 278.4 (M+1)+; Retention time: 1.2 minutes. LC method A. Step 6: 2-Dimethoxyphosphoryl-1-[5-isopropyl-7-(2-methoxyethyl)pyrrolo[2,3- b]pyrazin-3-yl]156xtracte [00342] A flask was flame-dried then cooled down to room temperature using a nitrogen stream, and charged with dimethyl methylphosphonate (0.22 mL, 2.057 mmol) and anhydrous THF (15.4 mL). The solution was cooled down to -78 °C and a solution of LiHMDS in THF (4.5 mL of 1 M, 4.500 mmol) was added dropwise. The reaction was stirred for 1 hour at -78 °C and a solution of methyl 5-isopropyl-7-(2-methoxyethyl)pyrrolo[2,3-b]pyrazine-3-carboxylate (401.7 mg, 1.448 mmol) in anhydrous THF (4.1 mL) was added dropwise. The reaction was then stirred at -78 °C for 1.5 h then slowly allowed to reach room temperature and stirred at room temperature for 30 minutes. The reaction was then quenched by the addition of a saturated aqueous solution of ammonium chloride (50 mL). The biphasic mixture was vigorously stirred for 30 minutes, then the layers were separated and the aqueous layer was extracted with ethyl acetate (5 x 15 mL). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by flash chromatography on silica gel using 0-20% MeOH/DCM over 20 minutes to give 2-dimethoxyphosphoryl-1-[5-isopropyl-7-(2-methoxyethyl)pyrrolo[2,3-b]pyrazin-3- yl]acetaldehyde (485.9 mg, 91%) as an orange oil.1H NMR (400 MHz, CDCl3) δ 9.19 (s, 1H), 7.75 (s, 1H), 5.11 (p, J = 6.8 Hz, 1H), 4.09 (s, 1H), 4.03 (s, 1H), 3.80 (s, 3H), 3.77 (s, 3H), 3.76 – 3.72 (m, 2H), 3.39 (s, 3H), 3.14 (t, J = 0.9 Hz, 2H), 1.60 (d, J = 6.8 Hz, 6H). ESI-MS m/z calc.369.14536, found 370.4 (M+1)+; Retention time: 1.11 minutes. LC method A. Example 30: Preparation of 1-[6-tert-Butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl]-2-dimethoxyphosphoryl-ethanone Step 1: 3-Bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine-7-carbaldehyde [00343] To a solution of 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine (1.0 g, 3.729 mmol) in DMF (12.0 mL) was added dropwise phosphorus oxychloride (1.05 mL, 11.27 mmol) at room temperature. The mixture was heated to 70 °C and stirred overnight. The mixture was cooled to room temperature and then it was quenched with water (10 mL) dropwise (exothermic). The solution was extracted with EtOAc (3 x 50 mL) then the combined organic phases were washed with aqueous saturated solution of sodium bicarbonate (50 mL), brine (50 mL) then concentrated under vacuum. The crude product was purified by flash chromatography on silica gel using 0-10% DCM/MeOH to provide 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazine-7-carbaldehyde (652.2 mg, 59%) as an orange solid.1H NMR (400 MHz, CDCl3) δ 10.70 (s, 1H), 8.65 (s, 1H), 4.11 (s, 3H), 1.72 (s, 9H). ESI-MS m/z calc.295.032, found 296.0 (M+1)+; Retention time: 1.41 minutes. LC method A. Step 2: 3-Bromo-6-tert-butyl-7-[(Z)-2-methoxyvinyl]-5-methyl-pyrrolo[2,3- b]pyrazine [00344] To a suspension of methoxymethyl(triphenyl)phosphonium chloride (1.27 g, 3.594 mmol) in THF (14.13 mL) at 0 °C was added NaHMDS in THF (4.41 mL of 1 M, 4.410 mmol) dropwise. The mixture was stirred at 0 °C for 45 min. A solution of 3-bromo-6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazine-7-carbaldehyde (652.2 mg, 2.202 mmol) in THF (8 mL) was added dropwise. The mixture was stirred at 0 °C for 1 hour. The mixture was partitioned between ethyl acetate (100 mL) and water (100 mL). The layers were separated, and the aqueous phase was extracted with ethyl acetate (2 x 100 mL). The combined organic layers were washed with brine (50 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel using 0-100% EtOAc/hexanes over 25 minutes to give 3-bromo-6-tert-butyl-7-[(Z)-2-methoxyvinyl]-5-methyl-pyrrolo[2,3- b]pyrazine (625.1 mg, 88%) as a yellow oil (mixture of Z and E isomers). ESI-MS m/z calc. 323.06332, found 324.1 (M+1)+; Retention time: 2.01 minutes. LC method A. Step 3: methyl 6-tert-butyl-7-[(Z)-2-methoxyvinyl]-5-methyl-pyrrolo[2,3- b]pyrazine-3-carboxylate [00345] To a suspension of 3-bromo-6-tert-butyl-7-[(Z)-2-methoxyvinyl]-5-methyl- pyrrolo[2,3-b]pyrazine (625.1 mg, 1.928 mmol) in methanol (2.0 mL) was added triethylamine (0.81 mL, 5.811 mmol). Nitrogen was bubbled into the mixture for 15 minutes and then Pd(dppf)Cl2.DCM (80.6 mg, 0.09870 mmol) was added. Nitrogen bubbling was then continued for 5 minutes. The mixture was then stirred at 100 °C under 50 psi carbon monoxide pressure for 18 hours. The mixture was then allowed to cool down to room temperature, was filtered on a Celite pad and concentrated. The resulting crude product was purified by silica gel chromatography (Gradient: 0-100% EtOAc in hexane over 25 minutes) to afford methyl 6-tert- butyl-7-[(Z)-2-methoxyvinyl]-5-methyl-pyrrolo[2,3-b]pyrazine-3-carboxylate (496.7 mg, 85%). ESI-MS m/z calc.303.1583, found 304.3 (M+1)+; Retention time: 1.37 minutes. LC method A. Step 4: Methyl 6-tert-butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3-b]pyrazine- 3-carboxylate [00346] A solution of methyl 6-tert-butyl-7-[(Z)-2-methoxyvinyl]-5-methyl-pyrrolo[2,3- b]pyrazine-3-carboxylate (496.7 mg, 1.637 mmol) in THF (31.1 mL) was bubbled with nitrogen over 5 minutes and then palladium on carbon (90.3 mg, 0.08485 mmol) was added. The resulting mixture was bubbled with hydrogen for 10 minutes and then was stirred at room temperature under balloon of hydrogen for 3 days. The crude was filtered through celite, washed with ethyl acetate, and concentrated under reduced pressure to provide crude methyl 6-tert- butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3-b]pyrazine-3-carboxylate (534.5 mg, 59%) as a yellow oil. ESI-MS m/z calc.305.17395, found 306.5 (M+1)+; Retention time: 1.46 minutes. LC method A. Step 5: 1-[6-tert-Butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]- 2-dimethoxyphosphoryl-ethanone [00347] A flask was flame-dried then cooled down to room temperature using a nitrogen stream and charged with dimethyl methylphosphonate (0.27 mL, 2.524 mmol) and anhydrous THF (21 mL). The solution was cooled down to -78 °C and a solution of LiHMDS in THF (5.3 mL of 1 M, 5.300 mmol) was added dropwise. The reaction was stirred for 1 hour at -78 °C and a solution of methyl 6-tert-butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3-b]pyrazine-3- carboxylate (534.5 mg, 1.750 mmol) in anhydrous THF (5.5 mL) was added dropwise. The reaction was then stirred at -78 °C for 1.5 h then slowly allowed to reach room temperature and stirred for 30 minutes. The reaction was then quenched by the addition of a saturated aqueous solution of ammonium chloride (50 mL). The biphasic mixture was vigorously stirred for 30 minutes, then the layers were separated, and the aqueous layer was extracted with ethyl acetate (5 x 15 mL). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by flash chromatography on silica gel using 0-20% MeOH/DCM over 20 minutes to give 1-[6-tert- butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-2-dimethoxyphosphoryl- ethanone (550.7 mg, 79%) as a orange oil. ESI-MS m/z calc.397.17667, found 398.2 (M+1)+; Retention time: 1.36 minutes. LC method A. VII. Synthesis of substituted alkyl aldehyde intermediates Example 31: Preparation of tert-Butyl (4S)-2,2-dimethyl-4-[(1S)-3-methyl-1-(3- oxopropoxy)butyl]oxazolidine-3-carboxylate Step 1: tert-Butyl (4S)-4-(1-hydroxy-3-methyl-butyl)-2,2-dimethyl-oxazolidine-3- carboxylate [00348] To a solution of tert-butyl (4S)-4-formyl-2,2-dimethyl-oxazolidine-3-carboxylate (18.9 g, 82.434 mmol) in anhydrous THF (300 mL) at -78 °C was added dropwise a THF solution of isobutylmagnesium chloride (80 mL of 2 M, 160.00 mmol). The reaction was let stirred in the dry-ice bath for 20 hours allowing the temperature to slowly warm to room temperature. To the reaction mixture at 0 °C was added a saturated aqueous solution of ammonium chloride (200 mL) and EtOAc (200 mL). To the bi-phasic solution was added brine (200 mL) and the phases were separated. The aqueous layer was extracted with EtOAc (2 x 100 mL) and the combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. Purification by silica gel chromatography (Gradient: 0-20% EtOAc in heptane) afforded tert-butyl (4S)-4-(1-hydroxy-3-methyl-butyl)-2,2- dimethyl-oxazolidine-3-carboxylate (19.7 g, 79%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 4.20 – 3.53 (overlapped with EtOAc, m, 4H), 2.00 – 1.77 (m, 1H), 1.61 (br. S., 4H), 1.50 (s, 12H), 1.42 – 1.32 (m, 1H), 1.22 – 1.08 (m, 1H), 0.99 – 0.94 (m, 3H), 0.92 (d, J = 6.6 Hz, 3H). ESI-MS m/z calc.287.2097, found 310.2 (M+23)+; Retention time: 1.89 minutes. LC method I. Step 2: tert-Butyl (4S)-4-[1-(3-tert-butoxy-3-oxo-propoxy)-3-methyl-butyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00349] To a solution of tert-butyl (4S)-4-(1-hydroxy-3-methyl-butyl)-2,2-dimethyl- oxazolidine-3-carboxylate (19.7 g, 65.119 mmol) in anhydrous THF (60 mL) was added portion- wise NaH (3.1 g, 60 %w/w, 77.507 mmol) at room temperature. To the suspension was added tert-butyl prop-2-enoate (19.250 g, 22 mL, 150.19 mmol) and the reaction was stirred for 7 hours. To the suspension was added water (100 mL) followed by EtOAc (200 mL) and brine (100 mL). The reaction mixture was stirred at room temperature for 18 hours then the phases were separated. The aqueous layer was extracted with EtOAc (3 x 100 mL) and the combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Purification by silica gel chromatography (Gradient: 0-20% EtOAc in heptane) afforded tert-butyl (4S)-4-[1-(3-tert-butoxy-3-oxo-propoxy)-3- methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate (21.8 g, 78%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 4.29 – 3.58 (m, 6H), 2.55 – 2.34 (m, 2H), 1.83 – 1.68 (m, 1H), 1.66 – 1.36 (m, 25H), 1.20 – 1.06 (m, 1H), 0.98 – 0.82 (m, 6H). ESI-MS m/z calc.415.2934, found 438.2 (M+23)+; Retention time: 2.29 minutes. LC method I. Step 3: tert-Butyl (4S)-4-[(1R)-1-(3-hydroxypropoxy)-3-methyl-butyl]-2,2- dimethyl-oxazolidine-3-carboxylate and tert-butyl (4S)-4-[(1S)-1-(3- hydroxypropoxy)-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00350] In a flask was added LAH (pellets form) (1.47 g, 38.731 mmol) and anhydrous THF (125 mL). The suspension was stirred at room temperature for 1 hour and was cooled in an ice- bath at 0 °C. To the suspension was added a solution of tert-butyl (4S)-4-[1-(3-tert-butoxy-3- oxo-propoxy)-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate (7.76 g, 18.263 mmol) in anhydrous THF (75 mL) slowly dropwise over 18 minutes in order to keep the internal temperature below 6 °C. The addition funnel was rinsed with anhydrous THF (25 mL) and the solution was rapidly added to the reaction. The reaction was stirred for 45 minutes, and water (1 mL) was slowly added. To the reaction mixture was added a 15% aqueous solution of NaOH (1 mL) followed by water (1 mL). To the slurry was added anhydrous magnesium sulfate (13 g) and the solution was stirred at room temperature for 1 hour. The suspension was filtered through Celite, the pad was rinsed with EtOAc (3 x 50 mL) and the combined filtrates were concentrated under reduced pressure. The crude residue was purified by silica gel chromatography (Gradient: 0-50 % EtOAc in heptane) and the mixed fractions were combined and concentrated under reduced pressure. The residue was purified by silica gel chromatography (Gradient: 0-50 % MTBE in heptane). The purest fractions of the less polar diastereomer of both purifications were combined and concentrated under reduced pressure to afford tert-butyl (4S)-4-[(1R)-1-(3- hydroxypropoxy)-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate (544 mg, 7%) as a colorless oil (99:1 mixture of of 4S,1R and 4S,1S isomers) ESI-MS m/z calc.345.2515, found 246.3 (M-99)+; Retention time: 2.23 minutes. The mixed fractions of the less polar and the more polar diastereomers were combined and concentrated under reduced pressure to afford tert-butyl (4S)-4-[1-(3-hydroxypropoxy)-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate (900 mg, 11%) as a colorless oil. Isolated as a mixture of the 4S,1S and 4S,1R diastereomers in a 78:22 ratio for the 4S,1S: ESI-MS m/z calc.345.2515, found 246.2 (M-99)+; Retention time: 3.77 minutes. The purest fractions of the more polar diastereomer were combined and concentrated under reduced pressure to afford tert-butyl (4S)-4-[(1S)-1-(3-hydroxypropoxy)-3- methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate (3.87 g, 59%) as a colorless oil. Isolated as a mixture of the 4S,1S and 4S,1R diastereomers in a 92:8 ratio for the 4S,1S: 1H NMR (400 MHz, CDCl3) δ 4.25 – 3.53 (m, 8H), 2.33 (br. S., 0.5H), 2.12 (br. S., 0.5H), 1.86 – 1.70 (m, 3H), 1.67 – 1.54 (overlapped with water, m, 3H), 1.53 – 1.37 (m, 13H), 1.31 – 1.20 (m, 1H), 1.00 – 0.82 (m, 6H). ESI-MS m/z calc.345.2515, found 246.3 (M-99)+; Retention time: 2.22 minutes. LC method O. Step 4: tert-Butyl (4S)-2,2-dimethyl-4-[(1S)-3-methyl-1-(3- oxopropoxy)butyl]oxazolidine-3-carboxylate [00351] To a solution of tert-butyl (4S)-4-[(1S)-1-(3-hydroxypropoxy)-3-methyl-butyl]-2,2- dimethyl-oxazolidine-3-carboxylate (3.87 g, 10.776 mmol) in dichloromethane (110 mL) cooled at 0 °C were added water (40 mL), sodium bicarbonate (1.45 g, 17.261 mmol), sodium bromide (225 mg, 2.1867 mmol) and TEMPO (265 mg, 1.6961 mmol). To the bi-phasic solution was added sodium hypochlorite (8 mL of 10 %w/v, 10.747 mmol) slowly dropwise over 16 minutes under vigorous stirring keeping the internal temperature below 5 °C. After being stirred for 30 minutes at 0 °C, a saturated aqueous solution of sodium bicarbonate (100 mL) was added. The phases were separated, and the aqueous layer was extracted with DCM (2 x 50 mL). The combined organic layers were washed with a saturated aqueous solution of sodium bicarbonate (2 x 50 mL), a 10% aqueous solution of Na2S2O3 (2 x 50 mL) and brine (50 mL), then dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert- butyl (4S)-2,2-dimethyl-4-[(1S)-3-methyl-1-(3-oxopropoxy)butyl]oxazolidine-3-carboxylate (4.1 g, 92%) as pale yellow oil. Rotamers in a (1:1) ratio were observed by 1H NMR.1H NMR (400 MHz, CDCl3) δ 9.79 (br. S., 0.5H), 9.77 (br. S., 0.5H), 4.26 – 3.68 (m, 6H), 2.76 – 2.52 (m, 2H), 1.78 – 1.67 (m, 1H), 1.66 – 1.56 (m, 3H), 1.53 – 1.43 (m, 12H), 1.42 – 1.37 (m, 1H), 1.28 – 1.18 (m, 1H), 1.02 – 0.78 (m, 6H). ESI-MS m/z calc.343.2359, found 366.4 (M+23)+; Retention time: 2.0 minutes. LC method I. Example 32: Preparation of tert-butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo- 2H-pyrrole-1-carboxylate Step 1: (5R)-5-[[tert-Butyl(diphenyl)silyl]oxymethyl]pyrrolidin-2-one [00352] To a stirred solution of (5R)-5-(hydroxymethyl)pyrrolidin-2-one (25.01 g, 212.89 mmol) in degassed dichloromethane (600 mL) were added imidazole (33.76 g, 490.95 mmol) and DMAP (2.8 g, 22.690 mmol) under nitrogen. The solution was then cooled down to 0 °C and tert-butyl(chloro)diphenylsilane (65.259 g, 63.000 mL, 232.68 mmol) was added dropwise over 20 minutes. The reaction was stirred at 0 °C for 30 minutes then allowed to reach room temperature and stirred 3 days at room temperature. The reaction was then partitioned between dichloromethane (350 mL) and water (300 mL). The layers were separated and the organic layer was washed with a saturated aqueous solution of ammonium chloride (250 mL), brine (200 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to afford (5R)-5- [[tert-butyl(diphenyl)silyl]oxymethyl]pyrrolidin-2-one (84.39 g, 100%) as a colorless oil that was directly used in the next step without further purification.1H NMR (400 MHz, CDCl3) δ 7.67 – 7.58 (m, 4H), 7.49 – 7.33 (m, 6H), 5.78 (br. S, 1H), 3.86 – 3.74 (m, 1H), 3.66 – 3.59 (m, 1H), 3.55 – 3.46 (m, 1H), 2.37 – 2.26 (m, 2H), 2.21 – 2.06 (m, 1H), 1.78 – 1.65 (m, 1H), 1.09 – 1.02 (m, 9H). ESI-MS m/z calc.353.1811, found 354.4 (M+1)+; Retention time: 1.98 minutes. LC method I. Step 2: tert-Butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo- pyrrolidine-1-carboxylate [00353] To a stirred solution of crude (5R)-5-[[tert-butyl(diphenyl)silyl]oxymethyl]pyrrolidin- 2-one (66.1 g, 171.45 mmol) in anhydrous acetonitrile (300 mL) were added DMAP (12.7 g, 103.96 mmol) and Boc2O (65 g, 297.83 mmol). The reaction was then stirred overnight at room temperature. The reaction was then concentrated under reduced pressure and dissolved in ethyl acetate (500 mL). The organic layer was washed with a saturated solution of ammonium chloride (3 x 250 mL), water (2 x 250 mL), brine (2 x 250 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to afford a bright orange solid. The crude solid was triturated overnight with pentane (150 mL), filtered, washed with pentane (5 x 20 mL), dried under high vacuum to afford tert-butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5- oxo-pyrrolidine-1-carboxylate (70.747 g, 90%) as a pale yellow powder that was directly used in the next step without further purification.1H NMR (400 MHz, CDCl3 ) δ 7.68 – 7.59 (m, 4H), 7.48 – 7.35 (m, 6H), 4.29 – 4.13 (m, 1H), 3.90 (dd, J = 10.5, 4.2 Hz, 1H), 3.71 (dd, J = 10.5, 2.4 Hz, 1H), 2.79 (dt, J = 17.6, 10.4 Hz, 1H), 2.44 (ddd, J = 17.6, 8.8, 3.2 Hz, 1H), 2.23 – 2.03 (m, 2H), 1.44 (s, 9H), 1.05 (s, 9H). ESI-MS m/z calc.453.2335, found 354.2 (M-99)+; Retention time: 1.98 minutes. LC method Q. Step 3: tert-Butyl (5R)-5-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-oxo-3- phenylselanyl-pyrrolidine-1-carboxylate [00354] A NaHMDS solution in THF (86 mL of 1 M, 86.000 mmol) was slowly added at - 78°C to a solution of tert-butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-pyrrolidine- 1-carboxylate (20 g, 41.883 mmol) in THF (100 mL) at a rate so that the temperature was maintained close to -78 °C. The reaction mixture was maintained at this temperature for 1 h then phenyl selenohypochlorite (8.8 g, 45.949 mmol) in THF (50 mL) was added dropwise at a rate to avoid the reaction internal temperature warming up above -78 °C. The reaction mixture was stirred at this temperature for 2 h then it was quenched with saturated aqueous ammonium chloride (100 mL) and left to warm up to room temperature. It was then left stirring at room temperature for 30 min. The phases were separated, and the aqueous phase was extracted with ethyl acetate (3 x 50 mL). The combined organics were dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was split in two equal portions which were then purified by reverse phase chromatography on a C18 column using a 70 to 100% gradient of acetonitrile in pure water to provide the desired tert-butyl (5R)-5-[[tert- butyl(diphenyl)silyl]oxymethyl]-2-oxo-3-phenylselanyl-pyrrolidine-1-carboxylate (21.1 g, 82%) as a yellow oil and as a 3:1 ratio of diastereomers (trans: cis) ESI-MS m/z calc.609.1814, found 510.2 (M-99)+; Retention time: 2.369 minutes; along with a small amount of bis addition product tert-butyl (5R)-5-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-oxo-3,3- bis(phenylselanyl)pyrrolidine-1-carboxylate (1 g, 3%) as a yellow oil ESI-MS m/z calc. 765.1292, found 666.0 (M-99)+; Retention time: 2.703 minutes. LC method R. Step 4: tert-Butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-2H- pyrrole-1-carboxylate [00355] Into an aqueous solution of H2O2 (14.69 g, 13.000 mL of 35 %w/w, 151.16 mmol) in THF (195 mL), a solution of tert-butyl (5R)-5-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-oxo-3- phenylselanyl-pyrrolidine-1-carboxylate (32.3 g, 49.668 mmol) in THF (65 mL) was slowly added at 15 °C over a period of 90 min. The reaction mixture was quenched cautiously with an aqueous solution of NaHSO3 (10%, 100 mL). The phases were separated and the organic layer was washed with a saturated aqueous solution of sodium bicarbonate (3 x 60 mL), water (60 mL), brine (60 mL), dried over anhydrous sodium sulfate and evaporated under vacuum to give tert-butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-2H-pyrrole-1-carboxylate (25.1 g, 101%) as a yellow solid.1H NMR (400 MHz, Chloroform-d) δ 7.64 – 7.59 (m, 4H), 7.48 – 7.36 (m, 6H), 7.29 – 7.27 (m, 1H), 6.18 (dd, J = 6.1, 1.7 Hz, 1H), 4.69 – 4.63 (m, 1H), 4.13 (dd, J = 9.8, 3.4 Hz, 1H), 3.84 (dd, J = 9.8, 6.6 Hz, 1H), 1.45 (s, 9H), 1.04 (s, 9H). ESI-MS m/z calc. 451.2179, found 352.4 (M-99)+; Retention time: 2.43 minutes. LC method O. Example 33: Preparation of tert-butyl (4R)-2,2-dimethyl-4-[(1R)-3-methyl-1-(2- oxoethyl)butyl]oxazolidine-3-carboxylate Step 1: tert-Butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-isobutyl-5- oxo-pyrrolidine-1-carboxylate [00356] To a suspension of copper iodide (42 g, 220.53 mmol) in anhydrous THF (600 mL) was added a solution of isobutyl magnesium chloride (in THF) (220 mL of 2 M, 440.00 mmol) at -78°C over 1 h. The reaction was stirred at -78°C for 1 h after which time a solution of tert- butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-2H-pyrrole-1-carboxylate (40 g, 88.568 mmol) in anhydrous THF (600 mL) was added dropwise over 3 hours keeping the internal temperature below -78 °C. The reaction was stirred in the dry-ice bath for 14 hours allowing the temperature to slowly reach room temperature. The reaction mixture was put in an ice-cold bath and was quenched by a saturated aqueous solution of ammonium chloride (500 mL) keeping the internal temperature below 20 °C. To the reaction mixture was added EtOAc (250 mL) and the bi-phasic solution was vigorously stirred at room temperature for 15 minutes. The organic layer was separated, and the aqueous phase was again extracted with EtOAc (2 x 250 mL). The combined organic layers were washed with brine (500 mL), dried over anhydrous sodium sulfate, and filtered. The solution was concentrated under reduced pressure and purified by column chromatography (ethyl acetate in heptane 0% to 15%). The fractions containing the desired product were combined and concentrated to afford tert-butyl (2R,3R)-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-3-isobutyl-5-oxo-pyrrolidine-1-carboxylate (43 g, 93%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 7.68 – 7.58 (m, 4H), 7.48 – 7.36 (m, 6H), 3.89 – 3.79 (m, 2H), 3.71 (dd, J = 9.8, 2.0 Hz, 1H), 2.88 (dd, J = 17.7, 8.9 Hz, 1H), 2.40 (q, J = 8.2 Hz, 1H), 2.14 (dd, J = 17.6, 2.0 Hz, 1H), 1.71 – 1.58 (m, 1H), 1.45 (s, 9H), 1.35 – 1.26 (m, 2H), 1.06 (s, 9H), 0.94 – 0.88 (m, 6H). ESI-MS m/z calc.509.2961, found 410.4 (M-99)+; Retention time: 2.52 minutes. LC method Q. Step 2: tert-Butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-hydroxy-3- isobutyl-pyrrolidine-1-carboxylate [00357] Lithium triethylborohydride (in THF) (31 mL of 1 M, 31.000 mmol) was slowly added to a solution of tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-isobutyl-5- oxo-pyrrolidine-1-carboxylate (16.6 g, 27.941 mmol) in anhydrous THF (170 mL) at -78 °C over 40 minutes keeping the internal temperature below -78 °C. The reaction was stirred at - 78°C for 1 hour and 40 minutes then a solution of lithium triethylborohydride (in THF) (8 mL of 1 M, 8.0000 mmol) was again added dropwise over 5 minutes keeping the temperature below - 78 °C. The reaction was stirred for 15 minutes then was quenched by the addition of a saturated aqueous solution of sodium bicarbonate (100 mL) allowing the temperature to reach -5 °C. To the reaction mixture was added an aqueous solution of hydrogen peroxide (in water) (1.6 g, 35 %w/w, 16.463 mmol) followed by DCM (200 mL) and the resulting bi-phasic solution was left stirring at room temperature for 15 min. The layers were separated, and the aqueous layer was extracted with DCM (3 x 100 mL). The combined organic layers were dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure to afford crude tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-hydroxy-3-isobutyl-pyrrolidine-1- carboxylate (17.4 g, 97%) as a clear yellow oil and as a mixture of diastereomers (approximate ratio of 55:45) that was used in the following step without further purification.1H NMR (400 MHz, CDCl3) δ 7.76 – 7.56 (m, 4H), 7.49 – 7.34 (m, 6H), 5.62 – 5.34 (m, 1H), 4.26 – 4.01 (m, 0.5H), 3.98 – 3.79 (m, 0.5H), 3.71 – 3.56 (m, 1H), 3.55 – 3.21 (m, 1H), 2.68 (br. S., 0.25H), 2.46 – 2.23 (m, 1.25H), 2.13 – 1.97 (m, 0.5H), 1.74 – 1.17 (m, 14H), 1.11 – 1.01 (m, 9H), 0.98 – 0.79 (overlapped with impurity, m, 6H). ESI-MS m/z calc.511.3118, found 394.4 (M-117)+; Retention time: 2.64 minutes. LC method S. Step 3: tert-Butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2- hydroxyethyl)-4-methyl-pentyl]carbamate [00358] To a stirred solution of tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5- hydroxy-3-isobutyl-pyrrolidine-1-carboxylate (31.4 g, 60.926 mmol) in methanol (650 mL) at 50 °C was added slowly NaBH4 (23 g, 607.94 mmol) during which the internal temperature reached 56 °C. The reaction was stirred at 50 oC for 1 hour and then more NaBH4 (12 g, 317.19 mmol) was added slowly. The resulting mixture was stirred at 50 °C for 30 minutes. To the reaction mixture was added water (500 mL) and the volatiles were concentrated under reduced pressure. To the resulting aqueous solution was added DCM (300 mL) and the phases were separated. The aqueous phase was extracted with DCM (3 x 150 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert-butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2- hydroxyethyl)-4-methyl-pentyl]carbamate (31 g, 98%) as a pale yellow oil that was used in the following step without further purification.1H NMR (400 MHz, CDCl3) δ 7.72 – 7.62 (m, 4H), 7.49 – 7.35 (m, 6H), 4.80 (d, J = 8.6 Hz, 1H), 3.87 – 3.74 (m, 1H), 3.74 – 3.54 (m, 4H), 2.04 – 1.96 (m, 1H), 1.91 (dq, J = 12.1, 6.2 Hz, 1H), 1.74 – 1.57 (m, 1H), 1.55 – 1.40 (m, 11H), 1.26 – 1.13 (m, 1H), 1.12 – 1.02 (m, 10H), 0.93 – 0.81 (m, 6H). ESI-MS m/z calc.513.3274, found 414.4 (M-99)+; Retention time: 2.54 minutes. LC method L. Step 4: tert-Butyl N-[(1R,2R)-2-(2-hydroxyethyl)-1-(hydroxymethyl)-4-methyl- pentyl]carbamate [00359] To a solution of tert-butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2- hydroxyethyl)-4-methyl-pentyl]carbamate (32 g, 59.729 mmol) and tert-butyl N-[(1R,2R)-1- [[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2-hydroxyethyl)-4-methyl-pentyl]carbamate (2.1 g, 3.7603 mmol) in anhydrous THF (320 mL) was added TBAF (in THF) (90 mL of 1 M, 90.000 mmol) at 0 °C. The reaction was stirred in the ice-cold bath for 3 hours. The reaction was quenched at 0 °C with a saturated aqueous sodium bicarbonate (500 mL) and EtOAc (250 mL). The biphasic solution was stirred for 1 hour at room temperature and the layers were separated. The aqueous layer was extracted with ethyl acetate (2 x 250 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified in two different batches of 17g by reversed-phase chromatography (Column: C18. Gradient: 5-100 % methanol in water). The fractions containing the desired product were combined and concentrated under reduced pressure to afford tert-butyl N-[(1R,2R)-2-(2-hydroxyethyl)-1-(hydroxymethyl)-4-methyl-pentyl]carbamate (15 g, 86%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 4.99 (d, J = 7.3 Hz, 1H), 3.84 – 3.72 (m, 2H), 3.70 – 3.59 (m, 3H), 2.82 – 2.70 (m, 1H), 2.45 – 2.32 (m, 1H), 1.97 – 1.83 (m, 1H), 1.73 – 1.56 (m, 2H), 1.55 – 1.40 (m, 10H), 1.31 – 1.18 (m, 1H), 1.18 – 1.06 (m, 1H), 0.94 – 0.83 (m, 6H). ESI- MS m/z calc.275.2097, found 220.4 (M-55)+; Retention time: 1.58 minutes. LC method S. Step 5: tert-Butyl (4R)-4-[(1R)-1-(2-hydroxyethyl)-3-methyl-butyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00360] To a solution of tert-butyl N-[(1R,2R)-2-(2-hydroxyethyl)-1-(hydroxymethyl)-4- methyl-pentyl]carbamate (6.27 g, 20.674 mmol) in anhydrous DCM (100 mL) was added 2,2- dimethoxypropane (42.350 g, 50 mL, 406.63 mmol) at room temperature. To the solution was added p-toluenesulfonic acid hydrate (457 mg, 2.4025 mmol) and the reaction mixture was stirred at room temperature for 3 hours. To the reaction mixture was added a saturated aqueous sodium bicarbonate solution (150 mL) and DCM (100 mL). The phases were separated, and the aqueous layer was extracted with DCM (3 x 100 mL). The combined organic layers were dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to afford crude tert-butyl (4R)-4-[(1R)-1-(2-hydroxyethyl)-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3- carboxylate (7.1 g, 98%) as a clear oil that was used in the following step without further purification.1H NMR (400 MHz, CDCl3) δ 4.15 – 3.60 (m, 5H), 2.15 – 2.03 (m, 1H), 1.70 – 1.53 (m, 6H), 1.52 – 1.42 (m, 12H), 1.38 – 1.25 (overlapped with impurity, m, 2H), 1.23 – 1.04 (m, 1H), 0.93 (d, J = 6.6 Hz, 3H), 0.85 (d, J = 6.4 Hz, 3H). ESI-MS m/z calc.315.241, found 338.4 (M+23)+; Retention time: 1.91 minutes. LC method I. Step 6: tert-Butyl (4R)-2,2-dimethyl-4-[(1R)-3-methyl-1-(2- oxoethyl)butyl]oxazolidine-3-carboxylate [00361] A stirred solution of oxalyl chloride (1.4550 g, 1.0 mL, 11.463 mmol) in anhydrous dichloromethane (15 mL) was cooled down to -78 °C and a solution of dimethylsulfoxide (1.5414 g, 1.4 mL, 19.728 mmol) in anhydrous dichloromethane (15 mL) was added dropwise. The reaction was stirred for 30 minutes at -78 °C after which time a solution of tert-butyl (4R)- 4-[(1R)-1-(2-hydroxyethyl)-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.49 g, 4.0291 mmol) in anhydrous dichloromethane (15 mL) was added. The reaction was stirred for 30 minutes at -78 °C after which time triethylamine (4.0656 g, 5.6 mL, 40.178 mmol) was added. The reaction was stirred for 30 minutes then allowed to reach room temperature and stirred for 30 minutes at room temperature. The reaction was quenched by the addition of a saturated aqueous solution of ammonium chloride (30 mL). The mixture was vigorously stirred at room temperature for 2 hours, then the layers were separated, and the aqueous layer was extracted with dichloromethane (4 x 15 mL). The combined organic layers were washed with water (20 mL), brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford tert-butyl (4R)-2,2-dimethyl-4-[(1R)-3-methyl-1-(2- oxoethyl)butyl]oxazolidine-3-carboxylate (1.34 g, 85%) as a yellow oil which was used without further purification.1H NMR (400 MHz, CDCl3) δ 9.78 (br. S., 1H), 3.98 – 3.87 (m, 2H), 3.81 (d, J = 8.1 Hz, 2H), 3.15 – 3.07 (m, 1H), 2.64 – 2.52 (m, 1H), 2.43 – 2.36 (m, 1H), 1.67 – 1.60 (overlapped with water, m, 2H), 1.51 – 1.45 (m, 15H), 0.95 – 0.88 (m, 6H). ESI-MS m/z calc. 313.2253, found 336.4 (M+23)+; Retention time: 2.0 minutes. LC method I. Example 34: Preparation of tert-butyl (4R)-4-[(1S)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate Step 1: tert-Butyl (4R)-4-[(E,1R)-5-ethoxy-1-isobutyl-5-oxo-pent-3-enyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00362] To a stirred solution of tert-butyl (4R)-2,2-dimethyl-4-[(1R)-3-methyl-1-(2- oxoethyl)butyl]oxazolidine-3-carboxylate (300 mg, 0.8614 mmol) in dry THF (2.4 mL) at 0 °C was added a solution of ethyl 2-diethoxyphosphorylacetate (384.20 mg, 0.34 mL, 1.7137 mmol) in dry THF (0.6 mL) followed by tetrabutylammonium iodide (48 mg, 0.1300 mmol) and potassium carbonate (238 mg, 1.7221 mmol). The mixture was stirred at 0 °C for 15 minutes and then at room temperature for 48 h. Water (5 mL) and EtOAc (5 mL) were added, and the mixture was extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (100 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. Purification by reverse-phase chromatography (Column: C18. Gradient: 5-100% MeCN in water with 0.1% formic acid) afforded tert-butyl (4R)-4-[(E,1R)-5-ethoxy-1-isobutyl-5-oxo-pent-3-enyl]-2,2- dimethyl-oxazolidine-3-carboxylate (24 mg, 7%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 7.05 – 6.88 (m, 1H), 5.83 (d, J = 15.4 Hz, 1H), 4.24 – 4.14 (m, 2H), 3.97 – 3.69 (m, 3H), 2.41 – 2.06 (m, 3H), 1.67 -1.56 (m, 4H), 1.51 – 1.41 (m, 12H), 1.32 – 1.25 (m, 4H), 1.16 – 1.08 (m, 1H), 0.97 – 0.82 (m, 6H). ESI-MS m/z calc.383.2672, found 284.5 (M-99)+; Retention time: 3.57 minutes. LC method 1C. Step 2: tert-Butyl (4R)-4-[(1S)-5-ethoxy-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00363] Palladium (on carbon) (12.5 g, 5 %w/w, 5.8730 mmol) was added to a solution of tert-butyl (4R)-4-[(E,1R)-5-ethoxy-1-isobutyl-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3- carboxylate (25 g, 65.120 mmol) in methanol (0.65 L) and hydrogen gas was bubbled into the suspension for 5 min, then the mixture was stirred under hydrogen atmosphere for 1 h. Celite (15 g) was added to the mixture which was stirred for 5 min, and then filtered on a compressed pad of Celite. The pad was washed with methanol and the solution was filtered again on a syringe filter to remove any remaining solid particles. The filtrate was concentrated under reduced pressure to provide crude tert-butyl (4R)-4-[(1S)-5-ethoxy-1-isobutyl-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate (24.5 g, 93%) as a clear oil.1H NMR (400 MHz, CDCl3) δ 4.22 – 3.75 (m, 5H), 2.41 – 2.21 (m, 2H), 2.17 – 1.91 (m, 1H), 1.81 – 1.58 (m, 6H), 1.53 – 1.42 (m, 12H), 1.41 – 1.23 (m, 5H), 1.20 – 0.98 (m, 2H), 0.96 – 0.75 (m, 6H). ESI-MS m/z calc. 385.2828, found 286.4 (M-99)+; Retention time: 2.21 minutes. LC method I. Step 3: tert-Butyl (4R)-4-[(1S)-5-hydroxy-1-isobutyl-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00364] Lithium aluminum hydride (in THF) (34 mL of 2 M, 68.000 mmol) was added to tert- butyl (4R)-4-[(1S)-5-ethoxy-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (24.5 g, 60.370 mmol) in THF (500 mL) at 0 °C and the reaction was stirred for 1 h. Water (7 mL) was added slowly to quench the excess of LAH, keeping the temperature close to 0 °C. Sodium sulfate (240 g) and Celite (70 g) where added to the mixture. The mixture was stirred for 30 min at room temperature then filtered, and the cake was washed with DCM (500 mL). The combined filtrates were concentrated under reduced pressure and the resulting residue was purified by reverse phase chromatography on a C18 column using a 5%, 50% (to elute the desired product) then 100% acetonitrile in pure water to provide tert-butyl (4R)-4-[(1S)-5- hydroxy-1-isobutyl-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (20.2 g, 97%) as a clear yellow oil.1H NMR (400 MHz, CDCl3) δ 4.05 – 3.78 (m, 3H), 3.70 – 3.59 (m, 2H), 2.14 – 1.94 (m, 1H), 1.69 – 1.51 (m, 8H), 1.49 – 1.44 (m, 12H), 1.42 – 1.20 (m, 3H), 1.18 – 0.99 (m, 2H), 0.95 – 0.79 (m, 6H). ESI-MS m/z calc.343.2723, found 366.4 (M+23)+; Retention time: 3.75 minutes. LC method Z. Step 4: tert-Butyl (4R)-4-[(1S)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00365] To a stirring mixture of tert-butyl (4R)-4-[(1S)-5-hydroxy-1-isobutyl-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate (1.1 g, 3.1991 mmol) and TEMPO (25 mg, 0.1600 mmol) in DCM (30 mL) at 0 °C was added a solution of sodium bromide (10 mg, 0.0972 mmol) in water (10 mL). This suspension was stirred at that temperature for 15 min then a cold solution (0 °C) prepared in another vial by mixing sodium hypochlorite (in water) (3.2 mL of 1.1 M, 3.5200 mmol) with sodium bicarbonate (376 mg, 4.4758 mmol) was added dropwise over 15 min, while avoiding to take the excess of solid sodium bicarbonate at the bottom of the vial. The mixture was stirred at 0 °C for 60 min, then it was diluted with distilled water (20 mL). The aqueous phase was separated, then extracted with DCM (2 x 20 mL). The combined organics were dried over sodium sulfate, filtered and concentrated under reduced pressure to provide crude tert-butyl (4R)-4-[(1S)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.05 g, 89%) as a clear oil, 1H NMR (400 MHz, CDCl3) δ 9.77 (br. S, 1H), 4.05 – 3.75 (m, 3H), 2.56 – 2.35 (m, 2H), 2.16 – 1.94 (m, 1H), 1.80 – 1.55 (m, 6H), 1.52 – 1.44 (m, 12H), 1.43 – 1.13 (m, 3H), 1.11 – 1.02 (m, 1H), 0.95 – 0.79 (m, 6H). ESI-MS m/z calc.341.2566, found 364.5 (M+23)+; Retention time: 2.91 minutes. LC method 1C. Example 35: Preparation of racemic-tert-butyl (4R)-4-[(1S)-1-isobutyl-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate Step 1: Racemic-tert-butyl (4R)-4-[(E,1R)-5-ethoxy-1-isobutyl-5-oxo-pent-3- enyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00366] Ethyl 2-diethoxyphosphorylacetate (29 g, 129.35 mmol) was added in portions at room temperature to a suspension of heptane-washed sodium hydride (in oil) (5.1 g, 60 %w/w, 127.51 mmol) in THF (200 mL). Once all the hydrogen had been liberated, that mixture was transferred to a solution of racemic-tert-butyl (4R)-2,2-dimethyl-4-[(1R)-3-methyl-1-(2- oxoethyl)butyl]oxazolidine-3-carboxylate (21.8 g, 43.818 mmol) in THF (125 mL) stirred at room temperature with a mechanical stirrer. The internal temperature was controlled by the submersion of the reaction flask in a water bath maintained at room temperature. The reaction mixture was stirred at this temperature for 30 min. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (500 mL) and extracted with ethyl acetate containing 20% heptane (200 mL) then with DCM (100 mL x 2). The combined organics were dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by reverse phase chromatography on a C18 column using a 5 to 100% gradient of acetonitrile in pure water to provide racemic-tert-tert-butyl (4R)-4-[(E,1R)-5-ethoxy-1-isobutyl- 5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (13.3 g, 79%) as a clear oil. ESI-MS m/z calc.383.2672, found 284.4 (M-99)+; Retention time: 2.2 minutes, and a fraction of lesser purity Racemic-tert-butyl (4R)-4-[(E,1R)-5-ethoxy-1-isobutyl-5-oxo-pent-3-enyl]-2,2-dimethyl- oxazolidine-3-carboxylate (910 mg, 5%) as a clear oil. ESI-MS m/z calc.383.2672, found 284.4 (M-99)+; Retention time: 2.2 minutes. LC method I. Step 2: Racemic-tert-butyl (4R)-4-[(1S)-5-ethoxy-1-isobutyl-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00367] Palladium (on carbon) (12.5 g, 5 %w/w, 5.8730 mmol) was added to a solution of racemic-tert-butyl (4R)-4-[(E,1R)-5-ethoxy-1-isobutyl-5-oxo-pent-3-enyl]-2,2-dimethyl- oxazolidine-3-carboxylate (25 g, 65.120 mmol) in methanol (0.65 L) and hydrogen gas was injected into the suspension for 5 min then left stirring under hydrogen atmosphere for 1 h. The reaction was complete by LCMS. Celite (15 g) was added to the mixture and the reaction was stirred for 5 min then filtered on a pad of Celite. The pad was washed with methanol and the solution was filtered again on a syringe filter to remove any remaining solid particles. The filtrate was concentrated under reduced pressure to provide crude racemic-tert-butyl (4R)-4- [(1S)-5-ethoxy-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (24.5 g, 93%) as a clear oil.1H NMR (400 MHz, CDCl3) δ 4.22 – 3.75 (m, 5H), 2.41 – 2.21 (m, 2H), 2.17 – 1.91 (m, 1H), 1.81 – 1.58 (m, 6H), 1.53 – 1.42 (m, 12H), 1.41 – 1.23 (m, 5H), 1.20 – 0.98 (m, 2H), 0.96 – 0.75 (m, 6H). ESI-MS m/z calc.385.2828, found 286.4 (M-99)+; Retention time: 2.21 minutes. LC method I. Step 3: Racemic-tert-butyl (4R)-4-[(1S)-5-hydroxy-1-isobutyl-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00368] Lithium aluminum hydride (in THF) (34 mL of 2 M, 68.000 mmol) was added to racemic-tert-butyl (4R)-4-[(1S)-5-ethoxy-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (24.5 g, 60.370 mmol) in THF (500 mL) at 0 °C and the reaction was stirred for 1 h. Water (7 mL) was added slowly to quench the excess of LAH, keeping the temperature close to 0 °C. Then sodium sulfate (240 g) and Celite (70 g) were added to the mixture. The mixture was stirred for 30 min at room temperature then filtered, and the cake was washed with DCM (500 mL). The combined filtrates were concentrated under reduced pressure and the resulting residue was purified by reverse phase chromatography on a C18 column using a 5%, 50% (to elute the desired product) then 100% acetonitrile in pure water to provide racemic-tert-butyl (4R)-4-[(1S)- 5-hydroxy-1-isobutyl-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (20.2 g, 97%) as a clear yellow oil.1H NMR (400 MHz, CDCl3) δ 4.05 – 3.78 (m, 3H), 3.70 – 3.59 (m, 2H), 2.14 – 1.94 (m, 1H), 1.69 – 1.51 (m, 8H), 1.49 – 1.44 (m, 12H), 1.42 – 1.20 (m, 3H), 1.18 – 0.99 (m, 2H), 0.95 – 0.79 (m, 6H). ESI-MS m/z calc.343.2723, found 366.4 (M+23)+; Retention time: 3.75 minutes. LC method Z. Step 4: Racemic-tert-butyl (4R)-4-[(1S)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00369] A stirred solution of oxalyl chloride (2.3280 g, 1.6 mL, 18.341 mmol) in anhydrous DCM (32 mL) was cooled down to -78 °C and a solution of dimethylsulfoxide (2.8626 g, 2.6 mL, 36.637 mmol) in anhydrous DCM (32 mL) was added dropwise. The reaction was stirred for 30 minutes at -78 °C and then a solution of racemic-tert-butyl (4R)-4-[(1S)-5-hydroxy-1- isobutyl-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (2.00 g, 5.8166 mmol) in anhydrous DCM (32 mL) was added. The reaction was stirred for 30 minutes at -78 °C and then TEA (6.0984 g, 8.4 mL, 60.267 mmol) was added. The reaction was stirred for 30 minutes then allowed to reach room temperature and stirred for 30 minutes at room temperature. The reaction was quenched by the addition of a saturated aqueous solution of ammonium chloride (30 mL). The mixture was vigorously stirred at room temperature for 30 minutes, then the layers were separated, and the aqueous layer was extracted with dichloromethane (2 x 75 mL). The combined organic layers were washed with brine (2 x 70 mL), dried over magnesium sulfate, filtered and concentrated under reduced pressure to afford crude racemic-tert-butyl (4R)-4-[(1S)- 1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (2.3 g, 96%) as a brownish oil that was directly used in the next step without further purification; 1H NMR (400 MHz, Chloroform-d) δ 9.77 (br s, 1H), 4.05 – 3.74 (m, 3H), 2.73 – 2.28 (m, 3H), 2.12 – 1.00 (overlapped with water, m, 22H), 0.96 – 0.81 (m, 6H). Example 36: Preparation of tert-butyl (4R)-4-[(1R)-3,3-dimethyl-1-(2-oxoethyl)butyl]-2,2- dimethyl-oxazolidine-3-carboxylate Step 1: tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-(2,2- dimethylpropyl)-5-oxo-pyrrolidine-1-carboxylate [00370] A flask containing copper iodide (3.18 g, 16.697 mmol) was flame-dried until bright yellow under high vacuum then cooled down to room temperature using a nitrogen stream and charged with anhydrous THF (53 mL). The heterogeneous mixture was cooled down to -78 °C and a solution of 2,2-dimethylpropylmagnesium chloride (in THF) (33 mL of 1 M, 33.000 mmol) was added dropwise over 15 min. The reaction was stirred at -78 °C for 30 min after which time a solution of tert-butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-2H- pyrrole-1-carboxylate (4.42 g, 8.3188 mmol) in anhydrous THF (53 mL) was added dropwise over 15 min. The reaction was stirred for 1 hour at -78 °C then allowed to reach room temperature overnight. The reaction was then quenched by the addition of a saturated aqueous solution of ammonium chloride (100 mL) and the mixture was vigorously stirred at room temperature for 15 minutes. The biphasic mixture was filtered on a pad of Celite while washing with EtOAc (150 mL). The layers were separated, and the aqueous layer was extracted with ethyl acetate (5 x 50 mL) and the combined organic layers were washed with water (2 x 50 mL) and brine (3 x 50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Purification by silica gel chromatography (Gradient: 0-20% EtOAc in heptanes) of the crude mixture afforded tert-butyl (2R,3R)-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-3-(2,2-dimethylpropyl)-5-oxo-pyrrolidine-1-carboxylate (2.8 g, 55%) as a yellow oil. ESI-MS m/z calc.523.3118, found 424.2 (M-99)+; Retention time: 2.3 minutes. LC method Q. Step 2: tert-Butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-(2,2- dimethylpropyl)-5-hydroxy-pyrrolidine-1-carboxylate [00371] A stirred solution of tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- (2,2-dimethylpropyl)-5-oxo-pyrrolidine-1-carboxylate (1.31 g, 2.4460 mmol) in anhydrous THF (16 mL) was cooled down to -78 °C then a solution of lithium triethylborohydride in THF (2.67 mL of 1 M, 2.6700 mmol) was added dropwise over 15 minutes. The reaction was then stirred at -78 °C for 1 hour then quenched by the addition of a saturated aqueous solution of sodium bicarbonate (50 mL) and slowly allowed to reach 0 °C after which time an aqueous solution of hydrogen peroxide (in water) (39.550 mg, 0.1 mL of 35 %w/w, 0.4070 mmol) was added dropwise. The reaction was stirred for 30 minutes at 0 °C then diluted with EtOAc (50 mL) and stirred for another 30 minutes. The layers were then separated and the organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-(2,2-dimethylpropyl)-5-hydroxy- pyrrolidine-1-carboxylate (1.27 g, 96%) as a pale yellow oil. ESI-MS m/z calc.525.3274, found 408.4 (M-117)+; Retention time: 2.28 minutes. LC method R. Step 3: tert-Butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2- hydroxyethyl)-4,4-dimethyl-pentyl]carbamate [00372] To a solution of tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-(2,2- dimethylpropyl)-5-hydroxy-pyrrolidine-1-carboxylate (1.27 g, 2.3453 mmol) in methanol (40 mL) was added sodium borohydride (3.1 g, 81.940 mmol). The reaction mixture was stirred at room temperature for 1 hour then heated at 50 °C for 1 hour. The reaction was then concentrated under reduced pressure and partitioned between water (100 mL) and dichloromethane (100 mL). The biphasic mixture was vigorously stirred at room temperature for 30 minutes. The layers were separated, and the aqueous layer was extracted with dichloromethane (3 x 100 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford tert-butyl N-[(1R,2R)-1-[[tert- butyl(diphenyl)silyl]oxymethyl]-2-(2-hydroxyethyl)-4,4-dimethyl-pentyl]carbamate (1.20 g, 95%) as a colorless oil that was directly used in the next step without further purification. ESI- MS m/z calc.527.3431, found 428.4 (M-99)+; Retention time: 2.03 minutes. LC method R. Step 4: tert-Butyl N-[(1R,2R)-2-(2-hydroxyethyl)-1-(hydroxymethyl)-4,4- dimethyl-pentyl]carbamate [00373] A stirred solution of tert-butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2- (2-hydroxyethyl)-4,4-dimethyl-pentyl]carbamate (1.2 g, 2.2394 mmol) in anhydrous THF (16 mL) under nitrogen atmosphere was cooled down to 0 °C and a solution of TBAF (in THF) (3.1 mL of 1 M, 3.1000 mmol) was added dropwise. The reaction was stirred at 0 °C for 1 hour then quenched by the addition of a saturated aqueous solution of ammonium chloride (50 mL) and diluted with EtOAc (50 mL). The mixture was stirred for 30 minutes at room temperature, then the layers were separated, and the aqueous layer was extracted with ethyl acetate (3 x 50 mL) and the combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude was purified by reverse phase chromatography (Column: C18. Gradient: 5-100% MeCN in water). The desired fractions were concentrated under reduced pressure to afford tert-butyl N-[(1R,2R)-2-(2-hydroxyethyl)-1-(hydroxymethyl)- 4,4-dimethyl-pentyl]carbamate (514 mg, 75%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 4.88 (br. S, 1H), 3.83 – 3.74 (m, 2H), 3.74 – 3.58 (m, 3H), 1.95 – 1.86 (m, 1H), 1.79 – 1.72 (m, 2H), 1.69 – 1.61 (m, 1H), 1.58 – 1.50 (m, 1H), 1.46 (s, 9H), 1.42 – 1.35 (m, 1H), 1.09 (dd, J = 14.2, 5.1 Hz, 1H), 0.93 (s, 9H). ESI-MS m/z calc.289.2253, found 234.2 (M-55)+; Retention time: 1.64 minutes. LC method I. Step 5: tert-Butyl (4R)-4-[(1R)-1-(2-hydroxyethyl)-3,3-dimethyl-butyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00374] To a stirred solution of tert-butyl N-[(1R,2R)-2-(2-hydroxyethyl)-1-(hydroxymethyl)- 4,4-dimethyl-pentyl]carbamate (514 mg, 1.6872 mmol) in anhydrous DCM (10 mL) were added 2,2-dimethoxypropane (3.4850 g, 4.1 mL, 33.462 mmol) and paratoluenesulfonic acid hydrate (32 mg, 0.1682 mmol). The reaction was stirred at room temperature for 90 minutes then concentrated under reduced pressure and partitioned between ethyl acetate (20 mL) and a saturated aqueous solution of sodium bicarbonate (20 mL). The mixture was vigorously stirred at room temperature for 20 minutes, then the layers were separated, and the aqueous layer was extracted with ethyl acetate (4 x 20 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude was purified by reverse phase chromatography on C18 (gradient: 5 to 100% acetonitrile in water; 12 CV). The desired fractions were concentrated under reduced pressure and the remaining water was co-evaporated with acetonitrile (4 x 5 mL), then dried under high vacuum to afford tert-butyl (4R)-4-[(1R)-1-(2-hydroxyethyl)-3,3-dimethyl-butyl]-2,2-dimethyl- oxazolidine-3-carboxylate (556 mg, 94%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 4.10 – 4.04 (overlapped with EtOAc, m, 1H), 3.99 – 3.92 (m, 1H), 3.87 – 3.75 (m, 2H), 3.74 – 3.59 (m, 2H), 1.94 – 1.74 (m, 2H), 1.69 – 1.55 (overlapped with water, m, 3H), 1.52 – 1.43 (m, 12H), 1.40 – 1.33 (m, 1H), 1.13 – 1.03 (m, 1H), 0.95 – 0.85 (m, 10H) ESI-MS m/z calc.329.2566, found 274.4 (M-55)+; Retention time: 1.96 minutes. LC method I.
Step 6: tert-Butyl (4R)-4-[(1R)-3,3-dimethyl-1-(2-oxoethyl)butyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00375] To a solution of tert-butyl (4R)-4-[(1R)-1-(2-hydroxyethyl)-3,3-dimethyl-butyl]-2,2- dimethyl-oxazolidine-3-carboxylate (449 mg, 1.3614 mmol) in dichloromethane (14 mL) cooled at 0 °C were added water (4.5 mL), sodium bicarbonate (181 mg, 2.1546 mmol), sodium bromide (24 mg, 0.2332 mmol) and TEMPO (32 mg, 0.2048 mmol). To the bi-phasic solution was added sodium hypochlorite (in water) (1.2 mL of 10 %w/v, 1.6120 mmol) slowly dropwise over 3 minutes under vigorous stirring. After being stirred for 4 hours at 0 °C, a saturated aqueous solution of sodium bicarbonate (10 mL) was added. The mixture was diluted with DCM (10 mL) and the phases were separated. The organic layer was washed with a saturated aqueous solution of sodium bicarbonate (2 x 20 mL), a 10% aqueous solution of Na2S2O3 (2 x 20 mL) and brine (20 mL), then dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert-butyl (4R)-4-[(1R)-3,3-dimethyl-1-(2-oxoethyl)butyl]-2,2- dimethyl-oxazolidine-3-carboxylate (425 mg, 91%) as pale orange oil. Mixture of rotamers: 1H NMR (400 MHz, CDCl3) δ 9.80 (br. S., 1H), 4.02 – 3.90 (m, 1.5H), 3.89 – 3.84 (m, 0.5H), 3.83 – 3.70 (m, 1.5H), 2.67 (br. S., 1H), 2.58 – 2.39 (m, 1.5H), 1.70 – 1.65 (overlapped with water, m, 1H), 1.51 – 1.44 (m, 14H), 1.17 – 1.06 (m, 1H), 0.95 – 0.90 (m, 10H). ESI-MS m/z calc. 327.241, found 350.4 (M+23)+; Retention time: 2.06 minutes. LC method I. Example 37: Preparation of tert-butyl (4R)-2,2-dimethyl-4-[(1S)-2-methyl-1-(2- oxoethyl)propyl]oxazolidine-3-carboxylate Step 1: tert-Butyl (2R,3S)-2-[[tert-Butyl(diphenyl)silyl]oxymethyl]-3-isopropenyl- 5-oxo-pyrrolidine-1-carboxylate [00376] A flask containing copper iodide (3.1 g, 16.277 mmol) was flame-dried under vacuum until bright yellow then cooled down to room temperature using a nitrogen stream and charged with anhydrous THF (80 mL). The mixture was cooled down-to -78 °C and a solution of isopropenylmagnesium bromide in THF (67 mL of 0.5 M, 33.500 mmol) was added dropwise over 45 minutes. The reaction was then stirred at -78 °C for 90 minutes after which time a solution of tert-butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-2H-pyrrole-1- carboxylate (5 g, 10.861 mmol) in anhydrous THF (80 mL) was added dropwise over 1 hour. The reaction was stirred at -78 °C then allowed to reach room temperature overnight. The reaction was cooled down to 0 °C and quenched by the dropwise addition of a saturated aqueous solution of ammonium chloride (150 mL) over one hour. The layers were then separated, and the aqueous layer was extracted with ethyl acetate (4 x 100 mL), dried over magnesium sulfate, filtered on a pad of celite and concentrated under reduced pressure. The crude was purified by flash chromatography on silica-gel (gradient: 0 to 10% ethyl acetate in heptanes, 10 CV). The desired fractions were concentrated under reduced pressure to afford tert-butyl (2R,3S)-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-3-isopropenyl-5-oxo-pyrrolidine-1-carboxylate (3.33 g, 62%) as a pale yellow oil.1H NMR (400 MHz, C6D6) δ 7.75 – 7.67 (m, 4H), 7.28 – 7.19 (m, 6H), 4.68 (br. S, 1H), 4.59 – 4.57 (m, 1H), 4.15 – 4.08 (m, 1H), 3.99 (dd, J = 10.3, 4.9 Hz, 1H), 3.70 (dd, J = 10.3, 2.4 Hz, 1H), 2.80 – 2.64 (m, 2H), 2.20 (dd, J = 17.1, 1.7 Hz, 1H), 1.44 (s, 12H), 1.13 (s, 9H). ESI-MS m/z calc.493.2648, found 394.4 (M-99)+; Retention time: 2.26 minutes. LC method L. Step 2: tert-Butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-isopropyl-5- oxo-pyrrolidine-1-carboxylate [00377] Nitrogen was bubbled for 15 minutes in a stirred solution of tert-butyl (2R,3S)-2- [[tert-butyl(diphenyl)silyl]oxymethyl]-3-isopropenyl-5-oxo-pyrrolidine-1-carboxylate (3.33 g, 6.7381 mmol) in anhydrous methanol (200 mL) after which time palladium on carbon (720 mg, 10 %w/w, 0.6766 mmol) was added. Hydrogen was then bubbled in the reaction for 10 minutes and the reaction was stirred overnight at room temperature under hydrogen atmosphere. Nitrogen was then bubbled through the reaction for 30 minutes and the mixture was concentrated under reduced pressure. The crude residue was then filtered on a pad of celite, washed with dichloromethane (2 x 50 mL), concentrated under reduced pressure then dried under high vacuum to afford crude tert-butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]- 3-isopropyl-5-oxo-pyrrolidine-1-carboxylate (3.107 g, 92%) as a pale yellow oil that was directly used in the next step without further purification.1H NMR (400 MHz, C6D6) δ 7.76 – 7.70 (m, 4H), 7.29 – 7.20 (m, 6H), 4.08 – 4.00 (m, 2H), 3.70 – 3.61 (m, 1H), 2.68 (dd, J = 17.9, 9.8 Hz, 1H), 2.05 (dd, J = 17.9, 2.2 Hz, 1H), 1.92 – 1.83 (m, 1H), 1.47 (s, 9H), 1.28 – 1.18 (m, 1H), 1.14 (s, 9H), 0.61 (d, J = 6.6 Hz, 3H), 0.60 (d, J = 6.6 Hz, 3H). ESI-MS m/z calc. 495.2805, found 396.4 (M-99)+; Retention time: 2.02 minutes. LC method T. Step 3: tert-Butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-hydroxy-3- isopropyl-pyrrolidine-1-carboxylate [00378] A stirred solution of crude tert-butyl (2R,3S)-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-3-isopropyl-5-oxo-pyrrolidine-1-carboxylate (3.107 g, 6.1986 mmol) in anhydrous THF (35 mL) was cooled down to -78 °C and a solution of lithium triethylborohydride (i.e. Super-H) in THF (6.9 mL of 1 M, 6.9000 mmol) was added dropwise over 5 minutes. The reaction was stirred at -78 °C for 90 min. The reaction was then quenched by the addition of a saturated aqueous solution of sodium bicarbonate (30 mL) and the mixture was slowly allowed to reach 0 °C after which time an aqueous solution of hydrogen peroxide (971.25 mg, 2.5 mL of 35 %w/w, 9.9938 mmol) was added dropwise. The mixture was then vigorously stirred for 30 minutes, then the layers were separated, and the aqueous layer was extracted with dichloromethane (3 x 10 mL). The combined organic layers were washed with brine (60 mL), dried over magnesium sulfate, filtered and concentrated under reduced pressure to afford a crude diastereomeric mixture of tert-butyl (2R,3S)-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-5-hydroxy-3-isopropyl-pyrrolidine-1-carboxylate (2.98 g, 79%) as a colorless oil. ESI-MS m/z calc.497.2961, found 380.4 (M-117)+; Retention time: 2.11 minutes. LC method U. Step 4: tert-Butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2- hydroxyethyl)-3-methyl-butyl]carbamate [00379] A stirred solution of crude tert-butyl (2R,3S)-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-5-hydroxy-3-isopropyl-pyrrolidine-1-carboxylate (2.98 g, 4.8854 mmol) in methanol (30 mL) was heated to 50 °C and sodium borohydride (1.9 g, 50.221 mmol) was added portion-wise over 45 minutes. The reaction was then stirred at 50 °C for 1 hour. Additional sodium borohydride (1.9 g, 50.221 mmol) was added, and the reaction was stirred at 50 °C for 30 minutes. Additional sodium borohydride (2.9 g, 76.654 mmol) was added and the reaction was stirred at 50 °C for 30 minutes then cooled down to room temperature and concentrated under reduced pressure. Water (200 mL) and dichloromethane (200 mL) were added, and the biphasic mixture was stirred at room temperature for 2 days. The layers were separated and the aqueous layer was extracted with dichloromethane (4 x 100 mL) and the combined organic layers were washed with brine (2 x 250 mL), dried over magnesium sulfate, filtered and concentrated under reduced pressure then dried under high vacuum to afford crude tert-butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2-hydroxyethyl)-3-methyl- butyl]carbamate (2.392 g, 93%) as a colorless oil that was directly used in the next step without further purification.1H NMR (400 MHz, CDCl3 ) δ 7.71 – 7.63 (m, 4H), 7.48 – 7.34 (m, 6H), 3.81 – 3.73 (m, 1H), 3.72 – 3.53 (m, 4H), 1.78 – 1.69 (m, 1H), 1.65 – 1.51 (overlapped with water, m, 3H), 1.44 (s, 9H), 1.07 (s, 9H), 0.91 (d, J = 6.8 Hz, 3H), 0.84 (d, J = 6.8 Hz, 3H). (2H missing, labile H) ESI-MS m/z calc.499.3118, found 500.4 (M+1)+; Retention time: 1.91 minutes. LC method T. Step 5: tert-Butyl N-[(1R,2S)-2-(2-hydroxyethyl)-1-(hydroxymethyl)-3-methyl- butyl]carbamate [00380] A stirred solution of tert-butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2- (2-hydroxyethyl)-3-methyl-butyl]carbamate (2.392 g, 4.5470 mmol) in anhydrous THF (45 mL) was cooled down to 0 °C and a solution of tetrabutylammonium fluoride in THF (6.9 mL of 1 M, 6.9000 mmol) was added dropwise. The reaction was stirred at 0 oC for 2 hours after which time the reaction was quenched by the addition of a saturated aqueous sodium bicarbonate (50 mL) and brine (50 mL). The mixture was vigorously stirred for 30 minutes, then the layers were separated, and the aqueous layer was extracted with ethyl acetate (4 x 30 mL). The combined organic layers were washed with brine (150 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude was purified by reverse-phase chromatography on C18 (gradient: 5 to 100% acetonitrile in water; 12 CV). The desired fractions were concentrated under reduced pressure and the remaining water was co-evaporated with acetonitrile (3 x 10 mL) then dried under high vacuum to afford tert-butyl N-[(1R,2S)-2-(2- hydroxyethyl)-1-(hydroxymethyl)-3-methyl-butyl]carbamate (1.02 g, 86%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 3.85 – 3.75 (m, 2H), 3.71 – 3.57 (m, 3H), 3.15 (br. S, 2H), 1.82 – 1.72 (m, 1H), 1.65 – 1.52 (m, 3H), 1.46 (s, 9H), 1.50 – 1.41 (m, 9H), 0.95 (d, J = 6.6 Hz, 3H), 0.91 (d, J = 6.8 Hz, 3H). (1H missing, labile H). ESI-MS m/z calc.261.194, found 206.2 (M- 55)+; Retention time: 1.51 minutes. LC method I. Step 6: tert-Butyl (4R)-4-[(1S)-1-(2-hydroxyethyl)-2-methyl-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00381] To a stirred solution of tert-butyl N-[(1R,2S)-2-(2-hydroxyethyl)-1-(hydroxymethyl)- 3-methyl-butyl]carbamate (1.02 g, 3.8988 mmol) in anhydrous dichloromethane (15 mL) were added 2,2-dimethoxypropane (8.5000 g, 10 mL, 81.615 mmol) and paratoluenesulfonic acid hydrate (75 mg, 0.3943 mmol). The reaction was stirred at room temperature for 2 hours then concentrated under reduced pressure. The brown oil was dissolved in dichloromethane (30 mL) and quenched by the addition of a saturated aqueous solution of sodium bicarbonate (50 mL). The layers were separated, and the aqueous layer was extracted with dichloromethane (3 x 30 mL). The combined organic layers were washed with brine (50 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude was purified by reverse- phase chromatography on C18 (gradient: 5 to 100% acetonitrile in water, 12 CV). The desired fractions were concentrated under reduced pressure and the remaining water was co-evaporated with acetonitrile, then dried under high vacuum to afford the starting material tert-butyl N- [(1R,2S)-2-(2-hydroxyethyl)-1-(hydroxymethyl)-3-methyl-butyl]carbamate (450 mg, 45%) as a colorless oil and the desired compound tert-butyl (4R)-4-[(1S)-1-(2-hydroxyethyl)-2-methyl- propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (324 mg, 28%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 4.17 – 3.79 (m, 3H), 3.75 – 3.60 (m, 2H), 1.94 – 1.73 (m, 2H), 1.70 – 1.41 (overlapped with water m, 17H), 0.96 (d, J = 7.1 Hz, 3H), 0.89 (d, J = 6.8 Hz, 3H). (1H missing, labile H). ESI-MS m/z calc.301.2253, found 324.4 (M+23)+; Retention time: 1.83 minutes. LC method U. Step 7: tert-Butyl (4R)-2,2-dimethyl-4-[(1S)-2-methyl-1-(2- oxoethyl)propyl]oxazolidine-3-carboxylate [00382] A flask was flame-dried then cooled down to room temperature using a nitrogen stream and charges with oxalyl chloride (494.70 mg, 340 μL, 3.8975 mmol) and anhydrous dichloromethane (9 mL). The solution was cooled down to -78 °C and a solution of dimethylsulfoxide (605.55 mg, 550 μL, 7.7501 mmol) in anhydrous dichloromethane (9 mL) was added dropwise. The mixture was stirred at -78 °C for 45 minutes after which time a solution of tert-butyl (4R)-4-[(1S)-1-(2-hydroxyethyl)-2-methyl-propyl]-2,2-dimethyl- oxazolidine-3-carboxylate (600 mg, 1.9209 mmol) in anhydrous dichloromethane (9 mL) was added dropwise and the reaction was stirred at -78 °C for 45 minutes. Triethylamine (1.1616 g, 1.6 mL, 11.479 mmol) was added dropwise and the mixture was stirred at -78 °C for 30 minutes then allowed to warm to room temperature, and then stirred at room temperature for 30 minutes. The reaction was quenched by the addition of a saturated aqueous solution of ammonium chloride (50 mL) and the reaction was stirred at room temperature for 30 minutes. The layers were separated and the aqueous layer was extracted with dichloromethane (4 x 30 mL) and the combined organic layers were washed with brine (2 x 50 mL), dried over magnesium sulfate, filtered and concentrated under reduced pressure to afford crude tert-butyl (4R)-2,2-dimethyl-4- [(1S)-2-methyl-1-(2-oxoethyl)propyl]oxazolidine-3-carboxylate (643 mg, 100%) as a pale yellow oil that was directly used in the next step without further purification.1H NMR (400 MHz, CDCl3) δ 9.76 (s, 1H), 4.12 – 3.77 (m, 3H), 2.73 – 2.58 (m, 1H), 2.35 – 2.19 (m, 2H), 1.83 – 1.72 (m, 1H), 1.61 – 1.45 (overlapped with water, m, 15H), 0.96 (d, J = 6.8 Hz, 3H), 0.87 (d, J = 6.8 Hz, 3H). Example 38: Preparation of tert-butyl (4R)-4-[(1S)-2,2-dimethyl-1-(2-oxoethyl)propyl]-2,2- dimethyl-oxazolidine-3-carboxylate Step 1: (4S,5R)-4-tert-Butyl-5-[[tert-butyl(diphenyl)silyl]oxymethyl]pyrrolidin-2- one [00383] To a stirred solution of ZnCl2 (in THF solution) (130 mL of 0.5 M, 65.000 mmol) at room temperature was added tert-butylmagnesium chloride (in THF solution) (66 mL of 1 M, 66.000 mmol) dropwise. The suspension was stirred at room temperature for 30 minutes and was then cooled down to -30 °C. A solution of tert-butylmagnesium chloride (in THF solution) (66 mL of 1 M, 66.000 mmol) in dry THF (50 mL) was added dropwise over 15 minutes. The reaction was stirred at -30 °C for 45 minutes and was then quenched by the addition of a saturated solution of ammonium chloride (50 mL) and was allowed to reach room temperature over 30 minutes. Water (250 mL) and EtOAc (250 mL) were added, and the mixture was extracted with EtOAc (3 x 250 mL). The combined organic layers were washed with brine (500 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 75 to 100% of MeCN in water. The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (100 mL) was then added, and the mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (1 x 250 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford (4S,5R)-4-tert- butyl-5-[[tert-butyl(diphenyl)silyl]oxymethyl]pyrrolidin-2-one (2.96 g, 50%) as a white solid. ESI-MS m/z calc.409.2437, found 410.2 (M-99)+; Retention time: 1.88 minutes. LC method Q. Step 2: tert-Butyl (2R,3S)-3-tert-butyl-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5- oxo-pyrrolidine-1-carboxylate [00384] To a stirred solution of (4S,5R)-4-tert-butyl-5-[[tert- butyl(diphenyl)silyl]oxymethyl]pyrrolidin-2-one (2.96 g, 6.6334 mmol) in MeCN (25 mL) at room temperature was added DMAP (433.8 mg, 3.5508 mmol) followed by Boc2O (4.69 g, 21.489 mmol). The mixture was stirred at room temperature for 15 minutes. Water (100 mL) and EtOAc (100 mL) were added, and the mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (3 x 250 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 80 to 100% of MeCN in water to afford after evaporation tert-butyl (2R,3S)-3-tert-butyl-2-[[tert-butyl(diphenyl)silyl]oxymethyl]- 5-oxo-pyrrolidine-1-carboxylate (3.44 g, 100%) as a sticky orange oil.1H NMR (400 MHz, CDCl3) δ 7.68 – 7.58 (m, 4H), 7.48 – 7.35 (m, 6H), 4.09 – 4.02 (m, 1H), 3.90 (dd, J = 10.4, 4.3 Hz, 1H), 3.61 (dd, J = 10.4, 2.3 Hz, 1H), 2.79 (dd, J = 18.5, 10.1 Hz, 1H), 2.37 (dd, J = 18.5, 2.1 Hz, 1H), 2.03 – 1.98 (overlapped with MeCN, m, 1H), 1.46 (s, 9H), 1.06 (s, 9H), 0.87 (s, 9H). ESI-MS m/z calc.509.2961, found 410.4 (M-99)+; Retention time: 2.05 minutes. LC method U. Step 3: tert-Butyl (2R,3S)-3-tert-butyl-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5- hydroxy-pyrrolidine-1-carboxylate [00385] To a stirred solution of tert-butyl (2R,3S)-3-tert-butyl-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-5-oxo-pyrrolidine-1-carboxylate (2.85 g, 5.1996 mmol) in dry THF (40 mL) at -78 °C was added lithium triethylborohydride (in THF solution) (5.5 mL of 1 M, 5.5000 mmol) dropwise over 5 minutes. The reaction was then stirred at -78 °C for 2 hours. More lithium triethylborohydride (in THF solution) (6 mL of 1 M, 6.0000 mmol) was added dropwise over 5 minutes and the mixture was stirred at -78 °C for 2 hours. The mixture was then quenched by the addition of a saturated aqueous solution of sodium bicarbonate (25 mL) and slowly allowed to reach 0 °C after which time an aqueous solution of hydrogen peroxide (in water) (237.30 mg, 0.6 mL of 35 %w/w, 2.4417 mmol) was added dropwise. The reaction was stirred for 30 minutes at 0 °C then diluted with EtOAc (50 mL) and stirred for another 30 minutes. The layers were then separated and the organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford crude 2:1 diastereomer mixture tert-butyl (2R,3S)-3-tert-butyl-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-hydroxy-pyrrolidine-1- carboxylate (2.89 g, 100%) as a colourless oil.1H NMR (400 MHz, CDCl3) δ 7.74 – 7.60 (m, 4H), 7.49 – 7.33 (m, 6H), 6.62 – 6.54 (m, 0.25H), 6.51 – 6.43 (m, 0.25H), 5.57 – 5.34 (m, 0.25H), 5.05 – 4.98 (m, 0.25H), 4.97 – 4.88 (m, 0.25H), 4.09 – 4.04 (m, 0.5H), 3.98 – 3.80 (m, 1H), 3.77 – 3.71 (m, 0.3H), 3.67 – 3.60 (m, 0.7H), 3.60 – 3.52 (m, 0.5H), 2.75 – 2.67 (m, 0.25H), 2.63 – 2.55 (m, 0.25H), 2.38 – 2.23 (m, 0.25H), 1.53 – 1.32 (m, 9H), 1.11 – 1.03 (m, 9H), 0.93 – 0.76 (m, 11H). ESI-MS m/z calc.511.3118, found 394.4 (M-117)+; Retention time: 2.57 minutes and ESI-MS m/z calc.511.3118, found 394.4 (M-117)+; Retention time: 2.72 minutes. LC method S. Step 4: tert-Butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2- hydroxyethyl)-3,3-dimethyl-butyl]carbamate [00386] To a stirred solution of crude tert-butyl (2R,3S)-3-tert-butyl-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-5-hydroxy-pyrrolidine-1-carboxylate (2.89 g, 5.1784 mmol) in methanol (50 mL) at room temperature was added sodium borohydride (2.3 g, 60.794 mmol) portion-wise over 15 minutes. The reaction was stirred at room temperature for 1 hour and then at 75 °C for 1 hour. The mixture was then cooled down to room temperature and more sodium borohydride (2.3 g, 60.794 mmol) was added portion-wise over 15 minutes. The reaction was stirred at 75 °C for 2 hours. The mixture was then cooled down to room temperature and more sodium borohydride (2.15 g, 56.829 mmol) was added portion-wise over 10 minutes. The reaction was stirred at 75 °C for 2 hours and was then cooled down to room temperature and concentrated in vacuo. Water (250 mL) and EtOAc (250 mL) were added, and the mixture was extracted with EtOAc (3 x 250 mL). The combined organic layers were washed with brine (500 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford crude tert-butyl N- [(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2-hydroxyethyl)-3,3-dimethyl- butyl]carbamate (3 g, 84%) as a white sticky solid.1H NMR (400 MHz, CDCl3) δ 7.72 – 7.64 (m, 4H), 7.47 – 7.35 (m, 6H), 4.93 (br. S, 1H), 4.21 – 4.02 (overlapped with EtOAc, m, 1H), 3.75 – 3.61 (m, 1H), 3.60 – 3.52 (m, 2H), 3.51 – 3.40 (m, 1H), 2.25 – 1.88 (m, 2H), 1.67 – 1.57 (m, 1H), 1.55 – 1.47 (m, 1H), 1.47 – 1.38 (m, 9H), 1.09 – 1.04 (m, 9H), 0.97 – 0.86 (m, 9H). ESI-MS m/z calc.513.3274, found 414.4 (M-99)+; Retention time: 2.36 minutes. LC method S. Step 5: tert-Butyl N-[(1R,2S)-2-(2-hydroxyethyl)-1-(hydroxymethyl)-3,3-dimethyl- butyl]carbamate [00387] To a stirred solution of tert-butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]- 2-(2-hydroxyethyl)-3,3-dimethyl-butyl]carbamate (3 g, 4.3501 mmol) in dry THF (60 mL) at 0 °C was added a solution of tetrabutylammonium fluoride in THF (7 mL of 1 M, 7.0000 mmol) dropwise. The reaction was stirred at 0 °C for 2 hours and was then quenched by the addition of a saturated solution of sodium bicarbonate (50 mL) and brine (50 mL). The mixture was vigorously stirred for 10 minutes, then the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (1 x 250 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeCN in water. The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (100 mL) was then added, and the mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (1 x 250 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford tert-butyl N- [(1R,2S)-2-(2-hydroxyethyl)-1-(hydroxymethyl)-3,3-dimethyl-butyl]carbamate (1.23 g, 98%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 5.01 (br. S, 1H), 4.12 – 4.00 (m, 1H), 3.87 (dt, J = 10.0, 4.7 Hz, 1H), 3.69 – 3.52 (m, 3H), 2.42 (br. S, 2H), 1.67 – 1.57 (m, 2H), 1.52 – 1.40 (m, 10H), 0.95 (s, 9H).1H NMR (400 MHz, CDCl3) δ 5.01 (br. S, 1H), 4.12 – 4.00 (m, 1H), 3.87 (dt, J = 10.0, 4.7 Hz, 1H), 3.69 – 3.52 (m, 3H), 2.42 (br. S, 2H), 1.67 – 1.57 (m, 2H), 1.52 – 1.40 (m, 10H), 0.95 (s, 9H). ESI-MS m/z calc.275.2097, found 176.4 (M-99)+; Retention time: 1.59 minutes. LC method I. Step 6: tert-Butyl (4R)-4-[(1S)-1-(2-hydroxyethyl)-2,2-dimethyl-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00388] To a stirred solution of tert-butyl N-[(1R,2S)-2-(2-hydroxyethyl)-1-(hydroxymethyl)- 3,3-dimethyl-butyl]carbamate (1.22 g, 4.2087 mmol) in dry DCM (20 mL) were added 2,2- dimethoxypropane (8.5000 g, 10 mL, 81.615 mmol) and paratoluenesulfonic acid hydrate (83.1 mg, 0.4369 mmol). The reaction was stirred at room temperature for 90 minutes. The reaction was quenched with a saturated aqueous solution of sodium bicarbonate (3mL), dried over sodium sulfate, filtered on a pad of celite, washed with DCM (2 x 25 mL) then concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeCN in water. The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (50 mL) was then added, and the mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (1 x 100 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford tert-butyl (4R)-4-[(1S)-1-(2-hydroxyethyl)-2,2-dimethyl-propyl]-2,2-dimethyl- oxazolidine-3-carboxylate (408.7 mg, 29%) as a pale yellow oil.1H NMR (400 MHz, DMSO- d6) δ 4.45 – 4.25 (m, 1H), 4.07 – 3.94 (m, 1H), 3.90 – 3.70 (m, 1H), 3.64 (d, J = 8.6 Hz, 1H), 3.40 – 3.28 (overlapped with impurity, m, 2H), 1.56 – 1.33 (m, 18H), 0.87 (s, 9H). ESI-MS m/z calc.315.241, found 338.4 (M+23)+; Retention time: 1.88 minutes. LC method I. Step 7: tert-Butyl (4R)-4-[(1S)-2,2-dimethyl-1-(2-oxoethyl)propyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00389] To a stirred solution of tert-butyl (4R)-4-[(1S)-1-(2-hydroxyethyl)-2,2-dimethyl- propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (373.4 mg, 1.1245 mmol) in DCM (11 mL) cooled at 0 °C were added a solution of sodium bicarbonate (160 mg, 1.9046 mmol) and sodium bromide (28.9 mg, 0.2809 mmol) in water (3 mL) followed by TEMPO (35.5 mg, 0.2272 mmol). To the bi-phasic solution was added dropwise sodium hypochlorite (in water solution) (1.25 mL of 1.1 M, 1.3750 mmol) under vigorous stirring. After being stirred for 1 hour at 0 °C, a saturated aqueous solution of sodium bicarbonate (10 mL) was added. The mixture was diluted with DCM (10 mL) and the phases were separated. The organic layer was washed with a saturated aqueous solution of sodium bicarbonate (2 x 10 mL), a 10% aqueous solution of Na2S2O3 (2 x 10 mL) and brine (50 mL), then dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford crude tert-butyl (4R)-4-[(1S)-2,2-dimethyl-1-(2- oxoethyl)propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (371.2 mg, 90%) as an orange oil.1H NMR (400 MHz, CDCl3) δ 9.77 (s, 1H), 4.12 – 4.03 (m, 1H), 3.94 – 3.78 (m, 1H), 3.71 – 3.54 (m, 1H), 1.57 – 1.40 (m, 17H), 0.95 – 0.86 (m, 10H). Example 39: Preparation of tert-butyl (4R)-4-[(1R)-1-(cyclopropylmethyl)-3-oxo-propyl]- 2,2-dimethyl-oxazolidine-3-carboxylate Step 1: tert-Butyl (2R,3R)-3-allyl-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo- pyrrolidine-1-carboxylate [00390] To a solution of ZnCl2 (in Me-THF) (23 mL of 1.9 M, 43.700 mmol) in dry THF (80 mL) at 0 °C was added allyl(chloro)magnesium (22 mL of 2 M, 44.000 mmol) dropwise over 10 minutes. The suspension was stirred at 0 °C for 5 minutes and was then cooled down to -40 °C. A solution of tert-butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-2H-pyrrole-1- carboxylate (4.2 g, 9.2439 mmol) in dry THF (40 mL) was added dropwise over 15 minutes. The solution was stirred at -40 °C for 1 hour and was then quenched by the addition of a saturated solution of ammonium chloride (100 mL) and was stirred 10 minutes at -40 °C. Water (250 mL) and EtOAc (250 mL) were added, and the mixture was extracted with EtOAc (3 x 250 mL). The combined organic layers were washed with brine (500 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 80 to 100% of MeCN in water. The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (100 mL) was then added, and the mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (1 x 250 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford tert-butyl (2R,3R)-3-allyl-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-5-oxo-pyrrolidine-1-carboxylate (3.25 g, 71%) as an orange oil. 1H NMR (400 MHz, CDCl3) δ 7.66 – 7.57 (m, 4H), 7.48 – 7.36 (m, 6H), 5.73 (ddt, J = 17.0, 10.4, 6.9 Hz, 1H), 5.15 – 5.05 (m, 2H), 3.93 – 3.84 (m, 2H), 3.74 – 3.67 (m, 1H), 2.93 (dd, J = 17.7, 8.9 Hz, 1H), 2.44 – 2.36 (m, 1H), 2.28 – 2.12 (m, 3H), 1.44 (s, 9H), 1.05 (s, 9H). ESI-MS m/z calc.493.2648, found 394.2 (M-99)+; Retention time: 2.35 minutes. LC method S. Step 2: tert-Butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- (cyclopropylmethyl)-5-oxo-pyrrolidine-1-carboxylate [00391] A vial was charged with KOH (14.99 g, 267.17 mmol) and water (25 mL) was added. The mixture was stirred for 2 minutes until KOH was dissolved. Diethyl ether (125 mL) was added. The mixture was cooled to 0 °C and 1-methyl-1-nitroso-urea (12.22 g, 106.69 mmol) was added. The mixture was stirred at 0 °C for 30 minutes. Stirring was stopped and the mixture was cooled to -78 °C. Once the aqueous phase was frozen, the yellow ethereal layer was transferred by a plastic pipette to another vial with pellets (about 15.08 g) of KOH. Half of the diazomethane solution in diethyl ether (65 mL) was added to a solution of tert-butyl (2R,3R)-3- allyl-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-pyrrolidine-1-carboxylate (2.65 g, 5.3622 mmol) in THF (125 mL) at 0 °C with gentle nitrogen flow. A solution of Pd(OAc)2 (244.9 mg, 1.0908 mmol) in THF (25 mL) was added dropwise over 20 minutes. The resulting mixture was stirred at 0 °C for 15 minutes and then the remainder of the ethereal diazomethane solution (60 mL) was added slowly. The reaction was allowed to warm to room temperature and stirred at room temperature for 2 hours. The mixture was filtered on celite, washed with EtOAc (250 mL) and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 80 to 100% of MeCN in water. The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (100 mL) was then added, and the mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (1 x 250 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford tert-butyl (2R,3R)-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-3-(cyclopropylmethyl)-5-oxo-pyrrolidine-1-carboxylate (2.71 g, 99%) as an orange oil.1H NMR (400 MHz, CDCl3) δ 7.68 – 7.60 (m, 4H), 7.48 – 7.36 (m, 6H), 4.01 – 3.97 (m, 1H), 3.91 – 3.84 (m, 1H), 3.74 (dd, J = 10.3, 2.4 Hz, 1H), 2.94 (dd, J = 17.7, 8.9 Hz, 1H), 2.51 – 2.39 (m, 1H), 2.27 (dd, J = 17.7, 1.8 Hz, 1H), 1.47 – 1.39 (m, 10H), 1.35 – 1.26 (m, 1H), 1.06 (s, 9H), 0.71 – 0.61 (m, 1H), 0.54 – 0.43 (m, 2H), 0.15 – 0.01 (m, 2H). ESI-MS m/z calc.507.2805, found 408.4 (M-99)+; Retention time: 2.43 minutes. LC method S. Step 3: tert-Butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- (cyclopropylmethyl)-5-hydroxy-pyrrolidine-1-carboxylate [00392] To a stirred solution of tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- (cyclopropylmethyl)-5-oxo-pyrrolidine-1-carboxylate (2.71 g, 5.3001 mmol) in dry THF (40 mL) at -78 °C was added lithium triethylborohydride (in THF solution) (6 mL of 1 M, 6.0000 mmol) dropwise over 15 minutes. The reaction was then stirred at -78 °C for 90 minutes. The mixture was then quenched by the addition of a saturated aqueous solution of sodium bicarbonate (50 mL) and slowly allowed to reach 0 °C after which time an aqueous solution of hydrogen peroxide (in water) (276.85 mg, 0.7 mL of 35 %w/w, 2.8487 mmol) was added dropwise. The reaction was stirred for 30 minutes at 0 °C then diluted with EtOAc (50 mL) and stirred for another 30 minutes. The layers were then separated and the organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford after evaporation crude a 2:1 diastereomer mixture tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- (cyclopropylmethyl)-5-hydroxy-pyrrolidine-1-carboxylate (2.77 g, 98%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 7.71 – 7.61 (m, 4H), 7.47 – 7.35 (m, 6H), 5.59 – 5.33 (m, 1H), 4.08 (s, 0.5H), 3.93 – 2.63 (m, 4H), 2.49 – 2.39 (m, 0.5H), 2.37 – 2.25 (m, 1H), 1.75 (d, J = 13.0 Hz, 1H), 1.64 – 1.46 (m, 5H), 1.36 (s, 6H), 1.10 – 1.02 (m, 9H), 0.79 – 0.67 (m, 1H), 0.52 – 0.38 (m, 2H), 0.19 - -0.03 (m, 2H). ESI-MS m/z calc.509.2961, found 392.2 (M-117)+; Retention time: 2.55 minutes and ESI-MS m/z calc.509.2961, found 392.4 (M-117)+; Retention time: 2.63 minutes. LC method S. Step 4: tert-Butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2- (cyclopropylmethyl)-4-hydroxy-butyl]carbamate [00393] To a stirred solution of tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- (cyclopropylmethyl)-5-hydroxy-pyrrolidine-1-carboxylate (2.77 g, 5.1949 mmol) in methanol (80 mL) at room temperature was added portion-wise NaBH4 (4.6 g, 121.59 mmol). The reaction was stirred at 50 °C for 2 hours and was then cooled down to room temperature. The volatiles were removed in vacuo. Water (100 mL) and EtOAc (100 mL) were added, and the mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (250 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford crude tert-butyl N- [(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(cyclopropylmethyl)-4-hydroxy- butyl]carbamate (2.65 g, 99%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 7.70 – 7.61 (m, 4H), 7.48 – 7.36 (m, 6H), 4.81 (d, J = 8.3 Hz, 1H), 3.86 (dd, J = 9.2, 5.0 Hz, 1H), 3.77 – 3.56 (m, 4H), 2.00 – 1.89 (m, 1H), 1.89 – 1.81 (m, 1H), 1.64 – 1.57 (overlapped with water, m, 2H), 1.44 (s, 9H), 1.31 – 1.22 (m, 1H), 1.19 – 1.10 (m, 1H), 1.07 (s, 9H), 0.75 – 0.64 (m, 1H), 0.49 – 0.38 (m, 2H), 0.07 - -0.07 (m, 2H). ESI-MS m/z calc.511.3118, found 412.4 (M-99)+; Retention time: 2.31 minutes. LC method S. Step 5: tert-Butyl N-[(1R,2R)-2-(cyclopropylmethyl)-4-hydroxy-1- (hydroxymethyl)butyl]carbamate [00394] To a solution of tert-butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2- (cyclopropylmethyl)-4-hydroxy-butyl]carbamate (2.65 g, 5.1419 mmol) in dry THF (80 mL) was added TBAF (in THF) (11 mL of 1 M, 11.000 mmol) at 0 °C. The reaction was stirred at 0 °C for 2.5 hours. Water (100 mL) and EtOAc (100 mL) were added, and the mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (250 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeOH in water. The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (100 mL) was then added, and the mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (1 x 250 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford tert-butyl N- [(1R,2R)-2-(cyclopropylmethyl)-4-hydroxy-1-(hydroxymethyl)butyl]carbamate (1.45 g, 98%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 5.05 (d, J = 7.3 Hz, 1H), 3.91 – 3.73 (m, 2H), 3.72 – 3.59 (m, 3H), 2.76 (br. S, 2H), 2.00 – 1.88 (m, 1H), 1.74 – 1.62 (m, 1H), 1.62 – 1.50 (m, 1H), 1.45 (s, 9H), 1.36 – 1.18 (m, 2H), 0.77 – 0.64 (m, 1H), 0.52 – 0.41 (m, 2H), 0.10 - -0.01 (m, 2H). ESI-MS m/z calc.273.194, found 218.2 (M-55)+; Retention time: 1.6 minutes. LC method I. Step 6: tert-Butyl (4R)-4-[(1R)-1-(cyclopropylmethyl)-3-hydroxy-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00395] To a solution of tert-butyl N-[(1R,2R)-2-(cyclopropylmethyl)-4-hydroxy-1- (hydroxymethyl)butyl]carbamate (7.65 g, 26.585 mmol) in dichloromethane (75 mL) at room temperature was added 2,2-dimethoxypropane (13.600 g, 16 mL, 130.58 mmol) followed by p- toluenesulfonic acid (507.8 mg, 2.6696 mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction was quenched with a saturated aqueous solution of sodium bicarbonate (50 mL) and the mixture was stirred at room temperature for 5 minutes. The layers were separated, and the aqueous layer was extracted with DCM (3 x 50 mL). The combined organic layers were washed with brine (1 x 100 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeCN in water. The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (100 mL) was then added, and the mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (1 x 250 mL), dried over sodium sulfate, filtered, and concentrated in vacuo to afford tert-butyl (4R)-4-[(1R)-1-(cyclopropylmethyl)-3-hydroxy- propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (6.848 g, 78%) as a pale yellow oil.1H NMR (400 MHz, CDCl3) δ 4.16 – 3.67 (m, 5H), 2.66 (br. S, 1H), 2.20 – 2.07 (m, 1H), 1.95 – 1.82 (m, 1H), 1.64 – 1.53 (overlapped with water, m, 3H), 1.52 – 1.43 (m, 12H), 1.36 – 1.14 (m, 3H), 0.77 – 0.63 (m, 1H), 0.57 – 0.46 (m, 1H), 0.46 – 0.34 (m, 1H), 0.12 – 0.03 (m, 1H), 0.03-0.11 (m, 1H). ESI-MS m/z calc.313.2253, found 336.0 (M+23)+; Retention time: 1.93 minutes. LC method X. Step 7: tert-Butyl (4R)-4-[(1R)-1-(cyclopropylmethyl)-3-oxo-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00396] To a stirred solution of tert-butyl (4R)-4-[(1R)-1-(cyclopropylmethyl)-3-hydroxy- propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (6.4 g, 19.398 mmol) in DCM (200 mL) cooled at 0 °C were added a solution of sodium bicarbonate (2.8 g, 33.331 mmol) and sodium bromide (400 mg, 3.8875 mmol) in water (60 mL) followed by TEMPO (606 mg, 3.8785 mmol). To the bi-phasic solution was added sodium hypochlorite (in water solution) (21 mL of 1.1 M, 23.100 mmol) slowly dropwise under vigorous stirring. After being stirred for 1 hour at 0 °C, a saturated aqueous solution of sodium bicarbonate (50 mL) was added. The mixture was diluted with DCM (100 mL) and the phases were separated. The organic layer was washed with a saturated aqueous solution of sodium bicarbonate (2 x 100 mL), a 10% aqueous solution of Na2S2O3 (2 x 100 mL) and brine (100 mL), then dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford crude tert-butyl (4R)-4-[(1R)-1-(cyclopropylmethyl)-3-oxo- propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (6.35 g, 100%) as an orange oil.1H NMR (400 MHz, CDCl3) δ 9.82 (s, 1H), 4.06 – 3.76 (m, 3H), 2.73 – 2.26 (m, 3H), 1.63 – 1.39 (overlapped with water, m, 16H), 1.25 – 1.11 (m, 1H), 0.76 – 0.57 (m, 1H), 0.56 – 0.42 (m, 2H), 0.15 - -0.06 (m, 2H). ESI-MS m/z calc.311.2097, found 334.2 (M+23)+; Retention time: 1.95 minutes. LC method I. VIII. Synthesis of substituted alkoxy aldehyde intermediates Example 40: Preparation of tert-butyl (4S)-4-(1-isopropoxy-5-oxo-pentyl)-2,2-dimethyl- oxazolidine-3-carboxylate (~85:15 mixture of diastereomers, the major isomer having the 4S,1R configuration) Step 1: Bromo(but-3-enyl)magnesium [00397] To a suspension of activated magnesium turnings (7.2 g, 296.24 mmol) in THF (148 mL) was added dropwise at 0 °C 4-bromobut-1-ene (20 g, 148.15 mmol). The solution was then refluxed for 2 h. The reaction mixture was then allowed to cool to room temperature. The resulting solution of bromo(but-3-enyl)magnesium (23.6 g, 100%) was used without further purification. Step 2: tert-Butyl (4S)-4-(1-hydroxypent-4-enyl)-2,2-dimethyl-oxazolidine-3- carboxylate [00398] To a solution of tert-butyl (4S)-4-formyl-2,2-dimethyl-oxazolidine-3-carboxylate (15 g, 65.424 mmol) in THF (120 mL) was slowly added at -78 °C bromo(but-3-enyl)magnesium in THF (65 mL of 1 M, 65.000 mmol). The solution was stirred at this temperature for 2 hours, then the reaction mixture was allowed to warm up to room temperature and stirred for 16 h. The reaction was quenched using a saturated aqueous solution of ammonium chloride (150 mL). The product was extracted three times using ethyl acetate (75 mL x 3) and combined organic layers were washed using brine (200 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified on silica gel (gradient: 0 to 30 % of EtOAc in heptane) to afford tert-butyl (4S)-4-(1-hydroxypent-4-enyl)-2,2-dimethyl-oxazolidine-3- carboxylate (mixture of diastereomers, 17.4 g, 93%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 5.84 (ddt, J = 17.1, 10.3, 6.6 Hz, 1H), 5.10 – 4.94 (m, 2H), 4.25 – 3.60 (m, 5H), 2.43 – 2.24 (m, 1H), 2.23 – 2.06 (m, 1H), 1.62 – 1.56 (m, 3H), 1.53 – 1.45 (m, 14H). ESI-MS m/z calc. 285.194, found 230.2 (M-55)+; Retention time: 1.84 minutes. LC method I. Step 3: tert-Butyl (4S)-2,2-dimethyl-4-pent-4-enoyl-oxazolidine-3-carboxylate [00399] To a solution of tert-butyl (4S)-4-(1-hydroxypent-4-enyl)-2,2-dimethyl-oxazolidine-3- carboxylate (17.4 g, 60.911 mmol) in dichloromethane (150 mL) at 0 °C was added Dess-Martin periodinane (31 g, 73.089 mmol) and the reaction was stirred for 1 hour at 0 °C and then at room temperature for 1 hour. A mixture of aqueous saturated solution of Na2S2O3 (250 mL), saturated sodium bicarbonate (250 mL) and 1N NaOH (150 mL) (To reach pH = 9) was added and the reaction mixture was stirred for 45 minutes. The phases were separated and the aqueous one was extracted with DCM (2 x 100 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford tert-butyl (4S)-2,2- dimethyl-4-pent-4-enoyl-oxazolidine-3-carboxylate (15 g, 69%) as a crude dark-orange compound.1H NMR (400 MHz, CDCl3) δ 5.93 – 5.73 (m, 1H), 5.09 – 4.93 (m, 2H), 4.44 (dd, J = 7.3, 2.4 Hz, 0.4H), 4.31 (dd, J = 7.7, 2.6 Hz, 0.6H), 4.19 – 4.07 (m, 1H), 3.92 (ddd, J = 16.7, 9.4, 2.6 Hz, 1H), 2.69 – 2.52 (m, 2H), 2.34 (q, J = 7.0 Hz, 2H), 1.71 (s, 1.8H), 1.65 (s, 1.2H), 1.52 – 1.46 (m, 6.6H, overlapped with water peak), 1.41 (s, 5.4H). ESI-MS m/z calc.283.1784, found 184.4 (M-99)+; Retention time: 1.9 minutes. LC method I. Step 4: tert-Butyl (4S)-4-(1-hydroxypent-4-enyl)-2,2-dimethyl-oxazolidine-3- carboxylate ~85:15 mixture of diastereomers, the major isomer having the 4S,1R configuration [00400] Zinc chloride (5.55 g, 40.723 mmol) was placed in a 500 mL flask and the flask was flame dried 3 times and placed on a high vacuum for 3 h. Diethyl ether (100 mL) was added to the flask and the suspension was brought to reflux for 2 h. The supernatant was added to a solution of sodium borohydride (3.07 g, 81.147 mmol) in diethyl ether (250 mL) and stirred at rt for 3 d. (solution A). To a solution of tert-butyl (4S)-2,2-dimethyl-4-pent-4-enoyl-oxazolidine-3- carboxylate (12.2 g, 31.559 mmol) in diethyl ether (250 mL) was added solution A (200 mL, 0.116 M, 0.68 eq.) at -20 °C dropwise over 40 min. The resulting solution was stirred for 45 min before an aqueous solution of ammonium chloride (sat., 200 mL) was added. The resulting mixture was stirred for 1 h slowly being allowed to reach rt. The phases were separated and the aqueous one was extracted with EtOAc (3 x 100 mL). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and was then purified by flash-chromatography on a silica gel cartridge, eluting with a gradient of 0 to 30% of EtOAc in heptanes to afford after evaporation tert-butyl (4S)-4-(1-hydroxypent-4- enyl)-2,2-dimethyl-oxazolidine-3-carboxylate (6.32 g, 49%) as a colorless oil (~85:15 mixture of diastereomers, the major isomer having the 4S,1R configuration).1H NMR (400 MHz, CDCl3) δ 5.92 – 5.79 (m, 1H), 5.11 – 4.95 (m, 2H), 4.08 – 3.63 (m, 4H), 2.45 – 2.27 (m, 1H), 2.23 – 2.11 (m, 1H), 2.05, 1.64 – 1.55 (m, 3H), 1.50 (s, 15H) ESI-MS m/z calc.285.194, found 230.2 (M-55)+; Retention time: 1.84 minutes. LC method I. Step 5: tert-Butyl (4S)-4-(1-isopropoxypent-4-enyl)-2,2-dimethyl-oxazolidine-3- carboxylate [00401] To a solution of tert-butyl (4S)-4-(1-hydroxypent-4-enyl)-2,2-dimethyl-oxazolidine-3- carboxylate (19.2 g, 53.218 mmol) in 2-iodopropane (365.50 g, 215 mL, 2.1501 mol) was added silver (I) oxide (68 g, 293.44 mmol) at room temperature. The solution was stirred at 45 °C for 24 h. The reaction mixture was filtered through a pad of Celite and washed with EtOAc (3 x 25 mL). The crude mixture was put on a silica pad (50 g) and was washed with a mixture of heptanes/EtOAc (2 x 475 mL / 25 mL) to afford, after evaporation, tert-butyl (4S)-4-(1- isopropoxypent-4-enyl)-2,2-dimethyl-oxazolidine-3-carboxylate (13.6 g, 66%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 5.92 – 5.74 (m, 1H), 5.12 – 4.90 (m, 2H), 4.25 – 3.85 (m, 3H), 3.84 – 3.63 (m, 2H), 2.36 – 1.99 (m, 2H), 1.77 – 1.39 (m, 17H), 1.21 – 1.06 (m, 6H). ESI-MS m/z calc.327.241, found 228.4 (M-99)+; Retention time: 2.17 minutes. LC method I. Step 6: tert-Butyl (4S)-4-(5-hydroxy-1-isopropoxy-pentyl)-2,2-dimethyl- oxazolidine-3-carboxylate ~85:15 mixture of diastereomers, the major isomer having the 4S,1R configuration [00402] To a 0 °C solution of tert-butyl (4S)-4-(1-isopropoxypent-4-enyl)-2,2-dimethyl- oxazolidine-3-carboxylate (13.6 g, 35.344 mmol) in tetrahydrofuran (150 mL) was added borane tetrahydrofuran complex (141 mL of 1 M, 141.00 mmol) and the solution was stirred for 1 h at that temperature. Sodium hydroxide (solution in water) (150 mL of 1 M, 150.00 mmol) was slowly introduced followed by hydrogen peroxide (solution in water) (4.6620 g, 12 mL of 35 %w/w, 47.970 mmol). The solution was allowed to warm up to rt over 30 min at which point water (50 mL) was added. The aqueous phase was extracted with EtOAc (3 x 50 mL). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and was then purified in two batches by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeOH in water. The combined purified product was evaporated to afford tert-butyl (4S)-4-(5-hydroxy-1-isopropoxy- pentyl)-2,2-dimethyl-oxazolidine-3-carboxylate (10.5 g, 82%) as a yellow oil (~85:15 mixture of diastereomers, the major isomer having the 4S,1R configuration).1H NMR (400 MHz, CDCl3) δ 4.17 – 4.06 (m, 1H), 4.01 – 3.56 (m, 6H), 1.74 – 1.37 (m, 22H), 1.19 – 1.11 (m, 6H). ESI-MS m/z calc.345.2515, found 246.3 (M-99)+; Retention time: 4.48 minutes. LC method J. Step 7: tert-Butyl (4S)-4-(1-isopropoxy-5-oxo-pentyl)-2,2-dimethyl-oxazolidine-3- carboxylate mixture of diastereomers, the major isomer having the 4S,1R configuration [00403] To a 0 °C solution of tert-butyl (4S)-4-(5-hydroxy-1-isopropoxy-pentyl)-2,2- dimethyl-oxazolidine-3-carboxylate (2.75 g, 7.8805 mmol) in dichloromethane (55 mL) was added a solution of sodium bromide (230 mg, 2.2353 mmol) in water (25 mL) followed by sodium bicarbonate (1.2 g, 14.285 mmol) and TEMPO (66 mg, 0.4224 mmol). The mixture was stirred for 5 min and sodium hypochlorite (solution in water) (5.8 mL of 10 %w/v, 7.7913 mmol) was added dropwise and the reaction was stirred at 0 °C for 1.5 h. DCM (30 mL) was added and the organic phase was washed with an aqueous solution of sodium bicarbonate (sat., 2 x 30 mL), an aqueous solution of Na2S2O3 (10%, 2 x 30 mL) and brine (30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered. The crude mixture was concentrated in vacuo to afford crude tert-butyl (4S)-4-(1-isopropoxy-5-oxo-pentyl)-2,2-dimethyl-oxazolidine- 3-carboxylate (2.44 g, 84%) as an orange oil (~85:15 mixture of diastereomers, the major isomer having the 4S,1R configuration). ESI-MS m/z calc.343.2359, found 244.4 (M-99)+; Retention time: 1.95 minutes. LC method I. Example 41: Preparation of tert-butyl (4S)-2,2-dimethyl-4-[(1R)-5-oxo-1-[1,2,2,2- tetradeuterio-1-(trideuteriomethyl)ethoxy]pentyl]oxazolidine-3-carboxylate Step 1: tert-Butyl (4S)-4-hex-5-enoyl-2,2-dimethyl-oxazolidine-3-carboxylate [00404] To a stirring solution of 5-bromopent-1-ene (8.580 g, 57.57 mmol) in THF (67.78 mL) at room temperature was added magnesium turnings (1.515 g, 57.56 mmol) and the resulting mixture was stirred for 5 minutes, then sonicated for 2 minutes and stirred for 2 hours at room temperature. The resulting brown mixture with some residual magnesium was then slowly added to a -78 oC solution of tert-butyl (4S)-4-formyl-2,2-dimethyl-oxazolidine-3- carboxylate (12 g, 52.34 mmol) in THF (45.18 mL) leaving the solid material behind. The solution was stirred at this temperature for 5 min then allowed to warm to room temperature and stirred for 45 minutes. The reaction was quenched using a saturated aqueous solution of NH4Cl (300 mL). The product was extracted with ethyl acetate (400 mL) and the organic fraction was washed with brine (200 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to give crude tert-butyl (4S)-4-(1-hydroxyhex-5-enyl)-2,2-dimethyl-oxazolidine-3- carboxylate. To a 0 °C solution of this crude compound in DCM (158.8 mL) was added Dess- martin periodinane (26.64 g, 62.81 mmol). The cold bath was removed, and the mixture was stirred overnight. A mixture of an aqueous saturated solution of Na2S2O3 (75 mL), a saturated solution of NaHCO3 (75 mL) and a 1N NaOH solution (37.5 mL, to reach pH = 9) was added and the reaction mixture was stirred for 20 minutes. The phases were separated, and the aqueous phase was extracted with DCM (3 x 100 mL). The combined organic phases were dried over anhydrous magnesium sulfate and filtered. The crude product was purified by silica gel chromatography using a shallow gradient from 100% hexanes to 100% EtOAc providing as a clear oil, tert-butyl (4S)-4-hex-5-enoyl-2,2-dimethyl-oxazolidine-3-carboxylate (8.9 g, 57%).1H NMR (400 MHz, CDCl3) δ 5.86 – 5.71 (m, 1H), 5.09 – 4.95 (m, 2H), 4.48 – 4.28 (m, 1H), 4.22 – 4.09 (m, 1H), 4.00 – 3.85 (m, 1H), 2.60 – 2.46 (m, 2H), 2.14 – 2.06 (m, 2H), 1.74 – 1.72 (m, 2H), 1.56 – 1.49 (m, 9H), 1.43 (s, 6H). Step 2: (4S)-4-[(1R)-1-Hydroxyhex-5-enyl]-2,2-dimethyl-oxazolidine-3- carboxylate [00405] To a solution of tert-butyl (4S)-4-hex-5-enoyl-2,2-dimethyl-oxazolidine-3-carboxylate (14.73 g, 49.53 mmol) in THF (147.3 mL) was added magnesium chloride (4.716 g, 49.53 mmol). The resulting white suspension was stirred at room temperature for 10 min. Then borane (t-butylamine salt) (5.169 g, 59.44 mmol) was added in one portion. The reaction mixture was stirred at room temperature for 1 h. The reaction was quenched at room temperature with aqueous saturated NH4Cl (100 mL), the organics were then diluted with EtOAc (100 mL) and washed once with a 100 mL half brine solution. The organics were dried filtered and evaporated then purified using silica gel chromatography with a shallow gradient from 100% hexanes to 100% EtOAc to afford tert-butyl (4S)-4-[(1R)-1-hydroxyhex-5-enyl]-2,2-dimethyl-oxazolidine- 3-carboxylate (14.83 g, 100%) as a clear oil.1H NMR (400 MHz, CDCl3) δ 5.88 – 5.76 (m, 1H), 5.07 – 4.99 (m, 1H), 4.99 – 4.91 (m, 1H), 4.18 – 3.59 (m, 5H), 2.19 – 2.07 (m, 2H), 1.65 – 1.57 (m, 4H), 1.55 – 1.46 (m, 15H). Step 3: tert-butyl (4S)-2,2-dimethyl-4-[(1R)-1-[1,2,2,2-tetradeuterio-1- (trideuteriomethyl)ethoxy]hex-5-enyl]oxazolidine-3-carboxylate [00406] To a solution of tert-butyl (4S)-4-[(1R)-1-hydroxyhex-5-enyl]-2,2-dimethyl- oxazolidine-3-carboxylate (700 mg, 2.338 mmol) in 1,1,1,2,3,3,3-heptadeuterio-2-iodo-propane (2.8 mL, 28.06 mmol) was added silver(I) oxide (2.709 g, 11.69 mmol). The solution was stirred at 45 °C for 16 h. The mixture was then cooled to room temperature and filtered through a pad of Celite and washed with EtOAc. The filtrate was concentrated then purified by silica gel chromatography using a shallow gradient from 100% hexanes to 100% EtOAc giving as a clear oil, tert-butyl (4S)-2,2-dimethyl-4-[(1R)-1-[1,2,2,2-tetradeuterio-1- (trideuteriomethyl)ethoxy]hex-5-enyl]oxazolidine-3-carboxylate (215.6 mg, 26%) 1H NMR (400 MHz, CDCl3) δ 5.87 – 5.73 (m, 1H), 5.00 (d, J = 16.9 Hz, 1H), 4.94 (d, J = 10.5 Hz, 1H), 4.12 – 3.79 (m, 3H), 3.79 – 3.61 (m, 1H), 2.11 – 2.03 (m, 2H), 1.70 – 1.58 (m, 2H), 1.58 – 1.51 (m, 3H), 1.48 (s, 9H), 1.47 – 1.42 (m, 3H), 1.41 – 1.30 (m, 2H). ESI-MS m/z calc.348.30054, found 349.2 (M+1)+; Retention time: 0.892 minutes. LC method B. Step 4: tert-Butyl (4S)-2,2-dimethyl-4-[(1R)-5-oxo-1-[1,2,2,2-tetradeuterio-1- (trideuteriomethyl)ethoxy]pentyl]oxazolidine-3-carboxylate [00407] To a stirring suspension of the tert-butyl (4S)-2,2-dimethyl-4-[(1R)-1-[1,2,2,2- tetradeuterio-1-(trideuteriomethyl)ethoxy]hex-5-enyl]oxazolidine-3-carboxylate (211.1 mg, 0.6057 mmol) in the acetone (1.382 mL) and water (460.4 µL) at rt under nitrogen atmosphere was added 4-methylmorpholine N-oxide (85.14 mg, 0.7268 mmol) followed by osmium tetroxide (2.5 %wt solution in 2-methyl-2-propanol) (123.3 mg, 0.01212 mmol). The resulting suspension was stirred at rt for 30 minutes. Acetone was carefully removed by evaporation. The reaction was diluted with EtOAc, washed with water (1 x) and brine (1 x), dried (magnesium sulfate), filtered and concentrated to a thick syrup which was used directly in the next step. A solution of this crude product and sodium periodate (259.2 mg, 1.212 mmol) in THF (5.066 mL) and water (3.378 mL) was stirred for 30 minutes. The reaction mixture was partitioned between ethyl acetate and water. The organic layer was separated, and the aqueous phase was extracted with ethyl acetate once more. The combined organic layers were dried over sodium sulfate, filtered and concentrated then purified by silica gel chromatography using a shallow gradient from 100% hexanes to 100% EtOAc giving tert-butyl (4S)-2,2-dimethyl-4-[(1R)-5-oxo-1- [1,2,2,2-tetradeuterio-1-(trideuteriomethyl)ethoxy]pentyl]oxazolidine-3-carboxylate (212 mg, 100%) as a clear oil.1H NMR (400 MHz, CDCl3) δ 9.84 – 9.72 (m, 1H), 4.15 – 3.90 (m, 3H), 3.82 – 3.64 (m, 1H), 2.52 – 2.43 (m, 2H), 1.96 – 1.82 (m, 1H), 1.65 – 1.60 (m, 2H), 1.57 (s, 3H), 1.51 (d, J = 3.7 Hz, 9H), 1.49 – 1.44 (m, 3H), 1.43 – 1.36 (m, 1H). ESI-MS m/z calc.350.27982, found 351.5 (M+1)+; Retention time: 0.701 minutes. LC method B. Example 42: Preparation of tert-butyl (4S)-4-[(1R)-1-isopropoxy-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate Step 1: tert-Butyl (4S)-4-[(1R)-1-isopropoxyhex-5-enyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00408] To a solution of tert-butyl (4S)-4-[(1R)-1-hydroxyhex-5-enyl]-2,2-dimethyl- oxazolidine-3-carboxylate (14.83 g, 49.53 mmol) in 2-iodopropane (219.7 mL, 2.197 mol) was added silver (I) oxide (35.25 g, 152.1 mmol). The solution was stirred at 45 °C for 18 h. The mixture was cooled down to room temperature, filtered and to the filtrate was added additional silver (I) oxide (35.25 g, 152.1 mmol) and the resulting mixture was stirred at 45°C overnight. The mixture was then cooled down to room temperature and filtered. The reaction mixture was filtered through a pad of Celite and washed with EtOAc. The filtrate was concentrated then purified by silica gel chromatography using a shallow gradient from 100% hexanes to 100% EtOAc giving as a clear oil, tert-butyl (4S)-4-[(1R)-1-isopropoxyhex-5-enyl]-2,2-dimethyl- oxazolidine-3-carboxylate (5.620 g, 33%).1H NMR (400 MHz, CDCl3) δ 5.88 – 5.75 (m, 1H), 5.02 (d, J = 17.4 Hz, 1H), 4.96 (d, J = 9.5 Hz, 1H), 4.10 (s, 1H), 3.92 (s, 2H), 3.84 – 3.58 (m, 2H), 2.13 – 2.02 (m, 2H), 1.56 (s, 4H), 1.51 (d, J = 7.2 Hz, 15H), 1.16 (d, J = 6.3 Hz, 6H). Some starting material was also isolated to give tert-butyl (4S)-4-[(1R)-1-hydroxyhex-5-enyl]- 2,2-dimethyl-oxazolidine-3-carboxylate (6.64 g, 45%). Step 2: tert-Butyl (4S)-4-[(1R)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00409] To a stirring suspension of tert-butyl (4S)-4-[(1R)-1-isopropoxyhex-5-enyl]-2,2- dimethyl-oxazolidine-3-carboxylate (5.58 g, 16.34 mmol) in acetone (36.52 mL) and water (12.17 mL) at rt under nitrogen atmosphere was added 4-methyl-4-oxido-morpholin-4-ium (2.033 mL, 19.61 mmol) followed by osmium tetroxide (2.5 wt% in 2-methyl-2-propanol) (4.097 mL, 13.07 mmol). The resulting suspension was stirred at rt for 30 minutes. Acetone was carefully removed by evaporation. The reaction was diluted with EtOAc and washed with water (1 x) and brine (1 x), dried over magnesium sulfate, filtered, and concentrated to a thick syrup of crude tert-butyl (4S)-4-[(1R)-5,6-dihydroxy-1-isopropoxy-hexyl]-2,2-dimethyl-oxazolidine-3- carboxylate. A solution of the crude product and sodium periodate (1.809 mL, 32.69 mmol) in THF (133.9 mL) and water (89.28 mL) was stirred for 30 minutes. The reaction mixture was partitioned between ethyl acetate and water. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate once more. The combined organic layers were dried over sodium sulfate, filtered and concentrated, then purified by silica gel chromatography using a shallow gradient from 100% hexanes to 100% EtOAc giving as a clear oil, tert-butyl (4S)-4- [(1R)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (4.61 g, 82%) ESI- MS m/z calc.343.23587, found 344.4 (M+1)+; Retention time: 1.78 minutes. LC method A.1H NMR (400 MHz, CDCl3) δ 9.78 (s, 1H), 4.14 – 4.05 (m, 1H), 3.96 – 3.86 (m, 2H), 3.84 – 3.65 (m, 2H), 2.53 – 2.40 (m, 2H), 1.91 – 1.78 (m, 1H), 1.74 – 1.66 (m, 1H), 1.56 (s, 3H), 1.52 (s, 3H), 1.50 (s, 9H), 1.47 – 1.38 (m, 2H), 1.16 (d, J = 6.1 Hz, 6H). IX. Synthesis of CFTR Correctors Example 43: Preparation of Compound 8 Step 1: tert-Butyl (4S)-4-[(1S)-1-[3-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin- 3-yl)-3-hydroxy-propoxy]-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3- carboxylate [00410] To a solution of 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine (60 mg, 0.2224 mmol) in anhydrous THF (0.8 mL) at -78 °C was added dropwise a solution of n-BuLi (in hexanes) (0.15 mL of 2.5 M, 0.3750 mmol). The reaction mixture was stirred for 15 minutes then a solution of tert-butyl (4S)-2,2-dimethyl-4-[(1S)-3-methyl-1-(3-oxopropoxy)butyl] oxazolidine-3-carboxylate (97 mg, 0.2683 mmol) in anhydrous THF (0.8 mL) was added dropwise. The reaction was stirred in the dry-ice bath for 2 hours and methanol (5 mL) was added. The reaction was concentrated under reduced pressure and was directly purified by reversed-phase chromatography (Column: C18. Gradient: 5-100 % methanol in water). The purest fractions of the desired product were combined and concentrated under reduced pressure. The residue was co-evaporated with acetonitrile (3 x 10 mL) and was dried in vacuo to afford tert-butyl (4S)-4-[(1S)-1-[3-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-hydroxy- propoxy]-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate (76 mg, 64%) as a yellow oil. 1H NMR (400 MHz, CDCl3) δ 8.83 – 8.23 (m, 1H), 6.70 (br. S., 1H), 5.11 (br. S., 1H), 4.32 – 3.53 (m, 9H), 2.32 – 1.97 (m, 2H), 1.89 – 1.16 (m, 28H), 0.99 – 0.87 (overlapped with impurity, m, 6H). ESI-MS m/z calc.532.3625, found 533.4 (M+1)+; Retention time: 2.15 minutes. LC method I. Step 2: tert-Butyl (4S)-4-[(1S)-1-[3-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin- 3-yl)-3-oxo-propoxy]-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00411] To a solution of tert-butyl (4S)-4-[(1S)-1-[3-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-hydroxy-propoxy]-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate (820 mg, 1.5377 mmol) in anhydrous DCE (20 mL) was added MnO2 (activated) (2.98 g, 29.136 mmol). The suspension was heated to 70 °C for 45 minutes then the reaction was allowed to cool down to room temperature. The suspension was filtered through Celite, and the pad was rinsed with DCM (3 x 20 mL). The combined filtrates were concentrated in vacuo to afford 850 mg of the desired product (85 % purity 1H NMR, 89% yield) as a mixture of diastereomers. The mixture was submitted to chiral SFC separation (Flow rate: 100 mL/min, 20% MeOH, column: i-Amylose 3-30mm, temperature = 40 °C, outlet pressure: 100 bar, injection volume: 1000 µL). The fastest eluting peak by SFC (peak 1) afforded, after evaporation to dryness, tert-butyl (4S)- 4-[(1R)-1-[3-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-oxo-propoxy]-3-methyl- butyl]-2,2-dimethyl-oxazolidine-3-carboxylate (50 mg, 6%) as a colorless oil. Rotamers observed in a 1.5:1 ratio:1H NMR (400 MHz, CDCl3) δ 9.10 (br. S., 1H), 6.66 (br. S., 1H), 4.18 – 3.72 (m, 9H), 3.71 – 3.57 (m, 1H), 3.50 (t, J = 6.6 Hz, 0.6H), 3.45 (t, J = 6.5 Hz, 0.4H), 1.82 – 1.31 (m, 26H), 1.21 – 1.07 (m, 1H), 1.02 – 0.82 (m, 6H). ESI-MS m/z calc.530.3468, found 431.4 (M-99)+; Retention time: 2.3 minutes. The slowest eluting peak by SFC (peak 2) afforded, after evaporation to dryness, tert-butyl (4S)-4-[(1S)-1-[3-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-oxo-propoxy]-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate (529 mg, 61%) as a beige solid.1H NMR (400 MHz, CDCl3) δ 9.12 (br. S., 1H), 6.62 (br. S., 1H), 4.39 – 3.75 (m, 9H), 3.69 – 3.36 (m, 2H), 1.84 – 1.31 (m, 26H), 1.30 – 1.11 (m, 1H), 1.00 – 0.75 (m, 6H). ESI-MS m/z calc.530.3468, found 431.4 (M-99)+; Retention time: 2.33 minutes. LC method I. Step 3: 3-[(1S)-1-[(1S)-1-Amino-2-hydroxy-ethyl]-3-methyl-butoxy]-1-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)propan-1-one [00412] To a solution of tert-butyl (4S)-4-[(1S)-1-[3-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-oxo-propoxy]-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate (380 mg, 0.6752 mmol) in Et2O (8 mL) and water (2 mL) at room temperature was added a solution of HCl (in diethyl ether) (5 mL of 2 M, 10.000 mmol). The reaction was stirred for 1 hour and HCl (in diethyl ether) (2 mL of 2 M, 4.00 mmol) was again added. The reaction was stirred for 1 hour and an aqueous saturated sodium bicarbonate solution (16 mL) was added until pH 7. The volatiles were removed under reduced pressure. The aqueous residue was diluted with DMSO (5 mL) and CH3CN (1 mL) and was purified by reversed-phase chromatography (Column: C18. Gradient: 5-100 % CH3CN/water with 0.1% formic acid). The fractions containing the product were combined and co-evaporated with water (3 x 10 mL) to afford 3-[(1S)-1-[(1S)-1-amino-2- hydroxy-ethyl]-3-methyl-butoxy]-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)propan-1- one (274 mg, 104%) as a yellow oil. ESI-MS m/z calc.390.2631, found 391.4 (M+1)+; Retention time: 1.4 minutes. LC method K. Step 4: [(2S,3S,5S)-5-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2- isobutyl-1,4-oxazepan-3-yl]methanol and [(2S,3S,5R)-5-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-2-isobutyl-1,4-oxazepan-3-yl]methanol [00413] To a solution of 3-[(1S)-1-[(1S)-1-amino-2-hydroxy-ethyl]-3-methyl-butoxy]-1-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)propan-1-one (264 mg, 0.6760 mmol) in anhydrous DCM (20 mL) at room temperature was added NaBH(Oac)3 (603 mg, 2.8451 mmol). The reaction was stirred at room temperature for 18 hours and a saturated aqueous solution of sodium bicarbonate (20 mL) was added. The phases were separated, and the aqueous solution was extracted with DCM (3 x 10 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography (Column: C18. Gradient: 5- 100 % acetonitrile in basic water (ammonium bicarbonate/ammonium hydroxide buffer pH = 10)). The purest fractions of the fastest eluting peak were combined, and the volatiles were concentrated under reduced pressure. To the aqueous solution was added brine (50 mL) and the solution was extracted with DCM (3 x 20 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford [(2S,3S,5S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2-isobutyl-1,4-oxazepan-3- yl]methanol (81 mg, 32%) as a white solid. ESI-MS m/z calc.374.2682, found 375.3 (M+1)+; Retention time: 3.56 minutes. The purest fractions of the slowest eluting peak were combined, and the volatiles were concentrated under reduced pressure. To the aqueous solution was added brine (50 mL) and the solution was extracted with DCM (3 x 20 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford [(2S,3S,5R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2-isobutyl- 1,4-oxazepan-3-yl]methanol (131 mg, 52%) as a white solid. ESI-MS m/z calc.374.2682, found 375.3 (M+1)+; Retention time: 3.68 minutes. LC method K. Step 5: 3-[[4-[[(2S,3S,5S)-5-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 2-isobutyl-1,4-oxazepan-3-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00414] Into a reaction vial was added 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (23.24 mg, 0.05562 mmol) and [(2S,3S,5S)-5-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-2-isobutyl-1,4-oxazepan-3-yl]methanol (25 mg, 0.06675 mmol) in anhydrous THF (0.28 mL). Sodium tert-butoxide (39.6 mg, 0.4120 mmol) was added to the reaction mixture at rt. The reaction was stirred for 1 h. The reaction was warmed to 40 °C and stirred for 1 h at that temperature. The reaction was cooled to room temperature and diluted with EtOAc (15 mL) and 1 M HCl (aq.) (15 mL). The two layers were separated, and the aqueous layer was extracted with EtOAc (5x15 mL). The combined organic layers were washed with brine (15 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3-[[4-[[(2S,3S,5S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2-isobutyl-1,4- oxazepan-3-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (37.7 mg, 86%) as a yellow solid. ESI-MS m/z calc.755.3465, found 756.2 (M+1)+; Retention time: 1.4 minutes. LC method A. Step 6: (4S,8S,9S)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-8-isobutyl-18,18-dioxo-7,11-dioxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one [00415] Under argon, 3-[[4-[[(2S,3S,5S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 2-isobutyl-1,4-oxazepan-3-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (37.7 mg, 0.04758 mmol) was combined with CDMT (12.6 mg, 0.07177 mmol) in DMF (4.8 mL) and cooled to 0 °C. N-methylmorpholine (15.7 µL, 0.1428 mmol) was added by syringe and the reaction was stirred at 0 °C for 30 minutes. The ice bath was then removed and stirring was continued for an additional 48 hours at room temperature. The reaction mixture was then partitioned between 25 mL 1M HCl and 25 mL ethyl acetate. The layers were separated, and the aqueous layer was extracted with an additional 25 mL ethyl acetate. The combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated. The crude product was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 25 min) to give (4S,8S,9S)-4-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-8-isobutyl-18,18-dioxo-7,11- dioxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (5.8 mg, 16%) as a yellow solid.1H NMR (400 MHz, CDCl3) δ 8.77 (t, J = 2.1 Hz, 1H), 8.49 (s, 1H), 7.95 (d, J = 7.9 Hz, 1H), 7.82 (dt, J = 7.6, 1.3 Hz, 1H), 7.61 (t, J = 7.8 Hz, 1H), 7.23 (t, J = 7.6 Hz, 1H), 7.08 (d, J = 7.6 Hz, 2H), 6.58 (s, 1H), 6.26 (s, 1H), 6.11 (dd, J = 13.2, 4.2 Hz, 1H), 5.79 (dd, J = 11.3, 4.9 Hz, 1H), 4.99 (t, J = 11.6 Hz, 1H), 4.23 (dt, J = 11.8, 4.7 Hz, 1H), 4.16 (s, 3H), 4.11 – 4.01 (m, 1H), 4.00 – 3.90 (m, 2H), 2.79 – 2.65 (m, 1H), 2.32 – 2.22 (m, 1H), 2.06 (s, 6H), 1.64 – 1.56 (m, 2H), 1.52 (s, 9H), 0.83 – 0.75 (m, 6H), 0.71 – 0.61 (m, 1H). ESI-MS m/z calc.737.33594, found 738.2 (M+1)+; Retention time: 1.89 minutes. LC method A. Example 44: Preparation of Compound 9 Step 1: 3-[[4-[[(2S,3S,5R)-5-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 2-isobutyl-1,4-oxazepan-3-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00416] Into a reaction vial was added 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (23.24 mg, 0.05562 mmol) and [(2S,3S,5R)-5-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-2-isobutyl-1,4-oxazepan-3-yl]methanol (25 mg, 0.06675 mmol) in anhydrous THF (0.28 mL). Sodium tert-butoxide (39.0 mg, 0.4058 mmol) was added to the reaction mixture at rt. The reaction was stirred for 1 h. The reaction was warmed to 40 °C and stirred at that temperature for 1 h. The reaction was cooled to room temperature and diluted with EtOAc (15 mL) and 1 M HCl (aq.) (15 mL). The two layers were separated, and the aqueous layer was extracted with EtOAc (5 x 15 mL). The combined organic layers were washed with brine (15 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3-[[4-[[(2S,3S,5R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2-isobutyl-1,4- oxazepan-3-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (38.8 mg, 88%) as a yellow solid. ESI-MS m/z calc.755.3465, found 756.2 (M+1)+; Retention time: 1.38 minutes. LC method A. Step 2: (4R,8S,9S)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-8-isobutyl-18,18-dioxo-7,11-dioxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one
[00417] Under argon, 3-[[4-[[(2S,3S,5R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 2-isobutyl-1,4-oxazepan-3-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (38.8 mg, 0.04897 mmol) was combined with CDMT (14.6 mg, 0.08316 mmol) in DMF (4.9 mL) and cooled to 0 °C. N-methylmorpholine (16.4 µL, 0.1492 mmol) was added by syringe and the reaction was stirred at 0 °C for 30 minutes. The ice bath was then removed and stirring was continued for an additional 16 hours at room temperature. The reaction mixture was then partitioned between 25 mL 1M HCl and 25 mL ethyl acetate. The layers were separated, and the aqueous layer was extracted with an additional 25 mL ethyl acetate. The combined organic layers were washed 2 x 25 mL with brine, dried over magnesium sulfate, filtered, and concentrated. The crude product was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 25 min) to give (4R,8S,9S)-4- (6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-8-isobutyl-18,18- dioxo-7,11-dioxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (6.4 mg, 17%) as a yellow solid.1H NMR (400 MHz, CDCl3) δ 8.74 (t, J = 1.8 Hz, 1H), 8.58 (s, 1H), 7.83 (d, J = 7.9 Hz, 1H), 7.76 (dt, J = 7.7, 1.4 Hz, 1H), 7.56 (t, J = 7.8 Hz, 1H), 7.26 – 7.21 (m, 1H), 7.11 (d, J = 7.6 Hz, 2H), 6.78 (s, 1H), 6.26 (s, 1H), 5.93 – 5.84 (m, 1H), 4.85 – 4.75 (m, 1H), 4.71 – 4.60 (m, 1H), 4.58 – 4.48 (m, 1H), 4.14 (s, 3H), 3.92 – 3.84 (m, 1H), 3.77 – 3.67 (m, 2H), 3.37 – 3.25 (m, 1H), 2.45 – 2.35 (m, 1H), 2.09 (s, 6H), 1.52 (s, 9H), 1.18 – 1.08 (m, 1H), 1.03 – 0.88 (m, 2H), 0.85 (d, J = 6.5 Hz, 3H), 0.73 (d, J = 6.7 Hz, 3H). ESI-MS m/z calc.737.33594, found 738.2 (M+1)+; Retention time: 1.86 minutes. LC method A. Example 45: Preparation of Compound 10 Step 1: tert-Butyl (4R)-4-[(1R)-5-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-1-isobutyl- 5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00418] To a solution of tert-butyl (4R)-2,2-dimethyl-4-[(1R)-3-methyl-1-(2- oxoethyl)butyl]oxazolidine-3-carboxylate (390 mg, 1.1721 mmol) and 1-(6-tert-butylfuro[2,3- b]pyrazin-2-yl)-2-dimethoxyphosphoryl-ethanone (378 mg, 1.1585 mmol) in isopropanol (10 mL) was added potassium carbonate (804 mg, 5.8174 mmol) at room temperature. The mixture was stirred at room temperature for 16 hours. An aqueous saturated solution of ammonium chloride (30 mL) was added to the mixture, which was extracted with DCM (3 x 20 mL). The combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude was purified by reverse phase chromatography (Column: C18. Gradient: 5-100% MeCN in water and by silica gel chromatography (Gradient: 0-20% EtOAc in heptanes) to afford tert-butyl (4R)-4-[(1R)-5-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-1-isobutyl-5- oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (201 mg, 31%) as a pale yellow oil (mixture of diastereomers). ESI-MS m/z calc.513.3203, found 414.4 (M-99)+; Retention time: 2.09 minutes. LC method R. Step 2: tert-Butyl (4R)-4-[(1S)-5-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-1-isobutyl- 5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00419] Nitrogen was bubbled through a stirred solution of tert-butyl (4R)-4-[(1R)-5-(6-tert- butylfuro[2,3-b]pyrazin-2-yl)-1-isobutyl-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3- carboxylate (201 mg, 0.3612 mmol) in methanol (40 mL) for 10 minutes after which time palladium on carbon (38 mg, 10 %w/w, 0.0357 mmol) was added. Nitrogen bubbling was continued for 10 minutes, followed by hydrogen bubbling. The reaction was then stirred under hydrogen atmosphere for 16 hours. Hydrogen was evacuated and the flask backfilled with nitrogen. The catalyst was filtered on Celite washing with EtOAc (250 mL) and the volatiles were removed under reduced pressure to afford tert-butyl (4R)-4-[(1S)-5-(6-tert-butylfuro[2,3- b]pyrazin-2-yl)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (142 mg, 71%) as a yellow oil. ESI-MS m/z calc.515.3359, found 416.4 (M-99)+; Retention time: 2.54 minutes. LC method R. Step 3: (5S)-5-[(1R)-1-Amino-2-hydroxy-ethyl]-1-(6-tert-butylfuro[2,3-b]pyrazin- 2-yl)-7-methyl-octan-1-one [00420] To a solution of tert-butyl (4R)-4-[(1S)-5-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-1- isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (220 mg, 0.3494 mmol) in 1,4- dioxane (2.5 mL) and water (0.2 mL) at room temperature was slowly added a solution of HCl (in 1,4-dioxane) (0.73 mL of 4 M, 2.92 mmol). The reaction was stirred for 2 hours and the reaction mixture was concentrated under reduced pressure. The residue was co-evaporated with water (3 x 10 mL) and MeCN (10 mL) to afford (5S)-5-[(1R)-1-amino-2-hydroxy-ethyl]-1-(6- tert-butylfuro[2,3-b]pyrazin-2-yl)-7-methyl-octan-1-one (hydrochloride salt) (143 mg, 99%) as a yellow solid. ESI-MS m/z calc.375.2522, found 376.4 (M+1)+; Retention time: 1.46 minutes. LC method S. Step 4: [(2R,3S)-7-(6-tert-Butylfuro[2,3-b]pyrazin-2-yl)-3-isobutyl-azepan-2- yl]methanol [00421] To a solution of (5S)-5-[(1R)-1-amino-2-hydroxy-ethyl]-1-(6-tert-butylfuro[2,3- b]pyrazin-2-yl)-7-methyl-octan-1-one (hydrochloride salt) (143 mg, 0.3468 mmol) in anhydrous DCM (7.5 mL) at room temperature was added NaBH(Oac)3 (245 mg, 1.156 mmol). The reaction mixture was stirred for 2 hours and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography (Column: C18. Gradient: 5-100% MeCN in water with 0.1% formic acid). The fractions containing both diastereomers were combined and the volatiles were removed under reduced pressure. The residue was co-evaporated with water (3 x 10 mL) and MeCN (10 mL) to afford [(2R,3S)-7-(6-tert-butylfuro[2,3-b]pyrazin-2- yl)-3-isobutyl-azepan-2-yl]methanol (23 mg, 17%) as a pale yellow solid. Isolated as a mixture of diastereomers: ESI-MS m/z calc.359.2573, found 360.3 (M+1)+; Retention time: 2.88 minutes. LC method J. Step 5: [(2R,3S,7R)-7-(6-tert-Butylfuro[2,3-b]pyrazin-2-yl)-3-isobutyl-azepan-2- yl]methanol and [(2R,3S,7S)-7-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-3-isobutyl- azepan-2-yl]methanol [00422] Three lots of diastereomeric mixtures [(2R,3S)-7-(6-tert-butylfuro[2,3-b]pyrazin-2- yl)-3-isobutyl-azepan-2-yl]methanol (16 mg, 0.0422 mmol), [(2R,3S)-7-(6-tert-butylfuro[2,3- b]pyrazin-2-yl)-3-isobutyl-azepan-2-yl]methanol (25 mg, 0.0503 mmol) and [(2R,3S)-7-(6-tert- butylfuro[2,3-b]pyrazin-2-yl)-3-isobutyl-azepan-2-yl]methanol (23 mg, 0.0606 mmol) were dissolved in MeCN (1 mL) and the reaction mixture was concentrated in vacuo. The diastereomeric mixture was submitted to chiral SFC separation (Flow rate: 75 mL/min, 20% MeOH, column: i-Amylose 3, 250 x 21.2 mm, temperature = 40 °C, outlet pressure: 100 bar, injection volume: 2000 µL). The earliest eluting peak by SFC (peak 1) gave after evaporation to dryness and lyophilization [(2R,3S,7R)-7-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-3-isobutyl- azepan-2-yl]methanol (16.4 mg, 56%) as pale yellow solid.1H NMR (400 MHz, Benzene-d6) δ 8.14 (s, 1H), 6.32 (s, 1H), 3.83 – 3.74 (m, 2H), 3.41 (t, J = 10.4 Hz, 1H), 2.50 (td, J = 9.5, 4.0 Hz, 1H), 2.15 – 2.05 (m, 1H), 1.83 – 1.74 (m, 1H), 1.63 – 1.40 (m, 4H), 1.23 – 1.11 (m, 3H), 1.10 (s, 9H), 0.96 – 0.91 (m, 2H), 0.84 (d, J = 6.6 Hz, 3H), 0.78 (d, J = 6.6 Hz, 3H), (1H missing, labile proton). ESI-MS m/z calc.359.2573, found 360.3 (M+1)+; Retention time: 2.88 minutes. The later eluting peak by SFC (peak 2) gave after evaporation to dryness and lyophilization [(2R,3S,7S)-7-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-3-isobutyl-azepan-2- yl]methanol (22.5 mg, 72%) as a pale yellow solid.1H NMR (400 MHz, Benzene-d6) δ 8.06 (s, 1H), 6.36 (s, 1H), 3.82 – 3.71 (m, 2H), 3.11 (t, J = 9.8 Hz, 1H), 2.52 – 2.42 (m, 1H), 2.04 – 1.93 (m, 1H), 1.83 – 1.71 (m, 1H), 1.70 – 1.61 (m, 1H), 1.60 – 1.52 (m, 1H), 1.51 – 1.24 (m, 5H), 1.17 – 1.13 (m, 1H), 1.11 (s, 9H), 0.94 – 0.88 (m, 1H), 0.85 (d, J = 6.6 Hz, 3H), 0.76 (d, J = 6.6 Hz, 3H), (1H missing, labile proton). ESI-MS m/z calc.359.2573, found 360.3 (M+1)+; Retention time: 2.88 minutes. LC method J. Step 6: 3-[[4-[[(2R,3S,7S)-7-(6-tert-Butylfuro[2,3-b]pyrazin-2-yl)-3-isobutyl- azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid [00423] In a 4 mL vial, 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (26 mg, 0.06222 mmol) and [(2R,3S,7S)-7-(6-tert-butylfuro[2,3- b]pyrazin-2-yl)-3-isobutyl-azepan-2-yl]methanol (18.4 mg, 0.05118 mmol) were combined in THF (250 µL). Solid sodium tert-butoxide (36 mg, 0.3746 mmol) was added in one portion. The reaction mixture was stirred at 35 °C for 2 hours. After cooling to room temperature, it was diluted with aqueous HCl (1 M, 50 mL) and extracted with a 5:1 mixture of DCM/iPrOH (2 × 50 mL). Organic layers were combined dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was redissolved in DCM and iPrOH (approximately 1 mL at 5:1) and triturated with hexanes (4 mL). Solids were collected to provide 3-[[4-[[(2R,3S,7S)- 7-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-3-isobutyl-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (44 mg, 91%). ESI- MS m/z calc.740.33563, found 741.4 (M+1)+; Retention time: 1.67 minutes. LC method A. Step 7: (4S,8S,9R)-4-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-14-(2,6- dimethylphenyl)-8-isobutyl-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one [00424] In a 4 mL vial, 3-[[4-[[(2R,3S,7S)-7-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-3-isobutyl- azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (44 mg, 0.05660 mmol) and CDMT (15 mg, 0.08544 mmol) were combined in DMF (0.5 mL). The mixture was cooled to 0 °C before the addition of NMM (19 µL, 0.1728 mmol). The reaction mixture was allowed to slowly warm to room temperature and stirred overnight. Purification by reverse phase HPLC (10-99% acetonitrile/5 mM aqueous HCl over 15 minutes) provided (4S,8S,9R)-4-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-14-(2,6-dimethylphenyl)- 8-isobutyl-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9] tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (9.5 mg, 23%) as a white solid.1H NMR (400 MHz, CDCl3) δ 8.66 (s, 1H), 8.29 (s, 1H), 8.06 (d, J = 7.9 Hz, 1H), 7.77 (d, J = 1.3 Hz, 1H), 7.60 (t, J = 7.8 Hz, 1H), 7.20 (t, J = 7.6 Hz, 1H), 7.04 (d, J = 7.6 Hz, 2H), 6.66 (s, 1H), 6.22 (s, 1H), 5.82 (dd, J = 12.3, 5.4 Hz, 1H), 5.76 (dd, J = 11.9, 5.3 Hz, 1H), 4.97 (t, J = 12.0 Hz, 1H), 4.40 – 4.31 (m, 1H), 2.30 – 2.15 (m, 2H), 1.99 (s, 6H), 1.89 – 1.64 (m, 4H), 1.41 (s, 9H), 1.30 – 1.18 (m, 3H), 0.84 – 0.74 (m, 1H), 0.69 (d, J = 2.4 Hz, 3H), 0.68 (d, J = 2.5 Hz, 3H). ESI-MS m/z calc.722.325, found 723.3 (M+1)+; Retention time: 2.48 minutes. LC method A. Example 46: Preparation of Compound 11 Step 1: tert-Butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00425] To a solution of 1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2- dimethoxyphosphoryl-ethanone (573.1 mg, 1.689 mmol) in isopropanol (30.0 mL) was added potassium carbonate (1.25 g, 9.045 mmol) and the reaction was left stirring for 5 min. This solution was then added to a solution of tert-butyl (4R)-2,2-dimethyl-4-[(1R)-3-methyl-1-(2- oxoethyl)butyl]oxazolidine-3-carboxylate (600 mg, 1.531 mmol) in isopropanol (30 mL). Palladium on carbon (1.75 g of 10 %w/w, 1.644 mmol) was added to the reaction mixture and the content was placed under hydrogen atmosphere using a balloon filled with hydrogen gas and left stirring overnight. The reaction mixture was filtered on a pad of Celite, the cake was washed with methanol, concentrated to give tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (809.5 mg, 100%) as an orange solid. ESI-MS m/z calc.528.36755, found 529.2 (M+1)+; Retention time: 2.49 minutes. LC method A. Step 2: (5S)-5-[(1R)-1-Amino-2-hydroxy-ethyl]-1-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-7-methyl-octan-1-one [00426] HCl (in dioxane) (14 mL of 4 M, 56.00 mmol) was added dropwise to a solution of tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-isobutyl-5-oxo- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (328 mg, 0.5707 mmol) in dioxane (12 mL) and water (4.8 mL) then left stirring at room temperature for 1 h. The reaction mixture was concentrated under reduced pressure and the resulting residue was purified by reverse phase chromatography on a C18 column using a 5 to 100% gradient of methanol in water to provide (5S)-5-[(1R)-1-amino-2-hydroxy-ethyl]-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-7- methyl-octan-1-one (160 mg, 72%) as a yellow solid 1H NMR (400 MHz, MeOH-d4) δ 8.97 (s, 1H), 6.63 (s, 1H), 4.13 (s, 3H), 3.83 (dd, J = 11.6, 4.0 Hz, 1H), 3.62 (dd, J = 11.5, 8.3 Hz, 1H), 3.38 (td, J = 6.8, 2.8 Hz, 2H), 3.35 – 3.32 (m, 1H), 1.92 – 1.77 (m, 3H), 1.75 – 1.63 (m, 1H), 1.57 (s, 9H), 1.55 – 1.40 (m, 2H), 1.30 – 1.22 (m, 2H), 0.98 – 0.91 (m, 6H), (3H, missing labile protons). ESI-MS m/z calc.388.2838, found 389.4 (M+1)+; Retention time: 1.43 minutes. LC method I. Step 3: [(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isobutyl-azepan-2-yl]methanol and [(2R,3S,7R)-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-3-isobutyl-azepan-2-yl]methanol [00427] A flask was flame-dried then cooled down to room temperature using a nitrogen stream and charged with (5S)-5-[(1R)-1-amino-2-hydroxy-ethyl]-1-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-7-methyl-octan-1-one (160 mg, 0.4114 mmol) dissolved in DCM (13 mL) and a solution of sodium triacetoxyborohydride (436 mg, 2.0572 mmol) in DCM (13 mL) was added at room temperature. The reaction mixture was stirred at this temperature for 2 h. The reaction mixture was quenched with a saturated sodium bicarbonate aqueous solution (25 mL) and this mixture was left stirring for 30 min. The aqueous phase was separated and extracted with DCM (3x 50 mL). The combined organics were dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by reverse phase chromatography on a C18 column using a 5 to 100% gradient of acetonitrile in pure water to provide a purified mixture of two products which were then separated from each other by reverse phase chromatography on a C18 column using a 0 to 70% gradient of acetonitrile in water to provide [(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isobutyl- azepan-2-yl]methanol (53 mg, 35%) as a pale yellow sticky solid 1H NMR (400 MHz, Benzene- d6) δ 8.59 (s, 1H), 6.60 (s, 1H), 4.27 (s, 1H), 3.87 (dd, J = 10.3, 4.4 Hz, 1H), 3.77 (dd, J = 10.4, 4.0 Hz, 1H), 3.51 (s, 3H), 3.12 (t, J = 10.0 Hz, 1H), 2.51 (td, J = 9.4, 4.0 Hz, 1H), 2.11 – 2.01 (m, 1H), 1.83 (dtd, J = 13.7, 10.4, 3.4 Hz, 1H), 1.74 – 1.64 (m, 1H), 1.63 – 1.56 (m, 1H), 1.53 – 1.25 (m, 5H), 1.20 – 1.14 (m, 1H), 1.12 (s, 9H), 0.97 – 0.89 (m, 1H), 0.86 (d, J = 6.6 Hz, 3H), 0.77 (d, J = 6.6 Hz, 3H). ESI-MS m/z calc.372.2889, found 373.2 (M+1)+; Retention time: 4.03 minutes and [(2R,3S,7R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isobutyl- azepan-2-yl]methanol (45 mg, 29%) as a pale yellow solid 1H NMR (400 MHz, Benzene-d6) δ 8.63 (s, 1H), 6.60 (s, 1H), 3.86 (dd, J = 11.2, 2.4 Hz, 1H), 3.81 (dd, J = 10.6, 4.3 Hz, 1H), 3.58 (s, 3H), 3.49 (t, J = 10.4 Hz, 1H), 2.55 (td, J = 9.5, 4.2 Hz, 1H), 2.21 – 2.10 (m, 1H), 1.86 – 1.74 (m, 1H), 1.61 – 1.44 (m, 3H), 1.28 – 1.06 (m, 12H), 1.04 – 0.88 (m, 3H), 0.85 (d, J = 6.8 Hz, 3H), 0.79 (d, J = 6.6 Hz, 3H), (one labile proton missing). ESI-MS m/z calc.372.2889, found 373.3 (M+1)+; Retention time: 4.44 minutes. LC method K. Step 4: 3-[[4-[[(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-isobutyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00428] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (20.39 mg, 0.04880 mmol) and [(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isobutyl-azepan-2-yl]methanol (20 mg, 0.05368 mmol) were combined in a flask and dissolved in THF (0.25 mL) followed by the addition of sodium tert-butoxide (29.1 mg, 0.3028 mmol). The reaction mixture was stirred for 30 minutes at room temperature then warmed to 40 °C for 2 hours. The reaction mixture was cooled to room temperature and diluted with EtOAc and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (5 x). The organic layers were combined, washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3-[[4-[[(2R,3S,7S)-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-3-isobutyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin- 2-yl]sulfamoyl]benzoic acid (24.0 mg, 65%) as a white solid. ESI-MS m/z calc.753.36725, found 754.2 (M+1)+; Retention time: 1.62 minutes. LC method A. Step 5: (4S,8S,9R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-8-isobutyl-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one [00429] 3-[[4-[[(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isobutyl- azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (24.0 mg, 0.03183 mmol) was azeotroped with toluene three times in a vial then put under vacuum for 30 minutes. Under argon, CDMT (9.2 mg, 0.0524 mmol) followed by DMF (3.2 mL) was added and the reaction mixture was cooled to 0 °C. N-methylmorpholine (10.5 µL, 0.0955 mmol) was added by syringe and the reaction was stirred at 0 °C for 30 minutes. The ice bath was then removed and stirring was continued for an additional 72 hours at room temperature. The reaction mixture was then partitioned between 25 mL 1M HCl and 25 mL ethyl acetate. The layers were separated, and the aqueous layer was extracted with an additional 25 mL of ethyl acetate. The combined organic layers were washed 2 x 25 mL with brine, dried over magnesium sulfate, filtered, and concentrated. The crude product was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 25 min) to give (4S,8S,9R)-4-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-8-isobutyl-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen- 2-one (3.7 mg, 15%) as a yellow solid.1H NMR (400 MHz, CDCl3) δ 8.83 (t, J = 1.8 Hz, 1H), 8.45 (s, 1H), 7.84 (d, J = 7.9 Hz, 1H), 7.79 (dt, J = 7.7, 1.4 Hz, 1H), 7.56 (t, J = 7.8 Hz, 1H), 7.23 (t, J = 7.6 Hz, 1H), 7.08 (d, J = 7.6 Hz, 2H), 6.56 (s, 1H), 6.27 (s, 1H), 5.94 – 5.86 (m, 2H), 4.88 (t, J = 11.8 Hz, 1H), 4.47 – 4.38 (m, 1H), 4.16 (s, 3H), 2.39 – 2.28 (m, 1H), 2.05 (s, 6H), 2.01 (s, 3H), 1.89 – 1.77 (m, 2H), 1.51 (s, 9H), 1.27 – 1.15 (m, 3H), 0.88 – 0.78 (m, 1H), 0.67 (dd, J = 6.3, 3.7 Hz, 6H). ESI-MS m/z calc.735.3567, found 736.4 (M+1)+; Retention time: 2.14 minutes. LC method A. Example 47: Preparation of Compound 16 Step 1: tert-Butyl (4S)-4-[5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-5- hydroxy-1-isopropoxy-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate ~85:15 mixture of diastereomers, the major isomer having the 4S,1R configuration [00430] The reaction was run in 3 small batches, the quantities indicated are for the total of the 3 reactions. To 8 mL of a solution of 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine (1.91 g, 6.6598 mmol) in tetrahydrofuran (24 mL) at -78 °C was added 0.94 mL of n-butyllithium (solution in hexanes) (2.82 mL of 2.5 M, 7.05 mmol) dropwise. As soon as the addition of n-BuLi was over, 8 mL of a solution of tert-butyl (4S)-4-(1-isopropoxy-5-oxo- pentyl)-2,2-dimethyl-oxazolidine-3-carboxylate (2.44 g, 6.6567 mmol) in tetrahydrofuran (24 mL) was added dropwise. The reaction was left stirring for 10 min and MeOH (5 mL) was added. The combined crude products were concentrated in vacuo and were then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 80% of MeCN in water to afford after evaporation tert-butyl (4S)-4-[5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-5-hydroxy-1-isopropoxy-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (2.29 g, 62%) as a yellow fluffy solid (mixture of isomers). ESI-MS m/z calc.532.3625, found 533.4 (M+1)+; Retention time: 2.08 minutes. LC method I. Step 2: tert-Butyl (4S)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate, tert- butyl (4S)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1- isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate, and tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-isopropoxy- 5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00431] To a solution of tert-butyl (4S)-4-[5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-5-hydroxy-1-isopropoxy-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (2.29 g, 4.144 mmol) in dichloroethane (50 mL) was added manganese (IV) oxide (activated) (8.32 g, 81.346 mmol). The resulting suspension was stirred at 70 °C for 1.5 h. The suspension was filtered on a pad of Celite and washed with DCM (2 x 50 mL). The crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeOH in acidic water (0.1% v/v of formic acid in water). The fractions containing the desired product were combined and the volatiles were removed under reduced pressure. To the remaining aqueous phase was added DCM (50 mL) and water (200 mL). The aqueous phase was extracted with DCM (2 x 50 mL). The combined organic phases were dried under anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford a dark semi-solid oil. The oil was purified by preparative SFC (Lux Column 5 µm, i-Amylose 3, 250 x 30 mm, 65.8 mg/mL, 52.7 mg/injection, flow rate 150 mL/min, eluant 50% methanol, outlet pressure 100 bar).The first peak afforded, after evaporation, tert-butyl (4S)-4-[(1S)-5-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (73.2 mg, 3%) as an orange oil.1H NMR (400 MHz, CDCl3) δ 9.13 (br. S., 1H), 6.59 (br. S., 1H), 4.28 – 4.03 (m, 4H), 4.03 – 3.88 (m, 2H), 3.87 – 3.61 (m, 2H), 3.42 – 3.15 (m, 2H), 2.15 – 1.92 (m, 1H), 1.83 – 1.34 (m, 27H), 1.28 – 1.04 (m, 6H). ESI-MS m/z calc.530.3468, found 431.4 (M-99)+; Retention time: 2.27 minutes. The second peak afforded, after evaporation, tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (364 mg, 16%) as an orange oil.1H NMR (400 MHz, CDCl3) δ 9.66 – 8.68 (m, 1H), 6.60 (br. S., 1H), 4.22 – 4.03 (m, 4H), 4.02 – 3.55 (m, 4H), 3.40 – 3.15 (m, 2H), 2.01 – 1.75 (m, 2H), 1.64 – 1.35 (m, 26H), 1.23 – 1.03 (m, 6H). ESI-MS m/z calc.530.3468, found 431.4 (M-99)+; Retention time: 2.28 minutes. The third peak afforded, after evaporation, the major isomer tert-butyl (4S)- 4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-isopropoxy-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate (1.45 g, 65%) as an orange oil.1H NMR (400 MHz, CDCl3) δ 9.11 (br. S., 1H), 6.68 (br. S., 1H), 4.20 – 4.05 (m, 3H), 4.03 – 3.65 (m, 4H), 3.79 – 3.64 (m, 1H), 3.29 (br. S., 2H), 2.01 – 1.76 (m, 2H), 1.67 – 1.42 (m, 26H), 1.21 – 1.07 (m, 6H). ESI-MS m/z calc.530.3468, found 431.4 (M-99)+; Retention time: 2.32 minutes. LC method I. Step 3: [(2S,3R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol and (5R,6S)-6-amino-1- (6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-7-hydroxy-5-isopropoxy- heptan-1-one [00432] To a solution of tert-butyl (4S)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.45 g, 2.7049 mmol) in 1,4-dioxane (30 mL) was added hydrochloric acid (solution in 1,4-dioxane) (13.5 mL of 4 M, 54.000 mmol) at rt. A drop of water was added, and the reaction was stirred for 18 h. The solvent was removed under reduced pressure and water (40 mL) and EtOAc (40 mL) were added. The phases were separated, and the aqueous one was extracted with EtOAc (2 x 40 mL) and DCM (3 x 40 mL). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeCN in water to afford (5R,6S)-6-amino-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 7-hydroxy-5-isopropoxy-heptan-1-one (892.2 mg, 80%) as a yellow crystalline solid.1H NMR (400 MHz, CDCl3) δ 9.16 – 9.05 (m, 1H), 8.32 – 8.04 (m, 2H), 7.11 – 7.04 (m, 1H), 4.31 – 4.22 (m, 2H), 4.05 – 3.82 (m, 3H), 3.81 – 3.70 (m, 1H), 3.66 – 3.57 (m, 1H), 3.42 – 3.28 (m, 2H), 2.01 – 1.82 (m, 2H), 1.79 – 1.67 (m, 4H), 1.65 – 1.55 (m, 9H), 1.32 – 1.11 (m, 6H). ESI-MS m/z calc.390.2631, found 391.4 (M+1)+; Retention time: 1.38 minutes. A second fraction was obtained to afford, after evaporation, [(2S,3R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-3-isopropoxy-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (215 mg, 21%) as a dark oil.1H NMR (400 MHz, CDCl3) δ 9.29 – 9.10 (m, 1H), 6.71 – 6.59 (m, 1H), 4.41 – 4.30 (m, 4H), 4.25 – 4.14 (m, 1H), 3.93 – 3.84 (m, 1H), 3.83 – 3.69 (m, 2H), 3.38 – 3.15 (m, 1H), 2.36 – 2.20 (m, 1H), 2.11 – 1.99 (m, 3H), 1.66 – 1.48 (m, 11H), 1.25 – 1.16 (m, 6H). ESI-MS m/z calc. 372.2525, found 373.4 (M+1)+; Retention time: 1.38 minutes. LC method I. Step 4: [(2S,3R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methanol [00433] To a solution of (5R,6S)-6-amino-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-7-hydroxy-5-isopropoxy-heptan-1-one (892.2 mg, 2.1681 mmol) in dichloromethane (35 mL) was added sodium triacetoxyborohydride (1.08 g, 5.0958 mmol). The reaction was stirred at rt for 0.5 h. A saturated aqueous solution of sodium bicarbonate (50 mL) was added. The phases were separated, and the aqueous one was extracted with DCM (2 x 50 mL). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 50% of MeCN in basic water (ammonium bicarbonate/ammonium hydroxide buffer pH = 10) to afford, after evaporation, [(2S,3R)-7-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methanol (690 mg, 79%) as a yellow oil. ESI-MS m/z calc.374.2682, found 375.4 (M+1)+; Retention time: 3.56 minutes. LC method K. Step 5: [(2S,3R,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methanol and [(2S,3R,7R)-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methanol [00434] A solution of a first batch of [(2S,3R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin- 3-yl)-3-isopropoxy-azepan-2-yl]methanol (690 mg, 1.7116 mmol) in acetonitrile (50 mL) and a solution of a second batch of [(2S,3R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methanol (208 mg, 0.4421 mmol) in acetonitrile (30 mL) were combined and concentrated under reduced pressure. The product was purified by preparative HPLC (Flow: 42,0 mL/min, Run time: 19 min, mobile phase conditions: initial 100% H2O (10 mM NH4CO3H) and 0% MeCN for 3 min, reach 30% MeCN over 1 min, reach 50% MeCN over 11 min and finally reach 95% MeCN over 2 min with a 2 min hold at 95%). Three fractions were obtained. The first fraction afforded, after evaporation and lyophilization, [(2S,3R,7S)-7-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methanol (342.6 mg, 42%) as a white solid.1H NMR (400 MHz, Benzene-d6) δ 8.54 (s, 1H), 6.58 (s, 1H), 4.07 – 3.97 (m, 2H), 3.48 (s, 3H), 3.44 – 3.29 (m, 2H), 3.20 – 3.11 (m, 1H), 2.99 (td, J = 8.3, 4.4 Hz, 1H), 2.08 – 1.96 (m, 1H), 1.93 – 1.68 (m, 4H), 1.67 – 1.54 (m, 1H), 1.12 (s, 9H), 1.08 (d, J = 6.1 Hz, 3H), 0.98 (d, J = 5.9 Hz, 3H) (2H missing, labile protons). ESI-MS m/z calc.374.2682, found 375.4 (M+1)+; Retention time: 3.55 minutes. The second fraction was a mixture of the two isomers. The third fraction afforded, after evaporation and lyophilization, [(2S,3R,7R)-7-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methanol (47.6 mg, 6%) as a yellow solid.1H NMR (400 MHz, Benzene-d6) δ 8.53 (s, 1H), 6.59 (s, 1H), 4.17 (dd, J = 10.3, 4.4 Hz, 1H), 3.82 (dd, J = 11.2, 3.9 Hz, 1H), 3.65 – 3.53 (m, 4H), 3.36 (sept, J = 6.0 Hz, 1H), 3.14 (td, J = 9.0, 4.5 Hz, 1H), 2.97 – 2.86 (m, 1H), 2.14 – 2.00 (m, 1H), 1.96 – 1.85 (m, 1H), 1.72 – 1.61 (m, 1H), 1.58 – 1.42 (m, 2H), 1.27 – 1.13 (m, 1H), 1.09 (s, 9H), 1.06 (d, J = 6.1 Hz, 3H), 1.00 (d, J = 6.1 Hz, 3H) (2 H missing, labile protons). ESI-MS m/z calc.374.2682, found 375.4 (M+1)+; Retention time: 3.66 minutes. LC method W. Step 6: 3-[[4-[[(2S,3R,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid
[00435] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (111.5 mg, 0.2668 mmol) and [(2S,3R,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methanol (100 mg, 0.2670 mmol) were combined in a flask and dissolved in THF (1.5 mL) followed by the addition of sodium tert-butoxide (155.5 mg, 1.618 mmol). The reaction mixture was stirred for 30 minutes at room temperature then warmed to 40 °C for 2 hours. The reaction mixture was cooled to room temperature and diluted with EtOAc and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (5 x). The organic layers were combined, washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3-[[4-[[(2S,3R,7S)-7-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (161.6 mg, 80%) as a white solid. ESI- MS m/z calc.755.3465, found 756.7 (M+1)+; Retention time: 1.48 minutes. LC method A. Step 7: (4S,8R,9S)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-8-isopropoxy-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one [00436] 3-[[4-[[(2S,3R,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (101.8 mg, 0.1347 mmol), azeotroped twice with toluene, was dissolved in NMP (6.2 mL) and TsCl (52.1 mg, 0.2733 mmol) was added. The reaction was stirred for 1 hour at room temperature. After 1 hour, NMM (59.3 µL, 0.5394 mmol) was added and the reaction was stirred at room temperature for 3 hours. Additional NMM (14.8 µL, 0.1346 mmol) was added and the reaction was stirred for 1 hour. Additional NMM (14.8 µL, 0.1346 mmol) was added and the reaction mixture was stirred for 1 hour. The reaction was poured into a mixture of ethyl acetate and 1 M HCl and the layers were separated. The aqueous layer was further extracted with ethyl acetate (2 x). The organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 25 min) to give (4S,8R,9S)-4- (6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-8-isopropoxy- 18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (18.1 mg, 18%) as a yellow solid.1H NMR (400 MHz, CDCl3) δ 8.79 (s, 1H), 8.47 (s, 1H), 7.91 – 7.84 (m, 2H), 7.58 (t, J = 7.8 Hz, 1H), 7.22 (t, 1H), 7.09 (s, 1H), 7.07 (s, 1H), 6.47 (s, 1H), 6.28 (s, 1H), 6.02 (dd, J = 12.8, 5.0 Hz, 1H), 5.85 – 5.75 (m, 1H), 4.81 – 4.69 (m, 2H), 4.13 (s, 3H), 3.58 (s, 1H), 3.40 – 3.30 (m, 1H), 2.45 – 2.35 (m, 1H), 2.14 – 1.93 (m, 9H), 1.91 – 1.82 (m, 1H), 1.80 – 1.71 (m, 1H), 1.50 (s, 9H), 0.99 (d, J = 6.0 Hz, 3H), 0.63 (d, J = 6.0 Hz, 3H). ESI-MS m/z calc.737.33594, found 738.2 (M+1)+; Retention time: 1.91 minutes. LC method A.
Example 48: Preparation of Compound 17 Step 1: 3-[[4-[[(2S,3R,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)-5-methyl-pyrimidin- 2-yl]sulfamoyl]benzoic acid [00437] Into a reaction vial was added [(2S,3R,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methanol (55 mg, 0.1469 mmol) and 3-[[4-chloro-6- (2,6-dimethylphenyl)-5-methyl-pyrimidin-2-yl]sulfamoyl]benzoic acid (65 mg, 0.1505 mmol) in anhydrous THF (2.2 mL). Then solid sodium tert-butoxide (72 mg, 0.7492 mmol) was added and the reaction was stirred for 3 hours at room temperature. The reaction was diluted with EtOAc (2 mL) and HCl (1.0 mL of 1 M, 1.00 mmol). The two layers were separated, and the aqueous layer was extracted with EtOAc (3 x1 mL). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give a crude light yellow solid of 3-[[4-[[(2S,3R,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)-5-methyl- pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (114 mg, 49%) ESI-MS m/z calc. 769.3622, found 770.547 (M+1)+; Retention time: 1.428 minutes. LC method A.
Step 2: (4S,8R,9S)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-8-isopropoxy-13-methyl-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one [00438] To a scintillation vial with stir bar was added 3-[[4-[[(2S,3R,7S)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)-5-methyl-pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (114 mg, 0.0721 mmol) and p-TsCl (34.3 mg, 0.1799 mmol) dissolved in NMP (2.0 mL). Then NMM (35 µL, 0.3183 mmol) was added and the reaction was stirred for 2 hours. Another aliquot of NMM (7.1 µL, 0.06458 mmol) was added and the reaction was stirred overnight for another 14 hours. The mixture was filtered and purified by reverse phase chromatography (20-99% MeCN-water with 5mM HCl modifier, 15 minute run) to give a light yellow solid of (4S,8R,9S)-4-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-8-isopropoxy-13-methyl- 18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (14.0 mg, 28%) 1H NMR (400 MHz, CDCl3) δ 8.79 (s, 1H), 8.47 (s, 1H), 8.01 (d, J = 8.0 Hz, 1H), 7.86 (d, J = 7.7 Hz, 1H), 7.62 (t, J = 7.8 Hz, 1H), 7.22 (t, J = 7.6 Hz, 1H), 7.12 (d, J = 7.6 Hz, 1H), 7.07 (d, J = 7.6 Hz, 1H), 6.65 (s, 1H), 6.01 (dd, J = 12.8, 4.9 Hz, 1H), 5.87 – 5.75 (m, 1H), 4.82 (d, J = 4.5 Hz, 2H), 4.18 (s, 3H), 3.51 (s, 1H), 3.30 (p, J = 6.0 Hz, 1H), 2.49 – 2.38 (m, 1H), 2.20 – 2.11 (m, 1H), 2.04 (s, 3H), 2.01 (s, 1H), 1.87 (s, 3H), 1.75 (s, 3H), 1.52 (s, 9H), 1.25 (s, 3H), 0.99 (d, J = 6.1 Hz, 3H), 0.60 (d, J = 6.0 Hz, 3H). ESI-MS m/z calc.751.35156, found 752.565 (M+1)+; Retention time: 1.977 minutes. LC method A. Example 49: Preparation of Compound 18 Step 1: 3-[[4-[[(2R,3S,7R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)-5-methoxy- pyrimidin-2-yl]sulfamoyl]benzoic acid [00439] 3-[[4-Chloro-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2-yl]sulfamoyl]benzoic acid (59.2 mg, 0.1322 mmol) and [(2S,3R,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-3-isopropoxy-azepan-2-yl]methanol (51.0 mg, 0.1362 mmol) were combined in a flask and dissolved in THF (1.4 mL) followed by the addition of sodium tert-butoxide (76.5 mg, 0.796 mmol). The reaction mixture was stirred for 1 hour at room temperature. The reaction mixture was diluted with EtOAc and 1 M HCl. The organic phase was separated and the aqueous layer was extracted with EtOAc (5 x). The organic layers were combined, washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3-[[4- [[(2R,3S,7R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2- yl]methoxy]-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (87.1 mg, 80%) as a yellow solid. ESI-MS m/z calc.785.35706, found 786.2 (M+1)+; Retention time: 1.57 minutes. LC method A. Step 2: (4S,8R,9S)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-8-isopropoxy-13-methoxy-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one
[00440] 3-[[4-[[(2R,3S,7R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)-5-methoxy-pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (87.1 mg, 0.1108 mmol) was dissolved in NMP (5.3 mL) and TsCl (43.6 mg, 0.2287 mmol) was added. The reaction was stirred for 10 minutes at room temperature. After 10 minutes, NMM (49.0 µL, 0.4457 mmol) was added and the reaction was stirred at room temperature for 3 hours. After 3 hours, additional NMM (12.2 µL, 0.1110 mmol) was added. After stirring an additional hour, more NMM (12.2 µL, 0.111 mmol) was added and the reaction was stirred overnight. The reaction was directly injected onto a reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 25 min) to give (4S,8R,9S)-4- (6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-8-isopropoxy-13- methoxy-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (11.1 mg, 12%) as a pale yellow solid.1H NMR (400 MHz, CDCl3) δ 8.76 (t, J = 1.7 Hz, 1H), 8.49 (s, 1H), 7.85 (t, J = 9.0 Hz, 2H), 7.57 (t, J = 7.7 Hz, 1H), 7.25 – 7.21 (m, 1H), 7.12 (d, J = 7.6 Hz, 1H), 7.06 (d, J = 7.5 Hz, 1H), 6.54 (s, 1H), 5.99 (d, J = 11.1 Hz, 1H), 5.82 (dd, J = 11.2, 5.3 Hz, 1H), 4.97 – 4.87 (m, 1H), 4.85 – 4.76 (m, 1H), 4.15 (s, 3H), 3.62 – 3.56 (m, 1H), 3.43 (s, 3H), 3.32 (hept, J = 6.0 Hz, 1H), 2.46 – 2.34 (m, 2H), 2.13 (s, 3H), 2.05 – 1.98 (m, 2H), 1.95 (s, 3H), 1.51 (s, 9H), 0.99 (d, J = 6.0 Hz, 3H), 0.91 – 0.82 (m, 2H), 0.64 (d, J = 5.9 Hz, 3H). ESI-MS m/z calc.767.3465, found 768.6 (M+1)+; Retention time: 1.92 minutes. LC method A. Example 50: Preparation of Compounds 20 and 21 Step 1: tert-Butyl (4R)-4-[(1R)-5-(7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin- 2-yl)-1-isobutyl-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00441] To a solution of tert-butyl (4R)-2,2-dimethyl-4-[(1R)-3-methyl-1-(2- oxoethyl)butyl]oxazolidine-3-carboxylate (202 mg, 0.4833 mmol) and 1-(7-cyclopropyl-6- isopropyl-furo[2,3-b]pyrazin-2-yl)-2-dimethoxyphosphoryl-ethanone (170 mg, 0.4825 mmol) in isopropanol (17 mL) was added potassium carbonate (333 mg, 2.409 mmol) at room temperature. The mixture was stirred at room temperature overnight. The crude material was filtered over celite, washed with methanol (20 mL) and carried forward as a solution of tert- butyl (4R)-4-[(1R)-5-(7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-1-isobutyl-5-oxo- pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (E and Z isomers mixture in solution, yield assumed to be quantitative) ESI-MS m/z calc.539.33594, found 540.3 (M+1)+; Retention time: 2.32 minutes. LC method C. Step 2: tert-Butyl (4R)-4-[(1S)-5-(7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin- 2-yl)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00442] A nitrogen flushed solution of tert-butyl (4R)-4-[(1R)-5-(7-cyclopropyl-6-isopropyl- furo[2,3-b]pyrazin-2-yl)-1-isobutyl-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate from the above step in isopropanol (17 mL) and methanol (20 mL) was stirred at room temperature, then palladium on carbon (103 mg of 10 %w/w, 0.09679 mmol) was added. The mixture was evacuated and then stirred under a hydrogen atmosphere using a hydrogen-filled balloon at room temperature for 5 hours. The mixture was filtered over a plug of celite, washed with methanol and concentrated. The crude was then purified by silica chromatography (40 g of silica) using a gradient eluent of 100% hexanes to 100% ethyl acetate over 20 minutes (product elutes at 32% EtOAc). The desired fractions were concentrated under reduced pressure, and dried under high vacuum to afford as a pale yellow oil tert-butyl (4R)-4-[(1S)-5-(7-cyclopropyl- 6-isopropyl-furo[2,3-b]pyrazin-2-yl)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (54 mg, 21%) ESI-MS m/z calc.541.35156, found 542.4 (M+1)+; Retention time: 2.42 minutes. LC method C. Step 3: [(2R,3S)-7-(7-Cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-3- isobutyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00443] To a stirred solution of tert-butyl (4R)-4-[(1S)-5-(7-cyclopropyl-6-isopropyl-furo[2,3- b]pyrazin-2-yl)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (54 mg, 0.09968 mmol) in a mixture of dioxane (1.5 mL) and water (750 µL) was added a solution of hydrogen chloride (4M in dioxane) (2.5 mL of 4 M, 10.0 mmol) dropwise. The reaction was stirred at room temperature for 16 hours, then the reaction was concentrated to dryness. The residual water was co-evaporated with acetonitrile (5 x 25 mL), then dried under high vacuum to afford crude [(2R,3S)-7-(7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-3-isobutyl-3,4,5,6- tetrahydro-2H-azepin-2-yl]methanol (38 mg, 99%) as an orange oil. ESI-MS m/z calc. 383.25726, found 384.4 (M+1)+; Retention time: 1.65 minutes. The product was carried forward without further purification. LC method A. Step 4: [(2R,3S)-7-(7-Cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-3- isobutyl-azepan-2-yl]methanol [00444] To a solution of [(2R,3S)-7-(7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-3- isobutyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (38 mg, 0.09908 mmol) in anhydrous dichloromethane (2 mL) at room temperature was added sodium triacetoxyborohydride (117 mg, 0.5520 mmol). The reaction was stirred at room temperature for 1 hour and a saturated aqueous solution of sodium bicarbonate (10 mL) was added. The phases were separated, and the aqueous solution was extracted with DCM (2 x). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude was then purified by silica chromatography (12 g of silica) using a gradient eluent of 100% dichloromethane to 20% methanol in DCM over 20 minutes (product eluted at 12% MeOH). The desired fractions were concentrated under reduced pressure, and dried under high vacuum to afford as a pale yellow solid [(2R,3S)-7-(7-cyclopropyl-6-isopropyl-furo[2,3- b]pyrazin-2-yl)-3-isobutyl-azepan-2-yl]methanol (29 mg, 76%), ESI-MS m/z calc.385.27292, found 386.4 (M+1)+; Retention time: 1.58 minutes. LC method A. Step 5: 3-[[4-[[(2R,3S)-7-(7-Cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-3- isobutyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00445] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (31 mg, 0.07419 mmol) and [(2R,3S)-7-(7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-3-isobutyl- azepan-2-yl]methanol (28 mg, 0.07262 mmol) were combined in a flask and dissolved in THF (500 µL) followed by the addition of sodium tert-butoxide (45 mg, 0.4682 mmol). The reaction mixture was stirred for 60 minutes at room temperature then warmed to 40 °C for 30 minutes then 50 °C for 1 hour. The reaction mixture was cooled to room temperature and diluted with EtOAc and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (5x). The organic layers were combined, washed with brine, and dried over sodium sulfate. The mixture was filtered, and then concentrated under reduced pressure. The crude was then purified by silica chromatography (12 g of silica) using a gradient eluent of 100% hexanes to 100% ethyl acetate over 20 minutes followed by a gradient of 100% dichloromethane to 20% methanol in dichloromethane. The desired fractions were concentrated under reduced pressure, and dried under high vacuum to give a mixture of diastereomers as a white solid of 3-[[4- [[(2R,3S)-7-(7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-3-isobutyl-azepan-2- yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (56 mg, 94%). ESI-MS m/z calc.766.35126, found 767.6 (M+1)+; Retention time: 1.79 minutes. LC method A. Step 6: (8S,9R)-4-(7-Cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)-14-(2,6- dimethylphenyl)-8-isobutyl-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one, diastereomer 1 (Compound 20), and (8S,9R)-4-(7-cyclopropyl-6- isopropyl-furo[2,3-b]pyrazin-2-yl)-14-(2,6-dimethylphenyl)-8-isobutyl-18,18- dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one, diastereomer 2 (Compound 21) diastereomer 1 diastereomer 2 [00446] Under argon, 3-[[4-[[(2R,3S)-7-(7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2-yl)- 3-isobutyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (56 mg, 0.07302 mmol), azeotroped twice with toluene, was dissolved in NMP (6 mL) and TsCl (28 mg, 0.1469 mmol) was added. The reaction was stirred for 30 minutes at room temperature. Then NMM (35 µL, 0.3183 mmol) was added and the reaction was stirred at room temperature overnight. The reaction still had starting material remaining so additional TsCl (21 mg, 0.1102 mmol) and NMM (35 µL, 0.3183 mmol) were added and the reaction was stirred an additional 2 hours. The crude mixture was purified directly by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 30 min) followed by reverse phase preparative chromatography (30-99% acetonitrile/5 mM aqueous HCl over 30 min) to give the first eluting peak 1, diastereomer 1, as a white solid (8S,9R)-4-(7-cyclopropyl-6-isopropyl- furo[2,3-b]pyrazin-2-yl)-14-(2,6-dimethylphenyl)-8-isobutyl-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen- 2-one (2.5 mg, 9%), 1H NMR (400 MHz, CDCl3) δ 8.72 (t, J = 1.7 Hz, 1H), 8.40 (s, 1H), 8.16 (s, 1H), 7.99 (dt, J = 8.0, 1.4 Hz, 1H), 7.75 (dt, J = 7.7, 1.3 Hz, 1H), 7.61 (t, J = 7.7 Hz, 1H), 7.26 (t, J = 7.6 Hz, 1H), 7.12 (d, J = 7.6 Hz, 2H), 6.33 (s, 1H), 5.79 (dd, J = 11.8, 4.2 Hz, 1H), 4.71 (t, J = 11.6 Hz, 1H), 4.51 (d, J = 11.5 Hz, 1H), 3.88 – 3.77 (m, 1H), 3.47 – 3.36 (m, 1H), 3.08 – 2.93 (m, 1H), 2.46 – 2.38 (m, 1H), 2.30 – 2.18 (m, 1H), 2.12 (s, 6H), 2.00 – 1.93 (m, 1H), 1.89 – 1.79 (m, 2H), 1.40 (dd, J = 10.2, 6.9 Hz, 6H), 1.37 – 1.32 (m, 3H), 1.29 (s, 1H), 1.18 – 1.11 (m, 1H), 1.08 – 0.99 (m, 1H), 0.86 – 0.70 (m, 5H), 0.64 (d, J = 6.6 Hz, 3H). ESI-MS m/z calc.748.3407, found 749.5 (M+1)+; Retention time: 2.09 minutes; and the second eluting peak 2, diastereomer 2, as a white solid (8S,9R)-4-(7-cyclopropyl-6-isopropyl-furo[2,3-b]pyrazin-2- yl)-14-(2,6-dimethylphenyl)-8-isobutyl-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (7.5 mg, 27%), 1H NMR (400 MHz, CDCl3) δ 8.64 (t, J = 1.8 Hz, 1H), 8.49 (s, 1H), 8.26 (s, 1H), 7.91 (dt, J = 8.0, 1.4 Hz, 1H), 7.81 (dt, J = 7.7, 1.4 Hz, 1H), 7.58 (t, J = 7.8 Hz, 1H), 7.24 (t, J = 7.6 Hz, 1H), 7.10 (d, J = 7.6 Hz, 2H), 6.33 (s, 1H), 5.82 (dd, J = 11.8, 5.5 Hz, 1H), 5.72 (dd, J = 13.0, 4.4 Hz, 1H), 5.06 (t, J = 12.0 Hz, 1H), 4.52 – 4.42 (m, 1H), 3.51 – 3.36 (m, 1H), 2.34 – 2.21 (m, 1H), 2.21 – 2.13 (m, 1H), 2.08 (s, 6H), 1.95 – 1.86 (m, 3H), 1.78 (t, J = 12.5 Hz, 1H), 1.71 – 1.65 (m, 1H), 1.40 (dd, J = 6.9, 3.9 Hz, 6H), 1.34 – 1.27 (m, 2H), 1.23 – 1.13 (m, 3H), 1.09 – 1.02 (m, 1H), 0.87 – 0.77 (m, 1H), 0.69 – 0.61 (m, 6H). ESI-MS m/z calc.748.3407, found 749.5 (M+1)+; Retention time: 2.14 minutes. LC method C. Example 51: Preparation of Compound 27 Step 1: 3-[[4-[[(2S,3R,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin- 2-yl]sulfamoyl]benzoic acid
[00447] A solution of 3-[[4-Chloro-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2- yl]sulfamoyl]benzoic acid (89 mg, 0.2042 mmol) and [(2S,3R,7S)-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methanol (69 mg, 0.1842 mmol) in THF (950 µL) was stirred for 5 minutes in a nitrogen-purged screwcap vial. Sodium tert-butoxide (120 mg, 1.249 mmol) was added and the reaction was stirred at rt for 90 minutes. The reaction mixture was then partitioned between 25 mL 1M HCl and 30 mL ethyl acetate. The layers were separated and the aqueous was extracted with an additional 20 mL ethyl acetate. The aqueous was then diluted with an equal volume of brine and extracted with an additional 2 x 40 mL ethyl acetate. The combined organics were washed with brine, dried over sodium sulfate. The crude material was purified by silica chromatography (40 g of silica) using a gradient of 100% dichloromethane to 20% methanol in dichloromethane. The desired fractions were concentrated under reduced pressure, and dried under high vacuum to give a light yellow solid 3-[[4- [[(2S,3R,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2- yl]methoxy]-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (134.6 mg, 90%). ESI-MS m/z calc.773.3371, found 774.6 (M+1)+; Retention time: 1.69 minutes. LC method A. Step 2: (4S,8R,9S)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-13-fluoro-8-isopropoxy-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one
[00448] 3-[[4-[[(2S,3R,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)-5-fluoro-pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (134.6 mg, 0.1739 mmol) was azeotroped twice with toluene under argon, and then was dissolved in NMP (5.5 mL). TsCl (86 mg, 0.4511 mmol) was added. The reaction was stirred for 15 minutes at room temperature. Then NMM (104 µL, 0.9459 mmol) was added and the reaction was stirred at room temperature for 20 h. The mixture was diluted with HCl (500 µL of 4 M, 2.000 mmol) followed by EtOAc, then the aqueous phase was extracted with EtOAc (2x). The organic layers were combined, washed with brine, and then concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 30 min) to give as an off-white solid (4S,8R,9S)-4- (6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-13-fluoro-8- isopropoxy-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (42.0 mg, 32%), 1H NMR (400 MHz, CDCl3) δ 8.75 (d, J = 1.8 Hz, 1H), 8.51 (s, 1H), 7.90 (t, J = 7.4 Hz, 2H), 7.63 (t, J = 7.8 Hz, 1H), 7.28 – 7.25 (m, 1H), 7.17 (d, J = 7.6 Hz, 1H), 7.11 (d, J = 7.6 Hz, 1H), 6.54 (s, 1H), 6.05 (dd, J = 12.9, 4.9 Hz, 1H), 5.78 (dd, J = 11.7, 5.5 Hz, 1H), 4.95 (t, J = 12.0 Hz, 1H), 4.79 – 4.69 (m, 1H), 4.15 (s, 3H), 3.68 – 3.62 (m, 1H), 3.46 – 3.36 (m, 1H), 2.45 – 2.36 (m, 1H), 2.18 (s, 3H), 2.15 – 2.01 (m, 2H), 2.01 (s, 3H), 1.99 – 1.79 (m, 3H), 1.53 (s, 9H), 1.02 (d, J = 6.1 Hz, 3H), 0.67 (d, J = 5.9 Hz, 3H) plus 1H exchangeable not observed. ESI- MS m/z calc.755.32654, found 756.6 (M+1)+; Retention time: 2.11 minutes. LC method A. Example 52: Preparation of Compound 31 Step 1: tert-Butyl (4R)-4-[(1S)-5-[6-tert-butyl-7-(2-methoxyethyl)-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl]-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate [00449] To a solution of 1-[6-tert-butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3-b]pyrazin- 3-yl]-2-dimethoxyphosphoryl-ethanone (358.4 mg, 0.9018 mmol) in isopropanol (15.0 mL) was added potassium carbonate (596.7 mg, 4.317 mmol) and left stirring for 5 min. This solution was then added to a solution of tert-butyl (4R)-2,2-dimethyl-4-[(1R)-3-methyl-1-(2- oxoethyl)butyl]oxazolidine-3-carboxylate (302.2 mg, 0.7713 mmol) in isopropanol (15 mL). Palladium on carbon (878.4 mg of 10 %w/w, 0.8254 mmol) was added to the reaction mixture and the content was placed under hydrogen atmosphere using a balloon filled with hydrogen gas and left stirring overnight. The reaction mixture was filtered on a pad of Celite, the cake was washed with methanol, concentrated to give tert-butyl (4R)-4-[(1S)-5-[6-tert-butyl-7-(2- methoxyethyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (452.6 mg, 100%) as an orange solid. ESI-MS m/z calc.586.4094, found 587.2 (M+1)+; Retention time: 2.28 minutes. LC method C. Step 2: [(2R,3S)-7-[6-tert-Butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl]-3-isobutyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00450] To a stirred solution of tert-butyl (4R)-4-[(1S)-5-[6-tert-butyl-7-(2-methoxyethyl)-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl]-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (452.6 mg, 0.7713 mmol) in a mixture of dioxane (81 mL) and water (41.0 mL) was added a solution of hydrogen chloride (19.3 mL of 4 M, 77.20 mmol) dropwise. After stirring for 2 hours, the reaction was not complete and additional hydrogen chloride (19.3 mL of 4 M, 77.20 mmol) was added. The reaction was stirred at room temperature overnight, then the reaction was concentrated to dryness. The residual water was co-evaporated with acetonitrile (5 x 25 mL), then dried under high vacuum to afford crude [(2R,3S)-7-[6-tert-butyl-7-(2- methoxyethyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-3-isobutyl-3,4,5,6-tetrahydro-2H-azepin-2- yl]methanol (330.9 mg, 100%) as an orange oil. ESI-MS m/z calc.428.31512, found 429.3 (M+1)+; Retention time: 0.69 minutes. LC method C. Step 3: [(2R,3S)-7-[6-tert-Butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl]-3-isobutyl-azepan-2-yl]methanol [00451] To a solution of [(2R,3S)-7-[6-tert-butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl]-3-isobutyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (330.9 mg, 0.7720 mmol) in anhydrous dichloromethane (17 mL) at room temperature was added sodium triacetoxyborohydride (820.4 mg, 3.871 mmol). The reaction was stirred at room temperature for 1.5 hours and a saturated aqueous solution of sodium bicarbonate (100 mL) was added. The phases were separated, and the aqueous solution was extracted with DCM (2 x). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 30 min) to give [(2R,3S)-7-[6-tert-butyl-7-(2- methoxyethyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-3-isobutyl-azepan-2-yl]methanol (50.2 mg, 15%) as a yellow oil. ESI-MS m/z calc.430.33078, found 431.6 (M+1)+; Retention time: 1.35 minutes. LC method A. Step 4: 3-[[4-[[(2R,3S)-7-[6-tert-Butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl]-3-isobutyl-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid
[00452] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (48.72 mg, 0.1166 mmol) and [(2R,3S)-7-[6-tert-butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl]-3-isobutyl-azepan-2-yl]methanol (50.2 mg, 0.1166 mmol) were combined in a flask and dissolved in THF (1.2 mL) followed by the addition of sodium tert-butoxide (68.8 mg, 0.7159 mmol). The reaction mixture was stirred for 30 minutes at room temperature then warmed to 40 °C for 1 hour. The reaction mixture was cooled to room temperature then diluted with EtOAc and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (5X). The organic layers were combined, washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3-[[4-[[(2R,3S)-7- [6-tert-butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-3-isobutyl-azepan-2- yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (72.9 mg, 74%) as a yellow solid. ESI-MS m/z calc.811.4091, found 812.3 (M+1)+; Retention time: 1.67 minutes. LC method A. Step 5: (8S,9R)-4-[6-tert-Butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl]-14-(2,6-dimethylphenyl)-8-isobutyl-18,18-dioxo-11-oxa-18λ6- thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one, diastereomer 2, compound 31
Diastereomer 2 [00453] 3-[[4-[[(2R,3S)-7-[6-tert-Butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3-b]pyrazin- 3-yl]-3-isobutyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (72.9 mg, 0.08977 mmol) was dissolved in NMP (4.4 mL) and TsCl (35.5 mg, 0.1862 mmol) was added. The reaction was stirred for 10 minutes at room temperature. After 10 minutes, NMM (39.5 µL, 0.3593 mmol) was added and the reaction was stirred at room temperature overnight. The reaction mixture was diluted with 1M HCl and extracted with ethyl acetate. The layers were separated, and the aqueous layer was further extracted with ethyl acetate (2 x). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to provide (8S,9R)-4-[6-tert-butyl-7-(2-methoxyethyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-14-(2,6- dimethylphenyl)-8-isobutyl-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one, diastereomer 2 (3.8 mg, 10%). The more polar and minor diastereomer 1 was not isolated.1H NMR (400 MHz, CDCl3) δ 8.77 (t, J = 1.8 Hz, 1H), 8.38 (s, 1H), 7.82 (d, J = 7.9 Hz, 1H), 7.73 (dt, J = 7.8, 1.4 Hz, 1H), 7.50 (t, J = 7.8 Hz, 1H), 7.15 (t, J = 7.6 Hz, 1H), 7.01 (d, J = 7.6 Hz, 2H), 6.20 (s, 1H), 5.89 – 5.77 (m, 2H), 4.84 (t, J = 11.9 Hz, 1H), 4.38 – 4.31 (m, 1H), 4.06 (s, 3H), 3.58 – 3.44 (m, 2H), 3.33 – 3.26 (m, 4H), 2.29 – 2.20 (m, 1H), 2.14 – 2.04 (m, 1H), 1.98 (s, 6H), 1.53 (s, 9H), 1.18 (s, 6H), 1.17 – 1.10 (m, 3H), 0.59 (t, J = 5.8 Hz, 6H). ESI-MS m/z calc. 793.39856, found 794.5 (M+1)+; Retention time: 2.27 minutes. LC method A. Example 53: Preparation of Compound 35 Step 1: tert-Butyl (4R)-4-[(E,1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin- 3-yl)-1-isopropyl-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00454] Nitrogen was bubbled through a stirred solution of 1-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-2-dimethoxyphosphoryl-ethanone (524 mg, 1.5427 mmol) in isopropanol (65 mL) for 15 minutes at which time grounded potassium carbonate (1.05 g, 7.5974 mmol) was added. The reaction was stirred at room temperature for 30 minutes at which time a solution of tert-butyl (4R)-2,2-dimethyl-4-[(1S)-2-methyl-1-(2- oxoethyl)propyl]oxazolidine-3-carboxylate (537 mg, 1.5425 mmol) in isopropanol (65 mL) was added dropwise. The reaction was stirred at room temperature for 178 hours at which time additional potassium carbonate (1.05 g, 7.5974 mmol) was added. The reaction was stirred overnight at room temperature, then filtered on a pad of celite, washed with ethyl acetate (3 x 25 mL) and concentrated under reduced pressure. The residue was dissolved in ethyl acetate (50 mL), washed with a saturated aqueous solution of ammonium chloride (2 x 50 mL), brine (50 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude was purified twice by reverse-phase chromatography on C18 (gradient: 5 to 100% acetonitrile in water, 15 CV) The desired fractions were concentrated under reduced pressure to afford tert-butyl (4R)-4-[(E,1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1- isopropyl-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (300 mg, 36%) as a bright yellow oil.1H NMR (400 MHz, CDCl3) δ 9.21 (s, 1H), 7.70 (d, J = 15.2 Hz, 1H), 7.34 – 7.23 (overlapped with CDCl3, m, 1H), 6.67 (s, 1H), 4.11 (s, 3H), 3.96 – 3.67 (m, 3H), 2.58 – 2.41 (m, 2H), 1.64 – 1.35 (overlapped with water, m, 26H), 1.02 (d, J = 7.1 Hz, 3H), 0.96 (d, J = 6.8 Hz, 3H). ESI-MS m/z calc.512.3363, found 513.4 (M+1)+; Retention time: 1.86 minutes. LC method U. Step 2: tert-Butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-1-isopropyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00455] Nitrogen was bubbled through a stirred solution of tert-butyl (4R)-4-[(E,1S)-5-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-isopropyl-5-oxo-pent-3-enyl]-2,2-dimethyl- oxazolidine-3-carboxylate (300 mg, 0.5559 mmol) in anhydrous methanol (10 mL) for 10 minutes at which time palladium on carbon (60 mg, 10 %w/w, 0.0564 mmol) was then added. Nitrogen was bubbled through the mixture for 10 minutes, followed by hydrogen for 15 minutes. The reaction was then stirred overnight at room temperature under hydrogen atmosphere. Nitrogen was bubbled through the mixture for 10 minutes and the mixture was filtered on a pad of celite, washed with ethyl acetate (5 x 10 mL) and concentrated under reduced pressure to afford a pale-yellow gel. The crude was purified by reverse-phase chromatography (gradient: 5 to 100% acetonitrile in water; 15 CV). The desired fractions were concentrated under reduced pressure, then dried under high vacuum to afford tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-isopropyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (127 mg, 42%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 9.11 (s, 1H), 6.69 (s, 1H), 4.21 – 3.63 (m, 7H), 3.37 – 3.18 (m, 2H), 1.93 – 1.73 (m, 4H), 1.66 – 1.39 (overlapped with water, m, 25H), 0.98 (d, J = 6.8 Hz, 3H), 0.90 (d, J = 6.8 Hz, 3H). ESI-MS m/z calc. 514.3519, found 515.4 (M+1)+; Retention time: 2.15 minutes. LC method Q. Step 3: [(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00456] To a stirred solution of tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-isopropyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (127 mg, 0.2344 mmol) in diethyl ether (2.5 mL) was added a solution of hydrogen chloride in diethyl ether (3 mL of 2 M, 6.0000 mmol) and water (0.650 mL). The reaction was stirred at room temperature overnight then neutralized with a saturated aqueous solution of sodium bicarbonate (5 mL) then concentrated under reduced pressure to remove the volatiles. The aqueous layer was extracted with dichloromethane (5 x 5 mL) and the combined organic layers were washed with brine (20 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure then dried under high vacuum. The crude was purified by reverse-phase chromatography on C18 (gradient: 5 to 100% methanol in water; 12CV). The desired fractions were concentrated under reduced pressure and the remaining water was co-evaporated with methanol (6 x 5 mL). Further drying under high vacuum afforded [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-3-isopropyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (43 mg, 49%) as a light yellow film. ESI-MS m/z calc.356.2576, found 357.4 (M+1)+; Retention time: 1.41 minutes. LC method I. Step 4: [(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropyl-azepan-2-yl]methanol, diastereomer 1, and [(2R,3S)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropyl-azepan-2-yl]methanol, diastereomer 2 Diastereomer 1 Diastereomer 2 [00457] To a stirred solution of [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-isopropyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (41 mg, 0.1093 mmol) in anhydrous DCM (5 mL) was added sodium triacetoxyborohydride (125 mg, 0.5898 mmol). The reaction was stirred overnight at room temperature then quenched by the addition of a saturated aqueous solution of sodium bicarbonate (10 mL). The biphasic mixture was vigorously stirred at room temperature for 30 minutes. The layers were then separated, and the aqueous layer was extracted with DCM (5 x 5 mL). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude was purified by reverse-phase chromatography on C18 (gradient: 5 to 100% acetonitrile in 10 mM ammonium bicarbonate buffer pH = 10; 25 CV). The desired fractions were concentrated under reduced pressure and the remaining water was co-evaporated with acetonitrile (5 x 5 mL) then freeze- dried to afford a white fluffy powder. The 1:1 diastereomeric mixture was subjected to SFC separation using the following conditions: a Cellulose 4 column (250 x 30 mm), 5 μM column at 40 °C, eluent: 40% methanol and 60% CO2 (0.1% diethylamine), flow rate: 150 mL/min, concentration : 4.6 mg/mL in methanol (no modifier), injection volume : 100 μL, pressure : 100 bars, wavelength: 210 nm. Evaporation of the solvent, followed by freeze-drying of the two fractions afforded the two diastereomers as follow: diastereomer 1, SFC Peak 1: [(2R,3S)-7-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropyl-azepan-2-yl]methanol (8.9 mg, 21%) as a brown fluffy powder.1H NMR (400 MHz, C6D6) δ 8.53 (s, 1H), 6.60 (s, 1H), 3.96 – 3.86 (m, 1H), 3.85 – 3.76 (m, 1H), 3.56 (s, 3H), 3.34 – 3.20 (m, 1H), 2.82 – 2.70 (m, 1H), 1.99 – 1.82 (m, 2H), 1.67 – 1.51 (m, 4H), 1.38 – 1.27 (m, 4H), 1.12 (s, 9H), 0.77 (d, J = 6.8 Hz, 3H), 0.69 (d, J = 6.8 Hz, 3H). ESI-MS m/z calc.358.2733, found 359.3 (M+1)+; Retention time: 3.84 minutes; and diastereomer 2, SFC Peak 2 : [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-isopropyl-azepan-2-yl]methanol (5.9 mg, 14%) as an off-white fluffy powder. 1H NMR (400 MHz, C6D6) δ 8.59 (br. S, 1H), 6.58 (s, 1H), 3.98 – 3.79 (m, 2H), 3.60 (s, 3H), 3.53 – 3.45 (m, 1H), 3.06 – 2.89 (m, 1H), 2.15 – 2.01 (m, 1H), 1.87 – 1.75 (m, 1H), 1.58 – 1.21 (m, 8H), 1.09 (s, 9H), 0.81 (d, J = 6.6 Hz, 3H), 0.70 (d, J = 6.4 Hz, 3H). ESI-MS m/z calc. 358.2733, found 359.3 (M+1)+; Retention time: 3.92 minutes. LC method V. Step 5: 3-[[4-[[(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid Diastereomer 1 [00458] [(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropyl-azepan-2- yl]methanol (diastereomer 1, 6.7 mg, 0.01869 mmol) was combined with 3-[[4-chloro-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (7.9 mg, 0.01891 mmol) in THF (0.2 mL) in a nitrogen-purged screwcap vial and stirred for 5 minutes at room temperature. Sodium tert-butoxide (12 mg, 0.1249 mmol) was added and the reaction was placed in an aluminum heating block at 35 °C for 3 hours. The reaction mixture was then partitioned between 15 mL 1M HCl and 15 mL ethyl acetate. The layers were separated and the aqueous was extracted with an additional 10 ml ethyl acetate. The aqueous was then diluted with an equal volume of brine and extracted with an additional 2 x 10 mL ethyl acetate. The combined organics were washed with brine, dried over sodium sulfate, and concentrated to give 3-[[4-[[(2R,3S)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropyl-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (diastereomer 1, 14.5 mg, 99%). ESI-MS m/z calc.739.35156, found 740.6 (M+1)+; Retention time: 0.63 minutes; LC method B. Step 6: (8S,9R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-8-isopropyl-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one, diastereomer 1, compound 35 Diastereomer 1 Diastereomer 1 [00459] 3-[[4-[[(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropyl- azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (diastereomer 1, 14.5 mg, 0.01868 mmol) was combined with Tosyl Chloride (approximately 10.68 mg, 0.05604 mmol) in anhydrous NMP (2 mL) in a nitrogen- purged vial. N-methylmorpholine (approximately 11.34 mg, 12.33 µL, 0.1121 mmol) was added and the reaction was stirred for 18 hours at room temperature. The reaction mixture was then partitioned between 1M HCl and ethyl acetate. The layers were separated, and the aqueous layer was extracted 3x with ethyl acetate. The combined organics were washed with water and brine, dried over sodium sulfate and concentrated. The resulting crude material was purified by preparative HPLC (10-99% I in water, HCl modifier, 20 minute run) to give (8S,9R)-4-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-8-isopropyl-18,18-dioxo- 11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (Diastereomer 1, 5.0 mg, 37%). ESI-MS m/z calc. 721.341, found 722.6 (M+1)+; Retention time: 2.04 minutes; LC method A.1H NMR (400 MHz, CDCl3) δ 8.84 (s, 1H), 8.47 (s, 1H), 8.06 (d, J = 8.0 Hz, 1H), 7.85 (d, J = 7.7 Hz, 1H), 7.65 (t, J = 7.8 Hz, 1H), 7.23 (t, J = 7.6 Hz, 1H), 7.10 (d, J = 7.6 Hz, 2H), 6.93 (s, 1H), 6.26 (s, 1H), 6.06 – 5.99 (m, 1H), 5.78 (dd, J = 11.4, 4.9 Hz, 1H), 4.73 (t, J = 11.3 Hz, 1H), 4.44 – 4.38 (m, 1H), 4.25 (s, 3H), 2.36 – 2.23 (m, 2H), 2.05 (s, 6H), 2.00 – 1.93 (m, 1H), 1.90 – 1.85 (m, 1H), 1.79 – 1.75 (m, 1H), 1.71 – 1.67 (m, 2H), 1.55 (s, 9H), 1.17 – 1.12 (m, 1H), 0.65 (d, J = 6.7 Hz, 3H), 0.44 (d, J = 6.7 Hz, 3H). Example 54: Preparation of Compound 36 Step 1: 3-[[4-[[(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid Diastereomer 2 [00460] [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropyl-azepan-2- yl]methanol (diastereomer 2, 4.5 mg, 0.01255 mmol) was combined with 3-[[4-chloro-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (5.3 mg, 0.01268 mmol) in THF (0.2 mL) in a nitrogen-purged screwcap vial and stirred for 5 minutes at room temperature. Sodium tert-butoxide (9 mg, 0.09365 mmol) was added and the reaction was placed in an aluminum heating block at 35 °C for 7 hours. The reaction mixture was then partitioned between 15 mL 1M HCl and 15 mL ethyl acetate. The layers were separated and the aqueous was extracted with an additional 10 ml ethyl acetate. The aqueous was then diluted with an equal volume of brine and extracted with an additional 2 x 10 mL ethyl acetate. The combined organics were washed with brine, dried over sodium sulfate, and concentrated to give 3-[[4-[[(2R,3S)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropyl-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (diastereomer 2, 9.5 mg, 97%). ESI-MS m/z calc.739.35156, found 740.6 (M+1)+; Retention time: 0.62 minutes; LC method B. Step 2: (8S,9R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-8-isopropyl-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one, diastereomer 2, compound 36 Diastereomer 2 Diastereomer 2 [00461] 3-[[4-[[(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropyl- azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (diastereomer 2, 9.5 mg, 0.01224 mmol) was combined with tosyl Chloride (approximately 7.000 mg, 0.03672 mmol) in anhydrous NMP (1.5 mL) in a nitrogen-purged vial. N-methylmorpholine (approximately 7.428 mg, 8.074 µL, 0.07344 mmol) was added and the reaction was stirred for 18 hours at room temperature. The reaction mixture was then partitioned between 1M HCl and ethyl acetate. The layers were separated, and the aqueous phase was extracted with 3x ethyl acetate. The combined organics were washed with water and brine, dried over sodium sulfate and concentrated. The resulting crude material was the purified by preparative HPLC (10-99% I in water, HCl modifier, 20 minute run) to give (8S,9R)-4-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-8-isopropyl-18,18- dioxo-11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (diastereomer 2, 1.6 mg, 18%). ESI-MS m/z calc. 721.341, found 722.6 (M+1)+; Retention time: 1.93 minutes; LC method A. Example 55: Preparation of Compound 41 Step 1: tert-Butyl (4R)-4-[(E,1S)-1-tert-butyl-5-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3- carboxylate [00462] Nitrogen was bubbled through a stirred solution of 1-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-2-dimethoxyphosphoryl-ethanone (265.7 mg, 0.7822 mmol) in isopropanol (30 mL) for 10 minutes at which time grounded potassium carbonate (538.1 mg, 3.8935 mmol) was added. The reaction was stirred at room temperature for 30 minutes at which time a solution of tert-butyl (4R)-4-[(1S)-2,2-dimethyl-1-(2-oxoethyl)propyl]-2,2-dimethyl- oxazolidine-3-carboxylate (295.6 mg, 0.7545 mmol) in isopropanol (30 mL) was added dropwise. The reaction was stirred at room temperature for 24 hours and was then concentrated in vacuo. Water (50 mL) and EtOAc (50 mL) were added, and the mixture was extracted with EtOAc (3 x 25 mL). The combined organic layers were washed with brine (50 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 50 to 100% of MeCN in acidic water (0.1% v/v of formic acid in water). The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (25 mL) was then added, and the mixture was extracted with EtOAc (3 x 25 mL). The combined organic layers were washed with brine (1 x 50 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford tert- butyl (4R)-4-[(E,1S)-1-tert-butyl-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-5-oxo- pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (Z/E mixture of isomers, 306.2 mg, 77%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 9.31 – 9.11 (m, 1H), 7.73 – 7.59 (m, 1H), 6.68 (s, 1H), 4.10 (s, 3H), 4.04 (dd, J = 8.8, 5.6 Hz, 1H), 3.82 (d, J = 8.6 Hz, 1H), 2.77 – 2.59 (m, 1H), 2.57 – 2.39 (m, 1H), 1.85 – 1.75 (m, 1H), 1.64 – 1.49 (m, 16H), 1.45 (d, J = 13.0 Hz, 9H), 1.00 (s, 9H). ESI-MS m/z calc.526.3519, found 527.4 (M+1)+; Retention time: 2.26 minutes. LC method T. Step 2: tert-Butyl (4R)-4-[(1S)-1-tert-butyl-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00463] Nitrogen was bubbled to a stirred solution of tert-butyl (4R)-4-[(E,1S)-1-tert-butyl-5- (6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-5-oxo-pent-3-enyl]-2,2-dimethyl- oxazolidine-3-carboxylate (301.8 mg, 0.5724 mmol) in dry methanol (10 mL) for 10 minutes, after which time palladium on carbon (68.7 mg, 10 %w/w, 0.0646 mmol) was added. Nitrogen was bubbled to the mixture for 10 minutes, followed by hydrogen for 15 minutes. The reaction was then stirred for 6 hours at room temperature under hydrogen atmosphere. Nitrogen was bubbled to the mixture for 10 minutes. The mixture was filtered on a pad of Celite, washed with MeOH (2 x 25 mL) and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 50 to 100% of MeCN in acidic water (0.1% v/v of formic acid in water). The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (25 mL) was then added, and the mixture was extracted with EtOAc (3 x 25 mL). The combined organic layers were washed with brine (1 x 50 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford tert- butyl (4R)-4-[(1S)-1-tert-butyl-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-5-oxo- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (140.2 mg, 43%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 9.10 (s, 1H), 6.61 (s, 1H), 4.16 – 3.98 (m, 5H), 3.75 (d, J = 8.1 Hz, 1H), 3.39 – 3.11 (m, 2H),1.96 – 1.74 (m, 2H), 1.59 – 1.29 (m, 27H), 0.95 (s, 9H). ESI-MS m/z calc. 528.3676, found 429.4 (M-99)+; Retention time: 2.3 minutes. LC method T. Step 3: (5S)-5-[(1R)-1-Amino-2-hydroxy-ethyl]-1-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-6,6-dimethyl-heptan-1-one [00464] To a solution of tert-butyl (4R)-4-[(1S)-1-tert-butyl-5-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (151.5 mg, 0.2722 mmol) in a mixture of dioxane (1.5 mL) and water (0.15 mL) at room temperature was added dropwise a solution of HCl (in 1,4-dioxane solution) (0.7 mL of 4 M, 2.8000 mmol). The reaction was stirred at room temperature for 1 hour. A saturated solution of sodium bicarbonate (5 mL, until pH = 7) was added. Water (10 mL) and DCM (10 mL) were added, and the mixture was extracted with DCM (3 x 10 mL). The combined organic layers were washed with brine (25 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeCN in acidic water (0.1% v/v of formic acid in water). The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (10 mL) was then added, and the mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with a saturated solution of sodium bicarbonate (1 x 25 mL) and brine (1 x 25 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford (5S)-5-[(1R)-1- amino-2-hydroxy-ethyl]-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-6,6-dimethyl- heptan-1-one (90.2 mg, 85%) as a yellow solid. ESI-MS m/z calc.388.2838, found 389.2 (M+1)+; Retention time: 1.42 minutes. LC method I. Step 4: (5S)-5-[(1R)-1-Amino-2-[tert-butyl(diphenyl)silyl]oxy-ethyl]-1-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-6,6-dimethyl-heptan-1-one [00465] To a stirred solution of (5S)-5-[(1R)-1-amino-2-hydroxy-ethyl]-1-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-6,6-dimethyl-heptan-1-one (89.8 mg, 0.2309 mmol) in DCM (1.5 mL) at 0 °C was added imidazole (49.1 mg, 0.7212 mmol) followed by tert-butyl-chloro- diphenyl-silane (103.88 mg, 0.1 mL, 0.3704 mmol). The mixture was stirred at 0 °C for 5 minutes and then at room temperature for 2 hours. Water (10 mL) and DCM (10 mL) were added, and the mixture was extracted with DCM (3 x 10 mL). The combined organic layers were washed with brine (20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeCN in water and then of 100% of MeOH in water to afford (5S)-5-[(1R)-1-amino-2-[tert-butyl(diphenyl)silyl]oxy-ethyl]-1-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-6,6-dimethyl-heptan-1-one (130.8 mg, 90%) as a yellow solid.1H NMR (400 MHz, CDCl3) δ 8.84 (s, 1H), 7.51 – 7.43 (m, 4H), 7.36 – 7.31 (m, 2H), 7.25 – 7.18 (m, 4H), 6.45 (s, 1H), 5.88 (d, J = 4.6 Hz, 1H), 5.58 (d, J = 5.4 Hz, 1H), 4.11 – 3.96 (m, 2H), 3.93 (s, 3H), 3.62 – 3.53 (m, 1H), 3.33 (dd, J = 9.9, 4.3 Hz, 1H), 2.52 (dt, J = 18.2, 6.5 Hz, 1H), 2.32 – 2.07 (m, 2H), 1.77 – 1.68 (m, 1H), 1.50 (s, 9H), 0.93 (s, 9H), 0.76 (s, 9H).2 missing labile protons. ESI-MS m/z calc.626.4016, found 627.4 (M+1)+; Retention time: 1.78 minutes. LC method I. Step 5: tert-Butyl-[[(2R,3S)-3-tert-butyl-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)azepan-2-yl]methoxy]-diphenyl-silane [00466] To a solution of (5S)-5-[(1R)-1-amino-2-[tert-butyl(diphenyl)silyl]oxy-ethyl]-1-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-6,6-dimethyl-heptan-1-one (127.4 mg, 0.2030 mmol) in DCM (10 mL) at room temperature was added NaBH(Oac)3 (195 mg, 0.9201 mmol). The mixture was stirred for 2 hours and a saturated aqueous solution of sodium bicarbonate (10 mL, until pH = 7) was added. The layers were separated, and the aqueous layer was extracted with DCM (3 x 25 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeCN in acidic water (0.1% v/v of formic acid in water). The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (10 mL) was then added, and the mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with a aqueous solution of sodium bicarbonate (1 x 25 mL) and brine (1 x 25 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford tert-butyl-[[(2R,3S)-3-tert-butyl-7-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)azepan-2-yl]methoxy]-diphenyl-silane (79.4 mg, 61%) as a yellow solid.1H NMR (400 MHz, CDCl3) δ 8.50 – 8.38 (m, 1H), 7.66 – 7.57 (m, 4H), 7.43 – 7.29 (m, 4H), 7.25 – 7.19 (m, 2H), 6.56 – 6.45 (m, 1H), 4.06 – 3.98 (m, 1H), 3.97 – 3.89 (m, 3H), 3.86 – 3.74 (m, 1H), 3.70 – 3.57 (m, 1H), 3.09 – 3.01 (m, 1H), 1.98 – 1.86 (m, 2H), 1.83 – 1.71 (m, 2H), 1.58 – 1.44 (m, 11H), 1.22 – 1.16 (m, 1H), 0.98 – 0.91 (m, 9H), 0.88 – 0.81 (m, 9H).1 missing labile proton. ESI-MS m/z calc.610.4067, found 611.4 (M+1)+; Retention time: 1.76 minutes. LC method I. Step 6: [(2R,3S,7S)-3-tert-Butyl-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin- 3-yl)azepan-2-yl]methanol [00467] To a stirred solution of tert-butyl-[[(2R,3S)-3-tert-butyl-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)azepan-2-yl]methoxy]-diphenyl-silane (74.8 mg, 0.1170 mmol) in dry THF (1.5 mL) at room temperature was added TBAF (in THF solution) (0.2 mL of 1 M, 0.2000 mmol). The mixture was stirred at room temperature for 2.5 hours. Water (10 mL) and EtOAc (10 mL) were added, and the mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (25 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeCN in acidic water (0.1% v/v of formic acid in water). The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (10 mL) was then added, and the mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with a saturated solution of sodium bicarbonate (1 x 25 mL) and brine (1 x 25 mL), dried over sodium sulfate, filtered and concentrated in vacuo to afford after lyophilization [(2R,3S,7S)-3-tert-butyl-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)azepan-2-yl]methanol (27.9 mg, 64%) as a white solid.1H NMR (400 MHz, Benzene-d6) δ 8.53 (s, 1H), 6.60 (s, 1H), 4.01 (dd, J = 11.1, 3.1 Hz, 1H), 3.78 (dd, J = 10.6, 4.3 Hz, 1H), 3.59 (s, 3H), 3.37 (t, J = 10.4 Hz, 1H), 3.05 (ddd, J = 10.2, 6.2, 4.4 Hz, 1H), 2.01 – 1.92 (m, 1H), 1.86 – 1.66 (m, 3H), 1.62 – 1.48 (m, 2H), 1.09 (s, 9H), 1.03 – 0.98 (m, 1H), 0.86 (s, 9H).2 missing labile protons. ESI-MS m/z calc.372.2889, found 373.3 (M+1)+; Retention time: 2.55 minutes. LC method J. Step 7: 3-[[4-[[(2R,3S,7S)-3-tert-Butyl-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00468] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (28 mg, 0.06701 mmol) and [(2R,3S,7S)-3-tert-butyl-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)azepan-2-yl]methanol (23.9 mg, 0.06415 mmol) in THF (300 µL) in a nitrogen-purged screwcap vial and stirred for 5 minutes followed by the addition of sodium tert-butoxide (40.5 mg, 0.4214 mmol) was added and the reaction was placed in an aluminum heating block at 35 °C for 3 h. An additional portion of sodium tert-butoxide (15.6 mg, 0.1623 mmol) was added, followed by heating at 45 °C for 1 additional hour. The reaction mixture was cooled and then partitioned between 15 mL 1M HCl and 15 mL ethyl acetate. The layers were separated and the aqueous was extracted with an additional 10 mL ethyl acetate. The aqueous was then diluted with an equal volume of brine and extracted an additional 2 x 10 mL ethyl acetate. The combined organics were washed with brine, dried over sodium sulfate, and concentrated to give a white solid 3-[[4-[[(2R,3S,7S)-3-tert-butyl-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (46 mg, 91%). ESI-MS m/z calc.753.36725, found 754.5 (M+1)+; Retention time: 1.58 minutes. LC method A. Step 8: (4S,8S,9R)-8-tert-Butyl-4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one [00469] Under argon 3-[[4-[[(2R,3S,7S)-3-tert-butyl-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (46 mg, 0.06101 mmol), azeotroped twice with toluene, was dissolved in NMP (2.0 mL) and then TsCl (30 mg, 0.1574 mmol) was added. The reaction was stirred for 15 minutes at room temperature. Then NMM (40 µL, 0.3638 mmol) was added and the reaction was stirred at room temperature for 18 h. The mixture was diluted with HCl (175 µL of 4 M, 0.7000 mmol) followed by EtOAc. Then the aqueous layer was extracted with EtOAc (2 x). The organic layers were combined, washed with brine, and then concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 30 min) to give as an off-white solid (4S,8S,9R)-8-tert-butyl-4-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen- 2-one (10.7 mg, 23%), 1H NMR (400 MHz, CDCl3) δ 8.98 (s, 1H), 8.44 (s, 1H), 8.02 (d, J = 7.8 Hz, 1H), 7.80 (d, J = 7.4 Hz, 1H), 7.59 (t, J = 7.7 Hz, 1H), 7.21 (d, J = 7.7 Hz, 1H), 7.09 (d, J = 7.5 Hz, 2H), 6.50 (s, 1H), 6.21 (s, 1H), 6.07 (dd, J = 11.9, 4.7 Hz, 1H), 5.92 (d, J = 13.0 Hz, 1H), 4.84 (t, J = 11.8 Hz, 1H), 4.53 – 4.45 (m, 1H), 4.17 (s, 3H), 2.41 – 2.33 (m, 1H), 2.24 – 2.18 (m, 1H), 2.05 (s, 6H), 1.92 – 1.88 (m, 1H), 1.83 – 1.80 (m, 1H), 1.71 – 1.68 (m, 1H), 1.49 (s, 9H), 1.40 (s, 1H), 1.26 (s, 1H), 0.55 (s, 9H)plus 1H exchangeable not observed. ESI-MS m/z calc.735.3567, found 736.6 (M+1)+; Retention time: 2.08 minutes. LC method A. Example 56: Preparation of Compound 43 Step 1: tert-Butyl (4S)-4-[(1R)-5-hydroxy-1-isopropoxy-5-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate ~85:15 mixture of diastereomers, the major isomer having the 4S,1R configuration [00470] To a solution of 3-bromo-5-methyl-6-[1-(trifluoromethyl)cyclopropyl] pyrrolo[2,3- b]pyrazine (3.69 g, 11.53 mmol) in tetrahydrofuran (40.0 mL) at -78 °C was added n- butyllithium (solution in hexanes) (5.4 mL of 2.5 M, 13.50 mmol) dropwise. As soon as the addition of n-BuLi was over, a solution of tert-butyl (4S)-4-[(1R)-1-isopropoxy-5-oxo-pentyl]- 2,2-dimethyl-oxazolidine-3-carboxylate (~85:15 mixture of diastereomers, the major isomer having the 4S,1R configuration, 3.3 g, 9.608 mmol) in tetrahydrofuran (40 mL) was added dropwise. Reaction was left stirring for 10 min and MeOH (5 mL) was added. The crude mixture was concentrated in vacuo and was then purified by flash chromatography on silica gel using 0-100% EtOAc in hexanes over 25 minutes to give tert-butyl (4S)-4-[(1R)-5-hydroxy-1- isopropoxy-5-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]pentyl]- 2,2-dimethyl-oxazolidine-3-carboxylate (3.37 g, 60%) as an orange-white solid. ESI-MS m/z calc.584.31854, found 585.2 (M+1)+; Retention time: 1.72 minutes. LC method A. Step 2: tert-Butyl (4S)-4-[(1R)-1-isopropoxy-5-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00471] To a solution of tert-butyl (4S)-4-[(1R)-5-hydroxy-1-isopropoxy-5-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (3.37 g, 5.764 mmol) in dichloroethane (75 mL) was added manganese (IV) oxide (activated) (12.5 g, 122.2 mmol). The resulting suspension was stirred at 70 °C for 30 minutes. The suspension was filtered on a pad of Celite and washed with DCM (2 x). The crude mixture was concentrated in vacuo and was then purified by flash chromatography on silica gel using 0- 100% EtOAc in hexanes over 30 minutes to provide tert-butyl (4S)-4-[(1R)-1-isopropoxy-5-[5- methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate (3.358 g, 100%) as a yellow-white foam. ESI-MS m/z calc. 582.3029, found 583.1 (M+1)+; Retention time: 1.94 minutes. LC method A. Step 3: [(2S,3R)-3-Isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H- azepin-2-yl]methanol [00472] To a solution of tert-butyl (4S)-4-[(1R)-1-isopropoxy-5-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (3.358 g, 5.763 mmol) in 1,4-dioxane (77.2 mL) was added hydrochloric acid (solution in 1,4-dioxane) (29.0 mL of 4 M, 116.0 mmol) at rt. Water (7 mL) was added and the reaction was stirred overnight. The solvent was removed under reduced pressure and the residual water was co-evaporated with acetonitrile (3 x), then dried under high vacuum to afford crude [(2S,3R)-3-isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2- yl]methanol (2.446 g, 100%) as a yellow oil. ESI-MS m/z calc.424.20862, found 425.2 (M+1)+; Retention time: 1.06 minutes. The crude material was taken forward without further purification. LC method A. Step 4: [(2S,3R)-3-Isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol, diastereomer 1, and [(2S,3R)-3-isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol, diastereomer 2 Diastereomer 1 Diastereomer 2 [00473] To a solution of [(2S,3R)-3-isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2- yl]methanol (2.446 g, 5.763 mmol) in anhydrous dichloromethane (127 mL) at room temperature was added sodium triacetoxyborohydride (24.48 g, 115.5 mmol). The reaction was stirred at room temperature for 1 hour and a saturated aqueous solution of sodium bicarbonate (100 mL) was added. The phases were separated, and the aqueous solution was extracted with DCM (2 x). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 25 min) to give the two main isomers, diastereomer 1, Peak 1: [(2S,3R)-3-isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (629.5 mg, 26%) as a yellow solid. ESI-MS m/z calc.426.22427, found 427.5 (M+1)+; Retention time: 1.19 minutes. And diastereomer 2, Peak 2: [(2S,3R)-3-isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (363.7 mg, 13%) as a yellow solid. ESI-MS m/z calc.426.22427, found 427.5 (M+1)+; Retention time: 1.23 minutes. LC method A. Step 5: 3-[[4-(2,6-Dimethylphenyl)-6-[[(2S,3R)-3-isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2- yl]methoxy]pyrimidin-2-yl]sulfamoyl]benzoic acid, diastereomer 1 Diastereomer 1 [00474] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (606.7 mg, 1.452 mmol) and [(2S,3R)-3-isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol, diastereomer 1 (619.4 mg, 1.452 mmol) were combined in a flask and dissolved in THF (16 mL) followed by the addition of sodium tert-butoxide (840.1 mg, 8.741 mmol). The reaction mixture was stirred for 1 hour then heated to 40 °C for 30 minutes then diluted with EtOAc and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (5 x). The organic layers were combined, washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3-[[4-(2,6-dimethylphenyl)-6-[[(2S,3R)-3- isopropoxy-7-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2- yl]methoxy]pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt), diastereomer 1 (1.170 g, 95%) as an orange- white solid. ESI-MS m/z calc.807.3026, found 808.6 (M+1)+; Retention time: 1.58 minutes. LC method A. Step 6: (8R,9S)-14-(2,6-Dimethylphenyl)-8-isopropoxy-4-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa- 18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one, diastereomer 1, compound 43
Diastereomer 1 Diastereomer 1 [00475] To a scintillation vial with a stir bar was added 3-[[4-(2,6-dimethylphenyl)-6- [[(2S,3R)-3-isopropoxy-7-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3- yl]azepan-2-yl]methoxy]pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) diastereomer 1 (45.9 mg, 0.01359 mmol) and p-TsCl (10.4 mg, 0.05455 mmol) dissolved in NMP (400 µL). Then NMM (12.0 µL, 0.1091 mmol) was added and the reaction was stirred overnight for 18 hours at 80 °C. The mixture was diluted with EtOAc, quenched with 1M HCl and the layers were separated. The mixture was extracted with additional EtOAc, the organic layers were combined, dried with sodium sulfate, concentrated, purified by reverse phase chromatography (10-99% MeCN-water w/ 5mM HCl modifier, 15 minute run) and dried via lyophilization to give a white solid of (8R,9S)-14-(2,6-dimethylphenyl)-8-isopropoxy-4-[5- methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa-18λ6- thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one, diastereomer 1 (1.1 mg, 10%).1H NMR (400 MHz, CDCl3) δ 8.78 (s, 1H), 8.56 (s, 1H), 7.86 (dd, J = 21.6, 7.7 Hz, 2H), 7.58 (t, J = 7.8 Hz, 1H), 7.23 (t, J = 7.6 Hz, 1H), 7.08 (d, J = 7.6 Hz, 2H), 6.78 (s, 1H), 6.28 (s, 1H), 6.00 (dd, J = 12.8, 4.7 Hz, 1H), 5.84 (d, J = 5.7 Hz, 1H), 4.75 (d, J = 5.8 Hz, 2H), 4.06 (s, 3H), 3.58 (s, 1H), 3.34 (p, J = 6.2 Hz, 1H), 2.41 (d, J = 14.9 Hz, 1H), 2.05 (s, 10H), 1.77 (t, J = 13.0 Hz, 1H), 1.58 (s, 2H), 1.24 (d, J = 17.0 Hz, 3H), 1.00 (d, J = 6.0 Hz, 3H), 0.65 (d, J = 5.9 Hz, 3H).19F NMR (376 MHz, CDCl3) δ -68.68. ESI- MS m/z calc.789.29205, found 790.644 (M+1)+; Retention time: 1.613 minutes. LC method C. Example 57: Preparation of Compound 50 Step 1: 3,5-Dibromo-1-[(4-methoxyphenyl)methyl]pyrazin-2-one [00476] To a solution of 3,5-dibromopyrazin-2-ol (40 g, 157.56 mmol) in a mixture of anhydrous DMF (100 mL) and THF (125 mL) was added portion-wise at 0 °C NaH in mineral oil (8.1 g, 60 %w/w, 202.52 mmol). The reaction mixture was stirred at this temperature for 30 min. Then, a solution of 4-methoxybenzyl chloride (29.2 g, 186.45 mmol) in THF (75 mL) was slowly introduced in the flask, followed by TBAI (2.93 g, 7.9325 mmol). The reaction was then allowed to warm up to rt and stirred overnight. A saturated aqueous solution of ammonium chloride (50 mL) and EtOAc (50 mL) were added, and the aqueous layer was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with water (3 x 70 mL), brine (2 x 60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was divided in two part and two purifications by silica gel chromatography (gradient: 0-40% EtOAc in heptane) were done. Both batches collected were combined together to afford 3,5-dibromo-1-[(4-methoxyphenyl)methyl]pyrazin-2-one (44 g, 71%) as a yellow semi-solid.1H NMR (400 MHz, Chloroform-d) δ 7.32 - 7.27 (m, 2H), 7.24 (s, 1H), 6.95 - 6.90 (m, 2H), 5.02 (s, 2H), 3.83 (s, 3H).ESI-MS m/z calc.371.91092, no ionization detected, Retention time: 1.75 minutes. LC method I. Step 2: 5-Bromo-3-(3,3-dimethylbut-1-ynyl)-1-[(4- methoxyphenyl)methyl]pyrazin-2-one [00477] To a stirred solution of 3,5-dibromo-1-[(4-methoxyphenyl)methyl]pyrazin-2-one (40 g, 102.13 mmol) in DMF (200 mL), was added triethylamine (72.600 g, 100 mL, 717.46 mmol). Nitrogen was bubbled into the mixture for 5 minutes. The solution was cooled to 0 °C, then 3,3- dimethylbut-1-yne (9.1801 g, 14 mL, 108.40 mmol), bis(triphenylphosphine) palladium(II) dichloride (6.5 g, 9.0494 mmol) and CuI (3.7 g, 19.039 mmol) were added. The mixture was stirred, under nitrogen, 10 minutes at 0 °C and 1 hour in a room temperature bath. More 3,3- dimethylbut-1-yne (655.72 mg, 1 mL, 7.7431 mmol) was added at 0 °C and the mixture was stirred for 30 minutes in a room temperature bath. More 3,3-dimethylbut-1-yne (327.86 mg, 0.5 mL, 3.8716 mmol) was added at 0 °C and the mixture was stirred for 30 minutes in a room temperature bath. Water (250 mL) and ethyl acetate (250 mL) were added at 0 °C and the mixture was extracted with ethyl acetate (3 x 250 mL). The combined organic layers were washed with water (2 x 100 mL), brine (50 mL), dried over sodium sulfate, filtered, concentrated in vacuo and co-evaporated with dichloromethane (3 x 25 mL), to afford 5-bromo- 3-(3,3-dimethylbut-1-ynyl)-1-[(4-methoxyphenyl)methyl]pyrazin-2-one (44.37 g, 71%) as an orange oil.1H NMR (400 MHz, CDCl3) δ 7.30 - 7.24 (m, overlapped with CHCl3, 2H), 7.18 - 7.16 (m, 1H), 6.93 - 6.88 (m, 2H), 4.98 (s, 2H), 3.83 - 3.79 (m, 3H), 1.37 - 1.34 (m, 9H). ESI- MS m/z calc.374.063, found 375.2 (M+1)+; Retention time: 1.93 minutes. LC method S. Step 3: 2-Bromo-6-tert-butyl-furo[2,3-b]pyrazine [00478] To a stirred solution of 5-bromo-3-(3,3-dimethylbut-1-ynyl)-1-[(4- methoxyphenyl)methyl]pyrazin-2-one (36 g, 47.583 mmol) in DCM (160 mL) was added AgOTf (1.3 g, 5.0596 mmol) followed by TFA (59.600 g, 40 mL, 522.70 mmol). The mixture was stirred 2 hours at room temperature and was then concentrated in vacuo. The resulting residue was purified by silica gel chromatography (Gradient: 0-30% EtOAc in heptanes) to afford 2-bromo-6-tert-butyl-furo[2,3-b]pyrazine (21.8 g, 99%) as a yellow solid.1H NMR (400 MHz, Chloroform-d) δ 8.26 (s, 1H), 6.58 (s, 1H), 1.43 (s, 9H). ESI-MS m/z calc.254.0055, found 255.2 (M+1)+; Retention time: 1.97 minutes. LC method I. Step 4: tert-butyl (4R)-4-[(1S)-5-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-1- cyclobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00479] 2-Bromo-6-tert-butyl-furo[2,3-b]pyrazine (10.75 g, 42.14 mmol) was dissolved in tetrahydrofuran (156 mL), then cooled to -78°C to give a suspension. n-BuLi (18 mL of 2.5 M, 45.00 mmol) was added, at a rate which maintained a temperature below -65 °C. Four minutes after the n-BuLi addition was complete, a solution of tert-butyl (4R)-4-[(1S)-1-cyclobutyl-5- [methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (12 g, 30.11 mmol) in tetrahydrofuran (48 mL) was added, at a rate which maintained a temperature below - 65 °C. Immediately when the addition was complete, the cold bath was removed and the reaction was allowed to warm up gradually to 5 °C, at which point ammonium chloride (26.9 mL of 18 %w/w, 90.52 mmol) was added. Three more equal batches were performed, then all four batches were combined. Most of the THF was evaporated, then the mixture was partitioned between MTBE (400 mL) and brine (200 mL). The layers were separated, the organic layer was dried with magnesium sulfate, then concentrated to give crude product. The crude product was purified by reverse phase preparative chromatography using a C18 column and a gradient eluent of 60% to 100% acetonitrile in water to give partially purified product. It was further purified by silica gel chromatography using a gradient eluent of 0% to 30% MTBE in heptane to give impure tert-butyl (4R)-4-[(1S)-5-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-1-cyclobutyl-5-oxo- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (40.2 g, 57%) ESI-MS m/z calc.513.32025, found 514.6(M+1)⁺; Retention time: 3.94 minutes., which was used without further purification. LC method A. LC method N. Step 5: (5S,6R)-6-Amino-1-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-5-cyclobutyl-7- hydroxy-heptan-1-one [00480] To a stirred solution of tert-butyl (4R)-4-[(1S)-5-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)- 1-cyclobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (50 mg, 0.0819 mmol) in diethyl ether (1 mL) and water (0.25 mL), was slowly added a solution of hydrogen chloride in diethyl ether (1 mL of 2 M, 2.0000 mmol). The reaction was stirred at room temperature for 16 hours, then neutralized using a saturated aqueous solution of sodium bicarbonate (1.5 mL) and concentrated under reduced pressure to remove the volatiles. The aqueous layer was extracted with dichloromethane (3 x 20 mL) and the combined organic layers were washed with brine (10 mL), dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude was purified by reverse-phase chromatography on C18 column (gradient 5 to 95% acetonitrile in water). The desired fractions were concentrated under reduced pressure to afford (5S,6R)-6- amino-1-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-5-cyclobutyl-7-hydroxy-heptan-1-one (10 mg, 19%) as a yellow semi-solid.1H NMR (400 MHz, CDCl3) δ 8.50 (s, 1H), 6.56 (s, 1H), 3.93 - 3.78 (m, 2H), 3.78 - 3.64 (m, 2H), 3.62 - 3.54 (m, 1H), 2.70 - 2.59 (m, 1H), 2.14 - 2.05 (m, 2H), 2.05 - 1.94 (m, 2H), 1.94 - 1.43 (m, overlapped with impurities, 10H), 1.43 - 1.39 (m, 9H). ESI- MS m/z calc.373.2365, found 374.2 (M+1)+; Retention time: 1.44 minutes. LC method S. Step 6: [(2R,3S,7S)-7-(6-tert-Butylfuro[2,3-b]pyrazin-2-yl)-3-cyclobutyl-azepan- 2-yl]methanol [00481] To a solution of (5S,6R)-6-amino-1-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-5- cyclobutyl-7-hydroxy-heptan-1-one (65 mg, 0.1340 mmol) in DCM (6 mL) at room temperature was added NaBH(OAc)3 (86 mg, 0.3977 mmol). The reaction mixture was stirred for 90 minutes and a saturated aqueous solution of sodium bicarbonate (10 mL) was added. The layers were separated, and the aqueous layer was extracted with DCM (3 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography on C18 column (gradient 5 to 70% methanol in 10 mM ammonium bicarbonate buffer: pH = 10). The fractions containing the products were combined and the volatiles were removed under reduced pressure. The residue was co-evaporated with water (3 x 10 mL), then with acetonitrile (2 x 15 mL). The resulting product was dried under high vacuum to afford [(2R,3S,7S)-7-(6-tert-butylfuro[2,3- b]pyrazin-2-yl)-3-cyclobutyl-azepan-2-yl]methanol (37.7 mg, 79%) as an off-white solid.1H NMR (400 MHz, Benzene-d6) δ 8.04 (s, 1H), 6.35 (s, 1H), 3.82 (dd, J = 9.0, 4.6 Hz, 1H), 3.65 (dd, J = 10.3, 4.4 Hz, 1H), 3.09 (t, J = 10.1 Hz, 1H), 2.57 - 2.49 (m, 1H), 2.10 - 2.00 (m, 1H), 2.00 - 1.92 (m, 1H), 1.92 - 1.76 (m, 2H), 1.76 - 1.64 (m, 2H), 1.64 - 1.53 (m, 3H), 1.53 - 1.38 (m, 3H), 1.30 - 1.20 (m, 1H), 1.11 (s, 9H), 1.09 - 1.02 (m, 1H). (2 H missing, labile protons). ESI-MS m/z calc.357.2416, found 358.2 (M+1)+; Retention time: 2.84 minutes. LC method J. Step 7: 3-[[4-[[(2R,3S,7S)-7-(6-tert-Butylfuro[2,3-b]pyrazin-2-yl)-3-cyclobutyl- azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid [00482] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (38 mg, 0.09094 mmol) and [(2R,3S,7S)-7-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-3-cyclobutyl-azepan-2- yl]methanol (31 mg, 0.08671 mmol) in THF (400 µL) in a nitrogen-purged screwcap vial and stirred for 5 minutes followed by the addition of sodium tert-butoxide (56 mg, 0.5827 mmol) was added and the reaction was placed in aluminum heating block at 35 °C for 3 h. The reaction mixture was cooled and then partitioned between 15 mL 1 M HCl and 15 mL ethyl acetate. The layers were separated and the aqueous was extracted with an additional 10 mL ethyl acetate. The aqueous was then diluted with an equal volume of brine and extracted an additional 2 x 10 mL ethyl acetate. The combined organics were washed with brine, dried over sodium sulfate. The crude material was purified by silica chromatography (12 g of silica) using a gradient of 100% dichloromethane to 20% methanol in dichloromethane. The desired fractions were concentrated under reduced pressure, and dried under high vacuum to give a white solid 3-[[4-[[(2R,3S,7S)-7- (6-tert-butylfuro[2,3-b]pyrazin-2-yl)-3-cyclobutyl-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (58 mg, 86%). ESI- MS m/z calc.738.31995, found 739.2(M+1)⁺; Retention time: 1.67 minutes. LC method A. Step 8: (4S,8S,9R)-4-(6-tert-Butylfuro[2,3-b]pyrazin-2-yl)-8-cyclobutyl-14-(2,6- dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one (Compound 50) [00483] Under argon 3-[[4-[[(2R,3S,7S)-7-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-3-cyclobutyl- azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (58 mg, 0.07850 mmol), azeotroped twice with toluene, was dissolved in NMP (2.5 mL) and then TsCl (39 mg, 0.2046 mmol) was added. The reaction was stirred for 15 minutes at room temperature. Then NMM (52 µL, 0.4730 mmol) was added and the reaction was stirred at room temperature for 20 h. The mixture was diluted with HCl (220 µL of 4 M in dioxane, 0.8800 mmol) followed by EtOAc (5 mL) and water (5 mL) and the two phases were separated. The aqueous phase was extracted with EtOAc (2x). The organic layers were combined, washed with brine, and then concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 30 min) to give as an off-white solid (4S,8S,9R)-4-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-8-cyclobutyl-14- (2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (12.8 mg, 22%), 1H NMR (400 MHz, CDCl3) δ 8.82 (d, J = 1.8 Hz, 1H), 8.28 (s, 1H), 8.01 (d, J = 7.9 Hz, 1H), 7.85 (d, J = 7.7 Hz, 1H), 7.57 (t, J = 7.8 Hz, 1H), 7.24 (t, J = 7.6 Hz, 1H), 7.10 (d, J = 7.6 Hz, 2H), 6.70 (s, 1H), 6.31 (s, 1H), 5.94 (dd, J = 11.8, 5.1 Hz, 1H), 5.85 - 5.76 (m, 1H), 4.96 (t, J = 12.0 Hz, 1H), 4.42 - 4.34 (m, 1H), 2.40 (q, J = 8.6 Hz, 1H), 2.26 - 2.17 (m, 2H), 2.09 (s, 6H), 1.99 - 1.87 (m, 2H), 1.83 - 1.74 (m, 4H), 1.64 - 1.54 (m, 4H), 1.43 (s, 9H), 0.95 - 0.87 (m, 1H) plus 1H exchangeable not observed. ESI-MS m/z calc.720.3094, found 721.5(M+1)⁺; Retention time: 2.40 minutes. LC method A. Example 58: Preparation of Compound 53 Step 1: tert-Butyl (4S)-4-[(1R)-5-hydroxy-1-isopropoxy-5-[5-methyl-6-(2,2,2- trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3-b]pyrazin-3-yl]pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate ~85:15 mixture of diastereomers, the major isomer having the 4S,1R configuration [00484] To a solution of 3-bromo-5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3- b]pyrazine (286.4 mg, 0.8891 mmol) in tetrahydrofuran (3.0 mL) at -78 °C was added n- butyllithium (solution in hexanes) (0.41 mL of 2.5 M, 1.025 mmol) dropwise. As soon as the addition of n-BuLi was over, a solution of tert-butyl (4S)-4-[(1R)-1-isopropoxy-5-oxo-pentyl]- 2,2-dimethyl-oxazolidine-3-carboxylate (~85:15 mixture of diastereomers, the major isomer having the 4S,1R configuration, 250 mg, 0.7279 mmol) in tetrahydrofuran (3 mL) was added dropwise. The reaction was left stirring for 10 min and MeOH (5 mL) was added. The crude mixture was concentrated in vacuo and was then purified by flash chromatography on silica gel using 0-100% EtOAc in hexanes over 25 minutes to give tert-butyl (4S)-4-[(1R)-5-hydroxy-1- isopropoxy-5-[5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3-b]pyrazin-3- yl]pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (217.6 mg, 51%) as an orange-white solid. ESI-MS m/z calc.586.3342, found 587.2 (M+1)+; Retention time: 1.94 minutes. LC method A. Step 2: tert-Butyl (4S)-4-[(1R)-1-isopropoxy-5-[5-methyl-6-(2,2,2-trifluoro-1,1- dimethyl-ethyl)pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00485] To a solution of tert-butyl (4S)-4-[(1R)-5-hydroxy-1-isopropoxy-5-[5-methyl-6- (2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3-b]pyrazin-3-yl]pentyl]-2,2-dimethyl-oxazolidine- 3-carboxylate (217.6 mg, 0.3709 mmol) and in dichloroethane (4.8 mL) was added manganese (IV) oxide (activated) (764.6 mg, 7.476 mmol). The resulting suspension was stirred at 70 °C for 30 minutes. The suspension was filtered on a pad of Celite and washed with DCM (2 x). The crude mixture was concentrated in vacuo and was then purified by flash chromatography on silica gel using 0-100% EtOAc in hexanes over 30 minutes to provide tert-butyl (4S)-4-[(1R)-1- isopropoxy-5-[5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3-b]pyrazin-3-yl]-5- oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (180.8 mg, 83%) as a yellow oil. ESI-MS m/z calc.584.31854, found 585.2 (M+1)+; Retention time: 2.17 minutes. LC method A. Step 3: [(2S,3R)-3-Isopropoxy-7-[5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl- ethyl)pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00486] To a solution of tert-butyl (4S)-4-[(1R)-1-isopropoxy-5-[5-methyl-6-(2,2,2-trifluoro- 1,1-dimethyl-ethyl)pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (180.8 mg, 0.3092 mmol) in 1,4-dioxane (4.2 mL) was added hydrochloric acid (solution in 1,4-dioxane) (1.55 mL of 4 M, 6.200 mmol) at rt. Five drops of water were added and the reaction was stirred overnight. The solvent was removed under reduced pressure and the residual water was co-evaporated with acetonitrile (3x), then dried under high vacuum to afford crude [(2S,3R)-3-isopropoxy-7-[5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3- b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (131.9 mg, 100%) as a yellow oil. ESI-MS m/z calc.426.22427, found 427.2 (M+1)+; Retention time: 1.18 minutes. The crude material was taken forward without further purification. LC method A. Step 4: [(2S,3R)-3-Isopropoxy-7-[5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl- ethyl)pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol, diastereomer 1, and [(2S,3R)-3-isopropoxy-7-[5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl- ethyl)pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol, diastereomer 2 Diastereomer 1 Diastereomer 2 [00487] To a solution of [(2S,3R)-3-isopropoxy-7-[5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl- ethyl)pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (131.9 mg, 0.3093 mmol) in anhydrous dichloromethane (6.9 mL) at room temperature was added sodium triacetoxyborohydride (398.4 mg, 1.880 mmol). The reaction was stirred at room temperature for 1 hour and a saturated aqueous solution of sodium bicarbonate (100 mL) was added. The phases were separated, and the aqueous solution was extracted with DCM (2 x). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 45 min) to give the two main isomers: diastereomer 1, Peak 1: [(2S,3R)-3-isopropoxy-7-[5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl- ethyl)pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (42.3 mg, 32%) as a white solid. ESI- MS m/z calc.428.2399, found 429.3 (M+1)+; Retention time: 1.15 minutes, and diastereomer 2, Peak 2: [(2S,3R)-3-isopropoxy-7-[5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3- b]pyrazin-3-yl]azepan-2-yl]methanol (27.8 mg, 21%) as a yellow solid. ESI-MS m/z calc. 428.2399, found 429.2 (M+1)+; Retention time: 1.1 minutes. LC method A.
Step 5: 3-[[4-(2,6-Dimethylphenyl)-6-[[(2S,3R)-3-isopropoxy-7-[5-methyl-6- (2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3-b]pyrazin-3-yl]azepan-2- yl]methoxy]pyrimidin-2-yl]sulfamoyl]benzoic acid, diastereomer 1 Diastereomer 1 [00488] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (41.25 mg, 0.09872 mmol) and [(2S,3R)-3-isopropoxy-7-[5-methyl-6-(2,2,2-trifluoro-1,1-dimethyl- ethyl)pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol, diastereomer 1 (42.3 mg, 0.09872 mmol) were combined in a flask and dissolved in THF (1.1 mL) followed by the addition of sodium tert-butoxide (581 mg, 6.045 mmol). The reaction mixture was stirred for 1 hour then heated to 40 °C for 30 minutes then diluted with EtOAc and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (5 x). The organic layers were combined, washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3-[[4-(2,6-dimethylphenyl)-6-[[(2S,3R)-3-isopropoxy-7-[5-methyl-6- (2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]pyrimidin- 2-yl]sulfamoyl]benzoic acid, diastereomer 1 (73.1 mg, 91%) as a white solid. ESI-MS m/z calc. 809.31824, found 810.2 (M+1)+; Retention time: 1.41 minutes. LC method A. Step 6: (8R,9S)-14-(2,6-Dimethylphenyl)-8-isopropoxy-4-[5-methyl-6-(2,2,2- trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa-
18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one, diastereomer 1 Diastereomer 1 Diastereomer 1 [00489] 3-[[4-(2,6-Dimethylphenyl)-6-[[(2S,3R)-3-isopropoxy-7-[5-methyl-6-(2,2,2-trifluoro- 1,1-dimethyl-ethyl)pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]pyrimidin-2- yl]sulfamoyl]benzoic acid, diastereomer 1 (73.1 mg, 0.09026 mmol) was dissolved in NMP (4.4 mL) and TsCl (38.4 mg, 0.2014 mmol) was added. Then NMM (40.0 µL, 0.3638 mmol) was added and the reaction was stirred at room temperature overnight. The reaction mixture was diluted with 1M HCl and extracted with ethyl acetate. The layers were separated, and the aqueous layer was further extracted with ethyl acetate (2 x). The combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 25 min) to give (8R,9S)-14-(2,6-dimethylphenyl)-8-isopropoxy-4-[5-methyl- 6-(2,2,2-trifluoro-1,1-dimethyl-ethyl)pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen- 2-one, diastereomer 1 (19.1 mg, 26%) as a white solid.1H NMR (400 MHz, CDCl3) δ 8.76 (s, 1H), 8.55 (s, 1H), 7.94 – 7.81 (m, 2H), 7.57 (t, J = 1.8 Hz, 1H), 7.22 (t, J = 7.6 Hz, 1H), 7.08 (d, J = 7.6 Hz, 2H), 6.75 (s, 1H), 6.27 (s, 1H), 6.00 (dd, J = 12.7, 4.8 Hz, 1H), 5.89 – 5.78 (m, 1H), 4.82 – 4.70 (m, 2H), 4.17 (s, 3H), 3.58 (s, 1H), 3.34 (hept, J = 6.0 Hz, 1H), 2.45 – 2.34 (m, 1H), 2.15 – 1.83 (m, 11H), 1.76 (d, J = 4.0 Hz, 6H), 1.00 (d, J = 6.1 Hz, 3H), 0.65 (d, J = 5.9 Hz, 3H). ESI-MS m/z calc.791.3077, found 792.2 (M+1)+; Retention time: 1.26 minutes. LC method C. Example 59: Preparation of Compound 55 Step 1: tert-Butyl (4S)-4-[5-[6-tert-butyl-5-(trideuteriomethyl)pyrrolo[2,3- b]pyrazin-3-yl]-5-hydroxy-1-isopropoxy-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate ~85:15 mixture of diastereomers, the major isomer having the 4S,1R configuration [00490] To a stirred solution of 3-bromo-6-tert-butyl-5-(trideuteriomethyl)pyrrolo[2,3- b]pyrazine (170 mg, 0.6269 mmol) in anhydrous tetrahydrofuran (3 mL) was added n- butyllithium (solution in hexanes) (300 µL of 2.5 M, 0.7500 mmol) (in hexanes) dropwise at -78 °C, under nitrogen. Immediately after the addition, a solution of tert-butyl (4S)-4-(1-isopropoxy- 5-oxo-pentyl)-2,2-dimethyl-oxazolidine-3-carboxylate (~85:15 mixture of diastereomers, the major isomer having the 4S,1R configuration, 200 mg, 0.5823 mmol) in anhydrous tetrahydrofuran (1 mL) was added dropwise. Stirring continued for another 15 min and methanol (2 mL) was added at that temperature. The reaction mixture was concentrated under reduced pressure and the residue was purified by reverse phase chromatography (10-99% acetonitrile in water (no modifier) over 15 min) to furnish tert-butyl (4S)-4-[5-[6-tert-butyl-5- (trideuteriomethyl)pyrrolo[2,3-b]pyrazin-3-yl]-5-hydroxy-1-isopropoxy-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate as a pale yellow semi-solid (112 mg, 36%) ESI-MS m/z calc. 535.3813, found 536.5 (M+1)+; Retention time: 1.86 minutes. LC method A. Step 2: tert-Butyl (4S)-4-[5-[6-tert-butyl-5-(trideuteriomethyl)pyrrolo[2,3- b]pyrazin-3-yl]-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate [00491] To a solution of tert-butyl (4S)-4-[5-[6-tert-butyl-5-(trideuteriomethyl)pyrrolo[2,3- b]pyrazin-3-yl]-5-hydroxy-1-isopropoxy-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (110 mg, 0.2053 mmol) in anhydrous dichloroethane (2.0 mL) was added activated manganese (IV) oxide (activated) (420 mg, 4.106 mmol) at ambient temperature under nitrogen.. The resulting dark suspension was stirred at 70 °C for 45 min. The reaction was allowed to cool to ambient temperature and the suspension was filtered on a pad of Celite and washed with dichloromethane (2 x 25 mL). The filtrate was concentrated under reduced pressure to furnish tert-butyl (4S)-4-[5-[6-tert-butyl-5-(trideuteriomethyl)pyrrolo[2,3-b]pyrazin-3-yl]-1-isopropoxy- 5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate as a brownish semi-solid (88 mg, 80%) ESI-MS m/z calc.533.36566, found 534.2 (M+1)+; Retention time: 2.3 minutes. LC method A. Step 3: [(2S)-7-[6-tert-Butyl-5-(trideuteriomethyl)pyrrolo[2,3-b]pyrazin-3-yl]-3- isopropoxy-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00492] To a stirred solution of tert-butyl (4S)-4-[5-[6-tert-butyl-5- (trideuteriomethyl)pyrrolo[2,3-b]pyrazin-3-yl]-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (109 mg, 0.2042 mmol) in 1,4-dioxane (2 mL) and was added water (0.1 mL), hydrochloric acid (in 1,4-dioxane) (2.0 mL of 4 M, 8.000 mmol) at ambient temperature, and the reaction was stirred at that temperature for 14 h. The volatiles were removed under reduced pressure, water (10 mL) was added to the residue and concentrated. The residual water was co-evaporated with acetonitrile (3 x 10 mL), followed by toluene (3 x 10 mL) then dried under high vacuum to furnish crude [(2S)-7-[6-tert-butyl-5- (trideuteriomethyl)pyrrolo[2,3-b]pyrazin-3-yl]-3-isopropoxy-3,4,5,6-tetrahydro-2H-azepin-2- yl]methanol (hydrochloride salt) as a yellow oil (80 mg, 95%) ESI-MS m/z calc.375.27136, found 376.3 (M+1)+; Retention time: 1.23 minutes. LC method A. Step 4: [(2S,3R)-7-[6-tert-Butyl-5-(trideuteriomethyl)pyrrolo[2,3-b]pyrazin-3- yl]-3-isopropoxy-azepan-2-yl]methanol, diastereomer 1 [00493] To a stirred solution of [(2S)-7-[6-tert-butyl-5-(trideuteriomethyl)pyrrolo[2,3- b]pyrazin-3-yl]-3-isopropoxy-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (hydrochloride salt) (80 mg, 0.1942 mmol) in anhydrous dichloromethane (800 µL) were added acetic-acid (25 µL, 0.4396 mmol) and DIPEA (100 µL, 0.5741 mmol) at 0 oC under nitrogen. After stirring 10 min, sodium triacetoxyborohydride (130 mg, 0.6134 mmol) was added. The reaction mixture was stirred at that temperature for 30 min and after quenching with methanol (1 mL) and water (0.5 mL), concentrated under reduced pressure. The crude was purified by reversed-phase chromatography (10-99% acetonitrile in water containing 5 mM HCl, over 20 min run) to give several isomers. The first isomer to elute was the major diastereomer 1, peak 1. The fractions containing the major diastereomer were concentrated under reduced pressure. Residual water was removed azeotropically with acetonitrile (5 x 10 mL), followed by toluene (3 x 10 mL) to furnish [(2S,3R)-7-[6-tert-butyl-5-(trideuteriomethyl)pyrrolo[2,3-b]pyrazin-3-yl]-3-isopropoxy- azepan-2-yl]methanol (hydrochloride salt), diastereomer 1, as a light-brown solid (41 mg, 51%). ESI-MS m/z calc.377.28702, found 378.3 (M+1)+; Retention time: 1.15 minutes. LC method A. Two minor diastereomers were concentrated separately and kept aside. Peak 2 (minor): [(2S,3R)-7-[6-tert-butyl-5-(trideuteriomethyl)pyrrolo[2,3-b]pyrazin-3-yl]-3-isopropoxy-azepan- 2-yl]methanol (hydrochloride salt) (4.0 mg, 5%) ESI-MS m/z calc.377.28702, found 378.3 (M+1)+; Retention time: 1.2 minutes; and Peak-3 (minor): [(2S,3S)-7-[6-tert-butyl-5- (trideuteriomethyl)pyrrolo[2,3-b]pyrazin-3-yl]-3-isopropoxy-azepan-2-yl]methanol (hydrochloride salt) (5.0 mg, 6%) ESI-MS m/z calc.377.28702, found 378.3 (M+1)+; Retention time: 1.2 minutes. LC method A. Step 5: 3-[[4-[[(2S,3R)-7-[6-tert-Butyl-5-(trideuteriomethyl)pyrrolo[2,3- b]pyrazin-3-yl]-3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid, diastereomer 1 diastereomer 1 [00494] A stirred solution of 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (41 mg, 0.09812 mmol) and [(2S,3R)-7-[6-tert-butyl-5- (trideuteriomethyl)pyrrolo[2,3-b]pyrazin-3-yl]-3-isopropoxy-azepan-2-yl]methanol (hydrochloride salt), diastereomer 1 (38 mg, 0.09179 mmol) in anhydrous tetrahydrofuran (500 µL) was sparged with nitrogen for 5 min. Then sodium tert-butoxide (55 mg, 0.5723 mmol) was added in a single portion at ambient temperature and stirring continued at that temperature for1 h, then at 40 °C for 1 h. The reaction mixture was allowed to cool to ambient temperature and diluted with ethyl acetate (20 mL) and hydrochloric acid (1.0 mL of 1.0 M, 1.000 mmol). The organic phase was separated, and the aqueous layer was extracted with ethyl acetate (2 x 15 mL). The combined organics were washed with brine (10 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude was purified by reversed-phase chromatography (10-99% acetonitrile in water containing 5 mM HCl, over 20 min run), and the fractions were concentrated under reduced pressure, co-evaporated with acetonitrile (2 x 10 mL) and toluene (2 x 10 mL) to furnish 3-[[4-[[(2S,3R)-7-[6-tert-butyl-5- (trideuteriomethyl)pyrrolo[2,3-b]pyrazin-3-yl]-3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt), diastereomer 1, as a white solid (48 mg, 66%). ESI-MS m/z calc.758.36536, found 759.2 (M+1)+; Retention time: 1.48 minutes. LC method A. Step 6: (8R,9S)-4-[6-tert-Butyl-5-(trideuteriomethyl)pyrrolo[2,3-b]pyrazin-3-yl]- 14-(2,6-dimethylphenyl)-8-isopropoxy-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(23),12,14,16(24),19,21-hexaen- 2-one, diastereomer 1, compound 55 diastereomer 1 diastereomer 1 [00495] To a stirred solution of 3-[[4-[[(2S,3R)-7-[6-tert-butyl-5- (trideuteriomethyl)pyrrolo[2,3-b]pyrazin-3-yl]-3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt), diastereomer 1 (45 mg, 0.05657 mmol) in anhydrous N-methylpyrrolidinone (3.0 mL) was added p-TsCl (40 mg, 0.2098 mmol) at ambient temperature under nitrogen, and the mixture was stirred for 5 min. Then N-methylmorpholine (50 µL, 0.4548 mmol) was added and the reaction was stirred for 18 h. Then more p-TsCl (20 mg, 0.1049 mmol) and N-methylmorpholine (25 µL, 0.2274 mmol) were added and the reaction was stirred for a total of 4 d. The crude reaction mixture was purified by preparative HPLC (25-99% acetonitrile in water, containing 5mM HCl as modifier, 15 min run). The mixed fractions were combined and evaporated, and repurified by HPLC (35- 99% acetonitrile in water, 20 min run, HCl modifier) and the pure fractions were combined with to furnish (8R,9S)-4-[6-tert-butyl-5-(trideuteriomethyl)pyrrolo[2,3-b]pyrazin-3-yl]-14-(2,6- dimethylphenyl)-8-isopropoxy-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo [17.3.1.112,16.03,9]tetracosa-1(23),12,14,16(24),19,21-hexaen-2-one as a white solid (1.6 mg, 4%), 1H NMR (400 MHz, MeOD) δ 8.47 (t, J = 1.9 Hz, 2H), 7.86 (dd, J = 7.7, 1.9 Hz, 1H), 7.61 (d, J = 7.7 Hz, 1H), 7.56 (t, J = 7.6 Hz, 1H), 7.19 (t, J = 7.6 Hz, 1H), 7.06 (d, J = 7.7 Hz, 2H), 6.52 (s, 1H), 6.21 (s, 1H), 5.86 (dd, J = 11.5, 5.4 Hz, 1H), 5.80 (dd, J = 12.8, 4.9 Hz, 1H), 4.94 (t, J = 12.0 Hz, 1H), 4.59 (dt, J = 12.3, 5.2 Hz, 1H), 3.63 (d, J = 4.8 Hz, 1H), 3.36 (hept, J = 6.1 Hz, 1H), 2.28 – 2.12 (m, 2H), 2.02 (s, 6H), 1.93 – 1.75 (m, 4H), 1.47 (s, 9H), 0.96 (d, J = 6.0 Hz, 3H), 0.64 (d, J = 6.0 Hz, 3H). ESI-MS m/z calc.740.3548, found 741.2 (M+1)+; Retention time: 1.88 minutes. LC method A. Example 60: Preparation of Compound 68 Step 1: 4-Benzyloxybutyl(bromo)magnesium [00496] Magnesium turnings (1.35 g, 55.544 mmol) were flamed dried under vacuum in a three-neck flask and, upon cooling, were suspended in dry tetrahydrofuran (100 mL). To the solution was added a small crystal of iodine (203 mg, 0.7998 mmol).4-Bromobutoxymethyl benzene (637.50 mg, 0.5 mL, 2.6219 mmol) was added to the mixture and the suspension was gently heated with a heat gun to initiate the reaction. The rest of the 4-bromobutoxymethyl benzene (8.9250 g, 7.0 mL, 36.707 mmol) was added dropwise at room temperature. After addition, the mixture was refluxed for 1 h, then was allowed to reach room temperature to afford a dark yellow solution of 4-benzyloxybutyl(bromo)magnesium (10.518 g, 107%) in THF. The suspension was used directly in the next step. Step 2: tert-Butyl (4S)-4-(5-benzyloxy-1-hydroxy-pentyl)-2,2-dimethyl- oxazolidine-3-carboxylate [00497] To a solution of tert-butyl (4S)-4-formyl-2,2-dimethyl-oxazolidine-3-carboxylate (248 mg, 1.0817 mmol) in tetrahydrofuran (10 mL) was added dropwise at -78 °C a solution of 4- benzyloxybutyl(bromo)magnesium (in THF) (1 g, 3.7391 mmol). The reaction was stirred for 1 h at that temperature and stirred for an additional 30 minutes while being allowed to warm to room temperature. A saturated aqueous solution of ammonium chloride (40 mL) was added, and the phases were separated. The aqueous phase was extracted with EtOAc (3 x 25 mL). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and was then purified by silica gel chromatography (Gradient: 0-30% EtOAc in Heptanes). The desired fractions were combined, concentrated under reduced pressure and purified by reversed-phase chromatography (Column: C18, Gradient : 5%-100% MeOH in water). The desired fractions were combined and concentrated under reduced pressure to afford a mixture of two diastereomers (75:25 ratio) of tert-butyl (4S)-4-(5- benzyloxy-1-hydroxy-pentyl)-2,2-dimethyl-oxazolidine-3-carboxylate (210 mg, 49%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 7.37 – 7.28 (m, 5H), 4.51 (s, 2H), 4.16 – 3.55 (m, 5H), 3.49 (t, J = 6.1 Hz, 2H), 1.74 – 1.62 (m, 3H), 1.62 – 1.55 (m,4H), 1.53 – 1.44 (m, 14H). ESI-MS m/z calc.393.2515, found 394.4 (M+1)+; Retention time: 1.96 minutes. LC method I. Step 3: tert-Butyl (4S)-4-(5-benzyloxy-1-methoxy-pentyl)-2,2-dimethyl- oxazolidine-3-carboxylate [00498] NaH (in mineral oil) (25 mg, 0.6251 mmol) was added to a solution of tert-butyl (4S)- 4-(5-benzyloxy-1-hydroxy-pentyl)-2,2-dimethyl-oxazolidine-3-carboxylate (175 mg, 0.3455 mmol) and methyl iodide (200 mg, 0.0877 mL, 1.4091 mmol) in DMF (3.7 mL) at 0 °C under nitrogen atmosphere. The solution was stirred under nitrogen for 30 minutes then was warmed to room temperature and stirred for 3 hours. More NaH (in mineral oil) (23 mg, 0.5751 mmol) was added at 0 °C followed by more methyl iodide (450 mg, 0.1974 mL, 3.1704 mmol). The reaction was stirred at 0 °C for 20 minutes, then at room temperature for 16 hours. The reaction was cooled down to 0 °C, quenched with water (25 mL) and the aqueous layer was extracted with EtOAc (3 x 15 mL). The combined organic layers were washed with water (20 mL), brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude material was purified by reversed-phase chromatography (Column: C18, Gradient: 5- 100 % MeOH in water). The desired fractions were combined and concentrated under reduced pressure to afford tert-butyl (4S)-4-(5-benzyloxy-1-methoxy-pentyl)-2,2-dimethyl-oxazolidine- 3-carboxylate (124 mg, 87%) as a colorless oil;1H NMR (400 MHz, CDCl3) as a mixture of diastereomers δ 7.38 – 7.27 (m, 5H), 4.51 (s, 2H), 4.11 – 3.99 (m, 1H), 3.95 – 3.86 (m, 1.5H), 3.83 – 3.57 (m, 1H), 3.53 – 3.45 (m, 2.5H), 3.45 – 3.38 (m, 3H), 1.71 – 1.37 (overlapped with impurities, m, 21H). ESI-MS m/z calc.407.2672, found 308.4 (M-99)+; Retention time: 2.14 minutes. LC method I. Step 4: tert-Butyl (4S)-4-(5-hydroxy-1-methoxy-pentyl)-2,2-dimethyl-oxazolidine- 3-carboxylate [00499] Nitrogen was bubbled through a stirred solution of tert-butyl (4S)-4-(5-benzyloxy-1- methoxy-pentyl)-2,2-dimethyl-oxazolidine-3-carboxylate (5.01 g, 11.777 mmol) in dry MeOH (110 mL) for 10 minutes, then palladium on carbon (2.54 g, 5 %w/w, 1.1934 mmol) was added. Nitrogen was bubbled through the mixture for 10 minutes, followed by hydrogen for 10 minutes. The reaction was then stirred for 18 hours, at room temperature, under hydrogen atmosphere. Nitrogen was bubbled through the mixture for 10 minutes, then the suspension was filtered through Celite. The cake was washed with EtOAc (2 x 40 mL) and the volatiles were removed under reduced pressure to afford the crude tert-butyl (4S)-4-(5-hydroxy-1-methoxy-pentyl)-2,2- dimethyl-oxazolidine-3-carboxylate (4.02 g, 100%) as a colorless oil;1H NMR (400 MHz, CDCl3) δ 4.25 – 4.19 (m, 0.2H), 4.09 – 3.99 (m, 1.1H), 3.97 – 3.85 (m, 1.4H), 3.84 – 3.74 (m, 0.3H), 3.70 – 3.56 (overlapped with impurities, m, 2.6H), 3.53 – 3.37 (overlapped with impurities, m, 3.4H), 1.70 – 1.37 (m, 22H). ESI-MS m/z calc.317.2202, found 340.2 (M+23)+; Retention time: 1.68 minutes. LC method I. Step 5: tert-Butyl (4S)-4-(1-methoxy-5-oxo-pentyl)-2,2-dimethyl-oxazolidine-3- carboxylate [00500] A flask was flame-dried and charged with oxalyl chloride (43.650 mg, 0.030 mL, 0.3439 mmol) and anhydrous dichloromethane (0.9 mL). The solution was cooled down to -78 °C and a solution of dimethylsulfoxide (55.050 mg, 0.050 mL, 0.7046 mmol) in anhydrous dichloromethane (0.9 mL) was added dropwise. The mixture was stirred at -78 °C for 45 minutes at which time a solution of tert-butyl (4S)-4-(5-hydroxy-1-methoxy-pentyl)-2,2- dimethyl-oxazolidine-3-carboxylate (60.5 mg, 0.1904 mmol) in anhydrous dichloromethane (0.9 mL) was added dropwise and the reaction was stirred at -78 °C for 45 minutes. Triethylamine (108.90 mg, 0.15 mL, 1.0762 mmol) was added dropwise and the mixture was stirred at -78 °C for 30 minutes then allowed to reach room temperature and stirred at room temperature for 30 minutes then quenched by the addition of a saturated aqueous solution of ammonium chloride (5 mL) and stirred at room temperature for 15 minutes. The layers were separated, and the aqueous layer was extracted with dichloromethane (3 x 5 mL) and the combined organic layers were washed with brine (5 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude material was purified by reversed phase chromatography (Column: C18, Gradient : 5%-100% MeCN in water). The desired fractions were combined and concentrated under reduced pressure to afford a mixture of two diastereomers and rotamers (unknown ratio) of tert-butyl (4S)-4-(1-methoxy-5-oxo-pentyl)-2,2-dimethyl-oxazolidine-3- carboxylate (38.6 mg, 61%) as a pale yellow oil; 1H NMR (400 MHz, CDCl3) δ 9.82 – 9.73 (m, 1H), 4.26 – 4.18 (m, 0.1H), 4.13 – 4.07 (m, 0.1H), 4.06 – 3.97 (m, 1H), 3.96 – 3.87 (m, 1.5H), 3.83 – 3.77 (m, 0.2H), 3.68 – 3.59 (m, 0.6H), 3.54 – 3.35 (m, 3.5H), 2.55 – 2.40 (m, 2H), 1.94 – 1.76 (m, 1H), 1.76 – 1.64 (m, 1H), 1.64 – 1.35 (overlapped with water, m, 17H). ESI-MS m/z calc.315.2046, found 338.2 (M+23)+; Retention time: 1.79 minutes. LC method I. Step 6: tert-Butyl (4S)-4-[5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-5- hydroxy-1-methoxy-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00501] To a solution of 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine (401 mg, 1.4206 mmol) in tetrahydrofuran (7.5 mL) at -78 °C was added n-butyllithium (solution in hexanes) (0.69 mL of 2.08 M, 1.4352 mmol) dropwise. As soon as the addition of n-Buli was complete, a solution of tert-butyl (4S)-4-(1-methoxy-5-oxo-pentyl)-2,2-dimethyl-oxazolidine-3- carboxylate (984 mg, 1.9156 mmol) in tetrahydrofuran (7.0 mL) was added dropwise. The reaction was stirred for 10 minutes and MeOH (10 mL) was added. The solvent was concentrated in vacuo and the crude product was purified by reversed phase chromatography (Column : C18, Gradient : 5 % MeCN in water, then 45-65% MeCN in water) to afford after evaporation tert-butyl (4S)-4-[5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-5-hydroxy- 1-methoxy-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (445 mg, 57%) as a yellow oil;1H NMR (400 MHz, CDCl3) δ 8.53 – 8.36 (m, 1H), 6.77 – 6.55 (m, 1H), 5.00 – 4.89 (m, 1H), 4.26 – 4.17 (m, 0.25H), 4.08 – 3.98 (m, 4H), 3.97 – 3.85 (m, 1.5H), 3.83 – 3.75 (m, 0.25H), 3.71 – 3.35 (m, 5H), 2.01 – 1.80 (m, 2H), 1.77 – 1.39 (overlapped with water, m, 28H). ESI-MS m/z calc.504.3312, found 505.4 (M+1)+; Retention time: 1.97 minutes. LC method I. Step 7: tert-Butyl (4S)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-1-methoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate and tert- butyl (4S)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1- methoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00502] To a solution of tert-butyl (4S)-4-[5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-5-hydroxy-1-methoxy-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (840 mg, 1.5779 mmol) in dichloroethane (24 mL) was added manganese (IV) oxide (activated) (3.38 g, 33.047 mmol). The resulting suspension was stirred at 80 °C for 2.5 hours. The suspension was filtered on a pad of Celite, the pad was washed with DCM (2 x 40 mL) and the solvent was evaporated under reduced pressure. The crude material was purified by reversed-phase chromatography (Gradient: 5% MeCN in water, then 60-90% MeCN in water). The desired fractions were combined and evaporated under reduced pressure to afford a yellow oil. The oil was purified by preparative SFC (Lux Column 5 µm, Cellulose 2, 250 x 30 mm, 51 mg/mL, 25.5 mg/injection) to afford the first diastereomers tert-butyl (4S)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-methoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (271 mg, 34%) as a yellow oil;1H NMR (400 MHz, CDCl3) δ 9.12 (s, 1H), 6.59 (s, 1H), 4.13 – 4.01 (m, 4H), 4.00 – 3.82 (m, 2H), 3.79 – 3.72 (m, 0.5H), 3.62 – 3.56 (m, 0.3H), 3.52 – 3.42 (m, 3.2H), 3.38 – 3.21 (m, 2H), 2.05 – 1.78 (m, 2H), 1.67 – 1.45 (m, 26H). ESI-MS m/z calc.502.3155, found 503.5 (M+1)+; Retention time: 2.14 minutes; and the second diastereomer to elute tert- butyl (4S)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-methoxy-5-oxo- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (160 mg, 20%) as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 9.12 (s, 1H), 6.59 (s, 1H), 4.28 – 4.19 (m, 0.5H), 4.13 – 4.02 (m, 4.5H), 3.98 – 3.88 (m, 1H), 3.72 – 3.62 (m, 0.5H), 3.58 – 3.52 (m, 0.5H), 3.51 – 3.39 (m, 3H), 3.39 – 3.21 (m, 2H), 2.10 – 1.96 (m, 1H), 1.89 – 1.75 (m, 1H), 1.72 – 1.42 (overlapped with water, m, 26H). ESI-MS m/z calc.502.3155, found 503.4 (M+1)+; Retention time: 2.13 minutes. LC method I. Step 8: [(2S,3R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-methoxy- 3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00503] To a solution of tert-butyl (4S)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-methoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (262 mg, 0.5207 mmol) in a mixture of 1,4-dioxane (4.6 mL) and water (0.5 mL) at room temperature was added dropwise a solution of hydrochloric acid (solution in 1,4-dioxane) (2.6 mL of 4 M, 10.400 mmol). The reaction was stirred for 1.5 hours, and the solvent was removed under reduced pressure. A saturated aqueous solution of sodium bicarbonate (10 mL, until pH = 7) was added, followed by water (20 mL) and DCM (25 mL). The mixture was extracted with DCM (3 x 15 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to afford 147 mg of crude compound. The aqueous layer was extracted with Me-THF (3-20 mL). The combined organic layers were washed with brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to afford 54.8 mg of the crude compound. The two crudes were combined and purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeCN in acidic water (0.1% v/v of formic acid in water). The fractions containing the desired product were combined and concentrated under reduced pressure to afford [(2S,3R)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-methoxy-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (208 mg, 100%) as a yellow oil; ESI-MS m/z calc.344.2212, found 345.4 (M+1)+; Retention time: 1.34 minutes. LC method I. Step 9: [(2S,3R,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- methoxy-azepan-2-yl]methanol and [(2S,3R,7R)-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-3-methoxy-azepan-2-yl]methanol [00504] To a solution of [(2S,3R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- methoxy-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (195 mg, 0.4880 mmol) in dichloromethane (5.6 mL) was added sodium triacetoxyborohydride (726 mg, 3.4255 mmol).The reaction was stirred at rt for 2 h. A saturated aqueous solution of sodium bicarbonate (25 mL) was added. The phases were separated and the aqueous one was extracted with DCM (3 x 25 mL). The combined organic phases were washed with brine (25 mL), dried over anhydrous sodium sulfate, and filtered. The crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 20 to 60% of MeCN in basic water (ammonium bicarbonate/ammonium hydroxide buffer pH = 10). The desired fractions for each diastereomer were combined and the volatiles were removed under reduced pressure. The resulting basic aqueous phase was extracted with DCM (3 x 20 mL), the combined organic extracts were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford separately, after freeze drying, the major diastereomer [(2S,3R,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- methoxy-azepan-2-yl]methanol (97.5 mg, 57%) as a light yellow solid; 1H NMR (400 MHz, Benzene-d6) δ 8.54 (s, 1H), 6.57 (s, 1H), 4.04 – 3.93 (m, 2H), 3.48 (s, 3H), 3.40 (dd, J = 10.3, 8.6 Hz, 1H), 3.07 – 2.99 (m, 4H), 2.98 – 2.91 (m, 1H), 2.05 – 1.96 (m, 1H), 1.89 – 1.70 (m, 4H), 1.63 – 1.52 (m, 1H), 1.11 (s, 9H), 2H missing, labile protons. ESI-MS m/z calc.346.2369, found 347.2 (M+1)+; Retention time: 3.36 minutes and the minor diastereomer [(2S,3R,7R)-7-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-methoxy-azepan-2-yl]methanol (37.8 mg, 22%) as a sticky yellow oil;1H NMR (400 MHz, Benzene-d6) δ 8.52 (s, 1H), 6.59 (s, 1H), 4.15 (dd, J = 10.3, 4.6 Hz, 1H), 3.78 (dd, J = 11.2, 3.9 Hz, 1H), 3.61 (d, J = 9.5 Hz, 1H), 3.58 (s, 3H), 3.18 – 3.10 (m, 1H), 3.06 (s, 3H), 2.74 – 2.66 (m, 1H), 2.09 – 2.00 (m, 1H), 1.99 – 1.90 (m, 1H), 1.71 – 1.62 (m, 1H), 1.57 – 1.46 (m, 1H), 1.43 – 1.31 (m, 2H), 1.26 – 1.11 (m, 2H), 1.09 (s, 9H). ESI- MS m/z calc.346.2369, found 347.2 (M+1)+; Retention time: 3.46 minutes. LC method W. Step 10: 3-[[4-[[(2S,3R,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-methoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid
[00505] In a 4 mL vial, 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (65 mg, 0.1556 mmol) and [(2S,3R,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-3-methoxy-azepan-2-yl]methanol (45 mg, 0.1299 mmol) were combined in THF (0.5 mL). Solid sodium tert-butoxide (75 mg, 0.7804 mmol) was added. The reaction mixture was stirred at 40 °C for 3 hours. It was diluted with EtOAc (50 mL) and washed with water (1× 50 mL). The aqueous layer was acidified to pH 3 with the addition of aqueous 1 M HCl and extracted with a 4:1 solution of DCM and isopropanol (3× 50 mL). All organic layers were combined, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The obtained crude solid was dissolved in 1:1 EtOAc/isopropanol (approximately 3 mL) and triturated with hexane (approximately 10 mL). Solids were filtered to provide 3-[[4-[[(2S,3R,7S)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-methoxy-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (79 mg, 91%) as a yellow solid. ESI-MS m/z calc.727.3152, found 728.3 (M+1)+; Retention time: 1.49 minutes. LC method A. Step 11: (4S,8R,9S)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14- (2,6-dimethylphenyl)-8-methoxy-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one [00506] In a round bottom flask, 4-methylbenzenesulfonyl chloride (52 mg, 0.2728 mmol) was added to a solution of 3-[[4-[[(2S,3R,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-3-methoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (79 mg, 0.1085 mmol) in NMP (5 mL). After stirring at room temperature for 15 minutes, 4-methylmorpholine (72 µL, 0.6549 mmol) was added. The reaction mixture was stirred overnight at room temperature. It was diluted with EtOAc (50 mL) and washed with HCl (2× 50 mL) and brine (2× 50 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated under reduced pressure. Purification by reverse phase HPLC (25-60% acetonitrile/5 mM aqueous HCl over 15 minutes) provided (4S,8R,9S)-4-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-8-methoxy-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen- 2-one (31.2 mg, 40%) as a yellow solid.1H NMR (400 MHz, CDCl3) δ 8.80 (s, 1H), 8.49 (s, 1H), 7.89 – 7.80 (m, 2H), 7.61 (t, J = 7.8 Hz, 1H), 7.25 (d, J = 7.6 Hz, 1H), 7.11 (d, J = 7.6 Hz, 2H), 6.61 (s, 1H), 6.26 (s, 1H), 6.07 (dd, J = 13.0, 4.4 Hz, 1H), 5.82 – 5.71 (m, 1H), 4.77 – 4.63 (m, 2H), 4.17 (s, 3H), 3.56 – 3.49 (m, 1H), 3.09 (s, 3H), 2.42 – 2.33 (m, 1H), 2.24 – 2.14 (m, 1H), 2.10 (s, 6H), 2.02 – 1.93 (m, 2H), 1.91 – 1.80 (m, 2H), 1.54 (s, 9H). ESI-MS m/z calc. 709.3046, found 710.4 (M+1)+; Retention time: 1.74 minutes. LC method A. Example 61: Preparation of Compounds 74 and 77 Step 1: tert-Butyl (4R)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-1-(cyclopropylmethyl)-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3- carboxylate [00507] To a solution of 1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-2- dimethoxyphosphoryl-ethanone (1.9 g, 5.5937 mmol) in isopropanol (150 mL) was added potassium carbonate (3 g, 21.707 mmol) and left stirring for 1 hour. This solution was then added to a solution of tert-butyl (4R)-4-[(1R)-1-(cyclopropylmethyl)-3-oxo-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate (1.45 g, 3.9577 mmol) in isopropanol (150 mL). The reaction was stirred at room temperature for 24 hours. The reaction mixture was quenched with water (50 mL) and then MeTHF (75 mL) was added. The phases were separated, and the aqueous phase was extracted with MeTHF (2X50 mL). The combined organic phase was washed with brine (100 mL), dried over sodium sulfate, filtered, concentrated to afford a crude. The resulting residue was purified by reverse phase chromatography on a C18 column using a 5 to 100% gradient of acetonitrile in pure water. The fractions containing the desired product were combined and concentrated under high vacuum to provide tert-butyl (4R)-4-[(1R)-5-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-(cyclopropylmethyl)-5-oxo-pent-3-enyl]-2,2- dimethyl-oxazolidine-3-carboxylate (1.9 g, 77%) as an off-white solid; ESI-MS m/z calc. 524.3363, found 525.4 (M+1)+; Retention time: 2.25 minutes. LC method S. Step 2: tert-Butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-1-(cyclopropylmethyl)-5-hydroxy-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate [00508] To a solution of tert-butyl (4R)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-(cyclopropylmethyl)-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3- carboxylate (1.9 g, 3.0563 mmol) in methanol (75 mL) was added Cobalt(II) chloride hexahydrate (70 mg, 0.2942 mmol) and the mixture was stirred for 20 minutes. To the mixture was added NaBH4 (1.1 g, 29.076 mmol) in DMF (10 mL) dropwise. The mixture was stirred for 2 hours at room temperature before being quenched with water (50 mL) slowly at room temperature. The mixture was extracted with dichloromethane (3 x 25 mL). The combined organic layer was washed with brine (75 mL), dried over sodium sulfate, and concentrated in vacuo. Purification of the residue by reverse phase (acetonitrile in water 0 to 95%). The fractions containing desired product were combined and concentrated under high vacuum to afford tert- butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-(cyclopropylmethyl)- 5-hydroxy-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.2 g, 68%) as a colorless oil; ESI- MS m/z calc.528.3676, found 529.4 (M+1)+; Retention time: 2.15 minutes. LC method S. Step 3: tert-Butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-1-(cyclopropylmethyl)-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00509] To a solution of tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-(cyclopropylmethyl)-5-hydroxy-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (1.2 g, 2.0653 mmol) in anhydrous DCE (60 mL) was added MnO2 (activated) (3.5 g, 40.259 mmol). The suspension was heated to 70 °C for 5 hours and the reaction was then cooled down to room temperature. The suspension was filtered through Celite, and the pad was rinsed with DCM (25 mL). The mixture was concentrated in vacuo to afford crude tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-(cyclopropylmethyl)-5-oxo- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.1 g, 66%) as a brownish oil; 1H NMR (400 MHz, CDCl3) δ 9.11 (s, 1H), 6.59 (s, 1H), 4.21 – 3.97 (m, 4H), 3.96 – 3.88 (m, 1H), 3.86 – 3.79 (m, 1H), 3.29 (t, J = 7.2 Hz, 1H), 2.22 – 2.02 (m, 1H), 1.93 – 1.79 (m, 1H), 1.74 – 1.52 (m, 12H), 1.51 – 1.41 (m, 13H), 1.39 – 1.09 (m, 5H), 0.78 – 0.65 (m, 1H), 0.54 – 0.33 (m, 2H), 0.15 - -0.10 (m, 2H). ESI-MS m/z calc.526.3519, found 527.3 (M+1)+; Retention time: 2.5 minutes. LC method Y. Step 4: (5S,6R)-6-Amino-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-5- (cyclopropylmethyl)-7-hydroxy-heptan-1-one [00510] A solution of tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin- 3-yl)-1-(cyclopropylmethyl)-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.1 g, 1.3596 mmol) in a mixture of HCl in dioxane (50 mL of 4 M, 200.00 mmol) and water (15 mL) was stirred at room temperature for 3 hours then concentrated under reduced pressure. The crude was purified by reverse-phase chromatography on C18 (gradient: 5 to 100% acetonitrile in water). The desired fractions were concentrated under reduced pressure, then co-evaporated with acetonitrile (5 x 5 mL) and dried under high vacuum to afford (5S,6R)-6-amino-1-(6-tert-butyl- 5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-5-(cyclopropylmethyl)-7-hydroxy-heptan-1-one (hydrochloride salt) (425 mg, 62%) as a yellow foam; 1H NMR (400 MHz, CDCl3) δ 9.01 (s, 1H), 8.16 (br. S., 3H), 6.51 (s, 1H), 4.04 (s, 3H), 4.01 – 3.94 (m, 1H), 3.93 – 3.83 (m, 1H), 3.81 – 3.67 (m, 1H), 3.42 – 3.20 (m, 2H), 2.43 – 2.21 (m, 1H), 2.10 – 1.97 (overlapped with acetonitrile peak, m, 1H), 1.91 – 1.77 (m, 2H), 1.76 – 1.66 (m, 2H), 1.57 – 1.42 (m, 10H), 1.38 – 1.27 (m, 1H), 0.79 – 0.65 (m, 1H), 0.51 – 0.34 (m, 2H), 0.21 - -0.01 (m, 2H). ESI-MS m/z calc. 386.2682, found 387.4 (M+1)+; Retention time: 1.38 minutes. LC method S. Step 5: [(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclopropylmethyl)azepan-2-yl]methanol and [(2R,3S,7R)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(cyclopropylmethyl)azepan-2-yl]methanol [00511] To a solution of (5S,6R)-6-amino-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-5-(cyclopropylmethyl)-7-hydroxy-heptan-1-one (425 mg, 0.9280 mmol) in anhydrous dichloromethane (35 mL) was added sodium triacetoxyborohydride (985 mg, 4.6475 mmol). The reaction mixture was stirred at room temperature for 2 hours then quenched by the addition of a saturated aqueous solution of sodium bicarbonate (25 mL) and stirred 30 minutes at room temperature. The layers were separated, and the aqueous layer was extracted with dichloromethane (3 x 25 mL) and the combined organic layers were washed with brine (50 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude was purified by reverse phase chromatography on C18 (gradient: 5 to 100% acetonitrile in water). The desired fractions were concentrated under reduced pressure to afford the first peak of diastereomer [(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclopropylmethyl)azepan-2-yl]methanol (130 mg, 37%) as an off-white solid; 1H NMR (400 MHz, CDCl3) δ 8.32 (s, 1H), 6.46 (s, 1H), 4.06 – 3.95 (m, 4H), 3.74 (dd, J = 10.3, 4.2 Hz, 1H), 3.13 (t, J = 10.1 Hz, 1H), 2.68 (td, J = 9.5, 4.2 Hz, 1H), 2.20 – 2.10 (m, 1H), 2.04 – 1.93 (m, overlapped with acetonitrile peak, 1H), 1.92 – 1.78 (m, 3H), 1.77 – 1.65 (m, 2H), 1.59 (qd, J = 9.2, 5.0 Hz, 1H), 1.51 (s, 9H), 1.44 – 1.34 (m, 1H), 1.16 (ddd, J = 13.9, 7.3, 3.9 Hz, 1H), 0.79 – 0.65 (m, 1H), 0.57 – 0.38 (m, 2H), 0.16 - -0.06 (m, 2H), (1H missing, labile proton). ESI-MS m/z calc.370.2733, found 371.4 (M+1)+; Retention time: 2.16 minutes and the second peak of diastereomer [(2R,3S,7R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclopropylmethyl)azepan-2-yl]methanol (125 mg, 35%) as an off-white solid; 1H NMR (400 MHz, CDCl3) δ 8.36 (s, 1H), 6.45 (s, 1H), 4.08 – 3.96 (m, 4H), 3.69 (dd, J = 10.8, 4.4 Hz, 1H), 3.44 (t, J = 10.5 Hz, 1H), 2.83 (td, J = 9.8, 4.4 Hz, 1H), 2.39 – 2.28 (m, 1H), 2.17 – 2.08 (m, 1H), 2.07 – 2.00 (m, overlapped with acetonitrile peak, 1H), 1.83 – 1.70 (m, 1H), 1.69 – 1.55 (m, 2H), 1.54 – 1.42 (m, 10H), 1.36 – 1.24 (m, 1H), 1.22 – 1.12 (m, 1H), 0.78 – 0.66 (m, 1H), 0.55 – 0.39 (m, 2H), 0.13 - -0.05 (m, 2H), (2H missing, labile proton). ESI-MS m/z calc. 370.2733, found 371.4 (M+1)+; Retention time: 2.17 minutes. LC method Z. Step 6: 3-[[4-[[(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-(cyclopropylmethyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00512] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (114 mg, 0.2728 mmol) and [(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclopropylmethyl)azepan-2-yl]methanol (66.9 mg, 0.1805 mmol) were combined in a vial and dissolved in THF (875 µL) followed by the addition of sodium tert-butoxide (112 mg, 1.165 mmol). The reaction mixture was stirred at 40 oC for 3 h. An additional 3 equivalents of base were added, and the reaction was continued for an additional 1 h. The reaction mixture was cooled down to room temperature and diluted with EtOAc and 1 M HCl (1 mL). The organic phase was separated, and the aqueous layer was extracted with EtOAc. The organic layers were combined, washed with brine, and dried over sodium sulfate. The mixture was filtered, and then concentrated under reduced pressure. The crude was carried forward to the next step as 3-[[4- [[(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(cyclopropylmethyl)azepan- 2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (170.8 mg, 120%). ESI-MS m/z calc.751.35156, found 752.2 (M+1)+; Retention time: 1.26 minutes. LC method A. Step 7: (4S,8S,9R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8-(cyclopropylmethyl)- 14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (Compound 74), and (4R,8R,9S)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8- (cyclopropylmethyl)-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (Compound 77) [00513] Under nitrogen 3-[[4-[[7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclopropylmethyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (179 mg, 0.2270 mmol), azeotroped 2 times with toluene, was dissolved in NMP (6 mL) and TsCl (172.5 mg, 0.9048 mmol) was added. The reaction was stirred for 15 minutes at room temperature. Then NMM (150 µL, 1.364 mmol) was added and the reaction was stirred at room temperature overnight. The mixture was neutralized with HCl (1M) (1ml) and diluted with EtOAc. The organic layers were collected, dried over magnesium sulfate, and concentrated. The crude material was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 30 min) to give impure (4S,8S,9R)-4-(6-tert-butyl- 5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8-(cyclopropylmethyl)-14-(2,6-dimethylphenyl)-18,18- dioxo-11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one as a yellow solid (33.5 mg, 19%). ESI-MS m/z calc. 733.341, found 734.5 (M+1)+; Retention time: 0.79 minutes. LC method B. Chiral SFC revealed the presence of about 20% of an enantiomeric isomer which was subsequently separated. [00514] Impure (4S,8S,9R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8- (cyclopropylmethyl)-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (33 mg, 0.04316 mmol) was subjected to chiral SFC using a ChiralPak AS column (21.2 x 250 mm; 5 μm) at 40 oC, a mobile phase (isocratic mode) constituted of 48% MeOH + 20 mM NH3, with a flow of 70 mL/min, an injection volume of 500 µL (concentration of solution 10.2 mg/mL in methanol), a 192 bar outlet pressure, and a detection wavelength of 210 nm. The collected fractions afforded pure peak 1 (first to elute) and a mixture containing peak 2 which was further purified by flash chromatography on silica gel (4 g, DCM: MeOH (0-10%)). Pure fractions of peak 1 were collected and concentrated to afford as a white ammonium salt (4S,8S,9R)-4-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8-(cyclopropylmethyl)-14-(2,6-dimethylphenyl)- 18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (ammonium salt) (12.3 mg, 54%) 1H NMR (400 MHz, CDCl3) δ 8.84 (s, 1H), 8.45 (s, 1H), 7.91 (d, J = 7.9 Hz, 1H), 7.80 (d, J = 7.6 Hz, 1H), 7.58 (t, J = 7.8 Hz, 1H), 7.25 – 7.21 (m, 1H), 7.09 (d, J = 7.6 Hz, 2H), 6.57 (s, 1H), 6.28 (s, 1H), 5.96 – 5.83 (m, 2H), 4.86 (t, J = 11.8 Hz, 1H), 4.54 – 4.44 (m, 1H), 4.16 (s, 3H), 2.38 – 2.28 (m, 1H), 2.25 – 2.15 (m, 1H), 2.08 (s, 6H), 2.00 – 1.92 (m, 2H), 1.90 – 1.80 (m, 2H), 1.55 (br s, 9H), 1.25 (s, 1H), 1.19 – 1.10 (m, 1H), 1.04 – 0.98 (m, 1H), 0.41 – 0.33 (m, 1H), 0.29 – 0.18 (m, 2H), -0.18 (m, 2H). ESI-MS m/z calc.733.341, found 734.4 (M+1)+; Retention time: 2.04 minutes. LC method A. Pure fractions of peak 2 gave after flash chromatography, as a white solid, (4R,8R,9S)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8- (cyclopropylmethyl)-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (2.4 mg, 25%).1H NMR (400 MHz, CDCl3) δ 8.84 (t, J = 1.8 Hz, 1H), 8.45 (s, 1H), 7.77 (d, J = 7.5 Hz, 1H), 7.72 (d, J = 7.9 Hz, 1H), 7.53 (t, J = 7.7 Hz, 1H), 7.26 – 7.22 (m, 1H), 7.09 (d, J = 7.6 Hz, 2H), 6.47 (s, 1H), 6.26 (s, 1H), 5.94 – 5.83 (m, 2H), 4.86 (t, J = 11.8 Hz, 1H), 4.51 – 4.43 (m, 1H), 4.14 (s, 3H), 2.38 – 2.29 (m, 1H), 2.25 – 2.13 (m, 1H), 2.08 (s, 5H), 2.00 – 1.93 (m, 2H), 1.88 – 1.81 (m, 2H), 1.50 (s, 9H), 1.26 (s, 1H), 1.17 – 1.09 (m, 1H), 1.05 – 0.99 (m, 1H), 0.90 – 0.82 (m, 1H), 0.36 (s, 1H), 0.30 – 0.18 (m, 2H), -0.16 - -0.27 (m, 2H). ESI-MS m/z calc. 733.341, found 734.4 (M+1)+; Retention time: 2.06 minutes. LC method A. Example 62: Alternative preparation of Compound 74 Step 1: tert-Butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-1-(cyclopropylmethyl)-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00515] Under nitrogen atmosphere, in a 500 mL round bottom flask equipped with an internal temperature probe, n-BuLi in hexanes (16.5 mL of 2.5 M, 41.25 mmol) was added to a solution of 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine (10.34 g, 38.56 mmol) in THF (130 mL) at -78 °C, at a rate which maintained the internal temperature below -67 °C (over 6 min). After 10 min, a solution of tert-butyl (4R)-4-[(1S)-1-(cyclopropylmethyl)-5- [methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (10.94 g, 27.45 mmol) in tetrahydrofuran (45 mL) was added, at a rate which maintained the internal temperature below -67 °C (over 10 min). When the addition was complete, the cold bath was removed and the reaction was allowed to warm up gradually to 7 °C, at which point aqueous ammonium chloride (100 mL of 18 %w/w, 336.5 mmol) was added all at once. The organic phase was separated from the aqueous phase. The aqueous phase was further extracted with EtOAc (2 x 100 mL). The combined organic phases were washed with brine (50 mL), dried over sodium sulfate and evaporated. The crude material was dissolved in a minimum volume of methanol, then purified by reverse phase chromatography using a C18 column and a gradient eluent of 40% to 100% acetonitrile in water). After evaporation of the solvents, the residue was taken in dioxane which was evaporated (operation run 2 x). Drying under vacuum provided tert- butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-(cyclopropylmethyl)- 5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (13.9 g, 89%) as a thick resin (92.4% purity by weight, contains 7.6% by weight of dioxane).1H NMR (400 MHz, CDCl3) δ 9.11 (s, 1H), 6.59 (s, 1H), 4.13 - 3.98 (m, 4H), 3.96 - 3.87 (m, 1H), 3.82 (d, J = 9.0 Hz, 1H), 3.29 (t, J = 7.3 Hz, 2H), 2.22 - 2.01 (m, 1H), 1.92 - 1.79 (m, 2H), 1.77 - 1.65 (m, 1H), 1.62 - 1.60 (m, 1H), 1.56 - 1.51 (m, 10H), 1.52 - 1.40 (m, 13H), 1.37 - 1.24 (m, 2H), 1.23 - 1.14 (m, 1H), 0.78 - 0.65 (m, 1H), 0.52 - 0.43 (m, 1H), 0.43 - 0.34 (m, 1H), 0.08 - 0.02 (m, 1H), -0.01 - -0.09 (m, 1H). ESI-MS m/z calc.526.3519, found 527.76(M+1)⁺; Retention time: 2.43 minutes. LC method A. Step 2: [(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclopropylmethyl)-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00516] Hydrochloric acid (50 mL of 4 M, 200.0 mmol) (4 M in dioxane) was added to a solution of tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1- (cyclopropylmethyl)-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (13.9 g, 24.38 mmol) in dioxane (100 mL) and water (1.5 mL). The solution was stirred at room temperature for 3 h. The solvent was removed under vacuum. Dioxane (50 mL) was added and evaporated (operation repeated twice). The residue was then dissolved in acetonitrile (200 mL) and concentrated to give as a yellow foamy solid [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-(cyclopropylmethyl)-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (hydrochloride salt) (11.7 g, 100%) 1H NMR (400 MHz, CDCl3) δ 9.10 (s, 1H), 8.13 (s, 3H), 6.90 (s, 1H), 4.20 (s, 3H), 3.96 - 3.86 (m, 1H), 3.84 - 3.73 (m, 1H), 3.68 - 3.56 (m, 1H), 3.42 - 3.24 (m, 2H), 2.08 - 1.98 (m, 1H), 1.95 - 1.75 (m, 2H), 1.75 - 1.62 (m, 2H), 1.58 (s, 9H), 1.50 - 1.39 (m, 1H), 1.37 - 1.24 (m, 1H), 0.77 - 0.61 (m, 1H), 0.50 - 0.32 (m, 2H), 0.17 - 0.03 (m, 2H). ESI-MS m/z calc.368.2576, found 369.51(M+1)⁺; Retention time: 1.28 minutes. LC method A. Step 3: [(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclopropylmethyl)azepan-2-yl]methanol [00517] Anhydrous acetonitrile (140 mL) was added to solid [(2R,3S)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(cyclopropylmethyl)-3,4,5,6-tetrahydro-2H-azepin-2- yl]methanol (hydrochloride salt) (11.7 g, 24.27 mmol), resulting in the heterogenous suspension of an oil. The mixture was sparged with nitrogen for 5 minutes. [[(1R,2R)-2-amino-1,2- diphenyl-ethyl]-(p-tolylsulfonyl)amino]-chloro-ruthenium;1-isopropyl-4-methyl-benzene (231 mg, 0.3631 mmol) was added followed by triethylamine (10 mL, 71.74 mmol) resulting in a suspension of solids. Formic acid (2.8 mL, 74.22 mmol) was added, and the mixture was stirred at 45 °C (internal temperature) for 14 h. The reaction was then stirred in an ice bath for an hour. The resulting solid was filtered and washed with a small volume of ice-cold acetonitrile to give a first crop of white solid (3.45 g) mostly constituted of a triethylamine HCl salt. Additional precipitation occurred in the cold filtrate upon scratching the inside of the flask with a spatula. The suspension was stirred in an ice bath for a few hours, and the solid was filtered and washed with ice-cold acetonitrile to give a different crop of tan solid (7.75 g) containing the expected product. This solid was dissolved in DCM (150 mL). The organic phase was washed with water (2 x 50 mL), aqueous 1 N NaOH (40 mL), dried over sodium sulfate and the solvents evaporated to give a solid. The solid was dissolved in a mixture of warm THF (20 mL) and 2Me-THF (50 mL) and the solvents were evaporated to give as a tan solid [(2R,3S,7S)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(cyclopropylmethyl)azepan-2-yl]methanol (7.47 g, 82%). 1H NMR (400 MHz, CDCl3) δ 8.31 (s, 1H), 6.45 (s, 1H), 4.04 - 3.94 (m, 4H), 3.73 (dd, J = 10.3, 4.1 Hz, 1H), 3.33 (broad s, 1H), 3.12 (t, J = 10.1 Hz, 1H), 2.67 (td, J = 9.5, 4.1 Hz, 1H), 2.20 - 2.09 (m, 1H), 2.05 - 1.92 (m, 1H), 1.91 - 1.63 (m, 5H), 1.63 - 1.54 (m, 1H), 1.50 (s, 9H), 1.44 - 1.32 (m, 1H), 1.24 - 1.10 (m, 1H), 0.77 - 0.65 (m, 1H), 0.56 - 0.39 (m, 2H), 0.14 - 0.05 (m, 1H), 0.04 - 0.01 (m, 1H). ESI-MS m/z calc.370.27325, found 371.55(M+1)⁺; Retention time: 1.23 minutes. LC method A. Step 4: 3-[[4-[[(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-(cyclopropylmethyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00518] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (136.5 mg, 0.3267 mmol) and [(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclopropylmethyl)azepan-2-yl]methanol (121.1 mg, 0.3268 mmol) were combined in a flask and dissolved in THF (2.0 mL) followed by the addition of NaOt-Bu (158.7 mg, 1.651 mmol). The reaction mixture was stirred for 1 hour at room temperature then warmed to 40 °C for 2 hours. The reaction mixture was cooled to room temperature and diluted with EtOAc and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (5 x). The organic layers were combined, washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3-[[4-[[(2R,3S,7S)-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-3-(cyclopropylmethyl)azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (201 mg, 78%) as a yellow solid. ESI-MS m/z calc.751.35156, found 752.3(M+1)⁺; Retention time: 2.15 minutes. LC method N. Step 5: (4S,8S,9R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8- (cyclopropylmethyl)-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (Compound 74) [00519] 3-[[4-[[(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclopropylmethyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (201 mg, 0.2549 mmol) was dissolved in NMP (4.1 mL) and Acetone (8.1 mL). TsCl (148.8 mg, 0.7805 mmol) was added followed by NMM (0.17 mL, 1.546 mmol) and the reaction was stirred at 55 °C for 22 hours. The reaction was poured into a mixture of ethyl acetate and 1 M HCl and the layers were separated. The aqueous layer was further extracted with ethyl acetate (2 x). The organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was taken up in DMSO/MeOH then injected into a reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 25 min). The pure fractions were extracted with EtOAc (3 x), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give (4S,8S,9R)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8-(cyclopropylmethyl)-14-(2,6- dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (82.0 mg, 42%) as a white solid.1H NMR (400 MHz, CDCl3) δ 8.85 (s, 1H), 8.45 (s, 1H), 7.77 (d, J = 7.6 Hz, 1H), 7.71 (d, J = 7.8 Hz, 1H), 7.52 (t, J = 7.7 Hz, 1H), 7.23 (d, J = 7.6 Hz, 1H), 7.08 (d, J = 7.7 Hz, 2H), 6.45 (s, 1H), 6.25 (s, 1H), 5.94 - 5.84 (m, 2H), 4.86 (t, J = 11.8 Hz, 1H), 4.53 - 4.42 (m, 1H), 4.13 (s, 3H), 2.38 - 2.27 (m, 1H), 2.26 - 2.13 (m, 1H), 2.08 (m, 7H), 2.00 - 1.90 (m, 2H), 1.89 - 1.82 (m, 2H), 1.49 (s, 9H), 1.18 - 1.08 (m, 1H), 1.05 - 1.02 (m, 1H), 0.39 - 0.31 (m, 1H), 0.31 - 0.17 (m, 2H), -0.16 - -0.29 (m, 2H). ESI-MS m/z calc.733.341, found 734.6(M+1)⁺; Retention time: 2.03 minutes. LC method A. Example 63: Preparation of Compounds 78 and 79 Step 1: Racemic-tert-butyl (4R)-4-[(4S)-8-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3- b]pyrrolizin-3-yl}-8-hydroxy-2-methyloctan-4-yl]-2,2-dimethyl-1,3-oxazolidine- 3-carboxylate [00520] To a solution of 3-bromo-8,8-dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizine (550 mg, 2.0377 mmol) in THF (14 mL) was slowly added at -100 °C n-BuLi (in hexane) (0.85 mL of 2.5 M, 2.1250 mmol) and the reaction was stirred for 1 min then a solution of racemic-tert-butyl (4R)-4-[(1S)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (850 mg, 2.0162 mmol) in THF (5 mL) was added dropwise at -100 °C. After 5 min at -100 °C a saturated aqueous solution of ammonium chloride (20 mL) and EtOAc (25 mL) were added. The mixture was allowed to warm to rt over 25 min and the phases were separated. The aqueous phase was extracted with EtOAc (3 x 35 mL) and the combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure. Purification by reversed-phase chromatography (C18 column. Gradient: 5-100 % MeCN in water). The volatiles were partially removed under reduced pressure. The aqueous layer was extracted with EtOAc (3 x 20 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford racemic-tert-butyl (4R)-4-[(4S)-8-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3- b]pyrrolizin-3-yl}-8-hydroxy-2-methyloctan-4-yl]-2,2-dimethyl-1,3-oxazolidine-3-carboxylate (520 mg, 48%) as a tan solid. ESI-MS m/z calc.528.3676, found 529.4 (M+1)+; Retention time: 2.45 minutes.1H NMR (400 MHz, Chloroform-d) δ 8.33 (br d, J = 8.6 Hz, 1H), 6.30 (s, 1H), 4.88 (br d, J = 4.4 Hz, 1H), 4.26 (t, J = 7.0 Hz, 2H), 4.03 – 3.74 (m, 3H), 3.56 – 3.40 (m, 1H), 2.43 (t, J = 6.8 Hz, 2H), 2.12 – 1.74 (m, 3H), 1.70 – 0.97 (overlapped with water, m, 28H), 0.91 – 0.79 (m, 6H). LC method O. Step 2: Racemic-tert-butyl (4R)-4-[(4S)-8-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3- b]pyrrolizin-3-yl}-2-methyl-8-oxooctan-4-yl]-2,2-dimethyl-1,3-oxazolidine-3- carboxylate [00521] To a solution of Racemic-tert-butyl (4R)-4-[(4S)-8-{8,8-dimethyl-6H,7H,8H- pyrazino[2,3-b]pyrrolizin-3-yl}-8-hydroxy-2-methyloctan-4-yl]-2,2-dimethyl-1,3-oxazolidine-3- carboxylate (520 mg, 0.9825 mmol) in anhydrous DCE (52 mL) was added MnO2 (activated) (2 g, 19.554 mmol). The suspension was heated to 70 °C for 1.5 hours and the reaction was then cooled down to room temperature. The suspension was filtered through Celite, and the pad was rinsed with DCM (55 mL). The mixture was concentrated in vacuo to afford crude racemic-tert- butyl (4R)-4-[(4S)-8-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3-yl}-2-methyl-8- oxooctan-4-yl]-2,2-dimethyl-1,3-oxazolidine-3-carboxylate (470 mg, 90%) as a yellow semi- solid; ESI-MS m/z calc.526.3519, found 527.3 (M+1)+; Retention time: 2.66 minutes.1H NMR (400 MHz, Chloroform-d) δ 9.09 (s, 1H), 6.39 (s, 1H), 4.32 (br t, J = 6.6 Hz, 2H), 4.11 – 3.98 (m, 1H), 3.98 – 3.89 (m, 1H), 3.88 – 3.78 (m, 1H), 3.31 – 3.14 (m, 2H), 2.48 (t, J = 6.8 Hz, 2H), 2.22 – 1.03 (overlapped with water, m, 29H), 0.97 – 0.89 (m, 3H), 0.87 – 0.81 (m, 3H). LC method O. Step 3: Racemic-(5S)-5-[(1R)-1-amino-2-hydroxyethyl]-1-{8,8-dimethyl- 6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3-yl}-7-methyloctan-1-one rac-4R,4S rac-5S,1R [00522] To a stirred solution of racemic-tert-butyl (4R)-4-[(4S)-8-{8,8-dimethyl-6H,7H,8H- pyrazino[2,3-b]pyrrolizin-3-yl}-2-methyl-8-oxooctan-4-yl]-2,2-dimethyl-1,3-oxazolidine-3- carboxylate (470 mg, 0.8807 mmol) in a mixture of dioxane (12 mL) and water (6.5 mL) was added a solution of hydrogen chloride in dioxane (20 mL of 4 M, 80.000 mmol) dropwise. The reaction was stirred at room temperature for 1 hour, then concentrated under reduced pressure. Residual water was co-evaporated with water (3 x 5 mL) and acetonitrile (5 x 10 mL), then dried under high vacuum to afford crude racemic-(5S)-5-[(1R)-1-amino-2-hydroxyethyl]-1-{8,8- dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3-yl}-7-methyloctan-1-one (hydrochloride salt) (436 mg, 100%) as an orange semi-solid that was directly used in the next step without further purification. ESI-MS m/z calc.386.2682, found 194.2 (M-192)+; Retention time: 1.92 minutes. 1H NMR (400 MHz, DMSO-d6) δ 8.91 (s, 1H), 7.88 (br s, 3H), 6.52 (s, 1H), 6.49 – 5.79 (m, 3H), 4.32 (t, J = 6.8 Hz, 2H), 3.67 – 3.59 (m, 2H), 3.21 (t, J = 7.3 Hz, 2H), 3.13 – 3.06 (m, 1H), 2.44 (t, J = 7.0 Hz, 2H), 1.81 – 1.74 (m, 1H), 1.71 – 1.57 (m, 3H), 1.44 (s, 6H), 1.42 – 1.22 (m, 3H), 1.13 – 1.04 (m, 1H), 0.86 (dd, J = 6.6, 3.9 Hz, 6H). LC method O. Step 4: Racemic-[(2R,3S,7S)-7-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3- b]pyrrolizin-3-yl}-3-(2-methylpropyl)azepan-2-yl]methanol and racemic- [(2R,3S,7R)-7-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3-yl}-3-(2- methylpropyl)azepan-2-yl]methanol rac-5S,1R rac-2R,3S,7S rac-2R,3S,7R [00523] To a solution of racemic-(5S)-5-[(1R)-1-amino-2-hydroxyethyl]-1-{8,8-dimethyl- 6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3-yl}-7-methyloctan-1-one (hydrochloride salt) (436 mg, 0.8783 mmol) in anhydrous dichloromethane (70 mL) was added sodium triacetoxyborohydride (940 mg, 4.4352 mmol). The reaction was stirred at room temperature for 3.5 hours then quenched by the addition of a saturated aqueous solution of sodium bicarbonate (60 mL) and stirred overnight at room temperature. The layers were separated, and the aqueous layer was extracted with dichloromethane (3 x 60 mL). The combined organic layers were washed with brine (70 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude was purified by reverse phase chromatography (C18 column. Gradient: 5 to 100% acetonitrile in water). The fractions containing the first eluting compound were concentrated under reduced pressure to afford impure product. The fractions containing the second eluting compound were concentrated under reduced pressure and lyophilized to afford racemic- [(2R,3S,7R)-7-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3-yl}-3-(2- methylpropyl)azepan-2-yl]methanol (64 mg, 18%) as a tan solid.1H NMR (400 MHz, Benzene- d6) δ 8.59 (s, 1H), 6.35 (s, 1H), 3.89 (dd, J = 11.2, 2.7 Hz, 1H), 3.84 (dd, J = 10.8, 4.2 Hz, 1H), 3.76 (t, J = 7.1 Hz, 2H), 3.52 (t, J = 10.4 Hz, 1H), 2.59 (td, J = 9.5, 4.2 Hz, 1H), 2.21 – 2.12 (m, 1H), 1.86 – 1.74 (m, 1H), 1.62 – 1.44 (m, 5H), 1.35 – 1.08 (m, 4H), 1.02 – 0.92 (m, 8H), 0.84 (d, J = 6.6 Hz, 3H), 0.78 (d, J = 6.4 Hz, 3H), (1H missing, labile proton). ESI-MS m/z calc. 370.2733, found 371.2 (M+1)+; Retention time: 5.82 minutes. LC method 1A. [00524] The impure first eluted compound was repurified by reversed-phase chromatography (C18 column. Gradient: 5-100 % MeCN in basic water (ammonium bicarbonate/ammonium hydroxide buffer pH = 10). The volatiles were removed under reduced pressure, the residue co- evaporated with water (2x 4 mL) and lyophilized to afford racemic-[(2R,3S,7S)-7-{8,8- dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3-yl}-3-(2-methylpropyl)azepan-2-yl]methanol (72 mg, 22%) as a white solid; 1H NMR (400 MHz, Benzene-d6) δ 8.54 (s, 1H), 6.36 (s, 1H), 3.87 (dd, J = 10.3, 4.4 Hz, 1H), 3.77 (dd, J = 10.3, 4.2 Hz, 1H), 3.73 – 3.67 (m, 2H), 3.11 (t, J = 10.0 Hz, 1H), 2.51 (td, J = 9.4, 4.0 Hz, 1H), 2.10 – 2.00 (m, 1H), 1.92 – 1.81 (m, 1H), 1.71 – 1.54 (m, 4H), 1.53 – 1.32 (m, 4H), 1.31 – 1.22 (m, 1H), 1.20 – 1.10 (m, 1H), 1.01 (d, J = 6.1 Hz, 6H), 0.94 – 0.83 (m, 4H), 0.75 (d, J = 6.6 Hz, 3H), (1H missing, labile proton). ESI-MS m/z calc.370.2733, found 371.2 (M+1)+; Retention time: 5.65 minutes. LC method 1A. Step 5: Racemic-3-[(4-{[(2R,3S,7S)-7-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3- b]pyrrolizin-3-yl}-3-(2-methylpropyl)azepan-2-yl]methoxy}-6-(2,6- dimethylphenyl)pyrimidin-2-yl)sulfamoyl]benzoic acid
[00525] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (74.4 mg, 0.1780 mmol) and racemic-[(2R,3S,7S)-7-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3- yl}-3-(2-methylpropyl)azepan-2-yl]methanol (60 mg, 0.1619 mmol) were combined in a flask and dissolved in THF (1 mL) followed by the addition of potassium tert-butoxide (109 mg, 0.9714 mmol). The reaction mixture was stirred for 17 hours at 40 °C. The reaction mixture was diluted with EtOAc and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (3 x). The organic layers were combined, washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give crude racemic-3- [(4-{[(2R,3S,7S)-7-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3-yl}-3-(2- methylpropyl)azepan-2-yl]methoxy}-6-(2,6-dimethylphenyl)pyrimidin-2-yl)sulfamoyl]benzoic acid (hydrochloride salt) (144 mg, 61%) ESI-MS m/z calc.751.35156, found 752.445 (M+1)+; Retention time: 2.013 minutes. LC method N. Step 6: 4-{8,8-Dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3-yl}-14-(2,6- dimethylphenyl)-8-(2-methylpropyl)-11-oxa-18λ6-thia-3,15,17,24- tetraazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(23),12(24),13,15,19,21- hexaene-2,18,18-trione, (4S,8S,9R) or (4R,8R,9S) configuration, enantiomer 1 (Compound 78), and 4-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3-yl}- 14-(2,6-dimethylphenyl)-8-(2-methylpropyl)-11-oxa-18λ6-thia-3,15,17,24- tetraazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(23),12(24),13,15,19,21- hexaene-2,18,18-trione, (4R,8R,9S) or (4S,8S,9R) configuration, enantiomer 2 (Compound 79)
rac-2R,3S,7S enantiomer 1 enantiomer 2 4S,8S,9R or 4R,8R,9S 4R,8R,9S or 4S,8S,9R [00526] Racemic-3-[(4-{[(2R,3S,7S)-7-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3- yl}-3-(2-methylpropyl)azepan-2-yl]methoxy}-6-(2,6-dimethylphenyl)pyrimidin-2- yl)sulfamoyl]benzoic acid (hydrochloride salt) (144 mg, 0.09863 mmol) was dissolved in NMP (2.0 mL) and TsCl (80.6 mg, 0.4228 mmol) was added. The reaction was stirred for 10 minutes at room temperature. After 10 minutes, NMM (59.9 mg, 0.5922 mmol) was added and the reaction was stirred at 40 °C for 2 hours and was purified by reverse phase HPLC preparative chromatography ((20-80% acetonitrile/5 mM aqueous HCl) over 20 min) which gave racemic- (4S,8S,9R)-4-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3-yl}-14-(2,6- dimethylphenyl)-8-(2-methylpropyl)-11-oxa-18λ6-thia-3,15,17,24- tetraazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(23),12(24),13,15,19,21-hexaene-2,18,18-trione (28.7 mg, 39%).1H NMR (400 MHz, CDCl3) δ 8.90 (s, 1H), 8.43 (s, 1H), 8.05 (d, J = 7.8 Hz, 1H), 7.83 (d, J = 7.6 Hz, 1H), 7.64 (t, J = 7.7 Hz, 1H), 7.24 (t, J = 7.6 Hz, 1H), 7.11 (d, J = 7.6 Hz, 2H), 6.52 (s, 1H), 6.31 (s, 1H), 5.97 – 5.88 (m, 1H), 4.81 (t, J = 11.9 Hz, 1H), 4.48 (t, J = 6.9 Hz, 2H), 4.45 – 4.38 (m, 1H), 2.50 (t, J = 6.9 Hz, 2H), 2.39 (m, 1H), 2.24 – 2.13 (m, 1H), 2.08 (s, 6H), 2.04 – 1.79 (m, 5H), 1.48 (d, J = 11.1 Hz, 6H), 1.30 – 1.17 (m, 3H), 0.89 – 0.82 (m, 1H), 0.69 (t, J = 6.1 Hz, 6H). ESI-MS m/z calc.733.341, found 734.398 (M+1)+; Retention time: 2.657 minutes. The enantiomers were separated by chiral SFC using a using a normal phase SFC-MS method utilizing a Phenomenex LUX-1 (21.2 x 250 mm, 5 μm) at 40 oC. Mobile phase was isocratic 22% MeOH (w/ 20 mM NH3), 70 % CO2 at a 70 mL/min flow with an outlet pressure of 186 bar, detection wavelength of 210 nm. The collected fractions afforded two peaks by chiral column analysis. The first peak to elute was SFC peak 1, which was concentrated to give 4-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3-yl}-14-(2,6-dimethylphenyl)-8-(2- methylpropyl)-11-oxa-18λ6-thia-3,15,17,24-tetraazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(23),12(24),13,15,19,21-hexaene-2,18,18-trione (ammonium salt), (4S,8S,9R) or (4R,8R,9S) configuration, enantiomer 1 (8.3 mg, 11%). ESI-MS m/z calc.733.341, found 734.783 (M+1)+; Retention time: 2.658 minutes. The second peak to elute was SFC peak 2 which was concentrated to give 4-{8,8-dimethyl-6H,7H,8H-pyrazino[2,3-b]pyrrolizin-3-yl}-14-(2,6- dimethylphenyl)-8-(2-methylpropyl)-11-oxa-18λ6-thia-3,15,17,24- tetraazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(23),12(24),13,15,19,21-hexaene-2,18,18-trione (ammonium salt) (4S,8S,9R) or (4R,8R,9S) configuration, enantiomer 2 (6.1 mg, 8%) ESI-MS m/z calc.733.341, found 734.398 (M+1)+; Retention time: 2.66 minutes. LC method N. Example 64: Preparation of Compounds 82 and 83 Step 1: tert-Butyl (4S)-4-[(1R)-5-hydroxy-1-isopropoxy-5-[5-methyl-6-[1- (trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate ~90:10 mixture of diastereomers, the major isomer having the 4S,1R configuration [00527] To a solution of 3-bromo-5-methyl-6-[1-(trifluoromethyl)cyclobutyl]pyrrolo[2,3- b]pyrazine (300.8 mg, 0.8912 mmol) in tetrahydrofuran (2.5 mL) at -78 °C was added n-butyllithium (solution in hexanes) (422 µL of 2.5 M, 1.055 mmol) dropwise. As soon as the addition of n-BuLi was over, a solution of tert-butyl (4S)-4-[(1R)-1-isopropoxy-5-oxo-pentyl]- 2,2-dimethyl-oxazolidine-3-carboxylate (~90:10 mixture of diastereomers, the major isomer having the 4S,1R configuration) (255 mg, 0.7424 mmol) in tetrahydrofuran (2.5 mL) was added dropwise. The reaction was stirred for 10 min and MeOH was added. The crude mixture was concentrated in vacuo and was then purified by flash chromatography on silica gel (40 g) using 0-100% EtOAc in hexanes within 25 minutes. The product came out at 55 and 75% of EtOAc within 15-25 min to afford tert-butyl (4S)-4-[(1R)-5-hydroxy-1-isopropoxy-5-[5-methyl-6-[1- (trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (235.8 mg, 53%) as a yellow solid. ESI-MS m/z calc.598.3342, found 599.3 (M+1)+; Retention time: 0.84 minutes. LC method B. Step 2: tert-Butyl (4S)-4-[(1R)-1-isopropoxy-5-[5-methyl-6-[1- (trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00528] Manganese dioxide (827 mg, 9.513 mmol) was added to a solution of tert-butyl (4S)- 4-[(1R)-5-hydroxy-1-isopropoxy-5-[5-methyl-6-[1-(trifluoromethyl)cyclobutyl]pyrrolo[2,3- b]pyrazin-3-yl]pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (235.8 mg, 0.3938 mmol) in 1,2- dichloroethane (2.3 mL). The reaction mixture was stirred at 70 oC for 3 h. More manganese dioxide (171 mg, 1.967 mmol) was added. After 5 h, the reaction was allowed to cool to room temperature and filtered through a pad of celite. The filtrate was concentrated under reduced pressure. Purification on silica gel (24 g) (0-100%) EtOAc/hexanes over 15 min, gave as an orange foam tert-butyl (4S)-4-[(1R)-1-isopropoxy-5-[5-methyl-6-[1- (trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (152 mg, 64%) ESI-MS m/z calc.596.31854, found 597.2 (M+1)+; Retention time: 0.94 minutes. LC method B. Step 3: [(2S,3R)-3-Isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H- azepin-2-yl]methanol [00529] To a solution of tert-butyl (4S)-4-[(1R)-1-isopropoxy-5-[5-methyl-6-[1- (trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (152 mg, 0.2547 mmol) in 1,4-dioxane (3.3 mL) was added hydrochloric acid (solution in 1,4-dioxane) (1.2 mL of 4 M, 4.800 mmol). A drop of water was added, and the reaction was stirred for 5 h at rt. The solvent was removed under reduced pressure and the residue was co-evaporated with toluene to afford as a yellow solid [(2S,3R)-3- isopropoxy-7-[5-methyl-6-[1-(trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6- tetrahydro-2H-azepin-2-yl]methanol (150.5 mg, 128%). ESI-MS m/z calc.438.22427, found 439.1 (M+1)+; Retention time: 1.32 minutes. LC method A. Step 4: [(2S,3R)-3-Isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol ~1:1 mixture of diastereomers [00530] To a solution of [(2S,3R)-3-isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2- yl]methanol (150.5 mg, 0.3261 mmol) in dichloromethane (12 mL) was added sodium triacetoxyborohydride (560 mg, 2.642 mmol). The reaction was stirred at rt for 3 h. Then a saturated aqueous solution of sodium bicarbonate was added. The phases were separated, and the aqueous layer was extracted with DCM. The combined organic phases were dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and then purified by flash chromatography on silica gel (24 g, eluting with a gradient of 0-10% of MeOH in DCM). [(2S,3R)-3-isopropoxy-7-[5-methyl-6-[1-(trifluoromethyl)cyclobutyl]pyrrolo[2,3- b]pyrazin-3-yl]azepan-2-yl]methanol (73.1 mg, 97%) was isolated as a yellow solid (~1:1 mixture of diastereomers). ESI-MS m/z calc.440.2399, found 441.2 (M+1)+; Retention time: 0.53 minutes. LC method B. Step 5: 3-[[4-(2,6-Dimethylphenyl)-6-[[(2S,3R)-3-isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2- yl]methoxy]pyrimidin-2-yl]sulfamoyl]benzoic acid
~1:1 mixture of diastereomers [00531] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (100 mg, 0.2393 mmol) and [(2S,3R)-3-isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (73.1 mg, 0.1576 mmol) were combined in a vial and dissolved in THF (920 µL) followed by the addition of sodium tert-butoxide (98 mg, 1.020 mmol). The reaction mixture was stirred at 40 C for 4 h. The reaction mixture was cooled down to room temperature and diluted with EtOAc and 1 M HCl (1 mL). The organic phase was separated, and the aqueous layer was extracted with EtOAc. The organic layers were combined, washed with brine, and dried over sodium sulfate. The mixture was filtered, and then concentrated under reduced pressure. The crude mixture of diastereomers was carried forward to the next step.3-[[4-(2,6-dimethylphenyl)-6-[[(2S,3R)-3-isopropoxy-7-[5- methyl-6-[1-(trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2- yl]methoxy]pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (170 mg, 126%). ESI- MS m/z calc.821.31824, found 822.2 (M+1)+; Retention time: 0.61 minutes. LC method B. Step 6: (8R,9S)-14-(2,6-Dimethylphenyl)-8-isopropoxy-4-[5-methyl-6-[1- (trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one, diastereomer 1 (Compound 82) and (8R,9S)-14-(2,6-dimethylphenyl)-8- isopropoxy-4-[5-methyl-6-[1-(trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18- dioxo-11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one, diastereomer 2 (Compound 83)
~1:1 mixture of diastereomers Diastereomer 1 Diastereomer 2 [00532] 3-[[4-(2,6-Dimethylphenyl)-6-[[(2S,3R)-3-isopropoxy-7-[5-methyl-6-[1- (trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (mixture of diastereomers, 170 mg, 0.1980 mmol) was dissolved in NMP (3 mL) and Acetone (7 mL). TsCl (138 mg, 0.7239 mmol) was added followed by NMM (192 µL, 1.746 mmol) and the reaction was stirred at 55 °C for 20 hours. The reaction was poured into a mixture of ethyl acetate and 1 M HCl and the layers were separated. The aqueous layer was further extracted with ethyl acetate (2 x). The organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was taken up in DMSO/MeOH then purified by reverse phase chromatography (1-99% acetonitrile/5 mM aqueous HCl over 25 min) to give a mixture of diastereomers. Additional purification by preparative HPLC (1-99% acetonitrile/5 mM aqueous HCl over 30 min) gave early peak, diastereomer 1, (8R,9S)-14-(2,6-dimethylphenyl)-8- isopropoxy-4-[5-methyl-6-[1-(trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18- dioxo-11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (11.9 mg, 15%) as a yellow solid.1H NMR (400 MHz, CDCl3) δ 8.81 (s, 1H), 8.65 (s, 1H), 7.75 (d, J = 7.7 Hz, 2H), 7.56 (t, J = 7.7 Hz, 1H), 7.27 – 7.23 (m, 1H), 7.11 (d, J = 7.6 Hz, 2H), 6.79 (s, 1H), 6.28 (s, 1H), 5.86 (s, 1H), 4.62 (br s, 1H), 4.45 (br s, 1H), 4.09 (br s, 1H), 3.91 (s, 3H), 3.77 (br s, 1H), 3.56 – 3.44 (m, 1H), 3.01 (br s, 1H), 2.88 (br s, 2H), 2.81 – 2.70 (m, 2H), 2.46 – 2.26 (m, 3H), 2.09 (s, 7H), 1.94 – 1.83 (m, 2H), 1.26 (s, 1H), 0.96 (d, J = 5.9 Hz, 3H), 0.89 (d, J = 5.9 Hz, 3H). ESI-MS m/z calc. 803.3077, found 804.4 (M+1)+; Retention time: 2.11 minutes; and late peak, diastereomer 2, (8R,9S)-14-(2,6-dimethylphenyl)-8-isopropoxy-4-[5-methyl-6-[1- (trifluoromethyl)cyclobutyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen- 2-one (17.5 mg, 22%) as a yellow solid.1H NMR (400 MHz, CDCl3) δ 8.74 (s, 1H), 8.55 (s, 1H), 7.85 – 7.82 (m, 1H), 7.57 (t, J = 7.7 Hz, 1H), 7.26 – 7.19 (m, 1H), 7.08 (d, J = 7.6 Hz, 2H), 6.73 (s, 1H), 6.26 (s, 1H), 6.06 – 5.95 (m, 1H), 5.86 – 5.78 (m, 1H), 4.73 (s, 1H), 3.95 (s, 3H), 3.59 (s, 1H), 3.40 – 3.30 (m, 1H), 2.88 (s, 2H), 2.84 – 2.72 (m, 2H), 2.54 – 2.23 (m, 3H), 2.21 – 1.86 (m, 11H), 1.84 – 1.65 (m, 2H), 1.01 (d, J = 6.0 Hz, 3H), 0.66 (d, J = 5.9 Hz, 3H). ESI-MS m/z calc.803.3077, found 804.4 (M+1)+; Retention time: 2.17 minutes. LC method A. Example 65: Preparation of Compound 88 Step 1: Tributyl-(6-tert-butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)stannane (n-Bu)3Sn Sn(n-Bu)3 + [00533] To a nitrogen flushed suspension of 6-tert-butyl-2-chloro-7-methyl-pyrrolo[2,3- d]pyrimidine (4.79 g, 21.263 mmol) in dioxane (75 mL) were added lithium chloride (6 g, 141.53 mmol), tricyclohexylphosphine (937.3 mg, 3.3424 mmol), tributyl(tributylstannyl)stannane (22.960 g, 20 mL, 39.579 mmol) and Pd2(dba)3 (956.1 mg, 1.0441 mmol). The resulting suspension was sealed in a pressure tube and heated at 110 °C overnight. The suspension was filtered through Celite. The filter cake was washed with EtOAc (50 mL) and an aqueous solution of 1M KF was added (100 mL) into the filtrate. The mixture was stirred for 30 min at rt. The resulting precipitate was filtered over Celite. The biphasic filtrate was transferred to a funnel, the phases were separated, and the organic phase was washed with an aqueous solution of 1M KF (2 x 50 mL), water (100 mL), brine (100 mL) and evaporated under reduced pressure. The crude was diluted with EtOAc (50 mL) and an aqueous solution of 1M KF (50 mL) was added. The mixture was stirred for 30 min at rt. The resulting precipitate was filtered over Celite. The biphasic filtrate was transferred to a funnel, the phases were separated, and the organic phase was washed with an aqueous solution of 1M KF (2 x 50 mL), water (50 mL), brine (50 mL), dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure. Purification by silica gel chromatography (Gradient 0 – 20% EtOAc in heptanes) afforded after evaporation of the volatiles tributyl-(6-tert-butyl-7-methyl- pyrrolo[2,3-d]pyrimidin-2-yl)stannane (4.08 g, 38%) as a yellow oil. ESI-MS m/z calc. 479.2322, found 480.3 (M+1)+; Retention time: 3.89 minutes.1H NMR (400 MHz, Chloroform- d) δ 8.83 (s, 1H), 6.25 (s, 1H), 4.01 (s, 3H), 1.70 – 1.59 (m, 6H), 1.48 (s, 9H), 1.43 – 1.32 (m, 6H), 1.23 (s, 6H), 0.90 (t, J = 7.3 Hz, 9H). LC method P. Step 2: tert-Butyl (4S)-4-[(1R)-5-(6-tert-butyl-7-methyl-pyrrolo[2,3-d]pyrimidin- 2-yl)-5-hydroxy-1-isopropoxy-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00534] To a solution of tributyl-(6-tert-butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)stannane (271 mg, 0.5407 mmol) in THF (2.6 mL) at -78°C was added n-butyllithium (281 µL of 2.5 M, 0.7025 mmol) dropwise. After 1 minute, a solution of tert-butyl (4S)-4-[(1R)-1-isopropoxy-5- oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (204 mg, 0.5940 mmol) in THF (2.6 mL) was added dropwise. The reaction was stirred for 30 min and a solution of ammonium chloride was added and the reaction was partitioned between water and EtOAc. The aqueous layer was extracted with EtOAc (3 x) and the organic layer was washed with brine, followed by drying over sodium sulfate. Evaporation of the solvents gave tert-butyl (4S)-4-[(1R)-5-(6-tert-butyl-7- methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-5-hydroxy-1-isopropoxy-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (316 mg, 45%). ESI-MS m/z calc.532.3625, found 530.899 (M+1)+; Retention time: 2.11 minutes. LC method N Step 3: tert-Butyl (4S)-4-[(1R)-5-(6-tert-butyl-7-methyl-pyrrolo[2,3-d]pyrimidin- 2-yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00535] To a solution of tert-butyl (4S)-4-[(1R)-5-(6-tert-butyl-7-methyl-pyrrolo[2,3- d]pyrimidin-2-yl)-5-hydroxy-1-isopropoxy-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (316 mg, 0.3796 mmol) in dichloroethane (4.4 mL) was added manganese dioxide (647 mg, 7.442 mmol). The resulting suspension was stirred at 70 °C for 1 h. The suspension was filtered on a pad of Celite and washed with DCM (2 x 50 mL). The crude mixture was concentrated in vacuo and was then purified by flash chromatography on silica gel (0-100% EtOAc in hexanes over 20 minutes)). This was then purified by reverse phase preparative HPLC ((20-100% acetonitrile/water) over 25 min) to give tert-butyl (4S)-4-[(1R)-5-(6-tert-butyl-7-methyl- pyrrolo[2,3-d]pyrimidin-2-yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (43.8 mg, 22%) ESI-MS m/z calc.530.3468, found 531.973 (M+1)+; Retention time: 2.432 minutes. LC method N. Step 4: [(2S,3R)-7-(6-tert-Butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-3- isopropoxy-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00536] To a stirred solution of tert-butyl (4S)-4-[(1R)-5-(6-tert-butyl-7-methyl-pyrrolo[2,3- d]pyrimidin-2-yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (43.8 mg, 0.08253 mmol) in a mixture of dioxane (1.3 mL) and water (636 µL) was added a solution of hydrogen chloride (2.1 mL of 4 M, 8.400 mmol) dropwise. The reaction was stirred at room temperature for 16 hours, then the reaction was concentrated to dryness. The residual water was co-evaporated with acetonitrile, then dried under high vacuum to afford crude [(2S,3R)-7-(6-tert- butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-3-isopropoxy-3,4,5,6-tetrahydro-2H-azepin-2- yl]methanol (90.4 mg, 247%). ESI-MS m/z calc.372.25253, found 373.455 (M+1)+; Retention time: 1.225 minutes. LC method N. Step 5: [(2S,3R)-7-(6-tert-Butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-3- isopropoxy-azepan-2-yl]methanol, diastereomer 1, and [(2S,3R)-7-(6-tert-butyl- 7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-3-isopropoxy-azepan-2-yl]methanol, diastereomer 2 [00537] To a solution of [(2S,3R)-7-(6-tert-butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-3- isopropoxy-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (90.4 mg, 0.2427 mmol) in anhydrous dichloromethane (4.8 mL) at room temperature was added sodium triacetoxyborohydride (311 mg, 1.467 mmol). The reaction was stirred at room temperature for 1 hour and a saturated aqueous solution of sodium bicarbonate was added. The phases were separated, and the aqueous solution was extracted with DCM (2 x). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (0-35% acetonitrile/5 mM aqueous HCl over 25 min) to give [(2S,3R)-7-(6-tert-butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-3-isopropoxy- azepan-2-yl]methanol (hydrochloride salt), diastereomer 1, peak 1 (5.2 mg, 4%). ESI-MS m/z calc.374.2682, found 375.491 (M+1)+; Retention time: 1.565 minutes, and [(2S,3R)-7-(6-tert- butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-3-isopropoxy-azepan-2-yl]methanol (hydrochloride salt) , diastereomer 2, peak 2 (2.3 mg, 2%). ESI-MS m/z calc.374.2682, found 375.436 (M+1)+; Retention time: 1.618 minutes. LC method N. Step 6: 3-[[4-[[(2S,3R)-7-(6-tert-Butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-3- isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid Diastereomer 1 [00538] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (6.4 mg, 0.01532 mmol) and [(2S,3R)-7-(6-tert-butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-3- isopropoxy-azepan-2-yl]methanol (hydrochloride salt), diastereomer 1 (5.2 mg, 0.01151 mmol) were combined in a vial and dissolved in THF (384 µL) followed by the addition of sodium tert- butoxide (9.4 mg, 0.08377 mmol). The reaction mixture was stirred for 30 minutes at 40 °C. The reaction mixture was diluted with EtOAc and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (3 x). The organic layers were combined, washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3-[[4-[[(2S,3R)-7-(6-tert-butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-3-isopropoxy- azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt), diastereomer 1 (18.8 mg, 46%) ESI-MS m/z calc.755.3465, found 756.572 (M+1)+; Retention time: 2.093 minutes. LC method N. Step 7: (8R,9S)-4-(6-tert-Butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-14-(2,6-dimethylphenyl)- 8-isopropoxy-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one Diastereomer 1 Diastereomer 1 [00539] 3-[[4-[[(2S,3R)-7-(6-tert-Butyl-7-methyl-pyrrolo[2,3-d]pyrimidin-2-yl)-3- isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt), diastereomer 1 (18.8 mg, 0.03635 mmol) was dissolved in NMP (576 µL) and acetone (1.2 mL) followed by the addition of TsCl (17.3 mg, 0.09075 mmol). Next, NMM (33.1 mg, 0.3272 mmol) was added, and the reaction was stirred at 55°C for 3 hours. The reaction was dried under reduced pressure to remove the acetone followed by dilution with DMSO and then reverse phase HPLC preparative chromatography ((20-70%, acetonitrile/5 mM aqueous HCl) over 25 minutes) which gave (8R,9S)-4-(6-tert-butyl-7-methyl-pyrrolo[2,3- d]pyrimidin-2-yl)-14-(2,6-dimethylphenyl)-8-isopropoxy-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen- 2-one (4.2 mg, 69%).1H NMR (400 MHz, CDCl3) δ 9.02 (s, 1H), 8.90 (s, 1H), 7.87 (d, J = 7.8 Hz, 1H), 7.82 (d, J = 7.5 Hz, 1H), 7.62 (t, J = 7.7 Hz, 1H), 7.32 – 7.23 (m, 1H), 7.12 (d, J = 7.6 Hz, 2H), 6.65 (s, 1H), 6.30 (s, 1H), 6.22 (d, J = 12.3 Hz, 1H), 5.96 (d, J = 7.4 Hz, 1H), 4.69 (s, 1H), 4.24 (s, 3H), 3.62 (s, 1H), 3.41 (p, J = 6.0 Hz, 1H), 2.33 – 2.14 (m, 1H), 2.09 (s, 6H), 1.99 (dd, J = 30.4, 14.1 Hz, 5H), 1.79 (t, J = 12.6 Hz, 1H), 1.53 (s, 9H), 1.07 (d, J = 5.9 Hz, 3H), 0.76 (d, J = 5.9 Hz, 3H). ESI-MS m/z calc.737.33594, found 738.635 (M+1)+; Retention time: 2.102 minutes. LC method N. Example 66: Preparation of Compound 89 Step 1: 4-Carboxybutyl(triphenyl)phosphonium bromide [00540] To a round-bottomed flask under nitrogen atmosphere containing 5-bromopentanoic acid (10 g, 54.135 mmol) in MeCN (150 mL) was added PPh3 (15 g, 56.618 mmol) and the reaction mixture was stirred at 80 °C for 48 hours. The volatiles were removed under vacuum and the residue was suspended in EtO2 (200 mL). The resulting white solid was filtered and washed with EtO2 (3 x 100 mL). The solid was dissolved in DCM and the volatiles were removed in vacuo to afford 4-carboxybutyl(triphenyl)phosphonium bromide (16.8 g, 69%) as a white solid.1H NMR (400 MHz, DMSO-d6) δ 13.07 – 11.15 (br s, 1H), 7.94 – 7.73 (m, 15H), 3.66 – 3.56 (m, 2H), 2.28 (t, J = 7.2 Hz, 2H), 1.69 (quin, J = 7.3 Hz, 2H), 1.63 – 1.50 (quin, J = 7.3 Hz, 2H). ESI-MS m/z calc.363.1514, found 363.1 (M)+; Retention time: 3.17 minutes. LC method X. Step 2: 6-[(4R)-3-tert-Butoxycarbonyl-2,2-dimethyl-oxazolidin-4-yl]hex-5-enoic acid [00541] To a solution of 4-carboxybutyl(triphenyl)phosphonium bromide (16.52 g, 35.821 mmol) in THF (50 mL) under argon atmosphere at -78 °C was added dropwise a solution of LHMDS (95 mL of 1 M, 95.000 mmol) in THF. The mixture was stirred for 30 minutes allowing it to warm to 0 °C at which time it was stirred for another 1 hour at room temperature. The mixture was cooled again to -78 °C and tert-butyl (4S)-4-formyl-2,2-dimethyl-oxazolidine- 3-carboxylate (7.23 g, 29.958 mmol) in THF (72 mL) was added dropwise. The mixture was stirred for 30 minutes at 0 °C and 2 hours at room temperature. The reaction was quenched with an aqueous saturated ammonium chloride solution (100 mL) and the volatiles were removed in vacuo. The product was extracted with EtOAc (250 mL), washed with water (250 mL) and brine (250 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. Purification by silica gel chromatography (Gradient: 0-80 % EtOAc in Heptanes) followed by purification by reversed-phase chromatography (C18 column. Gradient: 5-100 % MeCN in basic water (ammonium bicarbonate/ammonium hydroxide buffer pH = 10) afforded after evaporation of the volatiles 6-[(4R)-3-tert-butoxycarbonyl-2,2-dimethyl-oxazolidin-4-yl]hex-5-enoic acid (1.56 g, 17%) as a white solid. Z/E ratio: 88:12. ESI-MS m/z calc.313.1889, found 258.1 (M- 55)+; Retention time: 2.02 minutes. LC method O. Step 3: 6-[(4R)-3-tert-Butoxycarbonyl-2,2-dimethyl-oxazolidin-4-yl]hexanoic acid [00542] A round-bottom flask was purged with nitrogen and palladium on carbon (530 mg, 10 %w/w, 0.4980 mmol) was added. Then was added a solution of 6-[(4R)-3-tert-butoxycarbonyl- 2,2-dimethyl-oxazolidin-4-yl]hex-5-enoic acid (1.56 g, 4.9729 mmol) in MeOH (46 mL). The mixture was purged with nitrogen for 5 minutes followed by hydrogen for 5 minutes and was stirred at room temperature for 2 hours and 30 minutes under hydrogen atmosphere. The mixture was purged with nitrogen for 5 minutes, filtered through Celite, washed with MeOH (80 mL) and the volatiles were removed under reduced pressure. The residue was diluted in EtOAc (50 mL), filtered through a syringe filter (5.0 μm) and evaporated in vacuo to afford crude 6-[(4R)- 3-tert-butoxycarbonyl-2,2-dimethyl-oxazolidin-4-yl]hexanoic acid (1.6 g, 102%) as a clear oil. 1H NMR (400 MHz, DMSO-d6) δ 3.91 – 3.82 (m, 1H), 3.79 – 3.63 (m, 2H), 2.18 (t, J = 7.4 Hz, 2H), 1.67 – 1.14 (m, 23H), (1H missing, labile proton). ESI-MS m/z calc.315.2046, found 260.2 (M-55)+; Retention time: 2.05 minutes. LC method O. Step 4: tert-Butyl (4R)-4-[6-[methoxy(methyl)amino]-6-oxo-hexyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00543] To a stirred solution of 6-[(4R)-3-tert-butoxycarbonyl-2,2-dimethyl-oxazolidin-4- yl]hexanoic acid (1.6 g, 5.0678 mmol) in DCM (16 mL) at 0 °C was added CDI (1.3 g, 8.0173 mmol) in portions to maintain the reaction temperature below 20 °C. The mixture was stirred at room temperature for 1 hour, then triethylamine (1.3725 g, 1.9 mL, 13.496 mmol) and N- methoxymethanamine (hydrochloride salt) (995 mg, 9.9965 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. DCM (100 mL) and water were added (100 mL). The aqueous layer was extracted with DCM (3 x 100 mL) The layers were separated and the organic layer was washed with a 5% aqueous solution of citric acid (150 mL), a saturated aqueous solution of sodium bicarbonate (150 mL), water (300 mL) and brine (300 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by silica gel chromatography (Gradient: 0-100 % EtOAc in Heptanes) yielded after evaporation of the volatiles tert-butyl (4R)-4-[6- [methoxy(methyl)amino]-6-oxo-hexyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.6 g, 88%) as a clear oil.1H NMR (400 MHz, DMSO-d6) δ 3.87 (dd, J = 8.7, 5.7 Hz, 1H), 3.79 – 3.66 (m, 2H), 3.64 (s, 3H), 3.07 (s, 3H), 2.35 (br t, J = 7.3 Hz, 2H), 1.66 – 1.14 (m, 23H). ESI-MS m/z calc.358.2468, found 359.2 (M+1)+; Retention time: 2.15 minutes. LC method O. Step 5: tert-Butyl (4R)-4-[6-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-6- oxo-hexyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00544] To a solution of 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine (175 mg, 0.6102 mmol) in anhydrous THF (3 mL) at -78 °C was added dropwise a solution of n-BuLi (in hexanes) (0.4 mL of 2.5 M, 1.0000 mmol). The reaction mixture was stirred for 10 minutes then a solution of tert-butyl (4R)-4-[6-[methoxy(methyl)amino]-6-oxo-hexyl]-2,2-dimethyl- oxazolidine-3-carboxylate (218 mg, 0.6075 mmol) in anhydrous THF (3 mL) was added dropwise. The reaction was stirred in the dry-ice bath for 40 minutes and methanol (10 mL) was added. The reaction was slowly allowed to warm to room temperature and was concentrated under reduced pressure. Purification by silica gel chromatography (Gradient: 0-50 % EtOAc in Heptanes) followed by purification by reversed-phase chromatography (C18 column. Gradient: 5-100 % MeCN in water) yielded after evaporation of the volatiles tert-butyl (4R)-4-[6-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-6-oxo-hexyl]-2,2-dimethyl-oxazolidine-3- carboxylate (155 mg, 52%) as a yellow oil.1H NMR (400 MHz, DMSO-d6) δ 8.93 (s, 1H), 6.64 (s, 1H), 4.04 (s, 3H), 3.91 – 3.83 (m, 1H), 3.81 – 3.65 (m, 2H), 3.22 (t, J = 7.3 Hz, 2H), 1.74 – 1.64 (m, 2H), 1.53 – 1.43 (m, 14H), 1.37 (br d, J = 9.2 Hz, 16H). ESI-MS m/z calc.486.3206, found 487.3 (M+1)+; Retention time: 2.57 minutes. LC method O. Step 6: (7R)-7-Amino-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8- hydroxy-octan-1-one [00545] To a stirred solution of tert-butyl (4R)-4-[6-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-6-oxo-hexyl]-2,2-dimethyl-oxazolidine-3-carboxylate (155 mg, 0.3182 mmol) in a mixture of 1,4-dioxane (5.5 mL) and water (1.4 mL) was added a solution of hydrochloric acid (solution in 1,4-dioxane) (1 mL of 4 M, 4.0000 mmol) dropwise. The reaction was stirred at room temperature for 16 hours, then additional hydrochloric acid (solution 1,4-dioxane) (1 mL of 4 M, 4.0000 mmol) was added, and the mixture was stirred for an additional 20 hours. The mixture was concentrated under reduced pressure and co-evaporated with acetonitrile (2 x 2 mL). Purification by reversed-phase chromatography (C18 column. Gradient: 5-100 % MeCN in water with 0.1 % formic acid) afforded after evaporation of the volatiles (7R)-7-amino-1-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8-hydroxy-octan-1-one (93 mg, 84%) as a yellow oil.1H NMR (400 MHz, DMSO-d6) δ 8.94 (s, 1H), 8.38 (br s, 1H), 6.64 (s, 1H), 4.05 (s, 3H), 3.52 (dd, J = 11.4, 3.8 Hz, 1H), 3.36 (dd, J = 11.4, 6.4 Hz, 1H), 3.23 (t, J = 7.3 Hz, 2H), 2.99 – 2.89 (m, 1H), 1.74 – 1.63 (m, 2H), 1.52 – 1.47 (m, 11H), 1.47 – 1.25 (m, 6H). ESI-MS m/z calc. 346.2369, found 347.3 (M+1)+; Retention time: 1.65 minutes. LC method O. Step 7: [(2R,8S)-8-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)azocan-2- yl]methanol [00546] A solution of (7R)-7-amino-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8- hydroxy-octan-1-one (375 mg, 1.0813 mmol) in anhydrous DCE (105 mL) with molecular sieves was stirred at room temperature during 48 h. Then, NaBH(OAc)3 (241 mg, 1.1371 mmol) was added. The reaction was stirred at room temperature for 48 h and a saturated aqueous solution of ammonium chloride (20 mL) was added. The reaction media was filtered on a celite pad that was washed with DCM (2 x 50 mL). The phases were separated, and the aqueous solution was extracted with DCM (3 x 50 mL). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by silica gel chromatography (Gradient: 0-100 % EtOAc in Heptanes) yielded after evaporation of the volatiles and freeze-drying [(2R,8S)-8-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)azocan-2-yl]methanol (116.2 mg, 31%) as a yellow solid.1H NMR (400 MHz, Benzene-d6) δ 8.57 (s, 1H), 6.60 (s, 1H), 4.00 (dd, J = 8.3, 3.8 Hz, 1H), 3.51 (s, 3H), 3.48 (dd, J = 10.1, 4.6 Hz, 1H), 3.11 (t, J = 9.7 Hz, 1H), 2.84 – 2.76 (m, 1H), 2.05 – 1.96 (m, 1H), 1.80 – 1.64 (m, 3H), 1.62 – 1.49 (m, 2H), 1.45 – 1.18 (overlapping with grease, m, 4H), 1.10 (s, 10H), (1H missing, labile proton). ESI-MS m/z calc.330.242, found 331.2 (M+1)+; Retention time: 3.42 minutes. LC method W. Step 8: 3-[[4-[[(2R,8S)-8-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)azocan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00547] In a 4 mL vial, 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (76 mg, 0.1819 mmol) and [(2R,8S)-8-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)azocan-2-yl]methanol (50 mg, 0.1513 mmol) were combined in THF (0.5 mL). Sodium tert- butoxide (87 mg, 0.9053 mmol) was added. The reaction mixture was stirred at 55 °C for 1 hour. It was diluted with EtOAc (50 mL) and extracted with water (1 × 50 mL). The aqueous layer was acidified to pH 6 with the addition of 1 M HCl and extracted with EtOAc (2× 50 mL). The final organic layers were combined, dried over sodium sulfate, filtered and concentrated under reduced pressure to provide 3-[[4-[[(2R,8S)-8-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)azocan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (103 mg, 91%) as a slightly yellow solid. ESI-MS m/z calc.711.3203, found 712.3 (M+1)+; Retention time: 1.44 minutes. LC method A. Step 9: (4S,10R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-15-(2,6- dimethylphenyl)-19,19-dioxo-12-oxa-19λ6-thia-3,16,18,25- tetrazatetracyclo[18.3.1.113,17.03,10]pentacosa-1(23),13,15,17(25),20(24),21- hexaen-2-one [00548] In a 4 mL vial, 3-[[4-[[(2R,8S)-8-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)azocan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (20 mg, 0.02673 mmol) and 4-methylbenzenesulfonyl chloride (13 mg, 0.06819 mmol) were combined in acetone (0.8 mL) and NMP (0.4 mL), followed by the addition of 4-methylmorpholine (18 µL, 0.1637 mmol). The reaction mixture was stirred at 55 °C for 2 days. It was concentrated to about a third of its volume under reduced pressure. Purification by reverse phase HPLC (30-99% acetonitrile/5 mM aqueous HCl over 15 minutes) provided (4S,10R)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-15-(2,6- dimethylphenyl)-19,19-dioxo-12-oxa-19λ6-thia-3,16,18,25- tetrazatetracyclo[18.3.1.113,17.03,10]pentacosa-1(23),13,15,17(25),20(24),21-hexaen-2-one (4 mg, 20%) as a beige solid.1H NMR (400 MHz, CDCl3) δ 8.80 (s, 1H), 8.49 (s, 1H), 8.01 (d, J = 8.0 Hz, 1H), 7.93 (d, J = 7.6 Hz, 1H), 7.69 (t, J = 7.8 Hz, 1H), 7.20 (t, J = 7.7 Hz, 1H), 7.03 (d, J = 7.6 Hz, 2H), 6.66 (t, J = 9.6 Hz, 1H), 6.56 (s, 1H), 6.02 (s, 1H), 4.74 (dd, J = 10.9, 4.2 Hz, 1H), 4.25 – 4.15 (m, 1H), 4.12 (s, 3H), 3.33 (t, J = 11.3 Hz, 1H), 2.73 – 2.64 (m, 2H), 2.21 – 2.14 (m, 1H), 1.98 (s, 6H), 1.94 – 1.82 (m, 4H), 1.55 (s, 9H), 1.54 – 1.50 (m, 2H), 1.28 – 1.21 (m, 1H). ESI-MS m/z calc.693.30975, found 694.4 (M+1)+; Retention time: 1.96 minutes. LC method A. Example 67: Preparation of Compound 93 Step 1: tert-Butyl (4R)-4-[(E,1R)-1-(cyclopropylmethyl)-5- [methoxy(methyl)amino]-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3- carboxylate [00549] To a suspension of sodium hydride (suspension in oil) (1.31 g, 60 %w/w, 32.753 mmol) in dry THF (100 mL) was added 2-diethoxyphosphoryl-N-methoxy-N-methyl-acetamide (7.9713 g, 7 mL, 32.658 mmol) at room temperature over 10 minutes. The suspension was stirred at room temperature for 20 minutes and was then added to a solution of tert-butyl (4R)-4- [(1R)-1-(cyclopropylmethyl)-3-oxo-propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (6.35 g, 19.371 mmol) in dry THF (100 mL). The mixture was stirred at room temperature for 75 minutes. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (100 mL) and extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (1 x 250 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeCN in water. The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (100 mL) was then added, and the mixture was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (1 x 250 mL), dried over sodium sulfate, filtered, and concentrated in vacuo to afford tert-butyl (4R)-4-[(E,1R)-1-(cyclopropylmethyl)-5-[methoxy(methyl)amino]-5-oxo-pent-3- enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (7.5 g, 98%) as a yellow oil (Z or E configuration of the double bond unknown).1H NMR (400 MHz, CDCl3) δ 7.09 – 6.91 (m, 1H), 6.59 – 6.26 (m, 1H), 4.02 – 3.77 (m, 3H), 3.74 – 3.66 (m, 3H), 3.28 – 3.20 (m, 3H), 2.66 – 2.42 (m, 1H), 2.37 – 2.08 (m, 2H), 1.56 – 1.43 (m, 15H), 1.38 – 1.31 (m, 1H), 1.24 – 1.15 (m, 1H), 0.76 – 0.62 (m, 1H), 0.58 – 0.37 (m, 2H), 0.13 - -0.07 (m, 2H). ESI-MS m/z calc.396.2624, found 397.4 (M+1)+; Retention time: 1.97 minutes. LC method I. Step 2: tert-Butyl (4R)-4-[(1S)-1-(cyclopropylmethyl)-5-[methoxy(methyl)amino]- 5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00550] Nitrogen was bubbled through a stirred solution of tert-butyl (4R)-4-[(E,1R)-1- (cyclopropylmethyl)-5-[methoxy(methyl)amino]-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine- 3-carboxylate (7.5 g, 18.896 mmol) in dry methanol (375 mL) for 10 minutes, then 10% palladium on carbon (50% wet) (5.94 g, 5 %w/w, 2.7908 mmol) was added. Nitrogen was bubbled through the mixture for 10 minutes, followed by hydrogen for 10 minutes. The reaction was then stirred for 1 hour at room temperature under hydrogen atmosphere. Nitrogen was bubbled through the mixture for 10 minutes. The mixture was filtered on a pad of Celite, washed with MeOH (500 mL) and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 50 to 100% of MeCN in water to afford after evaporation tert-butyl (4R)-4-[(1S)-1-(cyclopropylmethyl)-5- [methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (5.3 g, 68%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 4.12 – 3.94 (m, 1H), 3.93 – 3.88 (m, 1H), 3.84 – 3.77 (m, 1H), 3.69 (s, 3H), 3.18 (s, 3H), 2.56 – 2.33 (m, 2H), 2.21 – 1.94 (overlapped with MeCN, m, 1H), 1.79 – 1.65 (m, 2H), 1.61 – 1.54 (overlapped with water, m, 4H), 1.51 – 1.43 (m, 12H), 1.35 – 1.11 (m, 3H), 0.76 – 0.63 (m, 1H), 0.54 – 0.44 (m, 1H), 0.43 – 0.35 (m, 1H), 0.10 – 0.01 (m, 1H), 0.00 - -0.09 (m, 1H). ESI-MS m/z calc.398.2781, found 399.4 (M+1)+; Retention time: 3.72 minutes. LC method 1B. Step 3: tert-Butyl (4R)-4-[(1S)-1-(cyclopropylmethyl)-5-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00551] To a solution of 3-bromo-5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazine (450.2 mg, 1.406 mmol) in tetrahydrofuran (7.25 mL) at -78 °C was added n- butyllithium (solution in hexanes) (0.53 mL of 2.5 M, 1.325 mmol) dropwise. The resulting solution was stirred for 5 minutes. A solution of tert-butyl (4R)-4-[(1S)-1-(cyclopropylmethyl)- 5-[methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (400 mg, 1.004 mmol) in tetrahydrofuran (2.85 mL) was added dropwise. The reaction was stirred for 15 min. The reaction mixture was diluted with EtOAc (20 mL) and ammonium chloride (8 mL) was added. The crude mixture was allowed to warm to room temperature and the phases were separated. The aqueous phase was extracted with EtOAc (2x) and the combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude material was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 15 min) to give tert-butyl (4R)-4-[(1S)-1-(cyclopropylmethyl)-5-[5-methyl-6- [1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (446.3 mg, 77%) as an orange foam. ESI-MS m/z calc.578.308, found 579.1 (M+1)+; Retention time: 2.4 minutes. LC method A. Step 4: [(2R,3S)-3-(Cyclopropylmethyl)-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H- azepin-2-yl]methanol [00552] To a solution of tert-butyl (4R)-4-[(1S)-1-(cyclopropylmethyl)-5-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (446.3 mg, 0.7712 mmol) in 1,4-dioxane (10 mL) was added hydrochloric acid (solution in 1,4-dioxane) (4.0 mL of 4 M, 16.00 mmol) at rt. Ten drops of water were added and the reaction was stirred for 18 hours. The solvent was removed under reduced pressure and the residual water was co-evaporated with acetonitrile (3 x), then dried under high vacuum to afford crude [(2R,3S)-3-(cyclopropylmethyl)-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2- yl]methanol (324.3 mg, 100%) as a yellow solid. ESI-MS m/z calc.420.21368, found 421.2 (M+1)+; Retention time: 1.28 minutes. The crude material was taken forward without further purification. LC method A. Step 5: [(2R,3S,7S)-3-(Cyclopropylmethyl)-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol, diastereomer 1, and [(2R,3S,7R)-3-(cyclopropylmethyl)-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol, diastereomer 2 [00553] To a solution of [(2R,3S)-3-(cyclopropylmethyl)-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2- yl]methanol (324.4 mg, 0.7715 mmol) in anhydrous dichloromethane (3.4 mL) at 0 °C was added sodium triacetoxyborohydride (983.9 mg, 4.642 mmol). The reaction was stirred at 0 °C for 1 hour then allowed to warm to room temperature. The reaction mixture was stirred for an additional 2 hours. A saturated aqueous solution of sodium bicarbonate was added. The phases were separated, and the aqueous solution was extracted with DCM (2 x). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 45 min) to give 211 mg of product. This material was subjected to chiral SFC separation using a Phenomenex LUX-4 (21.2 x 250 mm, 5 μm) column at 40 oC, a gradient of MeOH (20 mM NH3) in CO2 at 70 mL/min flow (26% of MeOH from 0 to 5.2 min, 32% of MeOH from 5.3 to 8.90 min, 26% of MeOH from 8.92 min to 13 min). Concentration of the sample was 23.4 mg/mL in MeOH. The injection volume was 520 μL with a variable outlet pressure and the detection wavelength was 224 nm. After evaporation of the solvents, each isomer was dissolved in DMSO (1 mL) and MeOH (1mL) and purified by reverse phase HPLC (10-99% acetonitrile/5 mM aqueous HCl over 15 min). The pure fractions were extracted with EtOAc (3x), washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give: diastereomer 1, Peak 1 [(2R,3S,7S)-3- (cyclopropylmethyl)-7-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3- yl]azepan-2-yl]methanol (82.4 mg, 25%) as a white solid.1H NMR (400 MHz, CDCl3) δ 8.62 (s, 1H), 6.74 (s, 1H), 4.40 (dd, J = 10.6, 4.7 Hz, 1H), 3.96 (s, 3H), 3.79 (d, J = 6.2 Hz, 2H), 3.20 – 3.04 (m, 1H), 2.51 – 2.29 (m, 3H), 2.14 – 2.00 (m, 2H), 1.98 – 1.83 (m, 2H), 1.63 – 1.59 (m, 2H), 1.44 – 1.33 (m, 1H), 1.28 – 1.23 (m, 3H), 0.73 – 0.62 (m, 1H), 0.58 – 0.43 (m, 2H), 0.14 – 0.02 (m, 2H). ESI-MS m/z calc.422.22934, found 423.2 (M+1)+; Retention time: 1.25 minutes; and diastereomer 2, Peak 2 [(2R,3S,7R)-3-(cyclopropylmethyl)-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (63.6 mg, 20%) as a yellow oil. ESI-MS m/z calc.422.22934, found 423.2 (M+1)+; Retention time: 1.25 minutes. LC method A. Step 6: 3-[[4-[[(2R,3S,7S)-3-(Cyclopropylmethyl)-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]- 6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid [00554] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (77.85 mg, 0.1863 mmol) and [(2R,3S,7S)-3-(cyclopropylmethyl)-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (78.7 mg, 0.1863 mmol) were combined in a flask and dissolved in THF (1.1 mL) followed by the addition of NaOtBu (89.6 mg, 0.9323 mmol). The reaction mixture was stirred for 1 hour at room temperature then warmed to 40 °C for 2 hours. The reaction mixture was cooled to room temperature and diluted with EtOAc and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (5 x). The organic layers were combined, washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3- [[4-[[(2R,3S,7S)-3-(cyclopropylmethyl)-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (136.1 mg, 64%) as a yellow-white solid. ESI-MS m/z calc.803.3077, found 804.2 (M+1)+; Retention time: 2.18 minutes. LC method N. Step 7: (4S,8S,9R)-8-(Cyclopropylmethyl)-14-(2,6-dimethylphenyl)-4-[5-methyl- 6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11- oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one [00555] 3-[[4-[[(2R,3S,7S)-3-(Cyclopropylmethyl)-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (136.1 mg, 0.1194 mmol) was dissolved in NMP (2.01 mL) and acetone (4.02 mL). TsCl (73.1 mg, 0.3834 mmol) was added followed by NMM (78.8 µL, 0.7167 mmol) and the reaction was stirred at 55 °C for 22 hours. The reaction was poured into a mixture of ethyl acetate and 1 M HCl and the layers were separated. The aqueous layer was further extracted with ethyl acetate (2 x). The combined organic layer was washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was taken up in DMSO/MeOH and purified by reverse phase HPLC (30-99% acetonitrile/5 mM aqueous HCl over 25 min). The pure fractions were extracted with EtOAc (3 x), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give (4S,8S,9R)-8-(cyclopropylmethyl)-14-(2,6-dimethylphenyl)-4-[5- methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa-18λ6- thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one (54.9 mg, 58%) as a white solid.1H NMR (400 MHz, CDCl3) δ 8.85 (s, 1H), 8.54 (s, 1H), 7.77 (t, J = 7.8 Hz, 2H), 7.54 (t, J = 7.7 Hz, 1H), 7.23 (d, J = 7.6 Hz, 1H), 7.08 (d, J = 7.6 Hz, 2H), 6.72 (s, 1H), 6.25 (s, 1H), 5.95 – 5.86 (m, 2H), 4.86 (t, J = 11.8 Hz, 1H), 4.51 – 4.43 (m, 1H), 4.05 (s, 3H), 2.33 (d, J = 15.5 Hz, 1H), 2.25 – 2.12 (m, 1H), 2.07 (s, 7H), 2.01 – 1.81 (m, 5H), 1.63 (s, 2H), 1.23 – 1.09 (m, 2H), 1.09 – 0.99 (m, 1H), 0.42 – 0.32 (m, 1H), 0.32 – 0.18 (m, 2H), -0.20 (q, J = 6.0 Hz, 2H).19F NMR (376 MHz, CDCl3) δ -68.74. ESI-MS m/z calc.785.2971, found 786.5 (M+1)+; Retention time: 2.2 minutes. LC method A. Example 68: Preparation of Compound 96 Step 1: tert-Butyl N-(6-bromo-5-fluoro-2-pyridyl)carbamate [00556] To a solution of 6-bromo-5-fluoro-pyridine-2-carboxylic acid (1 g, 4.5455 mmol) in anhydrous t-BuOH (45 mL) was added a scoop of sodium sulfate (150 mg) and triethylamine (1.0890 g, 1.5 mL, 10.762 mmol). The mixture was stirred for 15 minutes at room temperature, then diphenylphosphoryl azide (1.6601 g, 1.3 mL, 6.0323 mmol) was added. The resulting solution was refluxed at 85 °C for 16 hours. The mixture was cooled to room temperature, was diluted in EtOAc (50 mL) and was filtered. The volatiles were removed under reduced pressure and the crude material was purified by silica gel chromatography (gradient: 0-6 % EtOAc in heptane). The desired fractions were combined and concentrated under reduced pressure to afford tert-butyl N-(6-bromo-5-fluoro-2-pyridyl)carbamate (946 mg, 71%) as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 7.90 (dd, J = 8.8, 3.2 Hz, 1H), 7.41 (dd, J = 8.8, 7.1 Hz, 1H), 7.22 (br. s., 1H), 1.52 (s, 9H).19F NMR (377 MHz, CDCl3) δ -121.54 (s, 1F). ESI-MS m/z calc. 290.0066, found 235.0 (M-55)+; Retention time: 1.93 minutes. LC method I. Step 2: 6-Bromo-5-fluoro-pyridin-2-amine [00557] To a solution of tert-butyl N-(6-bromo-5-fluoro-2-pyridyl)carbamate (928 mg, 3.1845 mmol) in DCM (30 mL) was added TFA (3.7000 g, 2.5 mL, 32.450 mmol). The mixture was stirred at room temperature for 2 h, after which it was quenched at 0 °C with saturated aqueous sodium bicarbonate solution (100 mL). The aqueous phase was extracted with DCM (3 x 50 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 6-bromo-5-fluoro-pyridin-2- amine (582 mg, 96%) as a white solid.1H NMR (400 MHz, CDCl3) δ 7.22 (dd, J = 8.6, 7.1 Hz, 1H), 6.40 (dd, J = 8.8, 2.7 Hz, 1H), 4.46 (br. s., 2H).19F NMR (377 MHz, CDCl3) δ -128.81 (s, 1F). ESI-MS m/z calc.189.9542, found 191.2 (M+1)+; Retention time: 1.48 minutes. LC method I. Step 3: 6-Bromo-5-fluoro-3-iodo-pyridin-2-amine [00558] 6-Bromo-5-fluoro-pyridin-2-amine (574 mg, 3.0022 mmol) was dissolved in DMF (28 mL). TFA (384.80 mg, 0.26 mL, 3.3748 mmol) was added followed by NIS (827 mg, 3.6758 mmol). The reaction mixture was stirred for 2 hours, then more NIS (886 mg, 3.9380 mmol) was added. The reaction was left stirring for 1.5 hour, then was diluted with EtOAc (40 mL), saturated aqueous sodium bicarbonate solution (40 mL) and saturated aqueous Na2S2O3 solution (40 mL). The mixture was stirred for 5 min and the phases were separated. The organic phase was washed with 5% aqueous LiCl solution (2 x 100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel chromatography (gradient: 0-10 % EtOAc in heptane) to afford 6-bromo-5-fluoro-3- iodo-pyridin-2-amine (819 mg, 86%) as a light yellow solid; 1H NMR (400 MHz, CDCl3) δ 7.63 (d, J = 6.4 Hz, 1H), 4.93 (br. s., 2H).19F NMR (377 MHz, CDCl3) δ -126.43 (s, 1F). ESI-MS m/z calc.315.8508, found 317.0 (M+1)+; Retention time: 1.74 minutes. LC method I. Step 4: 6-Bromo-3-(3,3-dimethylbut-1-ynyl)-5-fluoro-pyridin-2-amine [00559] A solution of 6-bromo-5-fluoro-3-iodo-pyridin-2-amine (812 mg, 2.5598 mmol) in anhydrous THF (25 mL) was bubbled with nitrogen for 5 minutes, then triethylamine (529.98 mg, 0.73 mL, 5.2375 mmol), CuI (52 mg, 0.2730 mmol) and PdCl2(PPh3)2 (84 mg, 0.1197 mmol) were added. The reaction mixture was cooled to 0 °C and 3,3-dimethylbut-1-yne (233.45 mg, 0.35 mL, 2.8420 mmol) was slowly added dropwise. The reaction was stirred at 0 °C for 1 hour then more 3,3-dimethylbut-1-yne (600.30 mg, 0.9 mL, 7.3079 mmol) was added dropwise. The reaction was stirred at 0 °C for 40 minutes then more 3,3-dimethylbut-1-yne (600.30 mg, 0.9 mL, 7.3079 mmol) was added dropwise. The reaction was stirred at 0 °C for 30 minutes then was diluted with EtOAc (50 mL). The suspension was filtered through Celite, and the pad was rinsed with EtOAc (3 x 50 mL). The combined organic filtrates were concentrated under reduced pressure and purified by reversed phase chromatography (gradient: 5% MeCN in water with 0.1% formic acid, then 50-70 % MeCN in water with 0.1% formic acid). The fractions containing the product were combined and concentrated under reduced pressure to afford 6- bromo-3-(3,3-dimethylbut-1-ynyl)-5-fluoro-pyridin-2-amine (663 mg, 95%) as a yellow solid; 1H NMR (400 MHz, CDCl3) δ 7.24 (d, J = 7.6 Hz, 1H), 4.90 (br. s., 2H), 1.34 (s, 9H).19F NMR (377 MHz, CDCl3) δ -129.17 (s, 1F). ESI-MS m/z calc.270.0168, found 271.2 (M+1)+; Retention time: 1.98 minutes. LC method I. Step 5: 6-Bromo-2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridine [00560] To a solution of 6-bromo-3-(3,3-dimethylbut-1-ynyl)-5-fluoro-pyridin-2-amine (3.86 g, 14.223 mmol) in anhydrous DMF (15 mL) and Me-THF (75 mL), in a flame-dried flask, at 0 °C under nitrogen, was added NaH (60% w/w in oil) (3.4 g, 85.008 mmol). The mixture was stirred at 85 °C for 2 hours, then was cooled down to 0 °C. More NaH (60% w/w in oil) (3.4 g, 85.008 mmol) was added and the mixture was stirred at 85 °C for 1 hours, then was cooled down to 0 °C. More NaH (60% w/w in oil) (3 g, 75.007 mmol) was added and the mixture was stirred at 85 °C for 1.5 hours, then was cooled down to 0 °C. More NaH (60% w/w in oil) (2 g, 50.005 mmol) and more DMF (25 mL) were added, and the mixture was stirred at 85 °C for 1 hour, then was cooled down to 0 °C. A solution of dimethyl sulfate (4.5220 g, 3.4 mL, 35.851 mmol) in anhydrous DMF (25 mL) was added dropwise to the mixture at 0 °C. The mixture was stirred at room temperature for 16 hours. The mixture was cooled down to 0 °C and more dimethyl sulfate (3.9900 g, 3 mL, 31.634 mmol) was added. The mixture was stirred at room temperature for 1 hour. The mixture was added dropwise to a saturated aqueous solution of ammonium chloride (400 mL). Ethyl acetate (250 mL) was added, the layers were separated, and the aqueous layer was extracted with ethyl acetate (3 x 250 mL). The organic layers were combined, washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure. The excess of mineral oil was removed by decantation. The crude residue was purified by reversed-phase chromatography (C18 column. gradient 50-100% acetonitrile in water with 0.1% formic acid). The desired fractions were combined, concentrated under reduced pressure and co-evaporated with acetonitrile (3 x 10 mL) and THF (2 x 5 mL), to afford 6-bromo-2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridine (1.725 g, 42%) as a tan solid.1H NMR (400 MHz, CDCl3) δ 7.51 (d, J = 8.3 Hz, 1H), 6.22 (s, 1H), 3.98 (s, 3H), 1.47 (s, 9H).19F NMR (377 MHz, CDCl3) δ -126.24 (s, 1F). ESI-MS m/z calc.284.0324, found 285.2 (M+1)+; Retention time: 2.11 minutes. LC method I. Step 6: tert-Butyl (4S)-4-[5-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3- b]pyridin-6-yl)-5-hydroxy-1-isopropoxy-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate ~80:20 mixture of diastereomers the major isomer having the 4S,1R configuration [00561] To a solution of 6-bromo-2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridine (500 mg, 1.7517 mmol), in tetrahydrofuran (10 mL), at -78 °C, was added n-butyllithium (solution in hexanes) (0.95 mL of 1.75 M, 1.6625 mmol) dropwise. As soon as the addition of n-BuLi was over, a solution of tert-butyl (4S)-4-(1-isopropoxy-5-oxo-pentyl)-2,2-dimethyl-oxazolidine-3- carboxylate (~80:20 mixture of diastereomers, the major isomer having the 4S,1R configuration, 525 mg, 1.5072 mmol), in tetrahydrofuran (2.5 mL), was added dropwise. The reaction was left stirring 1 hour at -78 °C, a saturated aqueous solution of ammonium chloride (4 mL) and ethyl acetate (8 mL) were added. The mixture was slowly warmed to room temperature and stirred for 15 minutes. The layers were separated, and the aqueous one was extracted with ethyl acetate (3 x 15 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude was purified by reverse-phase chromatography on C18 column (gradient 70 to 90% acetonitrile in water). The desired fractions were concentrated under reduced pressure until dryness and co-evaporated with acetonitrile (3 x 10 mL), to afford tert-butyl (4S)-4-[5-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6-yl)- 5-hydroxy-1-isopropoxy-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (391 mg, 47%) as a pale yellow semi-solid.1H NMR (400 MHz, CDCl3) δ 7.45 (d, J = 9.8 Hz, 1H), 6.26 - 6.19 (m, 1H), 5.10 - 5.00 (m, 1H), 4.34 - 4.16 (m, 1H), 4.12 - 4.02 (m, 1H), 4.01 - 3.97 (m, 3H), 3.96 - 3.83 (m, 2H), 3.81 - 3.55 (m, 2H), 1.93 - 1.78 (m, 1H), 1.78 - 1.65 (m, 1H), 1.61 - 1.33 (m, overlapped with water, 28H), 1.15 - 1.01 (m, 6H).19F NMR (377 MHz, CDCl3) δ -140.63 (s, 1F). ESI-MS m/z calc.549.3578, found 550.4 (M+1)+; Retention time: 2.39 minutes. LC method S. Step 7: tert-Butyl (4S)-4-[5-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3- b]pyridin-6-yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate [00562] To a solution of tert-butyl (4S)-4-[5-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3- b]pyridin-6-yl)-5-hydroxy-1-isopropoxy-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (840 mg, 1.3218 mmol) in 1,2-dichloroethane (22 mL) was added manganese (IV) oxide (activated) (2.7 g, 26.398 mmol). The resulting suspension was stirred at 70 °C for 6 hours and at room temperature for 18 hours. More manganese (IV) oxide (activated) (1.3 g, 12.710 mmol) was added, and the suspension was stirred at 70 °C for 3 hours. More manganese (IV) oxide (activated) (0.7 g, 6.8441 mmol) was added, and the suspension was stirred at 70 °C for 4 hours. More manganese (IV) oxide (activated) (0.5 g, 4.8886 mmol) was added, and the suspension was stirred at 70 °C for 1 hour and at room temperature for 18 hours. The suspension was filtered on a pad of Celite and washed with DCM (3 x 25 mL). The crude mixture was concentrated in vacuo and was then purified by reverse-phase chromatography on a C18 cartridge, eluting with a gradient of 75 to 81% of acetonitrile in 10 mM ammonium bicarbonate buffer (pH = 10). The fractions containing the desired product were combined, concentrated under reduced pressure, and co-evaporated with acetonitrile (3 x 10 mL) and dichloromethane (15 mL). The impure fractions from the reverse-phase chromatography were combined, concentrated and purified again by reverse-phase chromatography on a C18 cartridge, eluting with a gradient of 80 to 88% of methanol in 10 mM ammonium bicarbonate buffer (pH = 10). The fractions containing the desired product were combined, concentrated under reduced pressure and co-evaporated with acetonitrile (3 x 10 mL) and dichloromethane (15 mL). Both pure samples were combined in dichloromethane (15 mL), to afford after evaporation tert-butyl (4S)-4-[5-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6-yl)-1-isopropoxy-5-oxo- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (435 mg, 59%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 7.51 (d, J = 11.5 Hz, 1H), 6.29 (s, 1H), 5.30 (s, 1H), 4.17 - 4.06 (m, 1H), 4.04 (s, 3H), 4.01 - 3.88 (m, 2H), 3.88 - 3.62 (m, 2H), 3.35 - 3.14 (m, 2H), 1.95 - 1.72 (m, 2H), 1.64 - 1.44 (m, overlapped with water, 25H), 1.21 - 1.02 (m, 6H).19F NMR (377 MHz, CDCl3) δ -133.54 (m, 1F). ESI-MS m/z calc.547.3421, found 448.2 (M-99)+; Retention time: 4.62 minutes. LC method W. Step 8: (6S)-6-Amino-1-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6- yl)-7-hydroxy-5-isopropoxy-heptan-1-one [00563] To a solution of tert-butyl (4S)-4-[5-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3- b]pyridin-6-yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (21 mg, 0.0290 mmol) in 1,4-dioxane (0.8 mL) and water (0.2 mL), was added hydrochloric acid (solution in 1,4-dioxane) (0.35 mL of 4 M, 1.4000 mmol). The reaction was stirred at room temperature for 20 hours, under nitrogen. The solvent was removed under reduced pressure and the mixture was co-evaporated with 1,4-dioxane (2 x 5 mL). The material was dissolved in 1,4- dioxane (3 mL) and a saturated aqueous solution of sodium bicarbonate (1 mL) was added. The mixture was stirred for 16 hours at room temperature. Water (2 mL) and EtOAc (10 mL) were added. The layers were separated, and the aqueous one was extracted with DCM (3 x 3 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by reversed-phase chromatography on C18 column (gradient 5% to 70% acetonitrile in 10 mM ammonium bicarbonate buffer : pH = 10). The fractions containing the product were combined and evaporated until dryness. The material was purified again by reversed-phase chromatography on C18 column (gradient 50% to 57% acetonitrile in 10 mM ammonium bicarbonate buffer (pH = 10)). The fractions containing the product were combined and evaporated until dryness to afford (6S)-6-amino-1-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6-yl)-7-hydroxy-5- isopropoxy-heptan-1-one (4 mg, 32%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 7.56 - 7.40 (m, 1H), 6.28 - 6.14 (m, 1H), 4.04 - 3.93 (m, 3H), 3.92 - 3.23 (m, 5H), 2.69 - 2.47 (m, 1H), 2.22 - 2.02 (m, 1H), 1.98 - 1.75 (m, 3H), 1.75 - 1.36 (m, 11H), 1.35 - 0.97 (m, 7H). (1H missing, labile proton).19F NMR (377 MHz, CDCl3) δ -133.44 (s, 1F). ESI-MS m/z calc.407.2584, found 390.4 (M-17)+; Retention time: 1.46 minutes. LC method S. Step 9: [(2S,3R,7S)-7-(2-tert-Butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6- yl)-3-isopropoxy-azepan-2-yl]methanol and [(2S,3R,7R)-7-(2-tert-butyl-5-fluoro- 1-methyl-pyrrolo[2,3-b]pyridin-6-yl)-3-isopropoxy-azepan-2-yl]methanol [00564] To a solution of (6S)-6-amino-1-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3- b]pyridin-6-yl)-7-hydroxy-5-isopropoxy-heptan-1-one (224 mg, 0.5029 mmol) in DCM (8 mL), at 0 °C, was added NaBH(OAc)3 (750 mg, 3.4679 mmol). The reaction mixture was stirred for 2 hours at room temperature and a saturated aqueous solution of sodium bicarbonate (10 mL) was added at 0 °C. The layers were separated, and the aqueous layer was extracted with DCM (3 x 15 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography on C18 column (gradient 40 to 65% acetonitrile in 10 mM ammonium bicarbonate buffer : pH = 10). The fractions containing the first eluting product were combined and the volatiles were removed under reduced pressure. The residue was co-evaporated with acetonitrile (4 x 10 mL). The resulting product was dried under high vacuum to afford [(2S,3R,7S)-7-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6-yl)-3-isopropoxy-azepan- 2-yl]methanol (80 mg, 39%) as a white solid.1H NMR (400 MHz, CDCl3) δ 7.40 (d, J = 10.3 Hz, 1H), 6.18 (s, 1H), 4.40 (ddd, J = 7.8, 5.4, 2.0 Hz, 1H), 3.99 (s, 3H), 3.91 (dd, J = 10.4, 4.8 Hz, 1H), 3.71 - 3.60 (m, 1H), 3.36 - 3.27 (m, 2H), 2.90 (td, J = 8.7, 4.9 Hz, 1H), 2.11 - 1.99 (m, 2H), 1.92 - 1.61 (m, 5H), 1.46 (s, 9H), 1.16 (dd, J = 14.3, 6.0 Hz, 6H). (1 H missing, labile proton).19F NMR (377 MHz, CDCl3) δ -139.48 (s, 1F). ESI-MS m/z calc.391.2635, found 392.2 (M+1)+; Retention time: 4.01 minutes. [00565] The fractions containing the mixed products from reversed-phase chromatography were combined and the volatiles were removed under reduced pressure. The mixture was purified by reversed-phase chromatography on C18 column (gradient 40 to 60% acetonitrile in 10 mM ammonium bicarbonate buffer : pH = 10). The fractions containing the mixed products were combined and the volatiles were removed under reduced pressure. The mixture was purified by reversed-phase chromatography on C18 column (isocratic 50% acetonitrile in 10 mM ammonium bicarbonate buffer : pH = 10). The residue was co-evaporated with acetonitrile (3 x 5 mL), then with CDCl3 for NMR purpose and with acetonitrile (1 mL). The resulting second-to- elute product was dried under high vacuum to afford [(2S,3R,7R)-7-(2-tert-butyl-5-fluoro-1- methyl-pyrrolo[2,3-b]pyridin-6-yl)-3-isopropoxy-azepan-2-yl]methanol (3 mg, 1%) as a pale yellow oil.1H NMR (400 MHz, CDCl3) δ 7.40 (d, J = 10.3 Hz, 1H), 6.17 (s, 1H), 4.33 - 4.23 (m, 1H), 3.99 (s, 3H), 3.92 (dd, J = 10.0, 4.4 Hz, 1H), 3.70 - 3.59 (m, 1H), 3.41 (t, J = 9.7 Hz, 1H), 3.26 (td, J = 9.1, 4.5 Hz, 1H), 3.22 - 3.13 (m, 1H), 2.19 - 2.08 (m, 2H), 2.01 - 1.91 (m, 1H), 1.89 - 1.74 (m, 2H), 1.71 - 1.51 (m, 3H), 1.46 (s, 9H), 1.16 (dd, J = 10.5, 6.1 Hz, 6H).19F NMR (377 MHz, CDCl3) δ -140.36 (s, 1F). ESI-MS m/z calc.391.2635, found 392.2 (M+1)+; Retention time: 4.15 minutes. LC method W. Step 10: 3-[[4-[[(2S,3R,7S)-7-(2-tert-Butyl-5-fluoro-1-methyl-pyrrolo[2,3- b]pyridin-6-yl)-3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid
[00566] Through a stirred solution of 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (86 mg, 0.2058 mmol) and [(2S,3R,7S)-7-(2-tert-butyl-5-fluoro-1- methyl-pyrrolo[2,3-b]pyridin-6-yl)-3-isopropoxy-azepan-2-yl]methanol (77 mg, 0.1900 mmol) in anhydrous tetrahydrofuran (1.8 mL), nitrogen was sparged for 5 min. Then sodium tert- butoxide (110 mg, 1.145 mmol) was added in a single portion at ambient temperature and stirring continued for 5 min, then the mixture was stirred at 40 °C for 2 h. The heating bath was removed and the reaction was allowed to cool down to ambient temperature. The reaction mixture was diluted with ethyl acetate (20 mL) and hydrochloric acid (2.5 mL of 1.0 M, 2.500 mmol). The organic phase was separated, and the aqueous layer was extracted with ethyl acetate (2 x 15 mL). The combined organics were washed with brine (10 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude was purified by reversed- phase chromatography (10-99% acetonitrile in water containing 5 mM HCl, over 20 min run), and the fractions were concentrated under reduced pressure, co-evaporated with acetonitrile (2 x 10 mL) and toluene (2 x 10 mL) to furnish 3-[[4-[[(2S,3R,7S)-7-(2-tert-butyl-5-fluoro-1-methyl- pyrrolo[2,3-b]pyridin-6-yl)-3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (dihydrochloride salt) (159 mg, 99%), ESI-MS m/z calc.772.34186, found 773.2(M+1)⁺; Retention time: 1.72 minutes. as a white solid. LC method A. Step 11: (4S,8R,9S)-4-(2-tert-Butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6- yl)-14-(2,6-dimethylphenyl)-8-isopropoxy-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (Compound 96)
[00567] To a stirred solution of 3-[[4-[[(2S,3R,7S)-7-(2-tert-butyl-5-fluoro-1-methyl- pyrrolo[2,3-b]pyridin-6-yl)-3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (dihydrochloride salt) (159 mg, 0.1880 mmol) in anhydrous NMP (3.6 mL) and acetone (7.2 mL) was added p-TsCl (145 mg, 0.7606 mmol) at ambient temperature under nitrogen and the reaction was stirred for 5 min. Then N- methylmorpholine (200 µL, 1.819 mmol) was added and the reaction was stirred at 35 oC for 2 h. The reaction was allowed to cool to ambient temperature and the volatiles were removed under reduced pressure. The remaining brownish solution was subjected to preparative HPLC (30-99% acetonitrile in water, containing 5 mM HCl as modifier, 15 min run). The fractions were combined and evaporated, then repurified by HPLC (40-99% acetonitrile in water, 30 min run, HCl modifier) to furnish product as a white solid. (4S,8R,9S)-4-(2-tert-butyl-5-fluoro-1- methyl-pyrrolo[2,3-b]pyridin-6-yl)-14-(2,6-dimethylphenyl)-8-isopropoxy-18,18-dioxo-11-oxa- 18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (45.70 mg, 32%), 1H NMR (400 MHz, CDCl3) δ 8.85 (t, J = 1.8 Hz, 1H), 7.82 (dt, J = 7.7, 1.4 Hz, 1H), 7.72 (d, J = 7.9 Hz, 1H), 7.53 (t, J = 7.7 Hz, 1H), 7.44 (d, J = 10.4 Hz, 1H), 7.23 (t, J = 7.6 Hz, 1H), 7.09 (d, J = 7.6 Hz, 2H), 6.29 (s, 1H), 6.18 (s, 1H), 6.14 - 6.03 (m, 2H), 4.81 (t, J = 11.9 Hz, 1H), 4.72 (dt, J = 12.3, 4.9 Hz, 1H), 4.16 (s, 3H), 3.61 - 3.54 (m, 1H), 3.32 (hept, J = 6.1 Hz, 1H), 2.40 (dd, J = 14.7, 6.4 Hz, 1H), 2.07 (s, 6H), 2.03 - 1.91 (m, 3H), 1.90 - 1.82 (m, 1H), 1.76 (t, J = 12.6 Hz, 1H), 1.46 (s, 9H), 0.99 (d, J = 6.0 Hz, 3H), 0.66 (d, J = 5.9 Hz, 3H)., ESI-MS m/z calc.754.33124, found 755.2(M+1)⁺; Retention time: 2.43 minutes. LC method A. Example 69: Preparation of Compound 98 Step 1: tert-Butyl 4-[5-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6- yl)-5-hydroxy-1-isobutyl-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate racemic mixture of diastereomer with anti configuration [00568] To a solution of 6-bromo-2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridine (302 mg, 1.0580 mmol) and TMEDA (155.00 mg, 0.2 mL, 1.3338 mmol) in toluene (4.5 mL) at -78 °C was added n-butyllithium (solution in hexanes) (0.72 mL of 1.6 M, 1.1520 mmol) dropwise. As soon as the addition of n-Buli was over, a solution of tert-butyl 4-(1-isobutyl-5-oxo-pentyl)- 2,2-dimethyl-oxazolidine-3-carboxylate (340 mg, 0.9508 mmol) in toluene (3.0 mL) was added dropwise. The reaction was left stirring for 30 minutes, then brought back to room temperature and quenched with a saturated solution of ammonium chloride (10 mL). EtOAc (10 mL) was added, the mixture was separated and the organic phase was washed with water (15 mL), brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford the crude tert-butyl 4-[5-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6-yl)-5- hydroxy-1-isobutyl-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (625 mg, 72%) as a yellow oil; ESI-MS m/z calc.547.3785, found 548.4 (M+1)+; Retention time: 2.59 minutes. LC method S. A solution of a second batch of tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-2-fluoro-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-5-hydroxy-1-isobutyl-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (191 mg, 0.1845 mmol) and the first batch of tert-butyl 4-[5-(2-tert-butyl-5-fluoro- 1-methyl-pyrrolo[2,3-b]pyridin-6-yl)-5-hydroxy-1-isobutyl-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (625 mg, 0.6879 mmol) in DMSO (3 mL) was injected on a C18 column and purified by reversed phase chromatography (Gradient: 5% MeCN in water, then 60%-100% MeCN in water). The desired fractions were concentrated under reduced pressure to provide tert-butyl 4- [5-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6-yl)-5-hydroxy-1-isobutyl-pentyl]- 2,2-dimethyl-oxazolidine-3-carboxylate (451 mg, 94%) as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 7.47 (d, J = 10.0 Hz, 1H), 6.24 (s, 1H), 5.10 - 5.01 (m, 1H), 4.36 - 4.19 (m, 1H), 4.01 (s, 3H), 3.97 - 3.84 (m, 2H), 3.82 - 3.74 (m, 1H), 2.14 - 1.78 (m, 3H), 1.78 - 1.58 (m, 4H), 1.55 - 1.33 (m, 23H), 1.31 - 1.15 (m, 2H), 1.12 - 0.95 (m, 2H), 0.93 - 0.84 (m, 3H), 0.80 (t, J = 6.8 Hz, 3H).19F NMR (377 MHz, CDCl3) δ -140.55 - -140.64 (m, 1F). ESI-MS m/z calc.547.3785, found 548.4 (M+1)+; Retention time: 2.59 minutes. LC method S. Step 2: tert-Butyl 4-[5-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6- yl)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate, enantiomer 1, and tert-butyl 4-[5-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6-yl)- 1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate, enantiomer 2 [00569] To a solution of tert-butyl 4-[5-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin- 6-yl)-5-hydroxy-1-isobutyl-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (449 mg, 0.8189 mmol) in dichloroethane (9 mL) was added manganese (IV) oxide (activated) (1.72 g, 16.817 mmol). The resulting suspension was stirred at 70 °C for 4.5 hours. The suspension was filtered on a pad of Celite, the pad was washed with DCM (2 x 30 mL) and the solvent was evaporated under reduced pressure. The residue was dissolved in dichloroethane (9 mL) and manganese (IV) oxide (activated) (441 mg, 4.3118 mmol) was added. The mixture was stirred at 70 °C for 2 hours, then more manganese (IV) oxide (activated) (345 mg, 3.3731 mmol) was added. The mixture was stirred for 2.5 hours at 70 °C and 16 hours at room temperature. More manganese (IV) oxide (activated) (392 mg, 3.8327 mmol) was added, and the mixture was stirred at 70 °C for 2.5 hours. The suspension was filtered on a pad of Celite, the pad was washed with DCM (2 x 50 mL) and the solvent was evaporated under reduced pressure. The crude product was purified by chiral SFC using 15% MeOH in CO2 as eluent. (Cellulose 2-30 mm column, outlet pressure 100 bar, injection time 10 s, 40 oC, 600 µL). Fractions containing the first peak, were combined and concentrated under reduced pressure to afford the first enantiomer tert-butyl 4-[5- (2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6-yl)-1-isobutyl-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate (138 mg, 58%) as a white foam; 1H NMR (400 MHz, CDCl3) δ 7.55 - 7.49 (m, 1H), 6.30 (s, 1H), 4.05 (s, 3H), 4.03 - 3.98 (m, 1H), 3.97 - 3.90 (m, 1H), 3.88 - 3.80 (m, 1H), 3.32 - 3.16 (m, 2H), 2.22 - 2.01 (m, 1H), 1.95 - 1.62 (m, 5H), 1.53 - 1.43 (m, 22H), 1.40 - 1.13 (m, 3H), 1.12 - 1.03 (m, 1H), 0.95 - 0.88 (m, 3H), 0.88 - 0.80 (m, 3H).19F NMR (377 MHz, CDCl3) δ -133.29 - -133.59 (m, 1F). ESI-MS m/z calc.545.3629, found 546.4 (M+1)+; Retention time: 2.64 minutes. Fractions containing the second peak were combined and concentrated under reduced pressure to afford the second enantiomer tert-butyl 4- [5-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6-yl)-1-isobutyl-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate (138 mg, 60%) as a white foam; 1H NMR (400 MHz, CDCl3) δ 7.57 - 7.48 (m, 1H), 6.30 (s, 1H), 4.05 (s, 3H), 4.03 - 3.97 (m, 1H), 3.97 - 3.90 (m, 1H), 3.89 - 3.79 (m, 1H), 3.33 - 3.15 (m, 2H), 2.21 - 2.00 (m, 1H), 1.95 - 1.62 (m, 5H), 1.54 - 1.40 (m, 22H), 1.39 - 1.12 (m, 3H), 1.12 - 1.03 (m, 1H), 0.96 - 0.88 (m, 3H), 0.87 - 0.79 (m, 3H).19F NMR (377 MHz, CDCl3) δ -133.32 - -133.59 (m, 1F). ESI-MS m/z calc.545.3629, found 546.4 (M+1)+; Retention time: 2.65 minutes. LC method S. Step 3: 5-(1-Amino-2-hydroxy-ethyl)-1-(2-tert-butyl-5-fluoro-1-methyl- pyrrolo[2,3-b]pyridin-6-yl)-7-methyl-octan-1-one [00570] To a solution of tert-butyl 4-[5-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin- 6-yl)-1-isobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (enantiomer 2, 135 mg, 0.2422 mmol) in a mixture of 1,4-dioxane (2.1 mL) and water (0.25 mL) at room temperature was added dropwise a solution of hydrochloric acid (solution in 1,4-dioxane) (1.3 mL of 4 M, 5.2000 mmol). The reaction was stirred for 2 hours, and the solvent was removed under reduced pressure. A saturated aqueous solution of sodium bicarbonate (20 mL) was added, followed by EtOAc (10 mL). The mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford 5-(1-amino-2-hydroxy-ethyl)-1-(2-tert-butyl-5- fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6-yl)-7-methyl-octan-1-one, enantiomer 2 (102 mg, 93%) as a yellow solid; 1H NMR (400 MHz, CDCl3) δ 7.52 (d, J = 11.5 Hz, 1H), 6.22 (s, 1H), 4.01 (s, 3H), 3.85 (dd, J = 7.3, 1.2 Hz, 1H), 3.79 - 3.73 (m, 1H), 3.67 - 3.62 (m, 1H), 2.67 - 2.58 (m, 1H), 1.94 - 1.78 (m, 3H), 1.78 - 1.59 (overlapped with water, m, 5H), 1.52 - 1.42 (m, 13H), 0.94 (d, J = 6.6 Hz, 6H).19F NMR (377 MHz, CDCl3) δ -132.84 (s, 1F). ESI-MS m/z calc.405.2792, found 406.4 (M+1)+; Retention time: 1.5 minutes. LC method I. Step 4: [7-(2-tert-Butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6-yl)-3-isobutyl- azepan-2-yl]methanol, diastereomer 1, and [7-(2-tert-butyl-5-fluoro-1-methyl- pyrrolo[2,3-b]pyridin-6-yl)-3-isobutyl-azepan-2-yl]methanol, diastereomer 2 [00571] To a solution of 5-(1-amino-2-hydroxy-ethyl)-1-(2-tert-butyl-5-fluoro-1-methyl- pyrrolo[2,3-b]pyridin-6-yl)-7-methyl-octan-1-one, enantiomer 2 (99.5 mg, 0.2196 mmol) in dichloromethane (2 mL) was added sodium triacetoxyborohydride (295 mg, 1.3919 mmol). The reaction was stirred at rt for 2 hours. The mixture was diluted with DCM (10 mL) and a saturated aqueous solution of sodium bicarbonate (5 mL) was added. The phases were separated, and the aqueous one was extracted with DCM (2 x 10 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude material was purified by reversed phase chromatography on a C18 cartridge, eluting with a gradient of 40 to 80% MeCN in basic water (ammonium bicarbonate/ammonium hydroxide buffer pH = 10). The desired fractions were combined, and the volatiles were removed under reduced pressure. The resulting basic aqueous phase was extracted with DCM (3 x 15 mL), the combined organic extracts were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford a mixture of two diastereomers as a light-yellow foam. The product was purified by preparative SFC (Lux Column 5 µm, Cellulose 4, 250 x 30 mm, 12.33 mg/mL, 12.33 mg/injection) using 30% EtOH with 0.1% DEA in CO2 as eluent. The fractions containing the first peak were concentrated under reduced pressure. The residue was purified by reversed phase chromatography on a C18 cartridge, eluting with a gradient of 0 to 100% MeCN in water. The desired fractions were combined and concentrated under reduced pressure to afford, after freeze drying, [7-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin- 6-yl)-3-isobutyl-azepan-2-yl]methanol, diastereomer 1 (41.9 mg, 49%) as a white solid; 1H NMR (400 MHz, Benzene-d6) δ 7.32 (d, J = 10.3 Hz, 1H), 6.01 (s, 1H), 4.48 (ddd, J = 9.0, 5.0, 1.8 Hz, 1H), 3.82 (dd, J = 10.3, 4.2 Hz, 1H), 3.61 (s, 3H), 3.14 (t, J = 10.0 Hz, 1H), 2.60 (td, J = 9.4, 4.2 Hz, 1H), 2.19 - 2.03 (m, 2H), 1.75 - 1.60 (m, 2H), 1.57 - 1.25 (m, 5H), 1.18 (s, 9H), 1.12 (ddd, J = 13.8, 9.8, 4.3 Hz, 1H), 0.91 (ddd, J = 13.4, 10.1, 3.2 Hz, 1H), 0.84 (d, J = 6.6 Hz, 3H), 0.75 (d, J = 6.6 Hz, 3H), (1H missing, labile proton).19F NMR (377 MHz, Benzene-d6) δ - 139.27 (s, 1F). ESI-MS m/z calc.389.2842, found 390.4 (M+1)+; Retention time: 2.63 minutes. The fractions containing the second peak were concentrated under reduced pressure. The residue was purified by reversed phase chromatography on a C18 cartridge, eluting with a gradient of 0 to 100% MeCN in water. The desired fractions were combined and concentrated under reduced pressure to afford, after freeze drying,[7-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin- 6-yl)-3-isobutyl-azepan-2-yl]methanol, diastereomer 2 (10.2 mg, 12%) as an off white solid; 1H NMR (400 MHz, Benzene-d6) δ 7.32 (d, J = 10.5 Hz, 1H), 6.00 (s, 1H), 4.43 (dt, J = 10.8, 2.2 Hz, 1H), 3.81 (dd, J = 10.3, 4.2 Hz, 1H), 3.70 (s, 3H), 3.50 (t, J = 10.3 Hz, 1H), 2.66 (td, J = 9.6, 4.0 Hz, 1H), 2.21 - 2.14 (m, 1H), 2.01 - 1.92 (m, 1H), 1.91 - 1.81 (m, 1H), 1.66 - 1.58 (m, 1H), 1.54 - 1.43 (m, 1H), 1.40 - 1.20 (m, 3H), 1.17 (s, 9H), 1.16 - 1.11 (m, 1H), 0.97 - 0.92 (m, 2H), 0.83 (d, J = 6.6 Hz, 3H), 0.76 (d, J = 6.4 Hz, 3H), (1H missing, labile proton).19F NMR (377 MHz, Benzene-d6) δ -139.79 (s, 1F). ESI-MS m/z calc.389.2842, found 390.2 (M+1)+; Retention time: 2.63 minutes. LC method 1B. Step 5: 3-[[4-[[7-(2-tert-Butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6-yl)-3- isobutyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid, diastereomer 1
diastereomer 1 [00572] Through a stirred solution of 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (45 mg, 0.1077 mmol) and [7-(2-tert-butyl-5-fluoro-1-methyl- pyrrolo[2,3-b]pyridin-6-yl)-3-isobutyl-azepan-2-yl]methanol (39 mg, 0.09971 mmol) in anhydrous tetrahydrofuran (1.0 mL), nitrogen was sparged for 5 min. Then sodium tert-butoxide (65 mg, 0.6763 mmol) was added in a single portion at ambient temperature and stirring was continued at that temperature for 5 min then at 40 °C for 2 h. The heating bath was removed, and the reaction was cooled down to ambient temperature. The reaction mixture was diluted with ethyl acetate (20 mL) and hydrochloric acid (1.2 mL of 1.0 M, 1.200 mmol). The organic phase was separated, and the aqueous layer was extracted with ethyl acetate (2 x 15 mL). The combined organics were washed with brine (10 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude was purified by reversed-phase chromatography (10-99% acetonitrile in water containing 5 mM HCl, over 20 min run), and the fractions were concentrated under reduced pressure, co-evaporated with acetonitrile (2 x 10 mL) and toluene (2 x 10 mL) to furnish 3-[[4-[[7-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6-yl)-3- isobutyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (dihydrochloride salt), diastereomer 1 (84 mg, 100%), ESI-MS m/z calc.770.36255, found 771.2(M+1)⁺; Retention time: 1.79 minutes. as a white solid. LC method A. Step 6: 4-(2-tert-Butyl-5-fluoro-1-methyl-pyrrolo[2,3-b]pyridin-6-yl)-14-(2,6- dimethylphenyl)-8-isobutyl-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one, diastereomer 1, (4R, 8R, 9S) or (4S, 8S, 9R) configuration (Compound 98).
diastereomer 1 diastereomer 1 [00573] To a stirred solution of 3-[[4-[[7-(2-tert-butyl-5-fluoro-1-methyl-pyrrolo[2,3- b]pyridin-6-yl)-3-isobutyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (dihydrochloride salt), diastereomer 1 (84 mg, 0.09954 mmol) in anhydrous NMP (2.0 mL) and acetone (4 mL) was added p-TsCl (76 mg, 0.3987 mmol) at ambient temperature under nitrogen and the reaction was stirred for 5 min. Then N- methylmorpholine (100 µL, 0.9095 mmol) was added dropwise and the reaction was stirred at room temperature for 18 h, then at 35 oC for 5 h. The reaction was allowed to cool to ambient temperature and the volatiles were removed under reduced pressure. The remaining brownish solution was subjected to preparative HPLC (30-99% acetonitrile in water, containing 5mM HCl as modifier, 15 min run), then purified a second time by HPLC (40-99% acetonitrile in water, 30 min run, HCl modifier) to furnish the product as a white solid.4-(2-tert-butyl-5-fluoro-1- methyl-pyrrolo[2,3-b]pyridin-6-yl)-14-(2,6-dimethylphenyl)-8-isobutyl-18,18-dioxo-11-oxa- 18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one, diastereomer 1, (4R, 8R, 9S) or (4S, 8S, 9R) configuration (31.5 mg, 41%), 1H NMR (400 MHz, CDCl3) δ 8.92 (t, J = 1.8 Hz, 1H), 7.81 - 7.71 (m, 2H), 7.53 (t, J = 7.8 Hz, 1H), 7.43 (d, J = 10.4 Hz, 1H), 7.23 (t, J = 7.6 Hz, 1H), 7.08 (d, J = 7.6 Hz, 2H), 6.30 (s, 1H), 6.17 (s, 1H), 6.12 (dd, J = 11.8, 5.4 Hz, 1H), 5.97 (dd, J = 13.2, 4.2 Hz, 1H), 4.96 (t, J = 11.9 Hz, 1H), 4.49 - 4.40 (m, 1H), 4.16 (s, 3H), 2.44 - 2.27 (m, 1H), 2.07 (s, 6H), 2.01 - 1.93 (m, 2H), 1.92 - 1.83 (m, 2H), 1.82 - 1.73 (m, 1H), 1.72 - 1.64 (m, 1H), 1.45 (s, 9H), 1.32 - 1.17 (m, 2H), 0.91 - 0.81 (m, 1H), 0.70 - 0.63 (m, 6H)., ESI-MS m/z calc.752.352, found 753.2(M+1)⁺; Retention time: 2.24 minutes. LC method C. Example 70: Preparation of Compound 107 Step 1: tert-Butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-cyclobutyl- 5-oxo-pyrrolidine-1-carboxylate [00574] To a -40 °C solution of cyclobutyl zinc bromide (solution in THF) (150 mL of 0.5 M, 75.0 mmol) was added dropwise over 3 h a solution of chlorotrimethylsilane (3.4240 g, 4 mL, 31.516 mmol) and tert-butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-2H-pyrrole-1- carboxylate (13.5 g, 29.892 mmol) in tetrahydrofuran (170 mL). The resulting mixture was stirred between at -40 °C for 41 h and additional chlorotrimethylsilane (684.80 mg, 0.8 mL, 6.3033 mmol) was added. A solution of saturated ammonium chloride in water (200 mL) was added followed by water (100 mL) and EtOAc (200 mL). The phases were separated, and the aqueous phase was extracted with EtOAc (2 x 100 mL). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 90% of MeCN in water to afford, after evaporation, tert-butyl (2R,3S)-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-3-cyclobutyl-5-oxo-pyrrolidine-1-carboxylate (7.05 g, 43%) as a sticky yellow oil.1H NMR (400 MHz, CDCl3) δ 7.68 - 7.59 (m, 4H), 7.46 - 7.36 (m, 6H), 3.88 - 3.81 (m, 1H), 3.79 - 3.74 (m, 1H), 3.68 (dd, J = 10.1, 2.3 Hz, 1H), 2.81 (dd, J = 17.9, 8.3 Hz, 1H), 2.32 - 2.17 (m, 2H), 2.15 - 2.03 (m, 3H), 1.93 - 1.78 (m, 2H), 1.70 - 1.58 (m, 2H), 1.44 (s, 9H), 1.06 (s, 9H). ESI-MS m/z calc.507.2805, found 408.2 (M-99)+; Retention time: 2.47 minutes. LC method S. Step 2: tert-Butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-cyclobutyl- 5-hydroxy-pyrrolidine-1-carboxylate [00575] A stirred solution of tert-butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- cyclobutyl-5-oxo-pyrrolidine-1-carboxylate (1.76 g, 2.9568 mmol) in anhydrous THF (21 mL) was cooled down to -78 °C, then a solution of lithium triethylborohydride in THF (4.5 mL of 1 M, 4.5000 mmol) was added dropwise over 30 minutes. The reaction was then stirred at -78 °C for 90 minutes, then quenched by the addition into a saturated aqueous solution of sodium bicarbonate (70 mL) at 0 °C. Then, an aqueous solution of hydrogen peroxide (in water) (118.65 mg, 0.3 mL of 35 %w/w, 1.2209 mmol) was added dropwise. The reaction was stirred for 15 minutes at room temperature, diluted with EtOAc (100 mL) and stirred for another 30 minutes. The layers were then separated, the aqueous layer was extracted with EtOAc (3 x 100 mL) and the combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert-butyl (2R,3S)-2- [[tert-butyl(diphenyl)silyl]oxymethyl]-3-cyclobutyl-5-hydroxy-pyrrolidine-1-carboxylate (1.862 g, 100%) as a pale yellow oil that was used in the next step without further purification. ESI-MS m/z calc.509.2961, found 392.2 (M-117)+; Retention time: 2.34 minutes. LC method Q. Step 3: tert-Butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2- cyclobutyl-4-hydroxy-butyl]carbamate [00576] To a solution of tert-butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- cyclobutyl-5-hydroxy-pyrrolidine-1-carboxylate (1.84 g, 2.9238 mmol) in methanol (65 mL) was slowly added sodium borohydride (3 g, 77.711 mmol), so that the internal temperature of the mixture was maintained around 50 °C from the exotherm. The reaction mixture was allowed to reach room temperature over 90 minutes, then heated at 70 °C for 5 minutes. More sodium borohydride (1 g, 25.904 mmol) was slowly added at that temperature and the mixture was heated at 70 °C for 1 hour. The reaction was then concentrated under reduced pressure and partitioned between water (100 mL) and dichloromethane (100 mL). The biphasic mixture was vigorously stirred at room temperature for 15 minutes. The layers were separated, and the aqueous layer was extracted with dichloromethane (2 x 250 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford tert-butyl N-[(1R,2S)-1-[[tert- butyl(diphenyl)silyl]oxymethyl]-2-cyclobutyl-4-hydroxy-butyl]carbamate (1.629 g, 100%) as a colorless oil that was directly used in the next step without further purification.1H NMR (400 MHz, CDCl3) δ 7.71 - 7.61 (m, 4H), 7.49 - 7.34 (m, 6H), 4.77 (br. s, 1H), 3.74 (br. s, 1H), 3.69 - 3.45 (m, 4H), 2.32 - 2.17 (m, 1H), 2.12 - 1.87 (m, 3H), 1.83 - 1.57 (m, 5H), 1.54 - 1.44 (m, 1H), 1.43 (s, 9H), 1.39 - 1.28 (m, 1H), 1.07 (s, 9H). ESI-MS m/z calc.511.3118, found 412.4 (M- 99)+; Retention time: 2.16 minutes. LC method Q. Step 4: tert-Butyl N-[(1R,2S)-2-cyclobutyl-4-hydroxy-1- (hydroxymethyl)butyl]carbamate [00577] To a 0 °C solution of tert-butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]- 2-cyclobutyl-4-hydroxy-butyl]carbamate (3.9 g, 7.4530 mmol) in tetrahydrofuran (40 mL) was added a solution of tetrabutylammonium fluoride (solution in THF) (12.5 mL of 1 M, 12.500 mmol) dropwise over 5 min. The solution was stirred at that temperature for 3 h. An aqueous solution of KHCO3 (5%, 40 mL) was added and the mixture was stirred for 1 h. The suspension was then filtered on a pad of sand which was washed with EtOAc (3 x 20 mL) and water (2 x 20 mL). The phases were separated, and the aqueous phase was extracted with EtOAc (2 x 30 mL). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and filtered. The crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 55% of MeCN in water to afford, after evaporation tert-butyl N-[(1R,2S)-2-cyclobutyl-4-hydroxy-1- (hydroxymethyl)butyl]carbamate (1.71 g, 84%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 5.15 - 4.92 (m, 1H), 3.82 - 3.71 (m, 2H), 3.70 - 3.54 (m, 3H), 2.52 - 2.41 (m, 1H), 2.39 - 2.24 (m, 1H), 2.14 - 2.04 (m, 2H), 2.03 - 1.93 (m, 1H), 1.89 - 1.65 (m, 5H), 1.54 - 1.50 (m, 2H), 1.46 (s, 9H). ESI-MS m/z calc.273.194, found 174.4 (M-99)+; Retention time: 1.54 minutes. LC method I. Step 5: tert-Butyl (4R)-4-[(1S)-1-cyclobutyl-3-hydroxy-propyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00578] To a solution of tert-butyl N-[(1R,2S)-2-cyclobutyl-4-hydroxy-1- (hydroxymethyl)butyl]carbamate (1.71 g, 6.2490 mmol) in dichloromethane (20 mL) were added 2,2-dimethoxypropane (12.750 g, 15 mL, 122.42 mmol) and p-toluenesulfonic acid monohydrate (140 mg, 0.7360 mmol) at room temperature. The solution was stirred at rt for 2 h. Acetone (10 mL) and an aqueous solution of sodium bicarbonate (saturated, 10 mL) were added. iPrOAc (50 mL) was added and removed under reduced pressure (3 x). To the resulting white liquid was added an aqueous solution of sodium bicarbonate (sat., 50 mL) and DCM (50 mL). The phases were separated, and the aqueous one was extracted with DCM (2 x 50 mL). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 80% of MeOH in water to afford, tert-butyl (4R)-4- [(1S)-1-cyclobutyl-3-hydroxy-propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.25 g, 64%) as a dark yellow oil.1H NMR (400 MHz, CDCl3) δ 4.15 - 4.01 (m, 1H), 3.96 - 3.62 (m, 4H), 2.61 - 2.42 (m, 1H), 2.12 - 1.92 (m, 3H), 1.90 - 1.64 (m, 5H), 1.63 - 1.58 (m, 2H), 1.58 - 1.52 (overlaps with water, m, 3H), 1.49 (s, 9H), 1.45 (s, 3H). ESI-MS m/z calc.313.2253, found 336.2 (M+23)+; Retention time: 1.87 minutes. LC method I. Step 6: tert-Butyl (4R)-4-[(1S)-1-cyclobutyl-3-oxo-propyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00579] To a 0 °C solution of tert-butyl (4R)-4-[(1S)-1-cyclobutyl-3-hydroxy-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate (1.25 g, 3.9841 mmol) in dichloromethane (25 mL) was added a solution of sodium bromide (140 mg, 1.3606 mmol) in water (12.5 mL) followed by sodium bicarbonate (600 mg, 7.1423 mmol) and TEMPO (28 mg, 0.1792 mmol). The mixture was vigorously stirred for 10 min and sodium hypochlorite (solution in water) (3.6 mL of 10 %w/v, 4.8360 mmol) was added over 1 h. The mixture was stirred for 1 h at 0 °C. MeOH (10 mL), water (50 ml) and DCM (50 mL) were added, the phases were separated, and the organic phase was washed with an aqueous solution of sodium bicarbonate (sat., 30 mL), an aqueous solution of Na2S2O3 (10%, 30 mL) and brine (30 mL). The organic phase was then dried over anhydrous sodium sulfate and filtered to afford crude tert-butyl (4R)-4-[(1S)-1-cyclobutyl-3- oxo-propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.3 g, 99%) as a pale-yellow oil. ESI-MS m/z calc.311.2097, found 334.2 (M+23)+; Retention time: 1.99 minutes 1H NMR (400 MHz, CDCl3) δ 9.85 - 9.74 (m, 1H), 4.02 - 3.69 (m, 3H), 2.54 - 2.25 (m, 4H), 2.13 - 1.99 (m, 1H), 1.97 - 1.78 (m, 3H), 1.78 - 1.64 (m, 2H), 1.63 - 1.53 (overlaps with water, m, 3H), 1.49 (s, 9H), 1.47 (s, 3H). LC method I. Step 7: tert-Butyl (4R)-4-[(1S)-1-cyclobutyl-5-[methoxy(methyl)amino]-5-oxo- pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00580] To a solution of heptanes washed sodium hydride (suspension in mineral oil) (498 mg, 60 %w/w, 12.451 mmol) in tetrahydrofuran (12.5 mL) was added a solution of 2- diethoxyphosphoryl-N-methoxy-N-methyl-acetamide (1.7 g, 7.1068 mmol) in tetrahydrofuran (12.5 mL) dropwise at room temperature. When the formation of hydrogen stopped (around 5 min) the reaction was stirred an additional 15 min and the resulting suspension was added to a solution of tert-butyl (4R)-4-[(1S)-1-cyclobutyl-3-oxo-propyl]-2,2-dimethyl-oxazolidine-3- carboxylate (1.3 g, 3.9574 mmol) in tetrahydrofuran (25 mL). The resulting solution was stirred for 1 h at room temperature and an aqueous solution of sodium bicarbonate (sat., 50 mL) was added followed by EtOAc (50 mL). The phases were separated, and the aqueous phase was extracted with EtOAc (2 x 50 mL). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 85% of MeCN in water to afford, after evaporation, tert-butyl (4R)-4-[(1S)-1-cyclobutyl-5- [methoxy(methyl)amino]-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.32 g, 82%) as a colorless oil. ESI-MS m/z calc.396.2624, found 397.4 (M+1)+; Retention time: 2.0 minutes.1H NMR (400 MHz, CDCl3) δ 7.06 - 6.88 (m, 1H), 6.54 - 6.33 (m, 1H), 4.03 - 3.76 (m, 3H), 3.73 - 3.65 (m, 3H), 3.27 - 3.19 (m, 3H), 2.55 - 2.25 (m, 2H), 2.12 - 1.91 (m, 4H), 1.89 - 1.67 (m, 4H), 1.56 (s, 3H), 1.52 - 1.47 (m, 9H), 1.47 - 1.40 (m, 3H). LC method I. Step 8: tert-Butyl (4R)-4-[(1S)-1-cyclobutyl-5-[methoxy(methyl)amino]-5-oxo- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00581] A solution of tert-butyl (4R)-4-[(1S)-1-cyclobutyl-5-[methoxy(methyl)amino]-5-oxo- pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (187 mg, 0.4711 mmol) in methanol (7 mL) was purged with nitrogen for 15 min and 10% palladium on carbon (50% wet) (175 mg, 5 %w/w, 0.0822 mmol) was added. Hydrogen was bubbled into the reaction for 15 min and the reaction was stirred for 45 min under hydrogen atmosphere. The reaction was filtered on Celite. The crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 90% of MeCN in water to afford, after evaporation, tert-butyl (4R)-4-[(1S)-1-cyclobutyl-5-[methoxy(methyl)amino]-5- oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (163.1 mg, 87%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 3.96 - 3.87 (m, 1H), 3.83 - 3.75 (m, 1H), 3.72 - 3.65 (m, 3H), 3.18 (s, 3H), 2.56 - 2.28 (m, 2H), 2.07 - 1.88 (m, 2H), 1.87 - 1.52 (m, 11H), 1.48 (s, 9H), 1.45 (s, 6H). ESI-MS m/z calc.398.2781, found 299.4 (M-99)+; Retention time: 2.03 minutes. LC method I. Step 9: tert-Butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-1-cyclobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00582] To a stirred yellow solution of 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine (450 mg, 1.569 mmol) in anhydrous tetrahydrofuran (8 mL) was added dropwise a solution of n- BuLi (in hexanes) (600 µL of 2.5 M, 1.500 mmol) over 2 min at -78 °C under nitrogen. The resulting red solution was stirred for 6 min at that temperature. Then a solution of tert-butyl (4R)-4-[(1S)-1-cyclobutyl-5-[methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine- 3-carboxylate (450 mg, 1.111 mmol) in anhydrous tetrahydrofuran (3 mL) was added slowly. After 15 min at that temperature, a saturated aqueous solution of ammonium chloride (10 mL) was added, followed by addition of EtOAc (25 mL). The mixture was allowed to warm to about 0 oC over 15 min and the phases were separated. The aqueous phase was extracted with EtOAc (3 x 15 mL) and the combined organics were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Purification was performed by reversed-phase chromatography (30-99 % MeCN in water over 20 min, no modifier). The volatiles were partially removed under reduced pressure. The aqueous layer was extracted with EtOAc (3 x 25 mL). The combined organics were dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to furnish tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-cyclobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (341 mg, 58%) as a yellow gum. ESI-MS m/z calc.526.3519, found 527.5 (M+1)+; Retention time: 2.04 minutes. LC method A. Step 10: [(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- cyclobutyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00583] To a stirred solution of tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-cyclobutyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (335 mg, 0.6360 mmol) in 1,4-dioxane (6.5 mL) and was added water (600 µL), hydrochloric acid (in 1,4- dioxane) (6.0 mL of 4 M, 24.00 mmol) at ambient temperature, and the reaction was stirred at that temperature for 16 h. The volatiles were removed under reduced pressure to dryness, and then water (10 mL) was added to the residue and concentrated. The residual water was co- evaporated with acetonitrile (3 x 10 mL), followed by toluene (3 x 10 mL) then dried under high vacuum to furnish crude [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- cyclobutyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (hydrochloride salt) (259 mg, 101%) as a yellow gum. ESI-MS m/z calc.368.2576, found 369.2 (M+1)+; Retention time: 1.28 minutes. LC method A. Step 11: [(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- cyclobutyl-azepan-2-yl]methanol and [(2R,3S,7R)-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-3-cyclobutyl-azepan-2-yl]methanol [00584] To a stirred solution of [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-cyclobutyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (hydrochloride salt) (259 mg, 0.6395 mmol) in anhydrous dichloromethane (3.0 mL) were added acetic-acid (80 µL, 1.407 mmol) and DIPEA (350 µL, 2.009 mmol) at ambient temperature under nitrogen. After stirring for 10 min, sodium triacetoxyborohydride (450 mg, 2.123 mmol) was added. The reaction mixture was stirred at rt for 10 min. After quenching with methanol (2.0 mL) and water (1.0 mL), the reaction was concentrated under reduced pressure. The crude was purified by reversed-phase chromatography (10-99% acetonitrile in water containing 5 mM HCl, over 20 min run) to furnish a diastereomeric mixture of [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-3-cyclobutyl-azepan-2-yl]methanol (hydrochloride salt) as a light-brown gum (227 mg, 87%). The diastereomers were separated by preparative SFC [AD-H (3 x 25 cm); 45% isopropanol (0.1% diethylamine)/CO2, 100 bar ; 70 mL/min, 240 nm.; Injection Vol. : 0.6 ml, 23 mg/ml methanol]. The individual peak materials were concentrated under reduced pressure and the residual water was removed azeotropically with acetonitrile (5 x 10 mL), followed by toluene (3 x 10 mL) to furnish the pure diastereomers as white gums. First to elute, SFC Peak-1 (major): [(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-cyclobutyl-azepan- 2-yl]methanol (118 mg, 50%), 1H NMR (400 MHz, CDCl3) δ 8.28 (s, 1H), 6.44 (s, 1H), 4.25 - 4.16 (m, 1H), 4.04 - 4.00 (m, 1H), 3.99 (s, 3H), 3.63 (dd, J = 10.3, 4.4 Hz, 1H), 3.12 (t, J = 10.2 Hz, 1H), 2.69 (ddd, J = 10.2, 8.3, 4.4 Hz, 1H), 2.32 (p, J = 8.6 Hz, 1H), 2.15 - 2.03 (m, 2H), 2.02 - 1.93 (m, 1H), 1.87 - 1.76 (m, 3H), 1.76 - 1.60 (m, 6H), 1.49 (s, 9H), 1.45 - 1.36 (m, 1H). ESI-MS m/z calc.370.27325, found 371.3 (M+1)+; Retention time: 1.22 minutes, and second to elute, SFC Peak-2 (minor): [(2R,3S,7R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- cyclobutyl-azepan-2-yl]methanol (16.0 mg, 7%), 1H NMR (400 MHz, CDCl3) δ 8.33 (s, 1H), 6.44 (d, J = 0.9 Hz, 1H), 4.27 - 4.15 (m, 1H), 4.02 (dd, J = 11.5, 2.8 Hz, 1H), 3.99 (s, 3H), 3.62 (dd, J = 10.7, 4.7 Hz, 1H), 3.44 (t, J = 10.6 Hz, 1H), 2.81 - 2.71 (m, 1H), 2.36 - 2.26 (m, 1H), 2.23 - 2.14 (m, 1H), 2.13 - 2.05 (m, 1H), 2.02 - 1.93 (m, 2H), 1.85 - 1.61 (m, 8H), 1.54 - 1.51 (m, 1H), 1.50 (s, 9H). ESI-MS m/z calc.370.27325, found 371.3 (M+1)+; Retention time: 1.21 minutes. LC method A. Step 12: 3-[[4-[[(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-cyclobutyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00585] A stirred solution of [(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-3-cyclobutyl-azepan-2-yl]methanol (107 mg, 0.2888 mmol) and 3-[[4-chloro-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (130 mg, 0.3111 mmol) in anhydrous tetrahydrofuran (2.5 mL) was sparged with nitrogen for 5 min. Then sodium tert-butoxide (175 mg, 1.821 mmol) was added in a single portion at ambient temperature and after stirring for 10 min, the reaction was allowed to stir at 42 °C for 3 h. The reaction was cooled down to room temperature. The reaction mixture was diluted with ethyl acetate (20 mL) and acidified with hydrochloric acid (3.0 mL of 1.0 M, 3.000 mmol). The organic phase was separated, and the aqueous layer was extracted with ethyl acetate (2 x 15 mL). The combined organics were washed with brine (10 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. Traces of water were removed azeotropically with acetonitrile (2 x 10 mL) and toluene (2 x 10 mL) to furnish 3-[[4-[[(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-cyclobutyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) as a yellow solid (111 mg, 49%). ESI-MS m/z calc.751.35156, found 752.2 (M+1)+; Retention time: 1.5 minutes. LC method A. Step 13: (4S,8S,9R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8- cyclobutyl-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one [00586] To a stirred solution of 3-[[4-[[(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-cyclobutyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (110 mg, 0.1395 mmol) in anhydrous NMP (2.2 mL) and acetone (4.4 mL) was added p-TsCl (120 mg, 0.6295 mmol) at ambient temperature under nitrogen and the reaction was stirred for 2 min. Then N-methylmorpholine (200 µL, 1.819 mmol) was added and the reaction was stirred at 52 oC for 3 h. The reaction was allowed to cool to ambient temperature and the volatiles were removed under reduced pressure. The remaining brownish solution was subjected to preparative HPLC (30-99% acetonitrile in water, containing 5 mM HCl as modifier, 30 min run). The desired fractions were concentrated under reduced pressure and dried to furnish as a yellow solid (4S,8S,9R)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-8-cyclobutyl-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen- 2-one (36 mg, 34%) 1H NMR (400 MHz, CDCl3) δ 8.85 (d, J = 2.0 Hz, 1H), 8.36 (s, 1H), 7.82 (d, J = 7.9 Hz, 1H), 7.77 (d, J = 7.7 Hz, 1H), 7.47 (t, J = 7.8 Hz, 1H), 7.16 (t, J = 7.9 Hz, 1H), 7.03 (d, J = 7.6 Hz, 2H), 6.43 (s, 1H), 6.25 (s, 1H), 5.97 (dd, J = 11.6, 5.1 Hz, 1H), 5.76 (dd, J = 13.2, 4.4 Hz, 1H), 4.84 (t, J = 11.8 Hz, 1H), 4.39 - 4.32 (m, 1H), 4.07 (s, 3H), 2.39 - 2.28 (m, 1H), 2.26 - 2.18 (m, 1H), 2.12 - 2.05 (m, 1H), 2.02 (s, 6H), 1.93 - 1.79 (m, 2H), 1.77 - 1.71 (m, 1H), 1.70 - 1.61 (m, 3H), 1.54 - 1.46 (m, 5H), 1.42 (s, 9H). ; ESI-MS m/z calc.733.341, found 734.9 (M+1)+; Retention time: 1.99 minutes. LC method A. Example 71: Preparation of Compound 108 Step 1: tert-Butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-3- propyl-pyrrolidine-1-carboxylate [00587] A flame-dried flask was charged with copper iodide (3.39 g, 17.800 mmol) in anhydrous THF (40 mL). The heterogeneous mixture was cooled down to -78 °C and a solution of propyl magnesium chloride (in Me-THF) (35 mL of 1 M, 35.000 mmol) was added dropwise over 30 minutes. The reaction was stirred at -78 °C for 1 h after which time a solution of tert- butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-2H-pyrrole-1-carboxylate (4.02 g, 8.9011 mmol) in anhydrous THF (20 mL) was added dropwise over 0.5 hour keeping the internal temperature below -78 °C. The dry-ice bath was then removed allowing the temperature to slowly reach room temperature over 30 minutes. The reaction mixture was put in an ice-cold bath and was quenched by a saturated aqueous solution of ammonium chloride (100 mL). EtOAc (100 mL) was added, the layers were separated, and the aqueous layer was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with water (2 x 100 mL) and brine (1 x 100 mL), dried over anhydrous sodium sulfate, filtered and the solvents were evaporated under reduced pressure to afford crude tert-butyl (2R,3R)-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-5-oxo-3-propyl-pyrrolidine-1-carboxylate (5.31 g, 72%) as a yellow oil. ESI-MS m/z calc.495.2805, found 396.2 (M-99)+; Retention time: 2.45 minutes. LC method S. Step 2: tert-Butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2- hydroxyethyl)pentyl]carbamate [00588] To a solution of tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-3- propyl-pyrrolidine-1-carboxylate (3.78 g, 7.0380 mmol) in ethanol (58 mL) at 50 °C was added sodium borohydride (2.72 g, 71.896 mmol) portion-wise over 2 minutes. The resulting mixture was stirred at this temperature for 1 hour then water (50 mL) was added, and the volatiles were removed under reduced pressure. The residual aqueous solution was diluted with EtOAc (50 mL) and the phases were separated. The aqueous solution was extracted with EtOAc (2 x 50 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert-butyl N- [(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2-hydroxyethyl)pentyl]carbamate (3.58 g, 81%) as a colorless oil that was used in the following step without further purification.1H NMR (400 MHz, DMSO-d6) δ 7.65 - 7.58 (m, 4H), 7.48 - 7.40 (m, 6H), 6.57 - 6.51 (m, 1H), 4.35 (t, J = 4.9 Hz, 1H), 3.73 - 3.63 (m, 1H), 3.56 (d, J = 6.1 Hz, 2H), 3.44 - 3.28 (overlapped with water, m, 2H), 1.61 - 1.53 (m, 1H), 1.49 - 1.09 (overlapped with EtOAc. m, 15H), 0.98 (s, 9H), 0.84 - 0.76 (m, 3H). ESI-MS m/z calc.499.3118, found 500.4 (M+1)+; Retention time: 2.34 minutes. LC method S. Step 3: tert-Butyl N-[(1R,2R)-2-(2-hydroxyethyl)-1- (hydroxymethyl)pentyl]carbamate [00589] To a solution of tert-butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(2- hydroxyethyl)pentyl]carbamate (3.58 g, 5.7093 mmol) in anhydrous THF (40 mL) at 0 °C was added a solution of TBAF (in THF) (10 mL of 1 M, 10.000 mmol). The reaction was stirred in the ice-cold bath for 1 hour and 30 minutes. The reaction was quenched at 0 °C with a saturated aqueous solution of sodium bicarbonate (50 mL) and EtOAc (50 mL). The phases were separated, and the aqueous layer was extracted with EtOAc (2 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography (Column: C18. Gradient: 5-80 % acetonitrile in water). The fractions containing the product were combined and the volatiles were removed under reduced pressure. The aqueous solution was diluted with brine (100 mL) and was extracted with DCM (4 x 50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford tert-butyl N-[(1R,2R)-2-(2-hydroxyethyl)-1- (hydroxymethyl)pentyl]carbamate (1.39 g, 85%) as a yellow oil.1H NMR (400 MHz, DMSO- d6) δ 6.39 (d, J = 9.3 Hz, 1H), 4.45 (t, J = 5.5 Hz, 1H), 4.36 (t, J = 5.0 Hz, 1H), 3.53 - 3.25 (overlapped with water, m, 5H), 1.61 - 1.44 (m, 2H), 1.41 - 1.08 (m, 14H), 0.83 (t, J = 6.8 Hz, 3H). ESI-MS m/z calc.261.194, found 206.4 (M-55)+; Retention time: 1.52 minutes. LC method I. Step 4: tert-Butyl (4R)-4-[(1R)-1-(2-hydroxyethyl)butyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00590] To a stirred solution of tert-butyl N-[(1R,2R)-2-(2-hydroxyethyl)-1- (hydroxymethyl)pentyl]carbamate (1.39 g, 4.8397 mmol) in anhydrous DCM (25 mL) were added 2,2-dimethoxypropane (10.200 g, 12 mL, 97.938 mmol) and p-toluenesulfonic acid hydrate (116 mg, 0.6098 mmol). The reaction was stirred at room temperature for 3 hours then concentrated under reduced pressure and partitioned between ethyl acetate (20 mL) and a saturated aqueous solution of sodium bicarbonate (20 mL). The mixture was vigorously stirred at room temperature for 20 minutes, then the layers were separated, and the aqueous layer was extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with brine (25 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert-butyl (4R)-4-[(1R)-1-(2-hydroxyethyl)butyl]-2,2-dimethyl-oxazolidine-3- carboxylate (1.56 g, 102%) as an orange oil that was used in the following step without further purification.1H NMR (400 MHz, DMSO-d6) δ 4.36 (t, J = 4.6 Hz, 1H), 3.92 - 3.81 (m, 2H), 3.76 (d, J = 8.1 Hz, 1H), 3.47 - 3.37 (m, 2H), 1.88 (br. s., 1H), 1.56 - 1.31 (m, 17H), 1.30 - 1.00 (m, 4H), 0.91 - 0.77 (m, 3H). ESI-MS m/z calc.301.2253, found 324.2 (M+23)+; Retention time: 1.84 minutes. LC method I. Step 5: tert-Butyl (4R)-2,2-dimethyl-4-[(1R)-1-(2-oxoethyl)butyl]oxazolidine-3- carboxylate [00591] To a solution of tert-butyl (4R)-4-[(1R)-1-(2-hydroxyethyl)butyl]-2,2-dimethyl- oxazolidine-3-carboxylate (1.46 g, 4.6064 mmol) in dichloromethane (40 mL) cooled at 0 °C were added water (15 mL), sodium bicarbonate (625 mg, 7.4399 mmol), sodium bromide (100 mg, 0.9719 mmol) and TEMPO (132 mg, 0.8448 mmol). To the bi-phasic solution was added a solution of sodium hypochlorite (in water) (4 mL of 10 %w/v, 5.3733 mmol) slowly dropwise over 5 minutes under vigorous stirring. After stirring for 1 hour at 0 °C, a saturated aqueous solution of sodium bicarbonate (50 mL) was added. The mixture was diluted with DCM (50 mL) and the phases were separated. The aqueous phase was extracted with DCM (2 x 20 mL) and the combined organic layers were washed with a 10% aqueous solution of Na2S2O3 (50 mL), a saturated aqueous solution of sodium bicarbonate (2 x 50 mL) and brine (50 mL). The organic solution was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert-butyl (4R)-2,2-dimethyl-4-[(1R)-1-(2-oxoethyl)butyl]oxazolidine-3- carboxylate (1.56 g, 96%) a pale orange oil.1H NMR (400 MHz, DMSO-d6) δ 9.66 (s, 1H), 3.92 - 3.67 (m, 3H), 2.44 - 2.26 (m, 2H), 1.60 - 1.04 (m, 20H), 0.84 (t, J = 6.7 Hz, 3H). ESI-MS m/z calc.299.2097, found 322.2 (M+23)+; Retention time: 1.95 minutes. LC method I. Step 6: tert-Butyl (4R)-4-[(1R)-5-[methoxy(methyl)amino]-5-oxo-1-propyl-pent- 3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00592] To a solution of sodium hydride (in mineral oil) (210 mg, 60 %w/w, 5.2505 mmol) in anhydrous THF (20 mL) was added dropwise 2-diethoxyphosphoryl-N-methoxy-N-methyl- acetamide (1.1630 g, 1 mL, 4.8619 mmol) while stirring the suspension at room temperature. Once the hydrogen gas evolution stopped, this solution was added dropwise to a solution of tert- butyl (4R)-2,2-dimethyl-4-[(1R)-1-(2-oxoethyl)butyl]oxazolidine-3-carboxylate (1.56 g, 4.4288 mmol) in anhydrous THF (20 mL) stirred at room temperature. The reaction mixture was stirred at room temperature for 1 hour and was quenched with a saturated aqueous solution of sodium bicarbonate (50 mL). This mixture was extracted with EtOAc (3 x 50 mL) and the combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert-butyl (4R)-4-[(1R)-5- [methoxy(methyl)amino]-5-oxo-1-propyl-pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (2 g, 100%) as an orange oil. Isolated as a mixture of E/Z stereoisomers, unknown ratio. ESI-MS m/z calc.384.2624, found 385.2 (M+1)+; Retention time: 1.97 minutes. LC method I. Step 7: tert-Butyl (4R)-4-[(1R)-5-[methoxy(methyl)amino]-5-oxo-1-propyl- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00593] A solution of tert-butyl (4R)-4-[(1R)-5-[methoxy(methyl)amino]-5-oxo-1-propyl- pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.43 g, 3.5331 mmol) in methanol (40 mL) was bubbled with nitrogen for 10 minutes then palladium on carbon (750 mg, 5 %w/w, 0.3524 mmol) was added. The suspension was sparged with hydrogen for 10 minutes then stirred under hydrogen atmosphere for 1 hour. The suspension was bubbled with nitrogen for 10 minutes, then was filtered through a short pad of Celite. The pad was rinsed with EtOAc (2 x 20 mL) and the combined organic filtrates were concentrated under reduced pressure to afford tert- butyl (4R)-4-[(1R)-5-[methoxy(methyl)amino]-5-oxo-1-propyl-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (1.36 g, 99%) as a cloudy white oil. ESI-MS m/z calc.386.2781, found 387.4 (M+1)+; Retention time: 2.04 minutes. LC method I. Step 8: tert-Butyl (4R)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-5-oxo-1-propyl-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00594] To a solution of 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine (291.9 mg, 1.089 mmol) in tetrahydrofuran (5.4 mL) at -78 °C was added n-butyllithium (solution in hexanes) (0.42 mL of 2.5 M, 1.050 mmol) dropwise. The resulting solution was stirred for 5 minutes. A solution of tert-butyl (4R)-4-[(1R)-5-[methoxy(methyl)amino]-5-oxo-1-propyl- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (300 mg, 0.7761 mmol) in tetrahydrofuran (2.1 mL) was added dropwise. The reaction was left stirring for 15 min. The reaction mixture was diluted with EtOAc (21 mL) and ammonium chloride (14 mL) was added. The crude mixture was allowed to warm to room temperature and the phases separated. The aqueous phases were extracted with EtOAc (2x) and the combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude material was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 20 min) to give tert-butyl (4R)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-5-oxo-1-propyl- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (180.0 mg, 45%) as an orange oil. ESI-MS m/z calc.514.3519, found 515.7(M+1)⁺; Retention time: 2.41 minutes. LC method A. Step 9: [(2R,3R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-propyl- 3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00595] To a solution of tert-butyl (4R)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-5-oxo-1-propyl-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (180.0 mg, 0.3497 mmol) in 1,4-dioxane (3.8 mL) was added hydrochloric acid (solution in 1,4-dioxane) (1.8 mL of 4 M, 7.200 mmol) at rt. Three drops of water were added and the reaction was stirred for 18 hours. The solvent was removed under reduced pressure and the residual water was co- evaporated with acetonitrile (3x), then dried under high vacuum to afford crude [(2R,3R)-7-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-propyl-3,4,5,6-tetrahydro-2H-azepin-2- yl]methanol (124.7 mg, 96%) as a yellow solid. ESI-MS m/z calc.356.2576, found 357.3(M+1)⁺; Retention time: 1.24 minutes. The crude material was taken forward without further purification. LC method A. Step 10: [(2R,3R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-propyl- azepan-2-yl]methanol, diastereomer 1 and [(2R,3R)-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-3-propyl-azepan-2-yl]methanol, diastereomer 2 [00596] To a solution of [(2R,3R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- propyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (124.7 mg, 0.3358 mmol) in anhydrous dichloromethane (8.1 mL) at room temperature was added sodium triacetoxyborohydride (430.9 mg, 2.033 mmol). After 2 h, additional sodium triacetoxyborohydride (36.9 mg, 0.1741 mmol) was added. The reaction was stirred at room temperature for an additional 2 h, diluted with DCM (10 mL), and a saturated aqueous solution of sodium bicarbonate (10 mL) was added. The phases were separated, and the aqueous solution was extracted with DCM (2 x). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 25 min) to give 57.2 mg which was submitted for SFC separation. This material was subjected to chiral SFC separation using a Phenomenex LUX-4 (21.2 x 250 mm, 5 μm) at 40 oC. The mobile phase was 32% MeOH (20 mM NH3) at 70 mL/min flow in gradient mode. Concentration of the sample was 19.1 mg/mL in MeOH. The injection volume was 500 μL with variable outlet pressure and the detection wavelength was 229 nm. Each isomer was dissolved in THF and concentrated down three times to give diastereomer 1, peak 1: [(2R,3R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-propyl-azepan-2- yl]methanol (24.8 mg, 21%) as a white solid ESI-MS m/z calc.358.27325, found 359.2(M+1)⁺; Retention time: 1.20 minutes. and diastereomer 2, peak 2: [(2R,3R)-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-3-propyl-azepan-2-yl]methanol (15.2 mg, 13%) as a white solid ESI-MS m/z calc.358.27325, found 359.3(M+1)⁺; Retention time: 1.20 minutes. LC method A. Step 11: 3-[[4-[[(2R,3R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- propyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid, diastereomer 1
diastereomer 1 [00597] 3-[[4-chloro-6-(2,6-Dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (28.90 mg, 0.06916 mmol) and [(2R,3R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- propyl-azepan-2-yl]methanol, diastereomer 1 (24.8 mg, 0.06917 mmol) were combined in a flask and dissolved in THF (0.43 mL) followed by the addition of NaOt-Bu (35.8 mg, 0.3725 mmol). The reaction mixture was stirred for 1 hour at room temperature then warmed to 40 °C for 2 hours. The reaction mixture was cooled to room temperature and diluted with EtOAc and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (5X). The organic layers were combined, washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3-[[4-[[(2R,3R)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-propyl-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (41.9 mg, 78%) as a yellow solid. ESI-MS m/z calc.739.35156, found 740.6 (M+1)⁺; Retention time: 1.54 minutes. LC method A. Step 12: (8R,9R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-18,18-dioxo-8-propyl-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12(24),13,15,19(23),20- hexaen-2-one, diastereomer 1 (Compound 108)
Diastereomer 1 Diastereomer 1 [00598] 3-[[4-[[(2R,3R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-propyl- azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt), diastereomer 1 (41.9 mg, 0.05397 mmol) was dissolved in NMP (0.84 mL) and acetone (1.7 mL). TsCl (32.4 mg, 0.1700 mmol) was added followed by NMM (35.6 µL, 0.3238 mmol) and the reaction was stirred at 55 °C for 22 hours. The reaction was poured into a mixture of ethyl acetate and 1 M HCl and the layers were separated. The aqueous layer was further extracted with ethyl acetate (2 x). The organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was taken up in DMSO/MeOH then injected into a reverse phase HPLC (30-99% acetonitrile/5 mM aqueous HCl over 25 min) to give (8R,9R)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 14-(2,6-dimethylphenyl)-18,18-dioxo-8-propyl-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12(24),13,15,19(23),20-hexaen-2-one diastereomer 1 (19.7 mg, 49%) as a tan solid.1H NMR (400 MHz, CDCl3) δ 8.82 (s, 1H), 8.45 (s, 1H), 7.80 (t, J = 6.6 Hz, 2H), 7.57 (t, J = 7.7 Hz, 1H), 7.23 (d, J = 7.6 Hz, 1H), 7.09 (s, 1H), 7.08 (s, 1H), 6.57 (s, 1H), 6.26 (s, 1H), 5.95 - 5.83 (m, 2H), 4.89 (t, J = 11.8 Hz, 1H), 4.49 - 4.41 (m, 1H), 4.16 (s, 3H), 2.35 - 2.27 (m, 1H), 2.08 (s, 6H), 1.96 - 1.87 (m, 3H), 1.87 - 1.78 (m, 3H), 1.50 (s, 9H), 1.40 - 1.23 (m, 4H), 0.76 (t, J = 7.0 Hz, 3H). ESI-MS m/z calc.721.341, found 722.9(M+1)⁺; Retention time: 2.02 minutes. LC method A. Example 72: Preparation of Compound 109 Step 1: tert-butyl (4S)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate, and tert- butyl (4S)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1- isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate
[00599] A crude mixture containing tert-butyl (4S)-4-[5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (339 mg, 0.5027 mmol) was mixed with another crude mixture containing tert-butyl (4S)-4-[5-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (4.0 g, 5.9469 mmol) in DMSO (10 mL). Both mixtures were constituted of two diastereomers in a ~90:10 ratio, the major isomer being in the 4S,1S configuration, and were obtained as impure minor chromatographic fractions from a large-scale (~100 g) synthesis of the 4S,1R diastereomer. The solution was directly purified by reversed- phase chromatography (column: C18; gradient: 5-90 % acetonitrile in water) to afford 2.92 g of the desired product as a mixture of diastereomers in an approximate 92:8 ratio. The mixture of diastereomers was submitted to chiral SFC separation (Flow rate: 100 mL/min, 30% MeOH, column: Cellulose 2, temperature = 40 °C, outlet pressure: 100 bar, injection volume: 1300 µL). The purest fractions containing the fastest eluting peak (SFC peak 1) were combined and concentrated under reduced pressure to afford tert-butyl (4S)-4-[(1S)-5-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (2.52 g, 72%) as a beige solid.1H NMR (400 MHz, CDCl3) δ 9.12 (s, 1H), 6.59 (s, 1H), 4.17 - 4.05 (m, 4H), 4.03 - 3.89 (m, 2H), 3.87 - 3.65 (m, 2H), 3.40 - 3.21 (m, 2H), 2.09 - 1.96 (m, 1H), 1.84 - 1.69 (m, 1H), 1.67 - 1.41 (overlapped with water, m, 26H), 1.22 - 1.09 (m, 6H). ESI-MS m/z calc.530.3468, found 531.4 (M+1)+; Retention time: 2.43 minutes. The purest fractions containing the slowest eluting peak (SFC peak 2) were combined and concentrated under reduced pressure to afford tert-butyl (4S)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (205 mg, 6%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 9.12 (s, 1H), 6.60 (s, 1H), 4.17 - 4.05 (m, 4H), 4.04 - 3.66 (m, 4H), 3.37 - 3.20 (m, 2H), 2.00 - 1.77 (m, 2H), 1.67 - 1.42 (overlapped with water, m, 26H), 1.21 - 1.08 (m, 6H). ESI-MS m/z calc.530.3468, found 431.2 (M-99)+; Retention time: 2.44 minutes. LC method U. Step 2: (5S,6S)-6-Amino-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-7- hydroxy-5-isopropoxy-heptan-1-one [00600] To a solution of tert-butyl (4S)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-isopropoxy-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (2.45 g, 4.4873 mmol) in toluene (20 mL) was added water (4.0000 g, 4 mL, 222.03 mmol) and a solution of HCl (in 1,4-dioxane) (12 mL of 4 M, 48.000 mmol) in one portion. The bi-phasic solution was stirred at room temperature for 20 hours and EtOAc (25 mL) was added to the mixture. Water (25 mL) was added and the phases were separated. The organic layer was washed with an aqueous solution of HCl 1 M (3 x 10 mL). The combined aqueous layers were cooled in an ice-bath and basified with an aqueous solution of NaOH 2 M (60 mL) until pH=12. The aqueous solution was diluted with brine (50 mL) and was extracted with DCM (5 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude, after freeze-drying, (5S,6S)-6-amino-1-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-7-hydroxy-5-isopropoxy-heptan-1-one (1.67 g, 95%) as a yellow solid. ESI-MS m/z calc.390.2631, found 391.4 (M+1)+; Retention time: 1.44 minutes. LC method U. Step 3: [(2S,3S,7R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methanol and [(2S,3S,7S)-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methanol
[00601] A solution of (5S,6S)-6-amino-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 7-hydroxy-5-isopropoxy-heptan-1-one (506 mg, 1.2944 mmol) in MeOH (20 mL) was heated at 65 °C for 18 hours. The reaction mixture was allowed to cool down to room temperature and the volatiles were removed under reduced pressure. The residue was diluted with anhydrous DCM (10 mL) and the solution was cooled to 0 °C. To the reaction mixture at 0 °C was added a solution of DIBAL (in toluene) (4 mL of 1 M, 4.0000 mmol) slowly dropwise. The reaction mixture was stirred for 2 hours and 20 minutes in the ice-cold bath and was quenched with a 10% aqueous solution of Rochelle salt (25 mL). The bi-phasic solution was stirred at room temperature for 20 hours. The phases were separated, and the aqueous solution was extracted with DCM (3 x 25 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography (Column: C18. Gradient: 5-100 % acetonitrile in basic water (ammonium bicarbonate/ammonium hydroxide buffer pH = 10)). The purest fractions containing the desired diastereomer were combined and the volatiles were removed under reduced pressure. The residual aqueous solution was diluted with brine (50 mL) and was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford, after freeze-drying, [(2S,3S,7R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methanol (80.1 mg, 17%) as a beige solid.1H NMR (400 MHz, Benzene-d6) δ 8.50 (s, 1H), 6.59 (s, 1H), 3.90 - 3.83 (m, 1H), 3.80 - 3.72 (m, 4H), 3.62 (dd, J = 9.5, 4.6 Hz, 1H), 3.34 - 3.25 (m, 1H), 3.22 - 3.12 (m, 2H), 2.15 - 2.06 (m, 1H), 1.87 - 1.79 (m, 1H), 1.75 - 1.64 (m, 2H), 1.51 - 1.28 (m, 3H), 1.09 (s, 9H), 1.04 (d, J = 6.1 Hz, 3H), 1.01 - 0.95 (m, 1H), 0.92 (d, J = 5.9 Hz, 3H). ESI-MS m/z calc.374.2682, found 375.2 (M+1)+; Retention time: 3.66 minutes. [00602] All fractions containing both diastereomers were combined and concentrated under reduced pressure. The residual aqueous solution was diluted with brine (50 mL) and was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (column: C18; gradient: 5-55 % acetonitrile in basic water (ammonium bicarbonate/ammonium hydroxide buffer pH = 10)). The purest fractions containing the (S,S,S)-diastereomer were combined and the volatiles were removed under reduced pressure. The residual aqueous solution was diluted with brine (50 mL) and was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (column: C18; gradient: 5- 45 % acetonitrile in water with 0.1% formic acid). The purest fractions containing the (S,S,S)- diastereomer were combined and the volatiles were removed under reduced pressure to afford, after freeze-drying, [(2S,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methanol (137.1 mg, 28%) as a beige solid.1H NMR (400 MHz, Benzene-d6) δ 8.45 (s, 1H), 6.56 (s, 1H), 4.38 - 4.31 (m, 1H), 3.92 - 3.83 (m, 1H), 3.73 (dd, J = 10.8, 4.9 Hz, 1H), 3.50 (s, 3H), 3.46 - 3.36 (m, 1H), 3.32 (t, J = 3.3 Hz, 1H), 2.90 (dd, J = 8.2, 5.0 Hz, 1H), 1.89 - 1.63 (m, 4H), 1.15 - 0.94 (m, 17H), (2H missing, labile protons). ESI-MS m/z calc.374.2682, found 375.2 (M+1)+; Retention time: 3.64 minutes. LC method W. Step 4: 3-[[4-[[(2S,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00603] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (163.8 mg, 0.3920 mmol) and [(2S,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methanol (146.8 mg, 0.3920 mmol) were combined in a flask and dissolved in THF (2.5 mL) followed by the addition of NaOt-Bu (204.7 mg, 2.130 mmol). The reaction mixture was stirred for 1 hour at room temperature then warmed to 40 °C for 1 hour. The reaction mixture was cooled to room temperature and diluted with EtOAc and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (5 x). The organic layers were combined, washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3-[[4-[[(2S,3S,7S)-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (235.0 mg, 76%) as a tan solid. ESI-MS m/z calc.755.3465, found 756.1(M+1)⁺; Retention time: 1.59 minutes. LC method A. Step 5: (4S,8S,9S)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-8-isopropoxy-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one (Compound 109) [00604] 3-[[4-[[(2S,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (235.0 mg, 0.2966 mmol) was dissolved in NMP (4.7 mL) and Acetone (9.5 mL). TsCl (171.5 mg, 0.8996 mmol) was added followed by NMM (0.2 mL, 1.819 mmol) and the reaction was stirred at 55 °C for 22 hours. The reaction was poured into a mixture of ethyl acetate and 1 M HCl and the layers were separated. The aqueous layer was further extracted with ethyl acetate (2 x). The organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was taken up in DMSO/MeOH then injected into a reverse phase HPLC (30-99% acetonitrile/5 mM aqueous HCl over 25 min). The fractions were extracted with EtOAc (3 x), washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give (4S,8S,9S)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-8- isopropoxy-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (111 mg, 49%) as a tan solid.1H NMR (400 MHz, CDCl3) δ 8.81 (s, 1H), 8.45 (s, 1H), 7.79 - 7.72 (m, 1H), 7.49 (d, J = 4.7 Hz, 2H), 7.26 - 7.23 (m, 1H), 7.08 (d, J = 7.6 Hz, 2H), 6.46 (s, 1H), 6.25 (s, 1H), 5.89 - 5.79 (m, 2H), 4.99 (t, J = 12.0 Hz, 1H), 4.87 - 4.80 (m, 1H), 4.13 (s, 3H), 3.31 (d, J = 11.3 Hz, 1H), 2.94 (hept, J = 6.2 Hz, 1H), 2.42 - 2.33 (m, 1H), 2.25 (t, J = 13.5 Hz, 1H), 2.19 - 2.03 (m, 7H), 2.00 (d, J = 12.0 Hz, 1H), 1.97 - 1.84 (m, 1H), 1.64 - 1.61 (m, 1H), 1.50 (s, 9H), 0.81 (d, J = 6.0 Hz, 3H), 0.29 (d, J = 6.1 Hz, 3H). ESI-MS m/z calc.737.33594, found 738.2(M+1)⁺; Retention time: 1.80 minutes. LC method A. Example 73: Preparation of Compound 110 Step 1: tert-Butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-cyclopropyl- 5-oxo-pyrrolidine-1-carboxylate [00605] To a solution of cyclopropyl magnesium bromide (solution in MeTHF) (24 mL of 1 M, 24.000 mmol) in tetrahydrofuran (75 mL) at -40 °C was added a suspension of copper iodide (1.93 g, 10.134 mmol) in tetrahydrofuran (10 mL) and the resulting suspension was stirred 1 h at that temperature. To the suspension at -40 °C was added a solution of tert-butyl (2R)-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-5-oxo-2H-pyrrole-1-carboxylate (5 g, 9.8421 mmol) in tetrahydrofuran (25 mL) dropwise over 1.25 hours. The reaction was stirred for 15 min and a saturated aqueous solution of ammonium chloride (100 mL) was added. Water (100 mL) was added and the aqueous phase was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by reversed phase chromatography (Column: C18, Gradient: 5-70 % MeCN in water) to afford tert-butyl (2R,3S)-2- [[tert-butyl(diphenyl)silyl]oxymethyl]-3-cyclopropyl-5-oxo-pyrrolidine-1-carboxylate (4.49 g, 89%) as an orange oil.1H NMR (400 MHz, CDCl3) δ 7.65 - 7.56 (m, 4H), 7.48 - 7.35 (m, 6H), 4.08 - 4.03 (m, 1H), 3.83 (dd, J = 10.4, 4.8 Hz, 1H), 3.69 (dd, J = 10.4, 2.6 Hz, 1H), 2.91 (dd, J = 17.6, 8.8 Hz, 1H), 2.34 (dd, J = 17.7, 1.6 Hz, 1H), 1.62 - 1.54 (overlapped with water, m, 1H), 1.45 (s, 9H), 1.04 (s, 9H), 0.95 - 0.84 (m, 1H), 0.58 - 0.49 (m, 2H), 0.21 - 0.13 (m, 2H). ESI-MS m/z calc.493.2648, found 516.2 (M+23)+; Retention time: 2.5 minutes. LC method U. 9H), 1.04 (s, 9H), 0.95 - 0.84 (m, 1H), 0.58 - 0.49 (m, 2H), 0.21 - 0.13 (m, 2H). Step 2: tert-Butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-cyclopropyl- 5-hydroxy-pyrrolidine-1-carboxylate [00606] Lithium triethylborohydride in THF (8 mL of 1 M, 8.0000 mmol) was added dropwise to a solution of tert-butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- cyclopropyl-5-oxo-pyrrolidine-1-carboxylate (982 mg, 1.8558 mmol) in anhydrous THF (20 mL) at - 78 °C. The reaction was then stirred at - 78 °C for 2 hours then quenched by the addition of a saturated aqueous solution of sodium bicarbonate (50 mL) and slowly allowed to reach 0 °C after which time an aqueous solution of hydrogen peroxide in water (435.05 mg, 1.1 mL, 4.4765 mmol) was added and the resulting solution was left stirring at this temperature for 30 min at 0 °C. Dichloromethane (50 mL) was added and the layers were separated. The aqueous solution was extracted with dichloromethane (2 x 50 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure to give crude tert-butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-cyclopropyl-5-hydroxy- pyrrolidine-1-carboxylate (894 mg, 95%) as a brown oil 1H NMR (400 MHz, Chloroform-d) δ 7.68 - 7.58 (m, 4H), 7.47 - 7.36 (m, 6H), 5.62 - 5.43 (m, 1H), 3.78 - 3.41 (m, 3H), 2.04 - 1.81 (m, 2H), 1.59 - 1.50 (m, 9H), 1.07 - 1.03 (m, 9H), 0.96 - 0.81 (m, 1H), 0.79 - 0.61 (m, 1H), 0.59 - 0.33 (m, 2H), 0.21 - -0.05 (m, 2H). ESI-MS m/z calc.495.2805, found 378.4 (M-117)+; Retention time: 2.7 minutes. LC method O. Step 3: tert-Butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2- cyclopropyl-4-hydroxy-butyl]carbamate [00607] To a stirred solution of tert-butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- cyclopropyl-5-hydroxy-pyrrolidine-1-carboxylate (7.978 g, 14.098 mmol) in methanol (80 mL) was added slowly NaBH4 (14.2 g, 375.34 mmol). The reaction mixture was diluted with methanol (80 mL). The reaction was heated at 75 °C for 2 h. To the reaction mixture was added water (50 mL) and the volatiles were concentrated under reduced pressure. To the resulting aqueous solution was added DCM (50 mL) and the phases were separated. The aqueous phase was extracted with DCM (3 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude product. The crude material was purified by reverse phase chromatography (gradient: 5 to 100 % MeOH in pure water, 16 CV) to afford tert-butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2- cyclopropyl-4-hydroxy-butyl]carbamate (4.167 g, 59%) as a yellow oil 1H NMR (400 MHz, CDCl3 ) δ 7.65 (td, J = 7.7, 1.5 Hz, 4H), 7.52 - 7.33 (m, 6H), 4.84 (m, 1H), 3.97 - 3.61 (m, 5H), 2.17 (m, 1H), 1.74 - 1.62 (overlapped with water, m, 2H), 1.44 (s, 9H), 1.07 (s, 10H), 0.69 - 0.53 (m, 1H), 0.52 - 0.33 (m, 2H), 0.30 - 0.16 (m, 1H), 0.15 - 0.02 (m, 1H). ESI-MS m/z calc. 497.2961, found 398.2 (M-99)+; Retention time: 2.29 minutes. LC method S. Step 4: tert-Butyl N-[(1R,2S)-2-cyclopropyl-4-hydroxy-1- (hydroxymethyl)butyl]carbamate [00608] To a solution of tert-butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2- cyclopropyl-4-hydroxy-butyl]carbamate (6.532 g, 12.349 mmol) in anhydrous THF (60 mL) was added TBAF in THF (40 mL of 1 M, 40.000 mmol) at 0 °C. The reaction was stirred in the ice- cold bath for 3 h. The reaction was quenched at 0 °C with a saturated aqueous sodium bicarbonate (50 mL) and EtOAc (50 mL). The biphasic solution was stirred for 1 h at RT and the layers were separated. The aqueous layer was extracted with EtOAc (2 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reverse phase chromatography C18 column eluting from 55-90% ACN in water. The fractions containing the desired product were combined and concentrated under reduced pressure to give tert-butyl N-[(1R,2S)-2-cyclopropyl- 4-hydroxy-1-(hydroxymethyl)butyl]carbamate (2.806 g, 83%) as a clear oil.1H NMR (400 MHz, DMSO-d6) δ 6.45 - 6.30 (m, 1H), 4.40 (t, J = 5.4 Hz, 1H), 4.31 (t, J = 5.0 Hz, 1H), 3.59 - 3.37 (m, 5H), 1.62 - 1.52 (m, 1H), 1.50 - 1.40 (m, 1H), 1.37 (s, 9H), 0.97 - 0.82 (m, 1H), 0.57 - 0.45 (m, 1H), 0.45 - 0.30 (m, 2H), 0.11 - 0.01 (m, 2H). Step 5: tert-Butyl (4R)-4-[(1S)-1-cyclopropyl-3-hydroxy-propyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00609] Tert-butyl N-[(1R,2S)-2-cyclopropyl-4-hydroxy-1-(hydroxymethyl)butyl]carbamate (726 mg, 2.5474 mmol) and 2,2-dimethoxypropane (297.50 mg, 350 μL, 2.8565 mmol) were dissolved in acetone (20 mL) at room temperature. To the solution was added para toluenesulfonic acid monohydrate (51 mg, 0.2681 mmol), and the orange solution was stirred at room temperature overnight. Then, 2,2-dimethoxypropane (297.50 mg, 350 μL, 2.8565 mmol) was added to the reaction mixture. The solution was stirred 5 h. The mixture was diluted with i- PrOAc (200 mL) and the reaction was quenched by addition of aqueous saturated sodium bicarbonate (200 mL). The mixture was stirred for 5 min at RT and layers were separated. The organic layer was washed with aqueous saturated sodium bicarbonate (100 mL) and brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude material. The crude was purified by reverse phase chromatography (gradient: 5 to 100 % MeCN in water, 16 CV). Fractions of desired product were combined, concentrated under reduced pressure, solubilized in DCM (20 mL), dried with anhydrous sodium sulfate, filtered and evaporated under reduced pressure to afford tert-butyl (4R)-4-[(1S)-1-cyclopropyl-3- hydroxy-propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (469 mg, 50%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 4.17 - 3.65 (m, 6H), 1.79 - 1.66 (m, 2H), 1.53 - 1.42 (m, 15H), 1.42 - 1.34 (m, 1H), 0.78 - 0.60 (m, 1H), 0.60 - 0.51 (m, 1H), 0.49 - 0.39 (m, 1H), 0.31 - 0.21 (m, 1H), 0.08 - -0.05 (m, 1H). ESI-MS m/z calc.299.2097, found 244.1 (M-55)+; Retention time: 2.04 minutes. LC method X. Step 6: tert-Butyl (4R)-4-[(1S)-1-cyclopropyl-3-oxo-propyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00610] A stirred solution of oxalyl chloride (2.9501 g, 2.0276 mL, 23.243 mmol) in anhydrous DCM (50 mL) was cooled down to - 78 °C and a solution of dimethylsulfoxide (3.6322 g, 3.2990 mL, 46.486 mmol) was added dropwise. The reaction was stirred for 30 min at - 78 °C and then a solution of tert-butyl (4R)-4-[(1S)-1-cyclopropyl-3-hydroxy-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate (2.355 g, 7.7476 mmol) in anhydrous DCM (50 mL) was added. The reaction was stirred for 30 minutes at - 78 °C and then triethylamine (7.8398 g, 10.799 mL, 77.476 mmol) was added. The reaction was stirred for 30 minutes then allowed to reach room temperature and stirred for 30 minutes at room temperature. The reaction was quenched by the addition of a saturated aqueous solution of ammonium chloride (20 mL). The mixture was vigorously stirred at room temperature for 30 minutes, then the layers were separated, and the aqueous layer was extracted with dichloromethane (2 x 20 mL). The combined organic layers were washed with brine (2 x 20 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert-butyl (4R)-4-[(1S)-1-cyclopropyl- 3-oxo-propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (2.63 g, 100%) as a yellow oil 1H NMR (400 MHz, CDCl3 ) δ 9.76 (t, J = 2.4 Hz, 1H), 4.22 - 3.77 (overlapped with impurity, m, 3H), 2.58 - 2.41 (overlapped with impurity, m, 1H), 1.86 (br. s, 1H), 1.74 - 1.60 (overlapped with water, m, 1H), 1.48 (s, 15H), 0.79 - 0.68 (m, 1H), 0.66 - 0.56 (m, 1H), 0.52 - 0.41 (m, 1H), 0.40 - 0.30 (m, 1H), 0.07 - -0.07 (m, 1H). The crude was directly used in the next step without purification and the yield will be calculated over two steps. Step 7: tert-Butyl (4R)-4-[(1S)-1-cyclopropyl-5-[methoxy(methyl)amino]-5-oxo- pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00611] To a suspension of sodium hydride (suspension in oil) (205 mg, 60 %w/w, 5.1255 mmol) in THF (15 mL) was added 2-diethoxyphosphoryl-N-methoxy-N-methyl-acetamide (1.27 g, 5.3092 mmol) at room temperature. The suspension was stirred at room temperature for 5 minutes and was then added to a solution of tert-butyl (4R)-4-[(1S)-1-cyclopropyl-3-oxo- propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.818 g, 5.8809 mmol) in THF (15 mL). The mixture was stirred at room temperature for 45 minutes. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (50 mL) and extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine (1 x 50 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude mixture was purified by reverse phase chromatography (gradient: 5 to 100 % MeCN in pure water, 16 CV). The fractions containing the desired product were combined and the organic solvent was evaporated. EtOAc (50 mL) was then added, and the mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (1 x 50 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to afford tert-butyl (4R)-4-[(1S)-1-cyclopropyl-5- [methoxy(methyl)amino]-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.6 g, 69%) as a pale-yellow oil.1H NMR (400 MHz, CDCl3 ) δ 7.11 - 6.87 (m, 1H), 6.54 - 6.32 (m, 1H), 4.13 - 3.85 (m, 3H), 3.70 (s, 3H), 3.24 (s, 3H), 2.54 - 2.35 (m, 1H), 2.34 -2.20 (m, 1H), 1.84 - 1.71 (m, 1H), 1.57 - 1.36 (m, 15H), 0.78 - 0.61 (m, 1H), 0.59 - 0.40 (m, 2H), 0.32 - 0.21 (m, 1H), 0.08 - -0.03 (m, 1H). ESI-MS m/z calc.382.2468, found 383.4 (M+1)+; Retention time: 1.91 minutes. LC method I. Step 8: tert-Butyl (4R)-4-[(1S)-1-cyclopropyl-5-[methoxy(methyl)amino]-5-oxo- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00612] Nitrogen was bubbled to a stirred solution of tert-butyl (4R)-4-[(1S)-1-cyclopropyl-5- [methoxy(methyl)amino]-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.712 g, 4.1894 mmol) in methanol (70 mL) for 10 minutes, then 10% palladium on carbon (50% wet) (940 mg, 5 %w/w, 0.4416 mmol) was added. Nitrogen was bubbled to the mixture for 10 minutes, followed by hydrogen for 10 minutes. The reaction was then stirred for 1 h at room temperature under hydrogen atmosphere. Nitrogen was bubbled to the mixture for 10 minutes. The mixture was filtered on a pad of Celite, washed with MeOH (50 mL) and concentrated under reduced pressure. The crude mixture was purified by reverse phase chromatography (gradient: 5 to 100 % MeCN in pure water, 16 CV). The fractions of desired product were concentrated under reduced pressure to remove MeCN and the aqueous residue was extracted with DCM (3 x 50 mL). The combined organic phases were dried with sodium sulfate, filtered and evaporated under reduced pressure to afford tert-butyl (4R)-4-[(1S)-1-cyclopropyl-5- [methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.245 g, 76%) as a pale yellow oil 1H NMR (400 MHz, CDCl3 ) δ 4.12 - 3.89 (m, 3H), 3.68 (s, 3H), 3.26 - 3.09 (m, 3H), 2.58 - 2.26 (m, 2H), 1.81 - 1.64 (m, 5H), 1.54 - 1.31 (m, 15H), 0.72 - 0.54 (m, 1H), 0.53 - 0.36 (m, 2H), 0.27 - 0.15 (m, 1H), 0.08 - -0.11 (m, 1H). ESI-MS m/z calc.384.2624, found 385.1 (M+1)+; Retention time: 2.03 minutes. LC method X. Step 9: tert-Butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-1-cyclopropyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00613] n-BuLi (in hexane) (0.16 mL of 1.6 M, 0.2560 mmol) was added to a solution of 3- bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine (66 mg, 0.2459 mmol) in anhydrous THF (1 mL) maintained at -78 °C and left stirring at this temperature for 15 min then a solution of tert-butyl (4R)-4-[(1S)-1-cyclopropyl-5-[methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (50 mg, 0.1235 mmol) in anhydrous THF (0.5 μL) was added at this temperature and then left stirring at -78 °C for 1 h. The reaction mixture was quenched with saturated aqueous ammonium chloride (5 mL) and left to warm-up to room temperature and stirred for 10 min. The mixture was extracted with MeTHF (3 x 10 mL) and the combined organics were dried over magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by reverse phase chromatography on a C18 column using a 5 to 100% gradient of acetonitrile in pure water to provide tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl- 5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-cyclopropyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (38 mg, 58%) as a clear oil 1H NMR (400 MHz, CDCl3) δ 9.29 - 9.00 (m, 1H), 6.98 - 6.76 (m, 1H), 4.28 - 3.91 (m, 6H), 3.35 - 3.18 (m, 2H), 1.97 - 1.21 (m, 29H), 0.70 - 0.42 (m, 3H), 0.30-0.18 (m, 1H), 0.11 - -0.04 (m, 1H). ESI-MS m/z calc.512.3363, found 513.3 (M+1)+; Retention time: 3.31 minutes. LC method 1C. Step 10: [(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- cyclopropyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00614] To a stirred solution of tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-1-cyclopropyl-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (138 mg, 0.2692 mmol) in 1,4-dioxane (2.5 mL) and was added water (250 µL), hydrochloric acid (in 1,4- dioxane) (2.5 mL of 4 M, 10.00 mmol) at ambient temperature, and the reaction was stirred at that temperature for 1 h. The volatiles were removed under reduced pressure to dryness and water (2 mL) was added to the residue and concentrated. The residual water was co-evaporated with acetonitrile (3 x 10 mL), followed by toluene (3 x 10 mL) then the product was dried under high vacuum to furnish crude [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- cyclopropyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (hydrochloride salt) (105 mg, 100%) ESI-MS m/z calc.354.24197, found 355.2(M+1)⁺; Retention time: 1.17 minutes. as a yellow gum. LC method A. Step 11: [(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- cyclopropyl-azepan-2-yl]methanol and [(2R,3S,7R)-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-3-cyclopropyl-azepan-2-yl]methanol [00615] To a stirred solution of [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-cyclopropyl-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (hydrochloride salt) (104 mg, 0.2660 mmol) in anhydrous dichloromethane (1.0 mL) were added acetic-acid (35 µL, 0.6155 mmol) and DIPEA (150 µL, 0.8611 mmol) at 0 oC under nitrogen. After stirring for 10 min, sodium triacetoxyborohydride (180 mg, 0.8493 mmol) was added. The reaction mixture was stirred at that temperature for 10 min and after quenching with methanol (1 mL) and water (0.5 mL), the mixture was concentrated under reduced pressure to produce crude material as a light-brown gum. It was purified by reverse-phase chromatography (10-99% acetonitrile in water containing 5 mM HCl, over 20 min run) to give [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-3-cyclopropyl-azepan-2-yl]methanol (hydrochloride salt) (70 mg, 67%) ESI-MS m/z calc. 356.2576, found 357.3(M+1)⁺; Retention time: 1.11 minutes. as a yellow gum. The diastereomers were further separated by preparative SFC [Preparative Method: Instrument- Teledyne Isco ACCQ Prep SFC, ChiralPak IC (21.2 x 250 mm, 5 um), 40 oC, isocratic, 36% MeOH + 20 mM NH3, flow 70 mL/min, conc.20 mg/mL (methanol, no modifier), inj. vol.: 0.5 ml, pressure 170 bar, wavelength 229 nm]. The individual peak materials were concentrated under reduced pressure and the residual water was removed azeotropically with acetonitrile (5 x 10 mL), followed by toluene (3 x 10 mL) to furnish pure diastereomers as white gums. SFC Peak-1 (major): [(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-cyclopropyl- azepan-2-yl]methanol (31 mg, 33%) 1H NMR (400 MHz, CDCl3) δ 8.31 (s, 1H), 6.45 (s, 1H), 4.15 (dd, J = 8.0, 5.4 Hz, 1H), 4.03 (dd, J = 10.3, 4.3 Hz, 1H), 4.00 (s, 3H), 3.18 (t, J = 10.2 Hz, 1H), 2.96 - 2.85 (m, 1H), 2.20 - 2.08 (m, 1H), 2.06 - 1.96 (m, 1H), 1.96 - 1.83 (m, 1H), 1.83 - 1.73 (m, 2H), 1.73 - 1.63 (m, 1H), 1.50 (s, 9H), 0.72 - 0.63 (m, 2H), 0.63 - 0.54 (m, 1H), 0.51 - 0.41 (m, 1H), 0.24 - 0.13 (m, 1H), 0.09 - 0.01 (m, 1H). ESI-MS m/z calc.356.2576, found 357.2(M+1)⁺; Retention time: 1.10 minutes.. SFC Peak-2 (minor): [(2R,3S,7R)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-cyclopropyl-azepan-2-yl]methanol (3 mg, 3%) 1H NMR (400 MHz, CDCl3) δ 8.33 (s, 1H), 6.45 (s, 1H), 4.08 (dd, J = 11.5, 3.5 Hz, 1H), 4.06 - 4.01 (m, 1H), 4.00 (s, 3H), 3.41 (t, J = 10.4 Hz, 1H), 3.09 - 2.99 (m, 1H), 2.33 (d, J = 13.9 Hz, 1H), 2.09 - 2.01 (m, 2H), 1.87 - 1.71 (m, 1H), 1.66 - 1.52 (m, 2H), 1.50 (s, 9H), 0.66 - 0.52 (m, 3H), 0.51 - 0.41 (m, 1H), 0.23 - 0.15 (m, 1H), 0.11 - 0.03 (m, 1H). ESI-MS m/z calc.356.2576, found 357.2(M+1)⁺; Retention time: 1.13 minutes. LC method A. Step 12: 3-[[4-[[(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-cyclopropyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00616] Through a stirred solution of [(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-cyclopropyl-azepan-2-yl]methanol (31 mg, 0.08695 mmol) and 3-[[4-chloro- 6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (38 mg, 0.09094 mmol) in anhydrous tetrahydrofuran (750 µL), nitrogen was sparged for 5 min. Then sodium tert-butoxide (60 mg, 0.6243 mmol) was added in a single portion at ambient temperature and after stirring for 10 min, the reaction was allowed to stir at 42 °C for 2 h. Then the heating source was removed and the reaction was allowed to cool down to ambient temperature. The reaction mixture was diluted with ethyl acetate (20 mL) and acidified with hydrochloric acid (1.5 mL of 1.0 M, 1.500 mmol). The organic phase was separated, and the aqueous layer was extracted with ethyl acetate (2 x 15 mL). The combined organics were washed with brine (10 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. Any remaining trace of water was removed azeotropically with acetonitrile (2 x 10 mL) and toluene (2 x 10 mL) to furnish 3-[[4- [[(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-cyclopropyl-azepan-2- yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (40 mg, 59%), ESI-MS m/z calc.737.33594, found 738.2 (M+1)⁺; Retention time: 1.46 minutes. as a white solid. LC method A. Step 13: (4S,8S,9R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8- cyclopropyl-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one (Compound 110) [00617] To a stirred solution of 3-[[4-[[(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-cyclopropyl-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (40 mg, 0.05165 mmol) in anhydrous NMP (800 µL) and acetone (1.6 mL) was added p-TsCl (35 mg, 0.1836 mmol) at ambient temperature under nitrogen and the reaction was stirred for 5 min. Then N-methylmorpholine (60 µL, 0.5457 mmol) was added and the reaction was stirred at 55 oC for 90 min. The reaction was allowed to cool to ambient temperature and the volatiles were removed under reduced pressure. The remaining brownish solution was subjected to preparative HPLC (30-99% acetonitrile in water, containing 5 mM HCl as modifier, 15 min run). The fractions were combined and evaporated, and repurified by HPLC (40-99% acetonitrile in water, 30 min run, HCl modifier) to furnish product as a yellow solid. (4S,8S,9R)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8- cyclopropyl-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (16 mg, 42%) 1H NMR (400 MHz, CDCl3) δ 8.77 (s, 1H), 8.48 (s, 1H), 7.81 - 7.74 (m, 2H), 7.56 (t, J = 7.7 Hz, 1H), 7.25 - 7.21 (m, 1H), 7.18 - 7.14 (m, 1H), 7.09 (d, J = 7.6 Hz, 2H), 6.79 (s, 1H), 6.22 (s, 1H), 5.99 - 5.85 (m, 2H), 4.87 (t, J = 11.8 Hz, 1H), 4.65 - 4.57 (m, 1H), 4.23 (s, 3H), 2.35 - 2.26 (m, 1H), 2.23 - 2.08 (m, 2H), 2.06 (s, 6H), 2.04 - 1.95 (m, 2H), 1.94 - 1.86 (m, 2H), 1.53 (s, 9H), 0.89 - 0.87 (m, 1H), 0.56 - 0.51 (m, 1H), 0.28 - 0.23 (m, 1H), 0.09 - -0.02 (m, 1H), -0.67 - -0.73 (m, 1H). ESI-MS m/z calc.719.3254, found 720.2(M+1)⁺; Retention time: 1.89 minutes. LC method A. Example 74: Preparation of Compound 112 Step 1: tert-Butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- (cyclobutylmethyl)-5-oxo-pyrrolidine-1-carboxylate [00618] To a -40 °C solution of bromo(cyclobutylmethyl)magnesium (solution in THF) (58 mL of 0.5 M, 29.000 mmol) in tetrahydrofuran (150 mL) was added copper iodide (652 mg, 3.4235 mmol) and the resulting suspension was stirred 25 min at that temperature. To the suspension was added a solution of tert-butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5- oxo-2H-pyrrole-1-carboxylate (10 g, 22.142 mmol) and chlorotrimethylsilane (2.7392 g, 3.2 mL, 25.213 mmol) in tetrahydrofuran (50 mL) dropwise over 50 min. The mixture was stirred for 30 min. An aqueous solution of ammonium chloride (sat., 25 mL) was added followed by EtOAc (50 mL) and water (50 mL). The phases were separated, and the aqueous one was extracted with EtOAc (2 x 25 mL). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 100% of MeCN in water to afford, after evaporation, crude tert-butyl (2R,3R)-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-3-(cyclobutylmethyl)-5-oxo-pyrrolidine-1-carboxylate (12.48 g, 98%) as a yellow oil with a 98.1% ee.1H NMR (400 MHz, CDCl3) δ 7.67 - 7.57 (m, 4H), 7.48 - 7.35 (m, 6H), 3.87 - 3.80 (m, 2H), 3.70 - 3.64 (m, 1H), 2.86 (dd, J = 17.7, 8.9 Hz, 1H), 2.38 - 2.21 (m, 2H), 2.13 (dd, J = 17.6, 2.0 Hz, 1H), 2.10 - 1.99 (overlaps with EtOAc, m, 2H), 1.95 - 1.75 (m, 2H), 1.70 - 1.59 (m, 2H), 1.58 - 1.54 (overlaps with water, m, 1H), 1.54 - 1.47 (m, 1H), 1.44 (s, 9H), 1.05 (s, 9H). ESI-MS m/z calc.521.2961, found 422.2 (M-99)+; Retention time: 2.57 minutes. LC method S. Step 2: tert-Butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2- (cyclobutylmethyl)-4-hydroxy-butyl]carbamate [00619] To a solution of tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- (cyclobutylmethyl)-5-oxo-pyrrolidine-1-carboxylate (6.25 g, 10.865 mmol) in ethanol (90 mL) at 50 °C was added sodium borohydride (4.12 g, 108.90 mmol) portion-wise over 5 minutes. The resulting mixture was stirred at this temperature for 1 hour then water (50 mL) was added, and the volatiles were removed under reduced pressure. The residual aqueous solution was diluted with EtOAc (50 mL) and the phases were separated. The aqueous solution was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert- butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-(cyclobutylmethyl)-4-hydroxy- butyl]carbamate (5.25 g, 82%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 7.69 - 7.63 (m, 4H), 7.48 - 7.36 (m, 6H), 3.75 - 3.56 (m, 4H), 2.40 - 2.27 (m, 1H), 2.05 - 1.96 (m, 2H), 1.90 - 1.73 (m, 3H), 1.71 - 1.64 (m, 4H), 1.61 - 1.48 (m, 3H), 1.44 (s, 9H), 1.35 - 1.22 (overlapped with EtOAc, m, 2H), 1.07 (s, 9H). ESI-MS m/z calc.525.3274, found 426.2 (M-99)+; Retention time: 2.41 minutes. LC method I. Step 3: tert-Butyl N-[(1R,2R)-2-(cyclobutylmethyl)-4-hydroxy-1- (hydroxymethyl)butyl]carbamate [00620] To a 0 °C solution of tert-butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]- 2-(cyclobutylmethyl)-4-hydroxy-butyl]carbamate (1.9 g, 3.1402 mmol) in tetrahydrofuran (15 mL) was added tetrabutylammonium fluoride (solution in THF) (6 mL of 1 M, 6.0000 mmol) dropwise over 1 min. The solution was stirred at that temperature for 3 h. An aqueous solution of potassium bicarbonate (5%, 40 mL) was added, and the mixture was stirred for 1 h. The suspension was then filtered on a pad of sand which was washed with EtOAc (3 x 20 mL) and water (2 x 20 mL). The phases were separated, and the aqueous one was extracted with EtOAc (2 x 30 mL). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 70% of MeCN in water to afford, after evaporation, tert-butyl N-[(1R,2R)-2-(cyclobutylmethyl)-4- hydroxy-1-(hydroxymethyl)butyl]carbamate (917.6 mg, 94%) as a colorless oil.1H NMR (400 MHz, DMSO-d6) δ 6.47 - 6.35 (m, 1H), 4.43 (t, J = 5.4 Hz, 1H), 4.37 (t, J = 5.0 Hz, 1H), 3.48 - 3.33 (overlaps with water, m, 4H), 3.31 - 3.26 (overlaps with water, m, 2H), 2.41 - 2.31 (overlaps with spinning sidebands of DMSO, m, 1H), 2.02 - 1.89 (m, 2H), 1.85 - 1.66 (m, 2H), 1.57 - 1.40 (m, 4H), 1.37 (s, 9H), 1.27 - 1.15 (m, 2H). ESI-MS m/z calc.287.2097, found 188.2 (M-99)+; Retention time: 1.61 minutes. LC method I. Step 4: tert-Butyl (4R)-4-[(1R)-1-(cyclobutylmethyl)-3-hydroxy-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00621] To a solution of tert-butyl N-[(1R,2R)-2-(cyclobutylmethyl)-4-hydroxy-1- (hydroxymethyl)butyl]carbamate (1.9 g, 0.3107 mmol) in acetone (1 mL) at room temperature were added 2,2-dimethoxypropane (170.00 mg, 0.2 mL, 1.6323 mmol) and p-toluenesulfonic acid monohydrate (8 mg, 0.0421 mmol). The reaction was stirred for 18 h. An aqueous solution of sodium bicarbonate (sat., 2 mL) was added and the mixture was stirred for 1 h. iPrOAc (20 mL) was added and removed under reduced pressure 3 times. iPrOAc (20 mL) and water (20 mL) were added, and the phase were separated. The aqueous phase was extracted with iPrOAc (2 x 20 mL). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 85% of MeCN in water to afford, after evaporation, tert-butyl (4R)-4-[(1R)-1-(cyclobutylmethyl)-3-hydroxy-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate (56.6 mg, 56%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 4.12 - 4.01 (m, 1H), 3.96 - 3.62 (m, 4H), 2.81 - 2.71 (m, 1H), 2.39 - 2.25 (m, 1H), 2.13 - 1.97 (m, 2H), 1.96 - 1.74 (m, 3H), 1.72 - 1.57 (m, 6H), 1.49 (s, 9H), 1.47 - 1.44 (m, 3H), 1.40 - 1.18 (m, 3H). ESI-MS m/z calc.327.241, found 350.2 (M+23)+; Retention time: 1.97 minutes. LC method I. Step 5: tert-Butyl (4R)-4-[(1R)-1-(cyclobutylmethyl)-3-oxo-propyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00622] To a solution of tert-butyl (4R)-4-[(1R)-1-(cyclobutylmethyl)-3-hydroxy-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate (1.02 g, 2.8096 mmol) in dichloromethane (28 mL) cooled at 0 °C were added water (10 mL), sodium bicarbonate (382 mg, 4.5473 mmol), sodium bromide (61 mg, 0.5928 mmol) and TEMPO (79 mg, 0.5056 mmol). To the bi-phasic solution was added a solution of sodium hypochlorite (in water) (2.42 mL of 10 %w/v, 3.2508 mmol) slowly dropwise over 5 minutes under vigorous stirring. After being stirred for 1 hour at 0 °C, sodium hypochlorite (in water) (0.6 mL of 10 %w/v, 0.8060 mmol) was added again slowly. After being stirred for another 30 minutes at 0 °C, a saturated aqueous solution of sodium bicarbonate (30 mL) was added. The mixture was diluted with DCM (30 mL) and the phases were separated. The aqueous phase was extracted with DCM (2 x 20 mL) and the combined organic layers were washed with a 10% aqueous solution of Na2S2O3 (30 mL), a saturated aqueous solution of sodium bicarbonate (2 x 30 mL) and brine (30 mL). The organic solution was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert- butyl (4R)-4-[(1R)-1-(cyclobutylmethyl)-3-oxo-propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (962 mg, 100%) as an orange oil.1H NMR (400 MHz, CDCl3) δ 9.74 (s, 1H), 3.96 - 3.89 (m, 1.5H), 3.81 (d, J = 7.8 Hz, 1.5H), 2.46 - 2.25 (m, 4H), 2.12 - 1.97 (m, 2.5H), 1.92 - 1.75 (m, 2.5H), 1.68 - 1.59 (overlapped with water, m, 6H), 1.49 (s, 9H), 1.48 (br. s., 3H). ESI-MS m/z calc.325.2253, found 348.2 (M+23)+; Retention time: 2.05 minutes. LC method I. Step 6: tert-Butyl (4R)-4-[(1R)-1-(cyclobutylmethyl)-5-[methoxy(methyl)amino]- 5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00623] To a solution of sodium hydride (in mineral oil) (132 mg, 60 %w/w, 3.3003 mmol) in anhydrous THF (12.5 mL) was added dropwise 2-diethoxyphosphoryl-N-methoxy-N-methyl- acetamide (788.80 mg, 0.68 mL, 3.2976 mmol) while stirring the suspension at room temperature. Once the hydrogen gas evolution stopped, this solution was added dropwise to a solution of tert-butyl (4R)-4-[(1R)-1-(cyclobutylmethyl)-3-oxo-propyl]-2,2-dimethyl- oxazolidine-3-carboxylate (962 mg, 2.8082 mmol) in anhydrous THF (12.5 mL) stirred at room temperature. The reaction mixture was stirred at room temperature for 1 hour and was quenched with a saturated aqueous solution of sodium bicarbonate (30 mL). This mixture was extracted with EtOAc (3 x 30 mL) and the combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert-butyl (4R)-4-[(1R)-1-(cyclobutylmethyl)-5-[methoxy(methyl)amino]-5-oxo-pent-3- enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.24 g, 100%) as an orange oil.1H NMR (400 MHz, CDCl3) δ 7.02 - 6.87 (m, 1H), 6.54 - 6.34 (m, 1H), 4.26 - 4.14 (m, 1H), 4.00 - 3.79 (m, 3H), 3.71 (s, 3H), 3.28 - 3.22 (m, 3H), 2.41 - 2.27 (m, 2H), 2.22 - 2.12 (m, 1H), 2.10 - 1.97 (overlapped with EtOAc, m, 4H), 1.91 - 1.75 (m, 2H), 1.70 - 1.58 (m, 5H), 1.48 (s, 9H), 1.46 (s, 3H). ESI-MS m/z calc.410.2781, found 411.4 (M+1)+; Retention time: 2.07 minutes. Isolated as a mixture of E/Z stereoisomers, unknown ratio. LC method I. Step 7: tert-Butyl (4R)-4-[(1S)-1-(cyclobutylmethyl)-5-[methoxy(methyl)amino]- 5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00624] A solution of tert-butyl (4R)-4-[(1R)-1-(cyclobutylmethyl)-5- [methoxy(methyl)amino]-5-oxo-pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (2.91 g, 6.1879 mmol) in methanol (80 mL) was bubbled with nitrogen for 10 minutes then palladium on carbon (1.25 g, 5 %w/w, 0.5873 mmol) was added. The suspension was sparged with hydrogen for 15 minutes then stirred under hydrogen atmosphere for 1.5 hours. The suspension was bubbled with nitrogen for 10 minutes, then was filtered through a short pad of Celite. The pad was rinsed with EtOAc (500 mL) and the combined organic filtrates were concentrated under reduced pressure. The residue was purified by reverse phase chromatography (Column: C18. Gradient: 5-100% MeCN in water) to afford tert-butyl (4R)-4-[(1S)-1-(cyclobutylmethyl)-5- [methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (2.129 g, 82%) as a pale-yellow oil.1H NMR (400 MHz, CDCl3) δ 4.03 - 3.87 (m, 2H), 3.85 - 3.77 (m, 1H), 3.69 (s, 3H), 3.18 (s, 3H), 2.50 - 2.36 (m, 2H), 2.35 - 2.25 (m, 1H), 2.10 - 1.95 (overlapped with MeCN, m, 2H), 1.86 - 1.76 (m, 2H), 1.73 - 1.52 (m, 9H), 1.50 - 1.43 (m, 12H), 1.42 - 1.33 (m, 1H), 1.29 - 1.18 (m, 1H), 1.16 - 1.01 (m, 1H). ESI-MS m/z calc.412.2937, found 413.4 (M+1)+; Retention time: 3.99 minutes. LC method 1B. Step 8: tert-Butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-1-(cyclobutylmethyl)-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00625] A solution of n-butyllithium (400 µL of 2.5 M, 1.000 mmol) in hexanes was added dropwise to a solution of 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine (273 mg, 1.018 mmol) in THF (5.4 mL) at -78 °C. After 5 minutes of stirring, a solution of tert-butyl (4R)-4- [(1S)-1-(cyclobutylmethyl)-5-[methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine- 3-carboxylate (300 mg, 0.7271 mmol) in THF (2.1 mL) as added. The reaction mixture was stirred at -78 °C for another 15 minutes. It was diluted with EtOAc (50 mL) and washed with saturated aqueous ammonium chloride solution (1× 50 mL) and brine (1× 50 mL). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. Purification by flash chromatography on silica gel (0-50% EtOAc/hexanes over 30 minutes) provided tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1- (cyclobutylmethyl)-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (103 mg, 26%) as a slightly yellow oil. ESI-MS m/z calc.540.36755, found 541.5 (M+1)⁺; Retention time: 2.57 minutes. LC method A. Step 9: [(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclobutylmethyl)-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00626] HCl in 1,4-dioxane (1 mL of 4 M, 4.000 mmol) was added to a solution of tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-1-(cyclobutylmethyl)-5-oxo- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (103 mg, 0.1905 mmol) in 1,4-dioxane (2 mL). One drop of water was added. The reaction mixture was stirred overnight at room temperature. Volatiles were removed under reduced pressure, and the crude product was azeotroped with toluene (4× 6 mL) to provide [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclobutylmethyl)-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (73 mg, 100%) as a golden yellow oil. ESI-MS m/z calc.382.27325, found 383.3(M+1)⁺; Retention time: 1.36 minutes. LC method A. Step 10: [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclobutylmethyl)azepan-2-yl]methanol, diastereomer 1, and [(2R,3S)-7-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(cyclobutylmethyl)azepan-2- yl]methanol, diastereomer 2 diastereomer 1 diastereomer 2 [00627] In a round bottom flask, sodium triacetoxyborohydride (202 mg, 0.9531 mmol) was added to [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(cyclobutylmethyl)- 3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (73 mg, 0.1908 mmol) in DCM (1.1 mL). The reaction mixture was stirred at room temperature for 1 hour. It was diluted with EtOAc and washed with water (1× 50 mL). The aqueous layer was acidified to pH 4 with the addition of 1 M HCl. It was extracted with EtOAc (1× 50 mL). All organic layers were combined, dried over sodium sulfate, filtered and concentrated under reduced pressure. Purification by reverse phase HPLC (10-99% acetonitrile/5 mM aqueous HCl over 15 minutes) provided [(2R,3S)-7-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(cyclobutylmethyl)azepan-2-yl]methanol (hydrochloride salt) (diastereomeric mixture, 66 mg, 82%) as a yellow foaming solid. The diastereomers were separated by chiral SFC using a ChiralPak IC column (21.2 x 250 mm, 5 µm) at 40 °C. Mobile phase was 32% MeOH (20 mM NH3), 68% CO2 at a 70 mL/min flow. This provided [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclobutylmethyl)azepan-2-yl]methanol (19.0 mg, 52%) as Peak 1, diastereomer 1, as a clear colorless oil. ESI-MS m/z calc.384.2889, found 385.3 (M+1)⁺; Retention time: 1.34 minutes. This also provided [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclobutylmethyl)azepan-2-yl]methanol (19.3 mg, 53%) as Peak 2, diastereomer 2, as a clear colorless oil. ESI-MS m/z calc.384.2889, found 385.3(M+1)⁺; Retention time: 1.34 minutes. LC method A. Step 11: 3-[[4-[[(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclobutylmethyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid, diastereomer 1
Diastereomer 1 [00628] In a 4 mL vial, solid sodium tert-butoxide (29 mg, 0.3018 mmol) was added to a mixture of 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (23 mg, 0.05504 mmol) and [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclobutylmethyl)azepan-2-yl]methanol, diastereomer 1 (19 mg, 0.04941 mmol) in THF (0.2 mL). The reaction mixture was stirred at 40 °C overnight. It was diluted with EtOAc (50 mL) and washed with aqueous HCl (1 M, 1× 50 mL). The aqueous layer was extracted with DCM (1× 50 mL). Organic layers were combined, dried over sodium sulfate, filtered and concentrated under reduced pressure. Purification by reverse phase HPLC (30-99% acetonitrile/5 mM aqueous HCl over 15 minutes) provided 3-[[4-[[(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-(cyclobutylmethyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (38 mg, 96%) as a clear yellow oil. ESI-MS m/z calc.765.36725, found 766.4(M+1)⁺; Retention time: 1.61 minutes. LC method A. Step 12: (8S,9R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8- (cyclobutylmethyl)-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one, diastereomer 1 (Compound 112)
Diastereomer 1 Diastereomer 1 [00629] In a round bottom flask, 4-methylbenzenesulfonyl chloride (23 mg, 0.1206 mmol) was added to a solution of 3-[[4-[[(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-(cyclobutylmethyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt), diastereomer 1 (38 mg, 0.04736 mmol) in acetone (2 mL) and NMP (1 mL).4-Methylmorpholine (31 µL, 0.2820 mmol) was added last. The reaction mixture was stirred at 55 °C for 2.5 hours. It was diluted with EtOAc (50 mL) and washed with aqueous HCl (1 M, 1× 50 mL). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. Purification by reverse phase HPLC (30-99% acetonitrile/5 mM aqueous HCl over 15 minutes) provided (8S,9R)-4-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-8-(cyclobutylmethyl)-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa- 18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one, diastereomer 1 (13.8 mg, 38%) as a yellow solid. 1H NMR (400 MHz, CDCl3) δ 8.85 (s, 1H), 8.48 (s, 1H), 7.84 (s, 1H), 7.82 (s, 1H), 7.61 (t, J = 7.6 Hz, 1H), 7.25 (d, J = 7.6 Hz, 1H), 7.12 (s, 1H), 7.10 (s, 1H), 6.58 (s, 1H), 6.29 (s, 1H), 5.90 (d, J = 6.4 Hz, 2H), 4.89 (t, J = 11.7 Hz, 1H), 4.49 - 4.41 (m, 1H), 4.18 (s, 3H), 2.38 - 2.28 (m, 1H), 2.10 (s, 6H), 1.99 - 1.92 (m, 2H), 1.89 - 1.81 (m, 4H), 1.74 - 1.65 (m, 4H), 1.53 (s, 9H), 1.49 - 1.35 (m, 4H), 1.15 - 1.07 (m, 1H). ESI-MS m/z calc.747.3567, found 748.5(M+1)⁺; Retention time: 2.10 minutes. LC method A. Example 75: Preparation of Compound 113 Step 1: 3-[[4-[[(2R,3S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclobutylmethyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid, diastereomer 2
Diastereomer 2 [00630] In a 4 mL vial, solid sodium tert-butoxide (29 mg, 0.3018 mmol) was added to a mixture of 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (23 mg, 0.05504 mmol) and [(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (cyclobutylmethyl)azepan-2-yl]methanol, diastereomer 2 (19.3 mg, 0.05019 mmol) in THF (0.2 mL). The reaction mixture was stirred at 40 °C overnight. It was diluted with EtOAc (50 mL) and washed with aqueous HCl (1 M, 1× 50 mL). The aqueous layer was extracted with DCM (1× 50 mL). Organic layers were combined, dried over sodium sulfate, filtered and concentrated under reduced pressure. Purification by reverse phase HPLC (30-99% acetonitrile/5 mM aqueous HCl over 15 minutes) provided 3-[[4-[[(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-(cyclobutylmethyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt), diastereomer 2 (38 mg, 94%) as a clear yellow oil. ESI-MS m/z calc.765.36725, found 766.4(M+1)⁺; Retention time: 1.58 minutes. LC method A. Step 2: (8S,9R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8- (cyclobutylmethyl)-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one, diastereomer 2
Diastereomer 2 Diastereomer 2 [00631] In a round bottom flask, 4-methylbenzenesulfonyl chloride (23 mg, 0.1206 mmol) was added to a solution of 3-[[4-[[(2R,3S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-(cyclobutylmethyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt), diastereomer 2 (38 mg, 0.04736 mmol) in acetone (2 mL) and NMP (1 mL).4-Methylmorpholine (31 µL, 0.2820 mmol) was added last. The reaction mixture was stirred at 55 °C for 2.5 hours. It was diluted with EtOAc (50 mL) and washed with aqueous HCl (1 M, 1× 50 mL). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. Purification by reverse phase HPLC (30-99% acetonitrile/5 mM aqueous HCl over 15 minutes) provided (8S,9R)-4-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-8-(cyclobutylmethyl)-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa- 18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one, diastereomer 2 (9.8 mg, 26%) as a yellow solid. ESI-MS m/z calc.747.3567, found 748.5(M+1)⁺; Retention time: 2.01 minutes. LC method A. Example 76: Preparation of Compound 115 Step 1: tert-Butyl (4R)-4-[(1S)-1-cyclobutyl-5-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00632] To a stirred yellow solution of 3-bromo-5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine (715 mg, 2.234 mmol) in anhydrous tetrahydrofuran (10 mL) was added dropwise a solution of n-BuLi (in hexanes) (800 µL of 2.5 M, 2.000 mmol) over 2 min at -78 °C under nitrogen. The resulting red solution was stirred for 6 min at that temperature. Then a solution of tert-butyl (4R)-4-[(1S)-1-cyclobutyl-5- [methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (600 mg, 1.481 mmol) in anhydrous tetrahydrofuran (4.0 mL) was added slowly. After 15 min at that temperature, saturated aqueous ammonium chloride (10 mL) was added, followed by addition of EtOAc (25 mL). The cooling bath was removed, and the mixture was allowed to warm to about 0 oC over 15 min and the phases were separated. The aqueous phase was extracted with EtOAc ( 3 x 15 mL) and the combined organics were concentrated under reduced pressure and purified by reversed-phase chromatography (30-99 % MeCN in water over 20 min, no modifier). The volatiles were removed under reduced pressure to furnish tert-butyl (4R)-4-[(1S)-1-cyclobutyl-5- [5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate (505 mg, 59%) ESI-MS m/z calc.578.308, found 579.2(M+1)⁺; Retention time: 2.03 minutes. as an orange film. LC method C. Step 2: [(2R,3S)-3-Cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H- azepin-2-yl]methanol [00633] To a stirred solution of tert-butyl (4R)-4-[(1S)-1-cyclobutyl-5-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (505 mg, 0.8727 mmol) in 1,4-dioxane (9.0 mL) was added water (900 µL), hydrochloric acid (in 1,4-dioxane) (8.0 mL of 4 M, 32.00 mmol) at ambient temperature, and the reaction was stirred at that temperature for 90 min. The volatiles were removed, water (4 mL) was added to the residue and removed under reduced pressure. The residual water was co-evaporated azeotropically with acetonitrile (3 x 10 mL), followed by toluene (3 x 10 mL) then dried under high vacuum to furnish crude [(2R,3S)-3-cyclobutyl-7-[5- methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H- azepin-2-yl]methanol (hydrochloride salt) (399 mg, 100%) ESI-MS m/z calc.420.21368, found 421.2(M+1)⁺; Retention time: 1.29 minutes. as a yellow gum. LC method A. Step 3: [(2R,3S,7S)-3-Cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol, and [(2R,3S,7R)-3-cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol [00634] To a stirred solution of [(2R,3S)-3-cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2- yl]methanol (hydrochloride salt) (399 mg, 0.8732 mmol) in anhydrous dichloromethane (4.0 mL) were added acetic-acid (100 µL, 1.758 mmol) and DIPEA (500 µL, 2.870 mmol) at ambient temperature nitrogen. After stirring for 15 min, sodium triacetoxyborohydride (600 mg, 2.831 mmol) was added at once. The reaction mixture was stirred at that temperature for 30 min, then quenched with methanol (5 mL) and water (2 mL), After concentrating under reduced pressure, the crude was purified by reversed-phase chromatography (10-99% acetonitrile in water containing 5 mM HCl, over 20 min run). The minor component was discarded and the major component (as a mixture of diastereomers) was concentrated under reduced pressure, to furnish [(2R,3S)-3-cyclobutyl-7-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazin-3-yl]azepan-2-yl]methanol (hydrochloride salt) (375 mg, 94%) ESI-MS m/z calc. 422.22934, found 423.2 (M+1)⁺; Retention time: 1.23 minutes. as an orange viscous oil. It was further purified by SFC [Preparative Method: Phenomenex LUX-4 (21.2 x 250 mm, 5 µm), 40 oC, gradient (28% to 34%, then to 28%), mobile phase 28% MeOH + 20 mM NH3, flow 70 mL/min, conc.16.2 mg/mL (methanol, with a drop of 7 N NH3 solution), inj. vol.: 0.5 ml, pressure variable, wavelength 210 nm] to separate the diastereomers (two fractions). The major fraction (Peak-1) [(2R,3S,7S)-3-cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (138.0 mg, 37%), 1H NMR (400 MHz, CDCl3) δ 8.39 (s, 1H), 6.73 (s, 1H), 4.05 (dd, J = 9.1, 4.6 Hz, 1H), 3.92 (s, 3H), 3.64 (dd, J = 10.4, 4.4 Hz, 1H), 3.15 (t, J = 10.3 Hz, 1H), 2.76 - 2.65 (m, 1H), 2.39 - 2.28 (m, 1H), 2.18 - 2.09 (m, 1H), 2.09 - 2.01 (m, 1H), 2.01 - 1.92 (m, 1H), 1.88 - 1.77 (m, 3H), 1.76 - 1.62 (m, 5H), 1.61 - 1.56 (m, 2H), 1.56 - 1.47 (m, 1H), 1.47 - 1.37 (m, 1H), 1.28 - 1.18 (m, 2H). ESI-MS m/z calc.422.22934, found 423.2 (M+1)⁺; Retention time: 1.25 minutes. was obtained as a pale-yellow viscous oil. The minor fraction (peak-2) [(2R,3S,7R)-3- cyclobutyl-7-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2- yl]methanol (42 mg, 11%) ESI-MS m/z calc.422.22934, found 423.2 (M+1)⁺; Retention time: 1.23 minutes. was obtained as a pale-yellow viscous oil. LC method A. Step 4: 3-[[4-[[(2R,3S,7S)-3-Cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]- 6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid [00635] Through a stirred solution of [(2R,3S,7S)-3-cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (138 mg, 0.3266 mmol) and 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (148 mg, 0.3542 mmol) in anhydrous tetrahydrofuran (3.0 mL), nitrogen was sparged for 5 min. Then sodium tert-butoxide (200 mg, 2.081 mmol) was added in a single portion at ambient temperature. After stirring for 10 min, the reaction was allowed to stir at 42 °C for 16 h. Then the heating source was removed, and the reaction cooled down to ambient temperature. The reaction mixture was diluted with ethyl acetate (20 mL) and acidified with hydrochloric acid (3.5 mL of 1.0 M, 3.500 mmol). The organic phase was separated, and the aqueous layer was extracted with ethyl acetate (2 x 15 mL). The combined organics were washed with brine (10 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. Traces of remaining water were removed azeotropically with acetonitrile (2 x 10 mL) and toluene (2 x 10 mL) to furnish 3-[[4-[[(2R,3S,7S)-3-cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (272 mg, 99%) ESI- MS m/z calc.803.3077, found 804.1(M+1)⁺; Retention time: 1.54 minutes. as a yellow solid. LC method A. Step 5: (4S,8S,9R)-8-Cyclobutyl-14-(2,6-dimethylphenyl)-4-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa- 18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (Compound 115) [00636] To a stirred solution of 3-[[4-[[(2R,3S,7S)-3-cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (272 mg, 0.3237 mmol) in anhydrous NMP (5.5 mL) and acetone (11 mL), were added p-TsCl (220 mg, 1.154 mmol) and N-methylmorpholine (360 µL, 3.274 mmol), in that order, at ambient temperature under nitrogen. The reaction was allowed to stir at 35 oC for 13 h. Then the reaction was allowed to cool to ambient temperature and the volatiles were removed under reduced pressure. The remaining brown solution was purified by preparative HPLC (30-99% acetonitrile in water, containing 5 mM HCl as modifier, 15 min run) to furnish (4S,8S,9R)-8-cyclobutyl-14-(2,6- dimethylphenyl)-4-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]- 18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (76 mg, 29%) as a pale-yellow solid.1H NMR (400 MHz, CDCl3) δ 8.90 (s, 1H), 8.53 (s, 1H), 7.87 - 7.77 (m, 2H), 7.52 (t, J = 7.7 Hz, 1H), 7.25 - 7.21 (m, 1H), 7.10 (d, J = 7.6 Hz, 2H), 6.81 (s, 1H), 6.30 (s, 1H), 6.06 (dd, J = 11.6, 5.0 Hz, 1H), 5.83 (d, J = 12.4 Hz, 1H), 4.90 (t, J = 11.7 Hz, 1H), 4.44 - 4.40 (m, 1H), 4.07 (s, 3H), 2.45 - 2.37 (m, 1H), 2.34 - 2.24 (m, 1H), 2.10 (s, 6H), 1.99 - 1.88 (m, 2H), 1.82 - 1.72 (m, 5H), 1.63 - 1.55 (m, 5H), 1.33 - 1.14 (m, 3H), 0.94 - 0.85 (m, 1H). ESI-MS m/z calc.785.2971, found 786.2(M+1)⁺; Retention time: 2.17 minutes. LC method A. Example 77: Preparation of Compound 121 Step 1: Triisopropyl(3-methylbut-3-en-1-ynyl)silane [00637] In a pressure tube, was added CuI (211 mg, 1.1079 mmol) and bis(triphenylphosphine)palladium(II) dichloride (390 mg, 0.5540 mmol) the mixture was placed under nitrogen atmosphere then THF (100 mL) and TEA (12.6 g, 124.52 mmol) were added followed by ethynyl(triisopropyl)silane (10.1 g, 55.379 mmol) and cold 2-bromoprop-1-ene (8.9 g, 73.568 mmol). The tube was closed then heated for 2 h at 50 °C in an oil bath. The reaction mixture was cooled to room temperature, partitioned between pentane (200 mL) and saturated aqueous ammonium chloride (300 mL) containing HCl (72 mL of 1 M, 72.000 mmol) and water (300 mL). The aqueous phase was separated and extracted with pentane (2 x 300 mL). The combined organics were dried on magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified on silica gel by elution in 100% pentane to provide triisopropyl(3-methylbut-3-en-1-ynyl)silane (12.85 g, 82%) as a clear oil 1H NMR (400 MHz, CDCl3) δ 5.36 - 5.33 (m, 1H), 5.24 (quin, J = 1.7 Hz, 1H), 1.94 - 1.89 (m, 3H), 1.10 - 1.07 (m, 21H). GCMS: EI-MS m/z calc.222.1804, found 222.1; Retention time: 8.32 minutes. [00638] GCMS method: Column HP 5 ms Ultra Inert 30 m x 250 µm x 0.25 µm. Control mode: Flow: 3.3 mL/min. Split ratio mode: 33:1. Split Flow: 165 mL/min. Carrier gas: He. Injector Temperature: 250 °C. FID Temperature: 300 °C, MSD transfer line 250 °C. Solvent delay: 3 min. Oven temperature: isotherm at 70 °C for 1 min, then linear heating at 10 °C/min until 140 °C, then 20 °C/min until 250 °C then isotherm 250 °C for 4 min. Run time 17.5 min. Step 2: 2-(2,2-Difluoro-1-methyl-cyclopropyl)ethynyl-triisopropyl-silane [00639] In a sealed tube, trimethyl(trifluoromethyl)silane (19.5 g, 137.14 mmol) was added to a suspension of triisopropyl(3-methylbut-3-en-1-ynyl)silane (12.85 g, 45.637 mmol) and sodium iodide (3.7 g, 24.684 mmol) in THF (125 mL). The tube was capped and heated at 70 °C for 3 h. Once cooled to room temperature, the reaction mixture was partitioned between saturated aqueous sodium bicarbonate (200 mL) with saturated aqueous sodium thiosulfate (50 mL) and pentane (200 mL). The layers were separated, and the aqueous layer was extracted again with pentane (2 x 100 mL). The combined organic layer was washed with brine (10 mL), dried over magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was dissolved in pentane then filtered on a thick pad of silica gel and the silica gel pad was washed with more pentane to provide upon concentration of the eluent, 2-(2,2-difluoro-1-methyl- cyclopropyl)ethynyl-triisopropyl-silane (12 g, 89%) as a clear oil 1H NMR (400 MHz, CDCl3) δ 1.60 (ddd, J = 12.7, 7.4, 5.4 Hz, 1H), 1.42 (dd, J = 2.9, 2.0 Hz, 3H), 1.29 (ddd, J = 12.6, 7.5, 5.1 Hz, 1H), 1.09 - 1.03 (m, 21H).19F NMR (377 MHz, CDCl3) δ -133.20 (d, J = 145.8 Hz, 1F), - 139.61 (d, J = 145.8 Hz, 1F). GCMS; EI-MS m/z calc.272.1772, found 272.0 ; Retention time: 8.94 minutes. [00640] GCMS method: Column HP 5 ms Ultra Inert 30 m x 250 µm x 0.25 µm. Control mode: Flow: 3.3 mL/min. Split ratio mode: 33:1. Split Flow: 165 mL/min. Carrier gas: He. Injector Temperature: 250 °C. FID Temperature: 300 °C, MSD transfer line 250 °C. Solvent delay: 3 min. Oven temperature: isotherm at 70 °C for 1 min, then linear heating at 10 °C/min until 140 °C, then 20 °C/min until 250 °C then isotherm 250 °C for 4 min. Run time 17.5 min. Step 3: 6-Bromo-3-[2-(2,2-difluoro-1-methyl-cyclopropyl)ethynyl]pyrazin-2- amine [00641] To a flame-dried flask were added 3,6-dibromopyrazin-2-amine (4.3 g, 17.003 mmol), Copper(I) iodide (967 mg, 5.0774 mmol), bis(triphenylphosphine)palladium(II) chloride (595 mg, 0.8477 mmol), triphenylphosphine (444 mg, 1.6928 mmol), triethyl amine (3.4848 g, 4.8 mL, 34.438 mmol), THF (45 mL) and TBAF (as solution in THF) (6.7 mL of 1 M, 6.7000 mmol). The mixture was degassed with nitrogen for 5 min and heated at 50 °C for 15 min. Then, 2-(2,2-difluoro-1-methyl-cyclopropyl)ethynyl-triisopropyl-silane (2.4 g, 8.1307 mmol) was added dropwise over 15 min and the reaction was stirred at 50 °C overnight. The mixture was cooled down to room temperature and quenched with water (100 mL). The two phases were separated and the aqueous was washed with Me-THF (100 mL) and further extracted with EtOAc (50 mL X 3). The combined organic layers were dried with anhydrous sodium sulfate, filtered and concentrated to afford a crude product (9.3 g) which was purified by reverse phase chromatography on C18 (gradient: 25-45% MeCN in water) to afford 6-bromo-3-[2-(2,2- difluoro-1-methyl-cyclopropyl)ethynyl]pyrazin-2-amine (1.11 g, 46%) as brown powder.1H NMR (400 MHz, Acetone-d6) δ 7.86 (s, 1H), 6.36 (br. s, 2H), 2.03 - 1.95 (m, 1H), 1.76 - 1.67 (m, 1H), 1.55 - 1.52 (m, 3H).19F NMR (377 MHz, Acetone-d6) δ -132.51 - -133.02 (m, 1F), - 140.16 (d, J = 147.1 Hz, 1F). ESI-MS m/z calc.286.987, found 288.0 (M+1)+; Retention time: 1.89 minutes. LC method U. Step 4: 3-Bromo-6-(2,2-difluoro-1-methyl-cyclopropyl)-5H-pyrrolo[2,3- b]pyrazine [00642] To a solution of 6-bromo-3-[2-(2,2-difluoro-1-methyl-cyclopropyl)ethynyl]pyrazin-2- amine (1.02 g, 3.4662 mmol) in anhydrous DMF (30 mL) at 0 °C under nitrogen, was added NaH (60% w/w in oil) (320 mg, 60 %w/w, 8.0008 mmol). Then, the mixture was stirred at 70 °C for 1 hour. After that, the mixture was allowed to cool down to 0 °C. A saturated aqueous solution of ammonium chloride (100 mL) was added, and the mixture was stirred at 0 °C for 1 hour. The resulting precipitate was filtered on Büchner and washed with distilled water (3 x 100 mL). The precipitate was dissolved in EtOAc (200 mL), dried over anhydrous sodium sulfate and the solvent was evaporated under reduced pressure to provide 3-bromo-6-(2,2-difluoro-1- methyl-cyclopropyl)-5H-pyrrolo[2,3-b]pyrazine (990 mg, 99%) as a brown powder.1H NMR (400 MHz, DMSO-d6) δ 12.42 (br. s, 1H), 8.48 (s, 1H), 6.69 (d, J = 2.0 Hz, 1H), 2.40 - 2.29 (m, 1H), 1.90 - 1.80 (m, 1H), 1.64 - 1.59 (m, 3H).. ESI-MS m/z calc.286.987, found 288.2 (M+1)+; Retention time: 1.82 minutes. LC method U. Step 5: 3-Bromo-6-(2,2-difluoro-1-methyl-cyclopropyl)-5-methyl-pyrrolo[2,3- b]pyrazine [00643] To a solution of 3-bromo-6-(2,2-difluoro-1-methyl-cyclopropyl)-5H-pyrrolo[2,3- b]pyrazine (995 mg, 3.4503 mmol) in DMF (25 mL) in a flame-dried flask (25 mL) was added cesium carbonate (2.36 g, 7.2433 mmol). The mixture was stirred in an ice-water bath for 15 minutes. Then, dimethyl sulfate (1.0374 g, 0.78 mL, 8.2247 mmol) was injected. After that, the mixture was allowed to warm up to room temperature and stirred for 3 hours. The reaction was quenched by adding water (200 mL) and brine (50 mL) and diluted by Me-THF (150 mL). The layers were separated, and the aqueous phase was extracted by Me-THF (50 mL X 3). The layers were separated, and the combined organic layers were washed with brine (50 mL). Then the organic solution was dried with anhydrous sodium sulfate, filtered, and evaporated under reduced pressure to afford a crude product which was purified by chromatography on silica gel (gradient: 2% to 30% ethyl acetate in heptane, 9 CV). The desired fractions were collected and evaporated to afford 3-bromo-6-(2,2-difluoro-1-methyl-cyclopropyl)-5-methyl-pyrrolo[2,3- b]pyrazine (718 mg, 68%) as a yellowish solid.1H NMR (400 MHz, CDCl3) δ 8.47 (s, 1H), 6.60 (s, 1H), 3.87 (s, 3H), 1.96 - 1.88 (m, 1H), 1.70 - 1.63 (m, 1H), 1.62 - 1.59 (m, 3H). ESI-MS m/z calc.301.0026, found 302.2 (M+1)+; Retention time: 3.31 minutes. LC method Z. Step 6: 3-Bromo-6-(2,2-difluoro-1-methyl-cyclopropyl)-5-methyl-pyrrolo[2,3- b]pyrazine, enantiomer 1, and 3-bromo-6-(2,2-difluoro-1-methyl-cyclopropyl)-5- methyl-pyrrolo[2,3-b]pyrazine, enantiomer 2 enantiomer 1 enantiomer 2 [00644] A racemic mixture of two batches of 3-bromo-6-(2,2-difluoro-1-methyl-cyclopropyl)- 5-methyl-pyrrolo[2,3-b]pyrazine (85 mg, 0.2813 mmol) and (718 mg, 2.3623 mmol) were combined and subjected to a SFC separation using the following conditions : i-Amylose 3 column (250 x 30 mm), 5 μM column at 40 °C, eluant : 20% methanol and 80% CO2, flow rate : 35.9 mL/min, concentration : 35 mg/mL in methanol (no modifier), injection volume : 500 μL, pressure : 100 bar, wavelength: 210 nm. Evaporation of the solvents afforded the two enantiomers as follows: enantiomer 1, SFC peak 1: 3-bromo-6-(2,2-difluoro-1-methyl- cyclopropyl)-5-methyl-pyrrolo[2,3-b]pyrazine (364 mg, 45%) as a yellow-white solid.1H NMR (400 MHz, CDCl3) δ 8.49 (br. s, 1H), 6.64 (s, 1H), 3.88 (s, 3H), 1.98 - 1.87 (m, 1H), 1.73 - 1.64 (m, 1H), 1.64 - 1.59 (m, 3H). ESI-MS m/z calc.301.0026, found 302.0 (M+1)+; Retention time: 3.31 minutes 19F NMR (377 MHz, CDCl3) δ -132.43 (d, J = 152.6 Hz, 1F), -138.31 (d, J = 152.6 Hz, 1F); and enantiomer 2, SFC peak 2: 3-bromo-6-(2,2-difluoro-1-methyl-cyclopropyl)- 5-methyl-pyrrolo[2,3-b]pyrazine (365 mg, 46%) as a yellow-white solid.1H NMR (400 MHz, CDCl3) δ 8.61 - 8.39 (m, 1H), 6.69 - 6.57 (m, 1H), 3.88 (s, 3H), 1.98 - 1.87 (m, 1H), 1.73 - 1.63 (m, 1H), 1.63 - 1.57 (m, 3H). ESI-MS m/z calc.301.0026, found 302.0 (M+1)+; Retention time: 3.31 minutes.19F NMR (377 MHz, CDCl3) δ -132.45 (d, J = 152.6 Hz, 1F), -138.32 (d, J = 152.6 Hz, 1F). LC method Z. Step 7: tert-butyl (4R)-4-[(1S)-1-(cyclopropylmethyl)-5-[6-(2,2-difluoro-1- methyl-cyclopropyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate enantiomer 1 [00645] To a stirred yellow solution of 3-bromo-6-(2,2-difluoro-1-methyl-cyclopropyl)-5- methyl-pyrrolo[2,3-b]pyrazine, enantiomer 1 (345 mg, 1.142 mmol) in anhydrous tetrahydrofuran (5 mL) was added dropwise a solution of n-BuLi (in hexanes) (450 µL of 2.5 M, 1.125 mmol) over 2 min at -78 °C under nitrogen. The resulting red solution was stirred for10 min at that temperature. Then a solution of tert-butyl (4R)-4-[(1S)-1-(cyclopropylmethyl)-5- [methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (300 mg, 0.7527 mmol) in anhydrous tetrahydrofuran (2 mL) was added slowly. After 30 min at that temperature, saturated aqueous ammonium chloride (10 mL) was added, followed by addition of EtOAc (25 mL). The mixture was allowed to warm to about 0 oC over 15 min and the phases were separated. The aqueous phase was extracted with EtOAc ( 3 x 15 mL) and the combined organics were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Purification was performed by reversed-phase chromatography (30-99 % MeCN in water over 20 min, no modifier). The volatiles were partially removed under reduced pressure. The aqueous layer was extracted with EtOAc (3 x 25 mL). The combined organics were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to furnish tert- butyl (4R)-4-[(1S)-1-(cyclopropylmethyl)-5-[6-(2,2-difluoro-1-methyl-cyclopropyl)-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (280 mg, 66%) ESI-MS m/z calc.560.3174, found 561.2(M+1)⁺; Retention time: 2.31 minutes. as a yellow film. LC method A. Step 8: [(2R,3S)-3-(Cyclopropylmethyl)-7-[6-(2,2-difluoro-1-methyl- cyclopropyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H- azepin-2-yl]methanol [00646] To a stirred solution of tert-butyl (4R)-4-[(1S)-1-(cyclopropylmethyl)-5-[6-(2,2- difluoro-1-methyl-cyclopropyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate (280 mg, 0.4994 mmol) in 1,4-dioxane (6.0 mL) was added water (0.6 mL), hydrochloric acid (in 1,4-dioxane) (5.0 mL of 4 M, 20.00 mmol) at ambient temperature, and the reaction was stirred at that temperature for 1 h. The volatiles were removed under reduced pressure to dryness and water (10 mL) was added to the residue and concentrated. The residual water was co-evaporated with acetonitrile (3 x 10 mL), followed by toluene (3 x 10 mL) then dried under high vacuum to furnish crude [(2R,3S)-3-(cyclopropylmethyl)-7-[6-(2,2- difluoro-1-methyl-cyclopropyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H- azepin-2-yl]methanol (hydrochloride salt) (219 mg, 100%) ESI-MS m/z calc.402.2231, found 403.2(M+1)⁺; Retention time: 1.10 minutes. as a yellow gum. LC method A. Step 9: [(2R,3S,7S)-3-(Cyclopropylmethyl)-7-[6-(2,2-difluoro-1-methyl- cyclopropyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol and [(2R,3S,7R)-3-(cyclopropylmethyl)-7-[6-(2,2-difluoro-1-methyl-cyclopropyl)-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol [00647] To a stirred solution of [(2R,3S)-3-(cyclopropylmethyl)-7-[6-(2,2-difluoro-1-methyl- cyclopropyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (hydrochloride salt) (219 mg, 0.4989 mmol) in anhydrous dichloromethane (2.0 mL) were added acetic-acid (60 µL, 1.055 mmol) and DIPEA (250 µL, 1.435 mmol) at 0 oC under nitrogen. After stirring for 15 min, sodium triacetoxyborohydride (340 mg, 1.604 mmol) was added and stirring was continued for another 20 min. The reaction was quenched with methanol (5 mL) and water (2 mL), then concentrated under reduced pressure. The crude was purified by reversed-phase chromatography (10-99% acetonitrile in water containing 5 mM HCl, over 20 min run). Separated peaks (diastereomers) were concentrated under reduced pressure, the residual water was removed azeotropically with acetonitrile (5 x 10 mL), followed by toluene (3 x 10 mL) to furnish products as light-brown gum. Peak 1 (major diastereomer): [(2R,3S,7S)-3- (cyclopropylmethyl)-7-[6-(2,2-difluoro-1-methyl-cyclopropyl)-5-methyl-pyrrolo[2,3-b]pyrazin- 3-yl]azepan-2-yl]methanol (hydrochloride salt) (165 mg, 75%) 1H NMR (400 MHz, CDCl3) δ 10.44 (s, 1H), 9.79 - 9.11 (m, 2H), 7.18 - 6.89 (m, 1H), 4.86 - 4.53 (m, 1H), 4.05 (s, 3H), 4.00 - 3.90 (m, 1H), 3.87 - 3.69 (m, 1H), 3.39 - 3.14 (m, 1H), 2.60 - 2.37 (m, 2H), 2.36 - 2.21 (m, 2H), 2.13 - 1.98 (m, 3H), 1.87 - 1.73 (m, 2H), 1.66 (s, 3H), 1.54 - 1.37 (m, 1H), 1.34 - 1.16 (m, 1H), 0.79 - 0.61 (m, 1H), 0.61 - 0.48 (m, 1H), 0.49 - 0.39 (m, 1H), 0.18 - -0.06 (m, 2H). ESI-MS m/z calc.404.23877, found 405.2(M+1)⁺; Retention time: 1.15 minutes. Peak 2 (minor diastereomer): [(2R,3S,7R)-3-(cyclopropylmethyl)-7-[6-(2,2-difluoro-1-methyl-cyclopropyl)-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (hydrochloride salt) (31 mg, 14%) ESI-MS m/z calc.404.23877, found 405.2(M+1)⁺; Retention time: 1.19 minutes. LC method A. Step 10: 3-[[4-[[(2R,3S,7S)-3-(Cyclopropylmethyl)-7-[6-(2,2-difluoro-1-methyl- cyclopropyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid
[00648] Through a stirred solution of [(2R,3S,7S)-3-(cyclopropylmethyl)-7-[6-(2,2-difluoro-1- methyl-cyclopropyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (hydrochloride salt) (155 mg, 0.3515 mmol) and 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (170 mg, 0.4068 mmol) in anhydrous tetrahydrofuran (3.0 mL), nitrogen was sparged for 5 min. Then sodium tert-butoxide (240 mg, 2.497 mmol) was added in a single portion at ambient temperature and after stirring for 10 min, the reaction was allowed to stir at 42 °C for 4 h. Then the heating source was removed and the reaction allowed to cool down to ambient temperature. The reaction mixture was diluted with ethyl acetate (20 mL) and acidified with hydrochloric acid (4.0 mL of 1.0 M, 4.000 mmol). The organic phase was separated, and the aqueous layer was extracted with ethyl acetate (2 x 15 mL). The combined organics were washed with brine (10 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. Traces of water were removed azeotropically with acetonitrile (2 x 10 mL) and toluene (2 x 10 mL) to furnish 3-[[4-[[(2R,3S,7S)-3-(cyclopropylmethyl)-7-[6-(2,2- difluoro-1-methyl-cyclopropyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]-6- (2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (265 mg, 92%) ESI-MS m/z calc.785.3171, found 786.2(M+1)⁺; Retention time: 1.47 minutes. as a yellow solid. LC method A. Step 11: (4S,8S,9R)-8-(Cyclopropylmethyl)-4-[6-(2,2-difluoro-1-methyl- cyclopropyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-14-(2,6-dimethylphenyl)- 18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one (Compound 121)
[00649] To a stirred solution of 3-[[4-[[(2R,3S,7S)-3-(cyclopropylmethyl)-7-[6-(2,2-difluoro- 1-methyl-cyclopropyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (265 mg, 0.3222 mmol) in anhydrous NMP (6 mL) and acetone (12 mL), was added p-TsCl (212 mg, 1.112 mmol) at ambient temperature under nitrogen and the reaction was stirred for 5 min. Then N- methylmorpholine (350 µL, 3.183 mmol) was added and the reaction was stirred at 35 oC for 4 h. The reaction was allowed to cool to ambient temperature and the volatiles were removed under reduced pressure. The remaining brownish solution was subjected to preparative HPLC (30-99% acetonitrile in water, containing 5mM HCl as modifier, 15 min run). The fractions were combined and evaporated, then purified a second time by HPLC (40-99% acetonitrile in water, 30 min run, HCl modifier) to furnish product as a yellow solid. (4S,8S,9R)-8- (cyclopropylmethyl)-4-[6-(2,2-difluoro-1-methyl-cyclopropyl)-5-methyl-pyrrolo[2,3-b]pyrazin- 3-yl]-14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (127 mg, 49%); 1H NMR (400 MHz, CDCl3) δ 9.08 (s, 1H), 8.70 (s, 1H), 7.96 (d, J = 7.0 Hz, 1H), 7.89 (d, J = 7.3 Hz, 1H), 7.79 - 7.66 (m, 1H), 7.43 (d, J = 7.5 Hz, 1H), 7.27 (d, J = 7.5 Hz, 2H), 6.80 (s, 1H), 6.43 (s, 1H), 6.25 - 6.09 (m, 1H), 6.09 - 5.93 (m, 1H), 5.11 - 4.84 (m, 1H), 4.78 - 4.59 (m, 1H), 4.19 (s, 3H), 4.03 - 3.84 (m, 1H), 2.59 - 2.28 (m, 3H), 2.26 (s, 6H), 2.19 - 2.11 (m, 2H), 2.10 - 2.02 (m, 3H), 1.79 (s, 3H), 1.40 - 1.27 (m, 1H), 1.27 - 1.16 (m, 1H), 0.67 - 0.49 (m, 1H), 0.50 - 0.32 (m, 2H), 0.11 - -0.14 (m, 2H). ; ESI-MS m/z calc.767.3065, found 768.2(M+1)⁺; Retention time: 2.04 minutes. LC method A. Example 78: Preparation of Compound 122 Step 1: 3-[[4-[[(2S,3S,7R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00650] [(2S,3S,7R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy- azepan-2-yl]methanol (36 mg, 0.09612 mmol) was combined with 3-[[4-chloro-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (40.5 mg, 0.09692 mmol) in anhydrous THF (300 µL) in a screwcap vial and stirred at room temperature for 5 minutes. sodium tert- butoxide (60 mg, 0.6243 mmol) was added and the reaction mixture was stirred at room temperature for 10 minutes, then warmed to 40 °C for 2 hours. The reaction was quenched into 1 M HCl (10 mL) and extracted 3 x 15 mL ethyl acetate. The aqueous layer was diluted with an equal volume of brine and extracted an additional 3 x 10 mL ethyl acetate. The combined organics were washed with brine, dried over sodium sulfate, and concentrated to give 3-[[4- [[(2S,3S,7R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-isopropoxy-azepan-2- yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (75 mg, 98%) ESI-MS m/z calc.755.3465, found 756.8 (M+1)⁺; Retention time: 0.63 minutes., which was used in the next step without additional purification. LC method B. Step 2: (4R,8S,9S)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-8-isopropoxy-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one (Compound 122)
[00651] 3-[[4-[[(2S,3S,7R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- isopropoxy-azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (75 mg, 0.09465 mmol) was dissolved in NMP (1.5 mL) and Acetone (3 mL). TsCl (55 mg, 0.2885 mmol) was added followed by 4-methylmorpholine (65 µL, 0.5912 mmol) and the reaction was stirred at 55 °C. After 2 hours, additional TsCl (25 mg, 0.1311 mmol) and 4-methylmorpholine (30 µL, 0.2729 mmol) were added. After a total reaction time of 20 hours the reaction mixture was cooled to room temperature and poured into a mixture of ethyl acetate and 1 M HCl, and the layers were separated. The aqueous layer was further extracted with ethyl acetate (2 x). The organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was taken up in DMSO/MeOH then injected into a reverse phase HPLC (30-99% acetonitrile/5 mM aqueous HCl over 25 min). The pure fractions were extracted with EtOAc (3 x), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give (4R,8S,9S)-4-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-8-isopropoxy-18,18- dioxo-11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (22.5 mg, 31%) as a white solid. ESI-MS m/z calc. 737.33594, found 738.9 (M+1)⁺; Retention time: 1.97 minutes.1H NMR (400 MHz, CDCl3) δ 8.69 (t, J = 2.0 Hz, 1H), 8.55 (s, 1H), 7.82 (d, J = 7.9 Hz, 1H), 7.72 (d, J = 7.7 Hz, 1H), 7.59 (t, J = 7.7 Hz, 1H), 7.33 - 7.29 (m, 1H), 7.13 (d, J = 7.6 Hz, 2H), 6.98 (s, 1H), 6.27 (s, 1H), 5.73 - 5.64 (m, 1H), 5.10 - 4.94 (m, 2H), 4.21 (s, 3H), 3.95 (d, J = 7.2 Hz, 2H), 3.72 (p, J = 6.1 Hz, 1H), 2.93 - 2.85 (m, 1H), 2.48 - 2.42 (m, 1H), 2.30 - 2.21 (m, 2H), 2.13 (s, 6H), 1.95 - 1.87 (m, 2H), 1.57 (s, 9H), 1.26 (d, J = 6.2 Hz, 3H), 1.21 (d, J = 6.0 Hz, 3H). LC method A. Example 79: Preparation of Compound 123 Step 1: 6-methoxy-3,3-dimethyl-hex-1-yne [00652] To a stirred solution of 3-methylbut-1-yne (1.0 g, 14.68 mmol) in anhydrous diethyl ether (5 mL) was added slowly a solution of n-butyllithium in hexanes (12 mL of 2.5 M, 30.00 mmol) at 0 °C under nitrogen. At the beginning, white precipitate started to form, but after halfway through the addition, reaction became homogeneous yellow. Then N,N,N',N'- tetramethylethane-1,2-diamine (2.2 mL, 14.58 mmol) was added slowly, and the reaction turned to a fine pale yellow slurry. The mixture was stirred at 65 °C (bath temperature, mild reflux) for 2 h. The red heterogeneous slurry was allowed to cool to ambient temperature before placed in a −78 °C bath. Then a solution of 1-bromo-3-methoxy-propane (2.24 g, 14.64 mmol) in cold anhydrous diethyl ether (2 mL) was added slowly. At the end of the addition, the red color turned to a brownish mixture. After 1 hour at that temperature, the bath was removed, and the reaction was placed in an ice-water bath (0-4 °C). Water (20 mL) and diethyl ether (25 mL) were added. The layers were separated. The aqueous phases were extracted with diethyl ether (3 × 15 mL) and the combined organics were washed with water (2 x 10 mL) and brine (10 mL), dried over anhydrous sodium sulfate, filtered and the solvent was carefully partially removed under reduced pressure (500 mbar, 20 °C bath temperature).to furnish 6-methoxy-3,3-dimethyl- hex-1-yne (1.050 g, 51%) 1H NMR (400 MHz, MeOD) δ 3.40 (t, J = 6.5 Hz, 2H), 3.33 (s, 3H), 2.35 (s, 1H), 2.12 - 2.02 (m, 2H), 1.45 - 1.40 (m, 2H), 1.19 (s, 6H). as a yellow liquid. Step 2: 6-Bromo-3-(6-methoxy-3,3-dimethyl-hex-1-ynyl)pyrazin-2-amine [00653] A stirred solution of 3,6-dibromopyrazin-2-amine (2.2 g, 8.699 mmol) in anhydrous THF (16.0 mL) was sparged with nitrogen for 10 min. Then triethylamine (1.4 mL, 10.04 mmol), copper (I) iodide (115 mg, 0.6038 mmol) and bis(triphenylphosphine)palladium(II) dichloride (212 mg, 0.3020 mmol) were added at ambient temperature. The heterogeneous yellow mixture was cooled to 0 °C and 6-methoxy-3,3-dimethyl-hex-1-yne (2.00 g of 51 %w/w, 7.274 mmol) was added dropwise over 5 min. After sitting for 2 hour at that temperature, the deep green mixture was diluted with ethyl acetate (100 mL). The suspension was filtered over a pad of silica gel and the filter cake was rinsed with ethyl acetate (2 x 20 mL). The orange filtrate was concentrated under reduced pressure and dry-loaded onto Celite. Purification by silica gel chromatography (330 g silica gel column, 0-20 % ethyl acetate in hexanes over 40 min, peak came around 15% ethyl acetate) furnished 6-bromo-3-(6-methoxy-3,3-dimethyl-hex-1- ynyl)pyrazin-2-amine (482 mg, 21%) as an orange-yellow solid.1H NMR (400 MHz, DMSO) δ 7.80 (d, J = 0.8 Hz, 1H), 6.83 (s, 2H), 3.33 - 3.31 (m, 2H), 3.23 (d, J = 0.8 Hz, 3H), 1.72 - 1.62 (m, 2H), 1.57 - 1.48 (m, 2H), 1.29 (s, 6H). ; ESI-MS m/z calc.311.06332, found 312.0(M+1)⁺; Retention time: 1.62 minutes. LC method A. Step 3: 3-Bromo-6-(4-methoxy-1,1-dimethyl-butyl)-5H-pyrrolo[2,3-b]pyrazine [00654] To a stirred orange-yellow slurry of 6-bromo-3-(6-methoxy-3,3-dimethyl-hex-1- ynyl)pyrazin-2-amine (480 mg, 1.537 mmol) in anhydrous tert-butanol (8.0 mL) was added solid potassium tert-butoxide (900 mg, 8.020 mmol) in one portion at ambient temperature under nitrogen. The reaction mixture was allowed to stir at 82 °C for 18 h. The reaction was allowed to cool down to ambient temperature. The volatiles were removed under reduced pressure and the residue was diluted with ice-water (50 mL), acidified with 1.0 M HCl (6.0 mL), extracted with ethyl acetate (3 x 25 mL), rinsed with brine (20 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to furnish product as a tan solid.3-bromo-6-(4- methoxy-1,1-dimethyl-butyl)-5H-pyrrolo[2,3-b]pyrazine (471 mg, 98%), ESI-MS m/z calc. 311.06332, found 312.0 (M+1)⁺; Retention time: 1.47 minutes. LC method A. Step 4: 3-Bromo-6-(4-methoxy-1,1-dimethyl-butyl)-5-methyl-pyrrolo[2,3- b]pyrazine [00655] To a stirred solution of 3-bromo-6-(4-methoxy-1,1-dimethyl-butyl)-5H-pyrrolo[2,3- b]pyrazine (471 mg, 1.509 mmol) in anhydrous DMF (8 mL) was added NaH (in mineral oil) (100 mg of 60 %w/w, 2.500 mmol) at 0 °C under nitrogen. After stirring the reaction mixture for 20 min, iodomethane (275 mg, 1.937 mmol) was added. After another 2 h, at that temperature, ice-water (100 mL) was added very carefully (caution! rapid effervescence), and the suspension was stirred for 10 min. The solid was filtered and washed with water (3 x 25 mL). The tan solid was collected and diluted with acetonitrile. The suspension was co-evaporated with acetonitrile (3 x 20 mL) and dried in vacuo to afford 3-bromo-5-methyl-6-(1- methylcyclopropyl)pyrrolo[2,3-b]pyrazine (978 mg, 98%) as a brown solid.1H NMR (400 MHz, CDCl3) δ 8.41 (s, 1H), 6.48 (s, 1H), 3.97 (s, 3H), 3.30 (t, J = 6.2 Hz, 2H), 3.27 (s, 3H), 1.92 - 1.86 (m, 2H), 1.48 (s, 6H), 1.38 - 1.33 (m, 2H). ESI-MS m/z calc.325.07898, found 326.1(M+1)⁺; Retention time: 1.75 minutes. LC method A. Step 5: tert-Butyl (4R)-4-[(1S)-1-(cyclopropylmethyl)-5-[6-(4-methoxy-1,1- dimethyl-butyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00656] To a stirred yellow solution of 3-bromo-6-(4-methoxy-1,1-dimethyl-butyl)-5-methyl- pyrrolo[2,3-b]pyrazine (335 mg, 1.027 mmol) in anhydrous tetrahydrofuran (4 mL) was added dropwise a solution of n-BuLi (in hexanes) (400 µL of 2.5 M, 1.000 mmol) at -78 °C under nitrogen. The resulting red solution was stirred for 6 min at -78 °C. Then a solution of tert-butyl (4R)-4-[(1S)-1-(cyclopropylmethyl)-5-[methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (260 mg, 0.6269 mmol) in anhydrous tetrahydrofuran (1.5 mL) was added slowly. After 15 min at that temperature, a saturated aqueous solution of ammonium chloride (10 mL) was added and the reaction was allowed to warm to about 0 oC over 15 min. The mixture was extracted with ethyl acetate (3 x 15 mL) and the combined organics were concentrated under reduced pressure, and the residue was purified by reversed-phase chromatography (30-99 % acetonitrile in water, no modifier). The volatiles were removed under reduced pressure and the product was further dried by azeotropically removing water with acetonitrile (2 x 15 mL) and toluene (2 x 10 mL) to furnish tert-butyl (4R)-4-[(1S)-1- (cyclopropylmethyl)-5-[6-(4-methoxy-1,1-dimethyl-butyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (95 mg, 26%) ESI-MS m/z calc. 584.3938, found 585.2 (M+1)⁺; Retention time: 2.38 minutes. as a yellow oil. LC method A. Step 6: [(2R,3S)-3-(Cyclopropylmethyl)-7-[6-(4-methoxy-1,1-dimethyl-butyl)-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol [00657] To a stirred solution of tert-butyl (4R)-4-[(1S)-1-(cyclopropylmethyl)-5-[6-(4- methoxy-1,1-dimethyl-butyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (95 mg, 0.1624 mmol) in anhydrous dioxane (1.2 mL) was added water (0.1 mL) water (549.4 µL), followed by hydrochloric acid (in 1,4-dioxane) (1.5 mL of 4.0 M, 6.000 mmol) at ambient temperature, and let the reaction stir at that temperature for 5 h. The volatiles were removed under reduced pressure to dryness and water (3 mL) was added to the residue and concentrated. Then the residual water was azeotropically removed with acetonitrile (3 x 10 mL), followed by toluene (3 x 10 mL). Upon further drying the crude was obtained as an orange-yellow gum. [(2R,3S)-3-(cyclopropylmethyl)-7-[6-(4-methoxy-1,1-dimethyl-butyl)-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (hydrochloride salt) (75 mg, 100%) ESI-MS m/z calc.426.29947, found 427.3 (M+1)⁺; Retention time: 1.27 minutes. LC method A. Step 7: [(2R,3S,7S)-3-(cyclopropylmethyl)-7-[6-(4-methoxy-1,1-dimethyl-butyl)- 5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol and [(2R,3S,7R)-3- (cyclopropylmethyl)-7-[6-(4-methoxy-1,1-dimethyl-butyl)-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl]azepan-2-yl]methanol
[00658] To a stirred solution of [(2R,3S)-3-(cyclopropylmethyl)-7-[6-(4-methoxy-1,1- dimethyl-butyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2- yl]methanol (hydrochloride salt) (75 mg, 0.1620 mmol) in anhydrous dichloromethane (700 µL) were added acetic acid (20 µL, 0.3517 mmol) and DIPEA (80 µL, 0.4593 mmol) at ambient temperature under nitrogen. After stirring for 15 min, sodium triacetoxyborohydride (110 mg, 0.5190 mmol) was added at once. The reaction mixture was stirred at that temperature for 15 min, then quenched with methanol (2 mL) and water (0.5 mL), After concentrating under reduced pressure, the crude was purified by reversed-phase chromatography (10-99% acetonitrile in water containing 5 mM HCl, over 20 min run) to give a mixture of diastereomers of [(2R,3S)-3-(cyclopropylmethyl)-7-[6-(4-methoxy-1,1-dimethyl-butyl)-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl]azepan-2-yl]methanol (hydrochloride salt) (65 mg, 86%) ESI-MS m/z calc. 428.31512, found 429.3 (M+1)⁺; Retention time: 1.24 minutes. as an orange viscous oil. The diastereomers were separated by SFC [column Phenomenex LUX-4 ((21.2 x 250mm, 5um), 40 oC, gradient (28% methanol + 20 mM NH3), 70 mL/min, inj. vol.: 0.5 ml, conc.21.4 mg/ml in methanol (no modifier), variable pressure, wavelength 229 nm.] to furnish two fractions. The major diastereomer (Peak-1) [(2R,3S,7S)-3-(cyclopropylmethyl)-7-[6-(4-methoxy-1,1-dimethyl- butyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (19 mg, 27%), 1H NMR (400 MHz, CDCl3) δ 8.32 (s, 1H), 6.45 (s, 1H), 4.03 (dd, J = 9.5, 4.6 Hz, 1H), 3.99 (s, 3H), 3.75 (dd, J = 10.5, 4.2 Hz, 1H), 3.30 (t, J = 6.1 Hz, 2H), 3.27 (s, 3H), 3.21 (t, J = 10.2 Hz, 1H), 2.71 (td, J = 9.5, 4.1 Hz, 1H), 2.21 - 2.13 (m, 1H), 2.05 - 1.97 (m, 1H), 1.95 - 1.79 (m, 5H), 1.77 - 1.69 (m, 1H), 1.66 - 1.59 (m, 1H), 1.48 (s, 6H), 1.42 - 1.33 (m, 3H), 1.22 - 1.13 (m, 1H), 0.75 - 0.65 (m, 1H), 0.55 - 0.40 (m, 2H), 0.14 - 0.05 (m, 1H), 0.05 - -0.03 (m, 1H). ; ESI-MS m/z calc. 428.31512, found 429.3(M+1)⁺; Retention time: 1.24 minutes. was obtained as a yellow viscous oil. The minor diastereomer (peak-2) [(2R,3S,7R)-3-(cyclopropylmethyl)-7-[6-(4-methoxy-1,1- dimethyl-butyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (6 mg, 8%) was obtained as a yellow viscous oil. LC method A. Step 8: 3-[[4-[[(2R,3S,7S)-3-(Cyclopropylmethyl)-7-[6-(4-methoxy-1,1-dimethyl- butyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid [00659] Through a stirred solution of [(2R,3S,7S)-3-(cyclopropylmethyl)-7-[6-(4-methoxy- 1,1-dimethyl-butyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (19 mg, 0.04433 mmol) and 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (22 mg, 0.05265 mmol) in anhydrous tetrahydrofuran (500 µL), nitrogen was sparged for 5 min. Then sodium tert-butoxide (30 mg, 0.3122 mmol) was added in a single portion at ambient temperature and after stirring for 5 min, the reaction was allowed to stir at 42 °C for 16 h. Then the heating source was removed and allowed to cool down to ambient temperature. The reaction mixture was diluted with ethyl acetate (20 mL) and acidified with hydrochloric acid (500 µL of 1.0 M, 0.5000 mmol). The organic phase was separated, and the aqueous layer was extracted with ethyl acetate (2 x 15 mL). The combined organics were washed with brine (10 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude was subjected to preparative HPLC (30-99% acetonitrile in water, containing 5 mM HCl as modifier, 15 min run). The combined fractions were concentrated under reduced pressure and the traces of water was removed azeotropically with acetonitrile (2 x 10 mL) and toluene (2 x 10 mL) to furnish 3- [[4-[[(2R,3S,7S)-3-(cyclopropylmethyl)-7-[6-(4-methoxy-1,1-dimethyl-butyl)-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (30 mg, 80%) ESI-MS m/z calc.809.39343, found 810.3 (M+1)⁺; Retention time: 0.60 minutes. as a yellow solid. LC method B. Step 9: (4S,8S,9R)-8-(Cyclopropylmethyl)-14-(2,6-dimethylphenyl)-4-[6-(4- methoxy-1,1-dimethyl-butyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo- 11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (Compound 123) [00660] To a stirred solution of 3-[[4-[[(2R,3S,7S)-3-(cyclopropylmethyl)-7-[6-(4-methoxy- 1,1-dimethyl-butyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (30 mg, 0.03544 mmol) in anhydrous NMP (0.6 mL) and acetone (1.2 mL), was added p-TsCl (25 mg, 0.1311 mmol) at ambient temperature under nitrogen, followed by the addition of N-methylmorpholine (30 mg, 0.2966 mmol). The reaction was allowed to stir at 35 oC for 2 h. The reaction was allowed to cool to ambient temperature and the volatiles were removed under reduced pressure. The remaining brownish solution was subjected to preparative HPLC (30-99% acetonitrile in water, containing 5 mM HCl as modifier, 15 min run). The product was repurified by HPLC (40-99% acetonitrile in water, 30 min run, HCl modifier) to furnish product as a yellow solid. (4S,8S,9R)-8-(cyclopropylmethyl)-14-(2,6-dimethylphenyl)-4-[6-(4-methoxy-1,1-dimethyl- butyl)-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (4.7 mg, 16%), 1H NMR (400 MHz, CDCl3) δ 8.84 (d, J = 2.0 Hz, 1H), 8.45 (s, 1H), 7.80 - 7.71 (m, 2H), 7.50 (t, J = 7.7 Hz, 1H), 7.19 (t, J = 7.6 Hz, 1H), 7.05 (d, J = 7.6 Hz, 2H), 6.44 (s, 1H), 6.17 (s, 1H), 5.94 - 5.85 (m, 2H), 4.85 (t, J = 11.8 Hz, 1H), 4.53 - 4.43 (m, 1H), 4.11 (s, 3H), 3.33 - 3.28 (m, 2H), 3.27 (s, 3H), 2.96 - 2.90 (m, 1H), 2.33 - 2.15 (m, 2H), 2.07 (s, 6H), 1.99 - 1.89 (m, 3H), 1.89 - 1.78 (m, 3H), 1.47 (s, 3H), 1.47 (s, 3H), 1.43 - 1.35 (m, 2H), 1.17 - 1.09 (m, 1H), 1.04 - 0.97 (m, 1H), 0.42 - 0.31 (m, 1H), 0.30 - 0.15 (m, 2H), -0.14 - -0.30 (m, 2H). ESI- MS m/z calc.791.3829, found 792.2 (M+1)⁺; Retention time: 1.98 minutes. LC method A. Example 80: Preparation of Compound 126 and Compound 127 Step 1: (5S)-5-[[tert-Butyl(diphenyl)silyl]oxymethyl]pyrrolidin-2-one [00661] To a stirred solution of (5S)-5-(hydroxymethyl)pyrrolidin-2-one (2 g, 17.372 mmol) in DCM (50 mL) were added imidazole (2.6 g, 38.192 mmol) and DMAP (212 mg, 1.7353 mmol). Then, tert-butyl(chloro)diphenylsilane (5.2 g, 18.919 mmol) was added dropwise over 15 min at 0 °C. The reaction was stirred at this temperature for 30 min then allowed to reach room temperature and stirred overnight at room temperature. The reaction was then partitioned between dichloromethane (75 mL) and water (50 mL). The layers were separated and the organic layer was washed with a saturated aqueous solution of ammonium chloride (5 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to afford (5S)-5- [[tert-butyl(diphenyl)silyl]oxymethyl]pyrrolidin-2-one (7.1 g, 98%) as a colorless oil which was used without further purification. ESI-MS m/z calc.353.1811, found 354.4 (M+1)+; Retention time: 1.52 minutes. LC method U. Step 2: tert-butyl (2S)-2-[[tert-Butyl(diphenyl)silyl]oxymethyl]-5-oxo- pyrrolidine-1-carboxylate [00662] To a stirred solution of (5S)-5-[[tert-butyl(diphenyl)silyl]oxymethyl]pyrrolidin-2-one (7.1 g, 17.071 mmol) in acetonitrile (35 mL) was added DMAP (1.25 g, 10.232 mmol). Boc2O (7 g, 32.074 mmol) was added to this solution. The resulting reaction mixture was stirred overnight at room temperature. The reaction was then concentrated under reduced pressure and the resulting residue was dissolved in ethyl acetate (100 mL), washed with a saturated aqueous solution of ammonium chloride (75 mL), brine (20 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude solid was triturated overnight with pentane, filtered, dried under vacuum to afford pure tert-butyl (2S)-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-5-oxo-pyrrolidine-1-carboxylate (6.61 g, 83%) as a pale yellow powder.1H NMR (400 MHz, CDCl3) δ 7.68 - 7.58 (m, 4H), 7.50 - 7.33 (m, 6H), 4.26 - 4.18 (m, 1H), 3.90 (dd, J = 10.4, 4.3 Hz, 1H), 3.71 (dd, J = 10.5, 2.4 Hz, 1H), 2.79 (dt, J = 17.5, 10.6 Hz, 1H), 2.44 (ddd, J = 17.6, 8.8, 3.2 Hz, 1H), 2.24 - 2.06 (m, 2H), 1.44 (s, 9H), 1.05 (s, 9H). ESI- MS m/z calc.453.2335, found 354.2 (M-99)+; Retention time: 2.08 minutes. LC method U. Step 3: tert-Butyl (5S)-5-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-oxo-3- phenylselanyl-pyrrolidine-1-carboxylate TBDPSO [00663] NaHMDS in THF (28 mL of 1 M, 28.000 mmol) was added slowly to a solution of tert-butyl (2S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-pyrrolidine-1-carboxylate (6.5 g, 13.984 mmol) in THF (68 mL) maintained at -78 °C. The reaction mixture was stirred for 1 h then phenyl selenohypochlorite (2.7 g, 14.098 mmol) was added dropwise dissolved in THF (34 mL) at -78 °C, then the reaction was left stirring for 2 h at this temperature and then quenched with saturated aqueous ammonium chloride (100 mL) and left to warm-up to room temperature. It was then left stirring at room temperature for 30 min. The aqueous solution was separated and washed with ethyl acetate containing 20% heptane (3 x 100 mL). The combined organics were dried over magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by reverse phase chromatography on a C18 column using a 70 to 100% gradient of acetonitrile in pure water to provide the desired product tert-butyl (5S)-5-[[tert- butyl(diphenyl)silyl]oxymethyl]-2-oxo-3-phenylselanyl-pyrrolidine-1-carboxylate (4.8 g, 55%) as a clear yellow oil as a 3:7 cis: trans mixture of isomers.1H NMR (400 MHz, CDCl3) δ 7.68 - 7.65 (m, 2H), 7.60 - 7.54 (m, 4H), 7.44 - 7.30 (m, 9H), 4.28 (t, J = 9.4 Hz, 1H), 4.03 - 3.98 (m, 1H), 3.91 (dd, J = 10.6, 3.5 Hz, 1H), 3.62 (dd, J = 10.5, 2.2 Hz, 1H), 2.56 - 2.46 (m, 1H), 2.21 (dt, J = 13.4, 9.3 Hz, 1H), 1.45 (s, 9H), 1.00 (s, 9H). ESI-MS m/z calc.609.1814, found 510.2 (M-99)+; Retention time: 2.4 minutes along with a product of bis addition tert-butyl (5S)-5- [[tert-butyl(diphenyl)silyl]oxymethyl]-2-oxo-3,3-bis(phenylselanyl)pyrrolidine-1-carboxylate (1.1 g, 10%) as a clear yellow oil 1H NMR (400 MHz, CDCl3) δ 7.68 - 7.65 (m, 2H), 7.60 - 7.54 (m, 4H), 7.44 - 7.30 (m, 9H), 4.28 (t, J = 9.4 Hz, 1H), 4.03 - 3.98 (m, 1H), 3.91 (dd, J = 10.6, 3.5 Hz, 1H), 3.62 (dd, J = 10.5, 2.2 Hz, 1H), 2.56 - 2.46 (m, 1H), 2.21 (dt, J = 13.4, 9.3 Hz, 1H), 1.45 (s, 9H), 1.00 (s, 9H). ESI-MS m/z calc.765.1292, found 664.0 (M-99)+; Retention time: 2.69 minutes. LC method U. Step 4: tert-Butyl (2S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-2H- pyrrole-1-carboxylate [00664] Hydrogen peroxide (in water) (3.7 g, 35 %w/w, 38.072 mmol) was added dropwise to a solution of tert-butyl (5S)-5-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-oxo-3-phenylselanyl- pyrrolidine-1-carboxylate (4.7 g, 7.5360 mmol) in THF (75 mL) maintained at 0 °C. This reaction mixture was stirred at this temperature for 60 min then left to warm up to room temperature and stirred for 2 h. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (100 mL) and this mixture was stirred for 30 min. The layers were separated, and the aqueous layer was extracted with ethyl acetate containing 10% heptane (3 x 100 mL). The organics were combined, dried over magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by reverse phase chromatography on a C18 column using a 5 to 100% gradient of acetonitrile in pure water to provide tert-butyl (2S)- 2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-2H-pyrrole-1-carboxylate (3.4 g, 96%) as a pale yellow solid 1H NMR (400 MHz, CDCl3) δ 7.61 (ddd, J = 9.7, 7.9, 1.2 Hz, 4H), 7.50 - 7.35 (m, 6H), 7.30 - 7.24 (overlap with chloroform, m, 1H), 6.18 (dd, J = 6.1, 1.5 Hz, 1H), 4.70 - 4.62 (m, 1H), 4.13 (dd, J = 9.7, 3.5 Hz, 1H), 3.84 (dd, J = 9.8, 6.4 Hz, 1H), 1.45 (s, 9H), 1.03 (s, 9H). ESI-MS m/z calc.451.2179, found 352.2 (M-99)+; Retention time: 2.01 minutes. LC method U. Step 5: tert-Butyl (2S,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-cyclobutyl- 5-oxo-pyrrolidine-1-carboxylate [00665] To a solution of cyclobutyl magnesium bromide (solution in THF) (100 mL of 0.5 M, 50.000 mmol) in tetrahydrofuran (300 mL) at -35 °C was added copper iodide (1.4 g, 7.3510 mmol) and the resulting suspension was stirred 0.5 hour at that temperature. To the suspension was added a solution of tert-butyl (2S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-2H- pyrrole-1-carboxylate (20 g, 41.848 mmol) in tetrahydrofuran (100 mL) dropwise over 1.5 h. The reaction was stirred at -35 °C for 30 min. An aqueous solution of ammonium chloride (sat., 100 mL) and water (100 mL) were added. The suspension was stirred for 1 h allowing it to reach rt. EtOAc (250 mL) was added, and the phases were separated. The organic phase was washed with brine (200 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford, after evaporation, crude tert-butyl (2S,3R)-2- [[tert-butyl(diphenyl)silyl]oxymethyl]-3-cyclobutyl-5-oxo-pyrrolidine-1-carboxylate (23.2 g, 81%) as a dark yellow oil. ESI-MS m/z calc.507.2805, found 408.4 (M-99)+; Retention time: 2.61 minutes.100 mg of product was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 90% of MeOH in water to afford tert-butyl (2S,3R)-2- [[tert-butyl(diphenyl)silyl]oxymethyl]-3-cyclobutyl-5-oxo-pyrrolidine-1-carboxylate (65 mg, 0%) as a colorless oil with an 99.9% ee.1H NMR (400 MHz, CDCl3) δ 7.76 - 7.57 (m, 4H), 7.51 - 7.31 (m, 6H), 3.91 - 3.81 (m, 1H), 3.81 - 3.74 (m, 1H), 3.69 (dd, J = 10.1, 2.1 Hz, 1H), 2.82 (dd, J = 17.9, 8.1 Hz, 1H), 2.33 - 2.19 (m, 2H), 2.17 - 2.01 (m, 3H), 2.01 - 1.78 (m, 2H), 1.74 - 1.52 (m, 2H), 1.44 (s, 9H), 1.07 (s, 9H). ESI-MS m/z calc.507.2805, found 408.2 (M-99)+; Retention time: 4.71 minutes. LC method U. Step 6: tert-Butyl N-[(1S,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2- cyclobutyl-4-hydroxy-butyl]carbamate TBD [00666] To a 50 °C solution of tert-butyl (2S,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- cyclobutyl-5-oxo-pyrrolidine-1-carboxylate (23.1 g, 33.804 mmol) in ethanol (200 mL) under nitrogen was added sodium borohydride (5 g, 132.16 mmol). The reaction was stirred for 3 hour at that temperature. Water (200 mL) was slowly introduced into the reaction under a nitrogen flow and the resulting solution was stirred for 1 h. The volatiles were evaporated under reduced pressure and EtOAc (200 mL) was added followed by brine (50 mL). The phases were separated and the aqueous one was extracted with EtOAc (200 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert-butyl N-[(1S,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-cyclobutyl-4-hydroxy- butyl]carbamate (21.9 g, 102%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 7.71 - 7.61 (m, 4H), 7.50 - 7.33 (m, 6H), 4.88 - 4.60 (m, 1H), 3.61 (m, 5H), 2.37 - 2.08 (m, 1H), 2.03 - 1.87 (m, 2H), 1.83 - 1.54 (m, 5H), 1.52 - 1.31 (m, 10H), 1.12 - 1.01 (m, 9H) (2H missing, labile proton). ESI-MS m/z calc.511.3118, found 412.4 (M-99)+; Retention time: 2.5 minutes. LC method U. Step 7: tert-Butyl N-[(1S,2R)-2-cyclobutyl-4-hydroxy-1- (hydroxymethyl)butyl]carbamate [00667] To a 0 °C solution of tert-butyl N-[(1S,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]- 2-cyclobutyl-4-hydroxy-butyl]carbamate (21.9 g, 31.667 mmol) in tetrahydrofuran (200 mL) was added tetrabutylammonium fluoride (solution in THF) (65 mL of 1 M, 65.000 mmol) dropwise over 15 min. The solution was stirred at that temperature for 2 h. An aqueous solution of potassium bicarbonate (5%, 200 mL) was added, and the mixture was stirred for 1 h. EtOAc (200 mL) was added, and the phases were separated, and the aqueous one was extracted with EtOAc (2 x 200 mL). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and was then purified in two lots by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 55% of MeOH in water. The volatiles were removed under reduced pressure. EtOAc (300 mL) was added followed by brine (400 mL). The phases were separated, and the aqueous one was extracted with EtOAc (2 x 300 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford the product (lot 02) as a yellow oil. To the aqueous phase from extraction was added 2-MeTHF (300 mL) and the phases were separated, and the aqueous one was extracted with 2-MeTHF (2 x 300 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford the product (lot 03) as a yellow oil. Mixed fractions were combined, and the volatiles were removed under reduced pressure. To the aqueous phase from extraction was added 2-MeTHF (300 mL) and the phases were separated, and the aqueous one was extracted with 2-MeTHF (2 x 300 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 55% of MeOH in water. The volatiles were removed under reduced pressure. To the aqueous phase from extraction was added 2-MeTHF (300 mL) and the phases were separated, and the aqueous one was extracted with 2-MeTHF (2 x 300 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford the product (lot 04) as a yellow oil. The three lots were combined in acetone (200 mL) and concentrated under reduced pressure to afford tert- butyl N-[(1S,2R)-2-cyclobutyl-4-hydroxy-1-(hydroxymethyl)butyl]carbamate (8.3 g, 71%) as a yellow oil.1H NMR (400 MHz, DMSO-d6) δ 6.34 (d, J = 9.0 Hz, 1H), 4.47 (br. s., 2H), 3.54 - 3.41 (m, 1H), 3.36 (t, J = 7.0 Hz, 2H), 3.30 - 3.22 (m, 2H), 2.27 - 2.10 (m, 1H), 2.04 - 1.84 (m, 2H), 1.78 - 1.49 (m, 5H), 1.47 - 1.32 (m, 10H), 1.31 - 1.20 (m, 1H). ESI-MS m/z calc.273.194, found 218.4 (M-55)+; Retention time: 1.63 minutes. LC method U. Step 8: tert-Butyl (4S)-4-[(1R)-1-cyclobutyl-3-hydroxy-propyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00668] To a solution of tert-butyl N-[(1S,2R)-2-cyclobutyl-4-hydroxy-1- (hydroxymethyl)butyl]carbamate (8.3 g, 22.559 mmol) in acetone (200 mL) were added 2,2- dimethoxypropane (14.450 g, 17 mL, 138.74 mmol) and p-toluenesulfonic acid monohydrate (434 mg, 2.2816 mmol) at room temperature. The solution was stirred at rt for 2 h. Acetone (10 mL) and an aqueous solution of sodium bicarbonate (sat., 10 mL) were added. iPrOAc (50 mL) was added and removed under reduced pressure (3 x). To the resulting liquid was added an aqueous solution of sodium bicarbonate (sat., 50 mL) and DCM (50 mL). The phases were separated, and the aqueous one was extracted with DCM (2 x 50 mL). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 80% of MeOH in water to afford crude tert-butyl (4S)- 4-[(1R)-1-cyclobutyl-3-hydroxy-propyl]-2,2-dimethyl-oxazolidine-3-carboxylate (10.5 g, 100%) as a dark yellow oil.1H NMR (400 MHz, DMSO-d6) δ 4.36 (br. s., 1H), 3.95 - 3.75 (m, 2H), 3.74 - 3.65 (m, 1H), 3.50 - 3.33 (m, 2H), 2.38 (br. s., 1H), 1.97 - 1.82 (m, 3H), 1.80 - 1.59 (m, 4H), 1.50 (br. s., 4H), 1.41 (s, 9H), 1.39 - 1.33 (m, 3H), 1.20 - 1.11 (overlaps with iPrOAc, m, 1H). ESI-MS m/z calc.313.2253, found 336.4 (M+23)+; Retention time: 2.0 minutes. LC method U. Step 9: tert-Butyl (4S)-4-[(1R)-1-cyclobutyl-3-oxo-propyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00669] To a 0 °C solution of tert-butyl (4S)-4-[(1R)-1-cyclobutyl-3-hydroxy-propyl]-2,2- dimethyl-oxazolidine-3-carboxylate (10.5 g, 22.613 mmol) in dichloromethane (200 mL) were added a solution of sodium bromide (695 mg, 6.7545 mmol) in water (100 mL) followed by sodium bicarbonate (3.91 g, 46.544 mmol) and TEMPO (365 mg, 2.3361 mmol). The mixture was vigorously stirred for 10 min and sodium hypochlorite (solution in water) (41.5 mL of 0.548 M, 22.742 mmol) was added over 5 min. The mixture was stirred for 30 min at 0 °C. MeOH (5 mL), water (30 mL) and DCM (30 mL) were added, the phases were separated, and the organic phase was washed with water (300 mL), an aqueous solution of Na2S2O3 (10%, 300 mL) and brine (300 mL). The organic phase was then dried over anhydrous sodium sulfate and filtered to afford crude tert-butyl (4S)-4-[(1R)-1-cyclobutyl-3-oxo-propyl]-2,2-dimethyl-oxazolidine-3- carboxylate (9.6 g, 90%) as a red oil. ESI-MS m/z calc.311.2097, found 334.2 (M+23)+; Retention time: 2.12 minutes 1H NMR (400 MHz, DMSO-d6) δ 9.67 (s, 1H), 3.97 - 3.59 (m, 3H), 2.45 - 2.16 (m, 3H), 2.03 - 1.56 (m, 7H), 1.54 - 1.45 (m, 3H), 1.45 - 1.34 (m, 12H). LC method U. Step 10: tert-Butyl (4S)-4-[(1R)-1-cyclobutyl-5-[methoxy(methyl)amino]-5-oxo- pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00670] To a solution of heptanes washed sodium hydride (suspension in mineral oil) (3.2 g, 60 %w/w, 80.008 mmol) in tetrahydrofuran (50 mL) was added a solution of 2- diethoxyphosphoryl-N-methoxy-N-methyl-acetamide (8.5 g, 35.534 mmol) in tetrahydrofuran (50 mL) dropwise at room temperature. When the formation of hydrogen stopped (around 15 min) the reaction was stirred for an additional 1 h and the resulting suspension was added to a solution of tert-butyl (4S)-4-[(1R)-1-cyclobutyl-3-oxo-propyl]-2,2-dimethyl-oxazolidine-3- carboxylate (9.6 g, 20.284 mmol) in tetrahydrofuran (100 mL) in small increments (around 10 mL). The resulting solution was stirred for 2 h at room temperature and an aqueous solution of sodium bicarbonate (sat., 100 mL) was added dropwise followed by EtOAc (100 mL). The phases were separated, and the aqueous one was extracted with EtOAc (2 x 100 mL). The combined organic phases were dried over anhydrous sodium sulfate and filtered. The crude mixture was concentrated in vacuo and was then purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 80% of MeOH in water to afford, after evaporation, tert-butyl (4S)-4-[(1R)-1-cyclobutyl-5-[methoxy(methyl)amino]-5-oxo-pent-3-enyl]-2,2- dimethyl-oxazolidine-3-carboxylate (7.35 g, 62%) as a red oil. ESI-MS m/z calc.396.2624, found 397.4 (M+1)+; Retention time: 2.14 minutes 1H NMR (400 MHz, DMSO-d6) δ 6.87 - 6.70 (m, 1H), 6.53 - 6.35 (m, 1H), 3.93 - 3.69 (m, 3H), 3.66 (s, 3H), 3.12 (s, 3H), 2.45 - 2.12 (m, 3H), 2.08 - 1.58 (m, 7H), 1.57 - 1.45 (m, 3H), 1.44 - 1.29 (m, 12H). LC method U. Step 11: tert-Butyl (4S)-4-[(1R)-1-cyclobutyl-5-[methoxy(methyl)amino]-5-oxo- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00671] A solution of tert-butyl (4S)-4-[(1R)-1-cyclobutyl-5-[methoxy(methyl)amino]-5-oxo- pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (7.35 g, 12.568 mmol) in methanol (75 mL) was purged with nitrogen for 10 min and palladium on carbon (10%, 50% wet) (742 mg, 5 %w/w, 0.3486 mmol) was added. The suspension was purged for 5 min with nitrogen and hydrogen (50 psi) was added. The reaction was stirred for 18 h. The reaction was purged with nitrogen for 5 min and filtered through a pad of Celite. The pad was washed with a mixture of EtOAc/heptanes (1:1) (2 x 100 mL). The crude mixture was concentrated in vacuo and was then purified by flash-chromatography on a silica gel cartridge, eluting with a gradient of 0 to 20% of EtOAc in heptanes. The volatiles were removed under reduced pressure. Anhydrous THF (30 mL) was added and removed under reduced pressure (repeated 3 times) to afford tert-butyl (4S)- 4-[(1R)-1-cyclobutyl-5-[methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (4.94 g, 96%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 4.12 - 3.72 (overlaps with THF, m, 3H), 3.69 (s, 3H), 3.18 (s, 3H), 2.58 - 2.28 (m, 3H), 2.17 - 1.53 (overlaps with THF, m, 10H), 1.52 - 1.34 (m, 15H), 1.20 - 0.97 (m, 1H). ESI-MS m/z calc.398.2781, found 299.4 (M-99)+; Retention time: 3.77 minutes. LC method 1B. Step 12: tert-Butyl (4S)-4-[(1R)-1-cyclobutyl-5-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00672] To a solution of 3-bromo-5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazine (5.8 g, 18.12 mmol) in tetrahydrofuran (91 mL) at -78 °C was added n-butyllithium (solution in hexanes) (6.5 mL of 2.5 M, 16.25 mmol) dropwise. The resulting solution was stirred for 5 minutes. A solution of tert-butyl (4S)-4-[(1R)-1-cyclobutyl-5- [methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (4.94 g, 12.40 mmol) in tetrahydrofuran (37 mL) was added dropwise. Reaction was left stirring for 15 min. The reaction mixture was diluted with EtOAc, and ammonium chloride was added. The crude mixture was allowed to warm to room temperature and the phases separated. The aqueous phases were extracted with EtOAc (2 x) and the combined organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude material was purified by reverse phase HPLC (1-99% acetonitrile/5 mM aqueous HCl over 15 min) to give tert-butyl (4S)-4-[(1R)-1-cyclobutyl-5-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (3.120 g, 43%) as an orange foam. ESI-MS m/z calc.578.308, found 579.9 (M+1)⁺; Retention time: 2.11 minutes. LC method C. Step 13: [(2S,3R)-3-Cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H- azepin-2-yl]methanol [00673] To a solution of tert-butyl (4S)-4-[(1R)-1-cyclobutyl-5-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (3.12 g, 5.392 mmol) in 1,4-dioxane (35 mL) was added hydrochloric acid (solution in 1,4-dioxane) (14 mL of 4 M, 56.00 mmol) at rt. Water (0.4 mL) was added and the reaction was stirred for 18 hours. The solvent was removed under reduced pressure and the residual water was co-evaporated with acetonitrile (3 x), then dried under high vacuum to afford crude [(2S,3R)-3-cyclobutyl-7-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2-yl]methanol (2.200 g, 97%) as a yellow solid. ESI-MS m/z calc.420.21368, found 421.2 (M+1)⁺; Retention time: 0.42 minutes. The crude material was taken forward without further purification. LC method A. Step 14: [(2S,3R,7S)-3-Cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol and [(2S,3R,7R)-3-cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol [00674] To a solution of [(2S,3R)-3-cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3,4,5,6-tetrahydro-2H-azepin-2- yl]methanol (2.2 g, 5.232 mmol) in anhydrous dichloromethane (22 mL) was added acetic acid (600 µL, 10.55 mmol) and DIPEA (2.8 mL, 16.07 mmol) in that order. The reaction was stirred for 20 minutes upon which sodium triacetoxyborohydride (3.7 g, 17.46 mmol) was added. The reaction was stirred for 15 minutes then quenched with MeOH and water. The volatiles were removed under reduced pressure. The crude product was purified by reverse phase HPLC (1- 99% acetonitrile/5 mM aqueous HCl over 30 min) to give 1.7 g of a diastereomeric mixture. This material was subjected to chiral SFC separation using a ChiralPak IA column (250 x 20 mm) at 30 oC. The mobile phase was 30% MeOH (0.1% diethylamine) at 50 mL/min flow in gradient mode, 100 bar outlet pressure and the detection wavelength was 22 nm. Each isomer was dissolved in THF and concentrated down three times to give: Isomer 1, Peak 1 [(2S,3R,7S)- 3-cyclobutyl-7-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan- 2-yl]methanol (502 mg, 23%) as a white solid ESI-MS m/z calc.422.22934, found 423.2 (M+1)⁺; Retention time: 1.20 minutes; and Isomer 2, Peak 2 [(2S,3R,7R)-3-cyclobutyl-7-[5- methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (1.092 g, 49%) as a yellow solid ESI-MS m/z calc.422.22934, found 423.2(M+1)⁺; Retention time: 1.21 minutes. LC method A. Step 15: 3-[[4-[[(2S,3R,7R)-3-Cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]- 6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid [00675] 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (1.080 g, 2.585 mmol) and [(2S,3R,7R)-3-cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (1.092 g, 2.585 mmol) were combined in a flask and dissolved in THF (16 mL) followed by the addition of NaOtBu (1.26 g, 13.11 mmol). The reaction mixture was stirred for 1 hour at room temperature then warmed to 40 °C for 2 hours. The reaction mixture was cooled to room temperature and diluted with EtOAc and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (5 x). The organic layers were combined, washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3-[[4-[[(2S,3R,7R)- 3-cyclobutyl-7-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan- 2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (2.09 g, 96%) as a yellow solid. ESI-MS m/z calc.803.3077, found 804.1 (M+1)⁺; Retention time: 1.50 minutes. LC method A. Step 16: 3-[[4-[[(2S,3R,7S)-3-Cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]- 6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid [00676] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (496.4 mg, 1.188 mmol) and [(2S,3R,7S)-3-cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methanol (502 mg, 1.188 mmol) were combined in a flask and dissolved in THF (7.4 mL) followed by the addition of NaOtBu (571.3 mg, 5.944 mmol). The reaction mixture was stirred for 1 hour at room temperature then warmed to 40 °C for 2 hours. The reaction mixture was cooled to room temperature and diluted with EtOAc and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (5 x). The organic layers were combined, washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 3- [[4-[[(2S,3R,7S)-3-cyclobutyl-7-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazin-3-yl]azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (856.5 mg, 86%) as a yellow solid. ESI-MS m/z calc.803.3077, found 804.1 (M+1)⁺; Retention time: 1.46 minutes. LC method A. Step 17: (4R,8R,9S)-8-Cyclobutyl-14-(2,6-dimethylphenyl)-4-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa- 18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (Compound 126)
[00677] 3-[[4-[[(2S,3R,7R)-3-Cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (2.09 g, 2.487 mmol) was dissolved in NMP (42 mL) and acetone (84 mL). TsCl (1.45 g, 7.606 mmol) was added followed by NMM (1.7 mL, 15.46 mmol) and the reaction was stirred at 55 °C for 22 hours. The reaction was poured into a mixture of ethyl acetate and 1 M HCl and the layers were separated. The aqueous layer was further extracted with ethyl acetate (2 x). The organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was taken up in DMSO/MeOH then injected into a reverse phase HPLC (10-99% acetonitrile/5 mM aqueous HCl over 25 min). The fractions were extracted with EtOAc, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give 850.6 mg of material that still contained an impurity. The crude product was taken up in DMSO/MeOH then injected into a reverse phase HPLC (10-99% acetonitrile/5 mM aqueous HCl over 25 min) and the desired fractions were concentrated to give (4R,8R,9S)-8-cyclobutyl- 14-(2,6-dimethylphenyl)-4-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin- 3-yl]-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (410 mg, 19%) as a tan solid.1H NMR (400 MHz, CDCl3) δ 8.91 (s, 1H), 8.52 (s, 1H), 7.86 (d, J = 7.8 Hz, 1H), 7.81 (d, J = 7.6 Hz, 1H), 7.53 (t, J = 7.8 Hz, 1H), 7.25 - 7.19 (m, 1H), 7.10 (d, J = 7.6 Hz, 2H), 6.79 (s, 1H), 6.31 (s, 1H), 6.06 (dd, J = 11.6, 5.1 Hz, 1H), 5.83 (dd, J = 13.2, 4.3 Hz, 1H), 4.90 (t, J = 11.8 Hz, 1H), 4.47 - 4.36 (m, 1H), 4.06 (s, 3H), 2.48 - 2.35 (m, 1H), 2.35 - 2.24 (m, 1H), 2.10 (s, 7H), 2.01 - 1.88 (m, 2H), 1.84 - 1.70 (m, 5H), 1.64 - 1.52 (m, 5H), 1.29 - 1.15 (m, 2H), 0.95 - 0.83 (m, 1H). ESI-MS m/z calc.785.2971, found 786.1(M+1)⁺; Retention time: 2.13 minutes. LC method A. Step 18: (4S,8R,9S)-8-Cyclobutyl-14-(2,6-dimethylphenyl)-4-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa- 18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (Compound 127) [00678] 3-[[4-[[(2S,3R,7S)-3-Cyclobutyl-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (856.5 mg, 1.019 mmol) was dissolved in NMP (17 mL) and Acetone (35 mL). TsCl (600 mg, 3.147 mmol) was added followed by NMM (700 µL, 6.367 mmol) and the reaction was stirred at 55 °C for 22 hours. The reaction was poured into a mixture of ethyl acetate and 1 M HCl and the layers were separated. The aqueous layer was further extracted with ethyl acetate (2 x). The organic layers were washed with brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The crude product was taken up in DMSO/MeOH then injected into a reverse phase HPLC (30-99% acetonitrile/5 mM aqueous HCl over 25 min). The pure fractions were extracted with EtOAc (3 x), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give (4S,8R,9S)-8-cyclobutyl-14-(2,6-dimethylphenyl)-4-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen- 2-one (160 mg, 19%) as a tan solid.1H NMR (400 MHz, CDCl3) δ 8.93 (s, 1H), 8.62 (s, 1H), 7.90 (d, 1H), 7.78 (d, J = 7.5 Hz, 1H), 7.57 (t, J = 7.7 Hz, 1H), 7.25 - 7.19 (m, 1H), 7.09 (d, J = 7.6 Hz, 2H), 6.71 (s, 1H), 6.32 (s, 1H), 5.77 (dd, J = 11.2, 4.3 Hz, 1H), 4.66 (dd, J = 12.0, 5.0 Hz, 1H), 4.52 (t, J = 11.4 Hz, 1H), 4.07 (dd, J = 11.3, 4.3 Hz, 1H), 4.00 (s, 3H), 3.08 - 2.95 (m, 1H), 2.43 - 2.31 (m, 1H), 2.32 - 2.19 (m, 1H), 2.05 (s, 6H), 1.91 - 1.71 (m, 6H), 1.61 - 1.35 (m, 5H), 1.28 - 1.11 (m, 2H), 0.90 - 0.77 (m, 1H), 0.77 - 0.64 (m, 1H). ESI-MS m/z calc.785.2971, found 786.8 (M+1)⁺; Retention time: 2.15 minutes. LC method A. Example 81: Preparation of Compound 128 Step 1: tert-Butyl (4S)-4-[(1R)-1-hydroxypent-4-enyl]-2,2-dimethyl-oxazolidine- 3-carboxylate [00679] To MeCN (175 mL) was added tert-butyl (4S)-2,2-dimethyl-4-pent-4-enoyl- oxazolidine-3-carboxylate (35 g, 113.64 mmol) at room temperature. Zinc trifluoromethanesulfonate (41.3 g, 113.61 mmol) was added in one portion and the reaction mixture was stirred for 45 min. Then Borane-tert-butylamine complex (11.9 g, 136.83 mmol) was added by portions over 20 min at room temperature in solution in MeCN (90 mL). The reaction was stirred for 1 h 30 min. The crude reaction mixture was cooled with an ice bath and quenched with acetone (35 mL) and left for stirring for 16 h at rt. Then the reaction mixture was diluted with MTBE (250 mL) and aqueous saturated ammonium chloride (200 mL). Phases were separated a first time and the organics were washed once with saturated aqueous Sodium bicarbonate (300 mL) solution, followed by 1 M aq NaOH (300 mL) and once with brine (350 mL), dried filtered and evaporated under reduced pressure to afford as a yellow oil tert-butyl (4S)-4-[(1R)-1-hydroxypent-4-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (33.7 g, 95%) ESI- MS m/z calc.285.194, found 230.2 (M-55)+; Retention time: 3.22 minutes.1H NMR (400 MHz, Chloroform-d) δ 5.82 (ddt, J = 17.0, 10.2, 6.7 Hz, 1H), 5.09 - 4.91 (m, 2H), 4.21 - 3.58 (m, 5H), 2.39 - 2.24 (m, 1H), 2.20 - 2.06 (m, 1H), 1.64 - 1.37 (m, 17H). LC method Z. Step 2: tert-Butyl (4S)-4-[(1R)-1-[tert-butyl(dimethyl)silyl]oxypent-4-enyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00680] To a solution of tert-butyl (4S)-4-[(1R)-1-hydroxypent-4-enyl]-2,2-dimethyl- oxazolidine-3-carboxylate (1.016 g, 3.2576 mmol) in anhydrous DCM (15 mL) at 0 °C were added imidazole (1.096 g, 16.099 mmol) followed by TBSCl (1.47 g, 9.7531 mmol) dropwise. The bath was removed, and the reaction as stirred at room temperature for 3 days. The reaction mixture was diluted with a saturated aqueous solution of sodium bicarbonate (50 mL) and extracted with DCM (2 x 50 mL). The combined organic layers were dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. Purification by reversed- phase chromatography (Column: C18. Gradient: 5-100 % acetonitrile in water then at 100% MeOH) afforded tert-butyl (4S)-4-[(1R)-1-[tert-butyl(dimethyl)silyl]oxypent-4-enyl]-2,2- dimethyl-oxazolidine-3-carboxylate (1.08 g, 83%) as a yellow oil.1H NMR (400 MHz, DMSO- d6) δ 5.90 - 5.71 (m, 1H), 5.07 - 4.89 (m, 2H), 4.19 - 3.72 (m, 4H), 2.13 - 1.96 (m, 2H), 1.53 - 1.34 (m, 17H), 0.93 - 0.78 (m, 9H), 0.12 - -0.05 (m, 6H). ESI-MS m/z calc.399.2805, found 300.4 (M-99)+; Retention time: 2.84 minutes. LC method 1D. Step 3: tert-Butyl (4S)-4-[(1R)-1-[tert-butyl(dimethyl)silyl]oxy-5-hydroxy- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate OTBDMS OTBDMS [00681] To a solution of tert-butyl (4S)-4-[(1R)-1-[tert-butyl(dimethyl)silyl]oxypent-4-enyl]- 2,2-dimethyl-oxazolidine-3-carboxylate (1.08 g, 2.6997 mmol) in anhydrous THF (7 mL) at 0 °C was added a solution of borane tetrahydrofuran complex (in THF) (5 mL of 1 M, 5.0000 mmol) dropwise. The reaction mixture was stirred at that temperature for 1 hour then a solution of sodium hydroxide (in water) (7 mL of 2 M, 14.000 mmol) was slowly added dropwise. The reaction mixture was stirred for 30 minutes then a solution of hydrogen peroxide (in water) (430 mg, 35 %w/w, 4.4246 mmol) was added dropwise. The bath was removed, and the reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was put in an ice- cold bath and a solution of sodium bisulfite (in water) (2.7 mL of 21 %w/v, 5.4487 mmol) was added dropwise when the internal temperature was about 5 °C. The bath was removed, and the bi-phasic mixture was stirred at room temperature for 15 minutes. The phases were separated, and the aqueous layer was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude tert-butyl (4S)-4-[(1R)-1-[tert- butyl(dimethyl)silyl]oxy-5-hydroxy-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.2 g, 99%) as a yellow oil that was used in the following step without further purification.1H NMR (400 MHz, CDCl3) δ 4.32 - 3.54 (overlapped with EtOAc, m, 6H), 1.71 - 1.33 (m, 22H), 0.95 - 0.83 (m, 9H), 0.14 - -0.04 (m, 6H). ESI-MS m/z calc.417.2911, found 318.4 (M-99)+; Retention time: 2.4 minutes. LC method U. Step 4: (5R)-5-[(4S)-3-tert-Butoxycarbonyl-2,2-dimethyl-oxazolidin-4-yl]-5-[tert- butyl(dimethyl)silyl]oxy-pentanoic acid [00682] To a solution of tert-butyl (4S)-4-[(1R)-1-[tert-butyl(dimethyl)silyl]oxy-5- hydroxy-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.2 g, 2.6721 mmol) in DCM (12 mL) and water (6 mL) at 0 °C were added NaBr (50 mg, 0.4859 mmol), sodium bicarbonate (458 mg, 5.4520 mmol) and TEMPO (20 mg, 0.1280 mmol). To the reaction mixture at 0 °C was added a solution of NaOCl (in water) (5 mL of 10 %w/v, 6.7166 mmol) dropwise. The reaction mixture was stirred for 1 hour then a solution of NaOCl (in water) (5 mL of 10 %w/v, 6.7166 mmol) was again added dropwise. The reaction was stirred for 30 min. Then, an aqueous solution of NaOH (2 mL of 2 M, 4.0000 mmol) was added followed by an aqueous 10 % w/w of Na2S2O3 (5 mL). The resulting white emulsion was extracted with Me-THF (5 x 50 mL) and the combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford crude (5R)-5-[(4S)-3-tert-butoxycarbonyl-2,2-dimethyl-oxazolidin-4- yl]-5-[tert-butyl(dimethyl)silyl]oxy-pentanoic acid (1.55 g, 101%) as a yellow oil that was used in the following step without further purification. ESI-MS m/z calc.431.2703, found 332.4 (M- 99)+; Retention time: 2.32 minutes. LC method U. Step 5: tert-Butyl (4S)-4-[(1R)-1-[tert-butyl(dimethyl)silyl]oxy-5- [methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00683] To a solution of (5R)-5-[(4S)-3-tert-butoxycarbonyl-2,2-dimethyl-oxazolidin-4-yl]-5- [tert-butyl(dimethyl)silyl]oxy-pentanoic acid (1.15 g, 2.6643 mmol) in anhydrous DMF (6 mL) at room temperature were added HATU (1.25 g, 3.2875 mmol) and 4-methylmorpholine (1.4720 g, 1.6 mL, 14.553 mmol). The mixture was stirred at room temperature for 5 minutes then N,O- dimethylhydroxylamine (hydrochloride salt) (415 mg, 4.2545 mmol) was added. The mixture was stirred at room temperature for 18 hours then was quenched with water (10 mL). The mixture was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with a saturated aqueous solution of sodium bicarbonate (20 mL) and brine (20 mL) then dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was first purified by reversed-phase chromatography (Column: C18. Gradient: 5-100 % acetonitrile in water) then by silica gel chromatography (Gradient: 0-100 % EtOAc in heptane) to afford tert-butyl (4S)-4-[(1R)-1-[tert-butyl(dimethyl)silyl]oxy-5-[methoxy(methyl)amino]-5- oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (626 mg, 49%) as a pale yellow oil.1H NMR (400 MHz, CDCl3) δ 4.30 - 4.05 (m, 2H), 3.97 - 3.79 (m, 2H), 3.72 - 3.64 (m, 3H), 3.18 (s, 3H), 2.53 - 2.33 (m, 2H), 1.76 - 1.42 (overlapped with water, m, 19H), 0.93 - 0.82 (m, 9H), 0.17 - -0.05 (m, 6H). ESI-MS m/z calc.474.3125, found 375.4 (M-99)+; Retention time: 2.46 minutes. LC method U. Step 6: tert-Butyl (4S)-4-[(1R)-1-hydroxy-5-[methoxy(methyl)amino]-5-oxo- pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00684] To a solution of tert-butyl (4S)-4-[(1R)-1-[tert-butyl(dimethyl)silyl]oxy-5- [methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (310 mg, 0.6472 mmol) in anhydrous THF (8 mL) at room temperature was added a solution of TBAF (in THF) (4 mL of 1 M, 4.0000 mmol). The reaction was stirred for 18 hours then was quenched with a saturated aqueous solution of sodium bicarbonate (20 mL) and EtOAc (20 mL). The phases were separated, and the aqueous layer was extracted with EtOAc (2 x 10 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reversed- phase chromatography (Column: C18. Gradient: 5-100 % acetonitrile in water) to afford, after co-evaporation with acetonitrile (5 x 10 mL), tert-butyl (4S)-4-[(1R)-1-hydroxy-5- [methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (195 mg, 82%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 4.17 - 3.62 (m, 7H), 3.18 (s, 3H), 2.60 - 2.39 (m, 2H), 1.98 - 1.71 (m, 2H), 1.68 - 1.35 (m, 18H). ESI-MS m/z calc.360.226, found 361.4 (M+1)+; Retention time: 1.72 minutes. LC method U. Step 7: tert-Butyl (4S)-4-[(1R)-5-[methoxy(methyl)amino]-5-oxo-1- (trifluoromethoxy)pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00685] In a flask under nitrogen containing tert-butyl (4S)-4-[(1R)-1-hydroxy-5- [methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (195 mg, 0.5329 mmol) were successively added AgOTf (444 mg, 1.7281 mmol), Selectfluor (320 mg, 0.9033 mmol) and KF (142 mg, 2.4442 mmol). The flask was purged with nitrogen 3 times under vacuum then to the solids were successively added trimethyl(trifluoromethyl)silane (245.60 mg, 0.2 mL, 1.7272 mmol), 2-fluoropyridine (169.20 mg, 0.15 mL, 1.7427 mmol) and anhydrous EtOAc (5 mL). The reaction was stirred for 24 hours at room temperature then trimethyl(trifluoromethyl)silane (245.60 mg, 0.2 mL, 1.7272 mmol) was again added. The reaction was stirred for 24 hours then was filtered through a short pad of Celite. The pad was rinsed with EtOAc (2 x 20 mL) and the combined filtrates were concentrated under reduced pressure. Purification by reversed-phase chromatography (Column: C18. Gradient: 5-100 % acetonitrile in water) afforded, after co-evaporation with acetonitrile (5 x 10 mL), tert-butyl (4S)-4-[(1R)-5-[methoxy(methyl)amino]-5-oxo-1-(trifluoromethoxy)pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (61 mg, 25%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 4.95 - 4.57 (m, 1H), 4.21 - 3.89 (m, 3H), 3.69 (s, 3H), 3.18 (s, 3H), 2.57 - 2.34 (m, 2H), 1.89 - 1.39 (m, 19H).19F NMR (377 MHz, CDCl3) δ -57.13 (s, 3F). ESI-MS m/z calc.428.2134, found 329.2 (M-99)+; Retention time: 2.06 minutes. LC method U. Step 8: tert-Butyl (4S)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-5-oxo-1-(trifluoromethoxy)pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00686] To a solution of 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine (30 mg, 0.1100 mmol) in anhydrous THF (0.5 mL) at -78 °C was slowly added dropwise a solution of n-BuLi (in hexanes) (0.1 mL of 1.6 M, 0.1600 mmol) followed by a solution of tert-butyl (4S)- 4-[(1R)-5-[methoxy(methyl)amino]-5-oxo-1-(trifluoromethoxy)pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (30 mg, 0.0696 mmol) in anhydrous THF (0.4 mL). The reaction was stirred for 10 minutes then methanol (1 mL) was added, and the mixture was concentrated under reduced pressure. The crude residue was purified first by reversed-phase chromatography (C18 column. Gradient: 5-80 % MeCN in water with 0.1 % formic acid) then by silica gel chromatography (Gradient: 0-40 % EtOAc in heptane) to afford tert-butyl (4S)-4-[(1R)-5-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-5-oxo-1-(trifluoromethoxy)pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate (10 mg, 22%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 9.12 (s, 1H), 6.60 (s, 1H), 4.96 - 4.61 (m, 1H), 4.16 - 3.95 (m, 6H), 3.41 - 3.21 (m, 2H), 2.03 - 1.63 (m, 4H), 1.62 - 1.39 (overlapped with water, m, 24H).19F NMR (377 MHz, CDCl3) δ - 57.07 (s, 3F). ESI-MS m/z calc.556.2873, found 557.4 (M+1)+; Retention time: 4.41 minutes. LC method 1B. Step 9: (5R,6S)-6-Amino-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-7- hydroxy-5-(trifluoromethoxy)heptan-1-one [00687] To a solution of tert-butyl (4S)-4-[(1R)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-5-oxo-1-(trifluoromethoxy)pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (772 mg, 1.3023 mmol) in 1,4-dioxane (8 mL) and water (0.1 mL) at room temperature was added a solution of HCl (in 1,4-dioxane) (4 mL of 4 M, 16.000 mmol). The reaction was stirred for 2 hours at room temperature then was co-evaporated with 1,4-dioxane (3 x 5 mL) followed by acetonitrile (3 x 10 mL) to afford crude (5R,6S)-6-amino-1-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-7-hydroxy-5-(trifluoromethoxy)heptan-1-one (hydrochloride salt) (660 mg, 100%) as an orange solid that was used in the following step without further purification. ESI- MS m/z calc.416.2035, found 399.2 (M-17)+; Retention time: 1.47 minutes. LC method U. Step 10: 3-[[4-[(2S,3R)-2-Amino-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin- 3-yl)-7-oxo-3-(trifluoromethoxy)heptoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00688] To a solution of (5R,6S)-6-amino-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-7-hydroxy-5-(trifluoromethoxy)heptan-1-one (hydrochloride salt) (540 mg, 1.0612 mmol) and 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (501 mg, 1.1989 mmol) in anhydrous THF (5 mL) was added sodium tert-butoxide (710 mg, 7.3879 mmol). The reaction was stirred at room temperature for 1 hour then was added an aqueous HCl 1M solution (20 mL) and the aqueous phase was extracted with pre-distilled Me-THF (5 x 20 mL). The combined organic layers were concentrated under reduced pressure to afford crude 3-[[4- [(2S,3R)-2-amino-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-7-oxo-3- (trifluoromethoxy)heptoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (996 mg, 67%) as an orange solid that was used in the following step without further purification. ESI-MS m/z calc.797.2819, found 798.2 (M+1)+; Retention time: 2.66 minutes. LC method 1B. Step 11: 3-[[4-[[(2S,3R)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (trifluoromethoxy)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid
[00689] To a solution of 3-[[4-[(2S,3R)-2-amino-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-7-oxo-3-(trifluoromethoxy)heptoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (995 mg, 0.7060 mmol) in anhydrous DCM (75 mL) at room temperature was added NaBH(OAc)3 (1.16 g, 5.4732 mmol). The reaction was stirred at room temperature for 30 minutes then an aqueous HCl 1M solution (100 mL) was added. The volatiles were removed under reduced pressure and the aqueous phase was extracted with pre-distilled Me-THF (5 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography (Column: C18. Gradient: 5-100 % acetonitrile in water with 0.1% HCl). The fractions containing the product were collected and the volatiles were removed under reduced pressure. The aqueous solution was diluted with brine (100 mL) and extracted with pre-distilled MeTHF (3 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford 3-[[4-[[(2S,3R)-7-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(trifluoromethoxy)azepan-2-yl]methoxy]-6- (2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (429 mg, 50%) as a yellow solid that was used in the following step without further purification. ESI-MS m/z calc.781.2869, found 782.2 (M+1)+; Retention time: 3.37 minutes. LC method 1B. Step 12: (8R,9S)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-18,18-dioxo-8-(trifluoromethoxy)-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one
[00690] To a solution of 3-[[4-[[(2S,3R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-(trifluoromethoxy)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (253 mg, 0.2068 mmol) in DCE (8 mL) was added a solution of T3P (in MeTHF) (1.5 mL of 50 %w/v, 2.3571 mmol) at room temperature. The mixture was stirred at 70 °C for 48 hours. The mixture was allowed to cool down to room temperature and was concentrated under reduced pressure. Purification by reversed-phase chromatography (Column: C18. Gradient: 20-80 % acetonitrile in water) afforded (8R,9S)-4-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-18,18-dioxo-8- (trifluoromethoxy)-11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one as a mixture of isomers (75 mg, 37%) as a yellow solid. ESI-MS m/z calc.763.2764, found 764.2 (M+1)+; Retention time: 2.07 minutes. LC method U. Step 13: (4S,8R,9S)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-18,18-dioxo-8-(trifluoromethoxy)-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one (Compound 128) [00691] Two lots of isomeric mixture (8R,9S)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin- 3-yl)-14-(2,6-dimethylphenyl)-18,18-dioxo-8-(trifluoromethoxy)-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (75 mg, 0.0769 mmol) and (32 mg, 0.0346 mmol) were combined in DMSO (3 mL) and the solution was purified by reversed-phase chromatography (Column: C18. Gradient: 30-70 % acetonitrile in water) to afford 60 mg of the desired product after freeze-drying. The solid was submitted to chiral SFC purification (Flow rate: 120 mL/min, 50% MeOH, column: Cellulose 5, temperature = 40 °C, outlet pressure: 100 bar, injection volume: 3000 μL). The fractions containing the major product were concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (Column: C18. Gradient: 50-70 % acetonitrile in water) to afford, after freeze- drying, (4S,8R,9S)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-18,18-dioxo-8-(trifluoromethoxy)-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen-2-one (20.1 mg, 23%) as an off-white solid.1H NMR (400 MHz, DMSO-d6) δ 8.54 (s, 1H), 8.27 (s, 1H), 7.93 (dd, J = 12.1, 7.7 Hz, 2H), 7.36 (t, J = 7.8 Hz, 1H), 7.08 - 6.99 (m, 1H), 6.86 (d, J = 7.6 Hz, 2H), 6.45 (s, 1H), 5.24 (t, J = 4.6 Hz, 1H), 5.16 - 5.04 (m, 2H), 4.99 - 4.84 (m, 1H), 4.60 (dd, J = 12.7, 10.8 Hz, 1H), 4.31 (dd, J = 12.8, 8.4 Hz, 1H), 3.83 (s, 3H), 2.45 - 2.34 (m, 2H), 2.26 - 2.16 (m, 1H), 1.98 - 1.85 (m, 1H), 1.73 - 1.62 (m, 1H), 1.54 (s, 6H), 1.47 (s, 9H), 1.34 - 1.22 (overlapped with grease, m, 1H), (1H missing, labile proton).19F NMR (377 MHz, DMSO- d6) δ -55.39 (s, 3F). ESI-MS m/z calc.763.2764, found 764.2 (M+1)+; Retention time: 3.08 minutes. LC method W. Example 82: Preparation of Compound 131 and Compound 132 Step 1: Ethyl 2-[(4R)-2-oxooxazolidin-4-yl]acetate [00692] To a stirred solution of benzyl N-[(3R)-5-oxotetrahydrofuran-3-yl]carbamate (22.5 g, 95.457 mmol) in ethanol (450 mL) was added sodium ethoxide (in EtOH solution) (90 mL of 2.65 M, 238.50 mmol) over 15 minutes and the reaction was stirred at room temperature for 2 hours. An aqueous solution of HCl (1 M, 350 mL) and DCM (350 mL) were added, and the mixture was extracted with DCM (3 x 150 mL). The combined organic layers were washed with brine (1 x 350 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude mixture was purified by flash-chromatography on a silica gel cartridge, eluting with a gradient of 0 to 100% of EtOAc in heptane to afford after evaporation ethyl 2-[(4R)-2-oxooxazolidin-4- yl]acetate (10 g, 57%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 6.02 (br. s., 1H), 4.56 (t, J = 8.6 Hz, 1H), 4.29 - 4.12 (m, 3H), 4.06 (dd, J = 8.8, 5.9 Hz, 1H), 2.73 - 2.54 (m, 2H), 1.27 (t, J = 7.2 Hz, 3H). ESI-MS m/z calc.173.0688, found 174.2 (M+1)+; Retention time: 1.1 minutes. LC method S. Step 2: Ethyl (2S)-4-methyl-2-[(4R)-2-oxooxazolidin-4-yl]pent-4-enoate [00693] To a solution of ethyl 2-[(4R)-2-oxooxazolidin-4-yl]acetate (7.7 g, 42.242 mmol) in THF (110 mL) at -78 °C was added NaHMDS (in THF solution) (84.484 mL of 1 M, 84.484 mmol) dropwise over 15 minutes. The solution was stirred 30 minutes. To this solution was added a solution of 3-bromo-2-methyl-prop-1-ene (12.051 g, 9 mL, 89.265 mmol) in THF (40 mL) dropwise at -78 °C. The mixture was stirred at -78 °C for 30 minutes and then at -40 °C for 3 hours. The reaction was quenched with a saturated aqueous solution of ammonium chloride (100 mL) and was then allowed to reach room temperature over 15 minutes. EtOAc (100 mL) were added, and the mixture was extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (100 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude mixture was purified by flash-chromatography on a silica gel cartridge, eluting with a gradient of 0 to 100% of EtOAc in heptane to afford after evaporation ethyl (2S)-4- methyl-2-[(4R)-2-oxooxazolidin-4-yl]pent-4-enoate (5.7 g, 51%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 6.00 (br. s., 1H), 4.81 (s, 1H), 4.74 (s, 1H), 4.49 (t, J = 8.7 Hz, 1H), 4.24 - 4.04 (m, 4H), 2.76 (td, J = 8.3, 5.5 Hz, 1H), 2.36 (dd, J = 14.2, 8.8 Hz, 1H), 2.28 - 2.19 (m, 1H), 1.74 (s, 3H), 1.26 (t, J = 7.2 Hz, 3H). ESI-MS m/z calc.227.1158, found 228.2 (M+1)+; Retention time: 1.53 minutes. LC method S. Step 3: (4R)-4-[(1S)-1-(Hydroxymethyl)-3-methyl-but-3-enyl]oxazolidin-2-one [00694] To a solution of oxazolidinone ethyl (2S)-4-methyl-2-[(4R)-2-oxooxazolidin-4- yl]pent-4-enoate (5.5 g, 22.992 mmol) and dry CaCl2 (17 g, 153.18 mmol) in dry ethanol (330 mL) at 0 °C was added slowly NaBH4 (13 g, 343.62 mmol) and the mixture was allowed to warm to room temperature. After 16 h, a saturated aqueous solution of CaCO3 (250 mL) and an aqueous solution of sodium potassium tartrate (450 mL, 1.0 M) were added. The mixture was extracted with EtOAc (2x 150 mL). The combined organic phases were dried over sodium sulfate and evaporated to afford (4R)-4-[(1S)-1-(hydroxymethyl)-3-methyl-but-3- enyl]oxazolidin-2-one (3.8 g, 81%) as a colorless oil;1H NMR (400 MHz, CDCl3) δ 6.76 (s, 1H), 4.82 (s, 1H), 4.75 (s, 1H), 4.50 (t, J = 8.7 Hz, 1H), 4.18 (dd, J = 8.6, 7.1 Hz, 1H), 3.92 (q, J = 8.0 Hz, 1H), 3.80 - 3.69 (m, 1H), 3.62 - 3.51 (m, 1H), 2.98 - 2.85 (m, 1H), 2.09 - 1.97 (m, 2H), 1.94 - 1.82 (overlapped with acetonitrile peak, m, 1H), 1.72 (s, 3H). ESI-MS m/z calc.185.1052, found 186.2 (M+1)+; Retention time: 1.35 minutes. The crude was used for next step without further purification. LC method 1D. Step 4: (4R)-4-[(1S)-1-[[tert-Butyl(diphenyl)silyl]oxymethyl]-3-methyl-but-3- enyl]oxazolidin-2-one [00695] To a stirred solution of (4R)-4-[(1S)-1-(hydroxymethyl)-3-methyl-but-3- enyl]oxazolidin-2-one (3.8 g, 18.711 mmol) in dichloromethane (150 mL) at 0 °C was added imidazole (7.5 g, 110.17 mmol) followed by tert-butyl-chloro-diphenyl-silane (7.9500 g, 7.5 mL, 28.924 mmol). The mixture was stirred at 0 °C for 15 minutes and then at room temperature for 4 hours. Water (75 mL) and DCM (75 mL) were added, and the mixture was extracted with DCM (3 x 50 mL). The combined organic layers were washed with brine (150 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography, eluting with a gradient of 5 to 90% acetonitrile in water. The fractions containing the desired product were combined and the organic solvent was evaporated to afford (4R)-4-[(1S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-methyl-but-3-enyl]oxazolidin-2-one (6.2 g, 71%) as a brownish oil; 1H NMR (400 MHz, CDCl3) δ 7.69 - 7.59 (m, 4H), 7.52 - 7.37 (m, 6H), 5.14 (s, 1H), 4.73 (s, 1H), 4.61 (s, 1H), 4.45 (t, J = 8.6 Hz, 1H), 4.14 (dd, J = 8.8, 7.1 Hz, 1H), 3.92 (q, J = 7.7 Hz, 1H), 3.66 (d, J = 4.4 Hz, 2H), 2.07 - 1.96 (m, 1H), 1.89 - 1.77 (m, 2H), 1.61 (s, 3H), 1.09 (s, 9H). ESI-MS m/z calc.423.223, found 424.2 (M+1)+; Retention time: 2.35 minutes. LC method 1D. Step 5: tert-Butyl (4R)-4-[(1S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-methyl- but-3-enyl]-2-oxo-oxazolidine-3-carboxylate [00696] To a solution of (4R)-4-[(1S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-methyl-but- 3-enyl]oxazolidin-2-one (6.2 g, 13.260 mmol) in dichloromethane (310 mL) was added triethylamine (2.6862 g, 3.7 mL, 26.546 mmol), DMAP (81 mg, 0.6630 mmol) followed by Boc2O (5.8 g, 6.1053 mL, 26.575 mmol) at room temperature. The mixture was stirred at room temperature for 2 hours. After evaporating all solvent, the obtained mixture was purified by reverse phase column chromatography (5 to 90% acetonitrile in water). The fraction containing the desired product were combined and concentrated to afford tert-butyl (4R)-4-[(1S)-1-[[tert- butyl(diphenyl)silyl]oxymethyl]-3-methyl-but-3-enyl]-2-oxo-oxazolidine-3-carboxylate (6.4 g, 92%) as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 7.68 - 7.59 (m, 4H), 7.50 - 7.36 (m, 6H), 4.72 (s, 1H), 4.66 (dt, J = 8.9, 3.4 Hz, 1H), 4.62 (s, 1H), 4.28 (dd, J = 9.3, 3.9 Hz, 1H), 4.19 - 4.12 (m, 1H), 3.73 (dd, J = 10.8, 3.9 Hz, 1H), 3.51 (dd, J = 11.0, 7.1 Hz, 1H), 2.59 (qt, J = 7.0, 3.7 Hz, 1H), 2.03 (d, J = 7.1 Hz, 2H), 1.61 (s, 3H), 1.55 (s, 9H), 1.07 (s, 9H). ESI-MS m/z calc. 523.2754, found 424.2 (M-99)+; Retention time: 2.49 minutes. LC method 1D. Step 6: tert-Butyl N-[(1R,2S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-1- (hydroxymethyl)-4-methyl-pent-4-enyl]carbamate [00697] To a solution of tert-butyl (4R)-4-[(1S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- methyl-but-3-enyl]-2-oxo-oxazolidine-3-carboxylate (1.95 g, 3.4291 mmol) in methanol (40 mL) was added cesium carbonate (450 mg, 1.3811 mmol). The reaction mixture was stirred at room temperature overnight, quenched with saturated solution of ammonium chloride (25 mL), and methanol was removed. The mixture was extracted with dichloromethane (3 x 25 mL), dried over sodium sulfate. The combined organic phases were filtered and concentrated under high vacuum. The crude was purified by reverse phase (acetonitrile in water, 0 to 100%). The fractions containing desired product were combined and concentrated under high vacuum to afford tert-butyl N-[(1R,2S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-1-(hydroxymethyl)-4- methyl-pent-4-enyl]carbamate (1.25 g, 71%) as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 7.70 - 7.62 (m, 4H), 7.50 - 7.36 (m, 6H), 6.03 (br. s., 1H), 4.75 - 4.69 (m, 1H), 4.64 - 4.58 (m, 1H), 3.83 - 3.72 (m, 3H), 3.68 - 3.61 (m, 1H), 3.58 (dd, J = 11.0, 4.9 Hz, 1H), 2.29 - 2.20 (m, 1H), 2.19 - 2.12 (m, 1H), 2.10 - 2.03 (m, 1H), 1.94 - 1.86 (m, 1H), 1.61 (s, 3H), 1.45 (s, 9H), 1.10 (s, 9H). ESI-MS m/z calc.497.2961, found 398.4 (M-99)+; Retention time: 2.36 minutes. LC method S. Step 7: tert-Butyl (4R)-4-[(1S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-methyl- but-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00698] To a solution of tert-butyl N-[(1R,2S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-1- (hydroxymethyl)-4-methyl-pent-4-enyl]carbamate (3.4 g, 6.5781 mmol) in dichloromethane (75 mL) at room temperature was added 2,2-dimethoxypropane (6.8000 g, 8 mL, 65.292 mmol), followed by p-Toluenesulfonic acid (125 mg, 0.6571 mmol). The reaction mixture was stirred at room temperature for 3 hours. The reaction was concentrated under reduced pressure and then the crude was purified by reverse phase column chromatography (0 to 100% acetonitrile in water). The fractions containing the desired product were combined and concentrated to afford tert-butyl (4R)-4-[(1S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-methyl-but-3-enyl]-2,2- dimethyl-oxazolidine-3-carboxylate (1.1 g, 31%) as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 7.69 - 7.59 (m, 4H), 7.48 - 7.33 (m, 6H), 4.69 (s, 1H), 4.62 (s, 1H), 4.25 - 3.80 (m, 3H), 3.77 - 3.42 (m, 2H), 2.49 - 2.38 (m, 1H), 2.37 - 2.02 (m, 2H), 1.67 - 1.56 (m, 6H), 1.51 - 1.37 (m, 12H), 1.06 (s, 9H). ESI-MS m/z calc.537.3274, found 560.4 (M+23)+; Retention time: 2.96 minutes. LC method S. Step 8: tert-Butyl (4R)-4-[(1S)-1-(hydroxymethyl)-3-methyl-but-3-enyl]-2,2- dimethyl-oxazolidine-3-carboxylate [00699] To a solution of tert-butyl (4R)-4-[(1S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- methyl-but-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (4 g, 7.0063 mmol) in anhydrous THF (80 mL) was added TBAF (in THF) (25 mL of 1 M, 25.000 mmol) at 0 °C. The reaction was stirred in the ice-cold bath for 18 hours. The reaction was quenched at 0 °C with a saturated aqueous sodium bicarbonate solution (50 mL) and EtOAc (50 mL). The bi-phasic solution was stirred for 1 hour at room temperature and the layers were separated. The aqueous layer was extracted with ethyl acetate (2 x 25 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography (5 to 80 % acetonitrile in pure water). The fractions containing the desired product were combined and concentrated under reduced pressure to afford tert-butyl (4R)-4-[(1S)-1-(hydroxymethyl)-3-methyl-but-3-enyl]-2,2-dimethyl- oxazolidine-3-carboxylate (1.65 g, 75%) as a colorless oil.1H NMR (400 MHz, CDCl3) δ 4.87 - 4.77 (m, 2H), 4.00 - 3.83 (m, 4H), 3.50 (d, J = 5.4 Hz, 2H), 2.53 - 2.34 (m, 1H), 1.82 (d, J = 13.7 Hz, 1H), 1.74 (s, 3H), 1.71 - 1.63 (m, 1H), 1.59 - 1.52 (m, 6H), 1.51 (s, 9H). ESI-MS m/z calc.299.2097, found 186.2 (M-113)+; Retention time: 2.01 minutes. LC method 1D. Step 9: tert-Butyl (4R)-4-[(1S)-1-[[2-[methoxy(methyl)amino]-2-oxo- ethoxy]methyl]-3-methyl-but-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00700] To a solution of tert-butyl (4R)-4-[(1S)-1-(hydroxymethyl)-3-methyl-but-3-enyl]-2,2- dimethyl-oxazolidine-3-carboxylate (825 mg, 2.6342 mmol) and 2-chloro-N-methoxy-N-methyl- acetamide (1.9 g, 13.812 mmol) in anhydrous THF (25 mL) at room temperature was stirred for 10 min and then NaH (in mineral oil) (550 mg, 60 %w/w, 13.751 mmol) was added portion- wise. The mixture was stirred at room temperature for 4 hours and then more NaH (in mineral oil) (270 mg, 60 %w/w, 6.7506 mmol) was added and then the resulting mixture was stirred for 18 hours. The mixture was cooled to 0 °C and then water (25 mL) and EtOAc (25 mL) were added, and the mixture was extracted with EtOAc (2 x 25 mL). The combined organic layers were washed with brine (50 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by reversed-phase chromatography (Gradient: 5-90% MeCN in water). The fractions containing the desired product were combined and concentrated in vacuo to afford tert-butyl (4R)-4-[(1S)-1-[[2-[methoxy(methyl)amino]-2-oxo-ethoxy]methyl]-3-methyl-but-3- enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (520 mg, 46%) as a light brownish oil; ESI-MS m/z calc.400.2573, found 301.4 (M-99)+; Retention time: 2.09 minutes. LC method 1D. Step 10: tert-Butyl (4R)-4-[(1S)-1-[[2-[methoxy(methyl)amino]-2-oxo- ethoxy]methyl]-3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00701] A solution of tert-butyl (4R)-4-[(1S)-1-[[2-[methoxy(methyl)amino]-2-oxo- ethoxy]methyl]-3-methyl-but-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (1.1 g, 2.5652 mmol) in methanol (55 mL) was bubbled with nitrogen over 5 minutes and then palladium on carbon (545 mg, 5 %w/w, 0.2561 mmol) was added. The resulting mixture was bubbled with hydrogen for 3 minutes and then was stirred at room temperature under balloon of hydrogen for 4 hours. The crude was filtered through a pad of celite. The pad was washed with ethyl acetate (25 mL) and the combined organic phases were concentrated under reduced pressure to afford tert-butyl (4R)-4-[(1S)-1-[[2-[methoxy(methyl)amino]-2-oxo-ethoxy]methyl]-3-methyl-butyl]- 2,2-dimethyl-oxazolidine-3-carboxylate (1.05 g, 99%) as a colorless oil; 1H NMR (400 MHz, CDCl3) δ 4.37 - 4.12 (m, 3H), 3.99 (d, J = 5.4 Hz, 2H), 3.68 (s, 3H), 3.56 - 3.40 (m, 2H), 3.18 (s, 3H), 2.51 - 2.28 (m, 1H), 1.67 - 1.55 (m, 4H), 1.47 (s, 12H), 1.32 - 1.16 (m, 2H), 0.96 - 0.82 (m, 6H). ESI-MS m/z calc.402.273, found 403.2 (M+1)+; Retention time: 2.14 minutes. LC method 1D. Step 11: tert-Butyl (4R)-2,2-dimethyl-4-[(1S)-3-methyl-1-[[2-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-2-oxo- ethoxy]methyl]butyl]oxazolidine-3-carboxylate [00702] To a solution of 3-bromo-5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazine (700 mg, 2.1846 mmol) in THF (15 mL) at -78°C was added n-butyllithium (solution in hexanes) (1.4 mL of 1.75 M, 2.4500 mmol) dropwise. As soon as the addition of n-BuLi was over, a solution of tert-butyl (4R)-4-[(1S)-1-[[2-[methoxy(methyl)amino]-2-oxo-ethoxy]methyl]- 3-methyl-butyl]-2,2-dimethyl-oxazolidine-3-carboxylate (500 mg, 1.2099 mmol) in THF (5 mL) was added dropwise. The mixture was stirred for 1 hour at -78 °C. A saturated aqueous solution of ammonium chloride (25 mL) and ethyl acetate (25 mL) were added. The mixture was slowly warmed up to room temperature and stirred for 30 minutes. The layers were separated, and the aqueous layer was extracted with ethyl acetate (2 x 25 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude was purified by reversed-phase chromatography on C18 column (gradient 5 to 85% acetonitrile in water). The desired fractions were concentrated under reduced pressure, then co-evaporated with acetonitrile (5 x 5 mL), to afford tert-butyl (4R)-2,2- dimethyl-4-[(1S)-3-methyl-1-[[2-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazin-3-yl]-2-oxo-ethoxy]methyl]butyl]oxazolidine-3-carboxylate (550 mg, 77%) as an brownish oil; 1H NMR (400 MHz, CDCl3) δ 9.16 (s, 1H), 6.86 (s, 1H), 5.22 - 4.97 (m, 2H), 4.32 - 4.17 (m, 1H), 4.11 - 4.01 (m, 2H), 3.98 (s, 3H), 3.67 - 3.50 (m, 2H), 2.59 - 2.35 (m, 1H), 1.68 - 1.60 (m, 7H), 1.49 (s, 12H), 1.33 - 1.26 (m, 3H), 0.94 (d, J = 6.6 Hz, 3H), 0.91 - 0.85 (m, 3H). 19F NMR (377 MHz, CDCl3) δ -68.51 (br. s., 3F). ESI-MS m/z calc.582.3029, found 483.2 (M- 99)+; Retention time: 2.4 minutes. LC method 1D. Step 12: 2-[(2S)-2-[(1R)-1-Amino-2-hydroxy-ethyl]-4-methyl-pentoxy]-1-[5- methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]ethanone [00703] To a solution of tert-butyl (4R)-2,2-dimethyl-4-[(1S)-3-methyl-1-[[2-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-2-oxo- ethoxy]methyl]butyl]oxazolidine-3-carboxylate (550 mg, 0.9289 mmol) in dioxane (17 mL) and water (7 mL) was added hydrochloric acid (solution in 1,4-dioxane) (12 mL of 4 M, 48.000 mmol). The reaction was stirred at room temperature for 2 hours, under nitrogen. The solvents were removed under reduced pressure and the mixture was co-evaporated with 1,4-dioxane (5 x 10 mL). The material was dissolved in acetonitrile (20 mL) and concentrated. The residue was co-evaporated with diethyl ether (5 x 10 mL) and concentrated under reduced pressure to afford crude of 2-[(2S)-2-[(1R)-1-amino-2-hydroxy-ethyl]-4-methyl-pentoxy]-1-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]ethanone (hydrochloride salt) (444 mg, 100%) as an brownish foam; 1H NMR (400 MHz, DMSO-d6) δ 9.02 (s, 1H), 8.13 - 7.91 (m, 3H), 7.03 (s, 1H), 5.17 (s, 2H), 3.93 (s, 3H), 3.75 (dd, J = 11.5, 3.9 Hz, 1H), 3.71 - 3.65 (m, 2H), 3.64 - 3.58 (m, 2H), 3.52 - 3.43 (m, 1H), 3.29 - 3.19 (m, 1H), 2.10 - 1.99 (m, 1H), 1.68 - 1.61 (m, 2H), 1.54 - 1.48 (m, 2H), 1.31 - 1.24 (m, 2H), 0.91 - 0.86 (m, 6H).19F NMR (377 MHz, DMSO-d6) δ -67.10 (s, 3F). ESI-MS m/z calc.442.2192, found 443.2 (M+1)+; Retention time: 1.53 minutes. The crude was used for next step without further purification. LC method 1D. Step 13: [(5R,6S)-6-Isobutyl-3-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-1,4-oxazepan-5- yl]methanol [00704] To a solution of 2-[(2S)-2-[(1R)-1-amino-2-hydroxy-ethyl]-4-methyl-pentoxy]-1-[5- methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]ethanone (265 mg, 0.5983 mmol) in anhydrous dichloromethane (30 mL) was added NaBH(OAc)3 (510 mg, 2.4063 mmol). The reaction mixture was stirred at room temperature for 18 hours then quenched by the addition of a saturated aqueous solution of sodium bicarbonate (25 mL) and stirred 30 minutes at room temperature. The layers were separated, and the aqueous layer was extracted with dichloromethane (3 x 10 mL) Thecombined organic layers were washed with brine (25 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure. The crude was purified by reverse phase chromatography (gradient: 5 to 90% acetonitrile in water). The desired fractions were concentrated under reduced pressure to afford mixture of diastereomers [(5R,6S)- 6-isobutyl-3-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-1,4- oxazepan-5-yl]methanol (90 mg, 29%) as a white solid;1H NMR (400 MHz, CDCl3) δ 8.42 (s, 1H), 6.74 (s, 1H), 4.37 (dd, J = 9.3, 3.2 Hz, 1H), 4.26 (dd, J = 12.2, 3.4 Hz, 1H), 3.99 (dd, J = 12.5, 2.7 Hz, 1H), 3.91 (s, 3H), 3.78 - 3.54 (m, 4H), 3.43 (t, J = 10.5 Hz, 1H), 2.93 - 2.81 (m, 1H), 1.80 - 1.68 (m, 1H), 1.63 - 1.56 (m, 3H), 1.26 - 1.21 (m, 2H), 1.16 - 1.01 (m, 2H), 0.96 - 0.87 (m, 6H), 0.87 - 0.84 (m, 1H).19F NMR (377 MHz, CDCl3) δ -68.74 (s, 3F). ESI-MS m/z calc.426.2243, found 427.2 (M+1)+; Retention time: 1.95 minutes. LC method 1D. Step 14: 3-[[4-(2,6-Dimethylphenyl)-6-[[(5R,6S)-6-isobutyl-3-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-1,4-oxazepan-5- yl]methoxy]pyrimidin-2-yl]sulfamoyl]benzoic acid [00705] To a solution of [(5R,6S)-6-isobutyl-3-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-1,4-oxazepan-5-yl]methanol (130 mg, 0.3045 mmol) and 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (140 mg, 0.3350 mmol) in anhydrous THF (3.9 mL) was added sodium tert-butoxide (120 mg, 1.2487 mmol). The reaction was stirred at room temperature for 4 hour then an aqueous HCl 1 M solution (10 mL) was added. The aqueous phase was extracted with pre-distilled Me-THF (5 x 5 mL). The combined organic layers were concentrated under reduced pressure to afford crude 3-[[4-(2,6-dimethylphenyl)-6-[[(5R,6S)-6-isobutyl-3-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-1,4-oxazepan-5- yl]methoxy]pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (280 mg, 99%) as a white solid; 19F NMR (377 MHz, DMSO-d6) δ -67.32 (s, 3F). ESI-MS m/z calc.807.3026, found 808.2 (M+1)+; Retention time: 1.63 minutes. LC method 1D. Step 15: (4R,8S,9R)-14-(2,6-dimethylphenyl)-8-isobutyl-4-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-6,11- dioxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (Compound 131) and (4S,8S,9R)-14- (2,6-dimethylphenyl)-8-isobutyl-4-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-6,11- dioxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (Compound 132) [00706] 3-[[4-(2,6-Dimethylphenyl)-6-[[(5R,6S)-6-isobutyl-3-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-1,4-oxazepan-5- yl]methoxy]pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (250 mg, 0.2680 mmol) in a nitrogen-purged round bottom flask fitted with a nitrogen balloon, was dissolved in anhydrous NMP (10 mL) and acetone (20 mL). TsCl (160 mg, 0.8392 mmol) was added followed by a rapid dropwise addition of 4-methyl-morpholine (184.00 mg, 0.2 mL, 1.8191 mmol) and the reaction was stirred at 55 °C for 4 hours. The reaction was cooled to room temperature and partially concentrated under rotary evaporation, then was poured into a mixture of ethyl acetate and an aqueous solution of 1 M HCl and the layers were separated. The aqueous layer was further extracted with ethyl acetate (3 x 15 mL). The combined organic layers were washed with water (1 x 25 mL), brine (1 x 25 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reverse phase column chromatography (acetonitrile in pure water 5 to 90%). The pure fractions were concentrated by rotary evaporation (bath temperature 40 °C), azeotroping with additional acetonitrile to afford 140 mg of the desired product after freeze-drying. The solid was submitted to chiral SFC purification (Flow rate: 120 mL/min, 50% MeOH, column: Cellulose 1, temperature = 40 °C, outlet pressure: 100 bar, injection volume: 500 µL). The fractions containing the minor, first-to- elute product were concentrated under reduced pressure, after freeze-drying to afford (4R,8S,9R)-14-(2,6-dimethylphenyl)-8-isobutyl-4-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-6,11-dioxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen- 2-one (22 mg, 10%) as a white solid; 1H NMR (400 MHz, Benzene-d6) δ 9.23 (s, 1H), 8.58 (s, 1H), 7.67 (dd, J = 16.3, 7.7 Hz, 2H), 6.99 - 6.94 (m, 1H), 6.90 - 6.78 (m, 3H), 6.65 (s, 1H), 6.37 (dd, J = 12.0, 5.1 Hz, 2H), 5.83 (s, 1H), 4.92 - 4.80 (m, 1H), 4.76 - 4.66 (m, 1H), 4.10 (dd, J = 14.2, 5.4 Hz, 1H), 3.96 - 3.75 (m, 5H), 3.62 - 3.50 (m, 1H), 1.96 - 1.91 (m, 1H), 1.87 (s, 6H), 1.58 (dt, J = 13.6, 6.7 Hz, 1H), 1.42 - 1.17 (m, 3H), 0.95 - 0.83 (m, 2H), 0.60 (d, J = 2.4 Hz, 3H), 0.58 (d, J = 2.4 Hz, 3H), 0.55 - 0.50 (m, 1H), 0.36 - 0.28 (m, 1H).19F NMR (377 MHz, Benzene-d6) δ -68.74 (s, 3F). ESI-MS m/z calc.789.292, found 790.1 (M+1)+; Retention time: 3.51 minutes and the fractions containing the major, second-to-elute product were concentrated under reduced pressure, after freeze-drying to afford (4S,8S,9R)-14-(2,6-dimethylphenyl)-8- isobutyl-4-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18- dioxo-6,11-dioxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (72 mg, 34%) as a white solid; 1H NMR (400 MHz, Benzene-d6) δ 9.26 (s, 1H), 8.74 (d, J = 2.0 Hz, 1H), 7.86 - 7.74 (m, 1H), 7.55 (d, J = 7.6 Hz, 1H), 6.98 - 6.94 (m, 1H), 6.92 - 6.76 (m, 3H), 6.70 (s, 1H), 5.97 - 5.85 (m, 2H), 5.14 (t, J = 10.5 Hz, 1H), 4.61 (d, J = 11.0 Hz, 1H), 4.50 (d, J = 9.8 Hz, 1H), 4.31 - 4.13 (m, 2H), 3.79 (d, J = 12.7 Hz, 1H), 3.68 (s, 3H), 3.51 (dd, J = 13.1, 6.7 Hz, 1H), 2.17 - 2.10 (m, 1H), 1.96 (br. s., 6H), 1.15 - 1.01 (overlapped with diethylamine, m, 3H), 0.97 - 0.78 (m, 3H), 0.72 (d, J = 5.9 Hz, 3H), 0.52 (d, J = 5.9 Hz, 3H), 0.48 - 0.40 (m, 1H), 0.27 - 0.17 (m, 1H).19F NMR (377 MHz, Benzene-d6) δ -68.76 (s, 3F). ESI-MS m/z calc.789.292, found 790.1 (M+1)+; Retention time: 3.45 minutes. LC method W. Example 83: Preparation of Compound 133 Step 1: (4S,8R,9R)-8-cyclobutyl-14-(2,6-dimethylphenyl)-4-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa- 18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (Compound 133) mixed with isomeric impurities [00707] The product was isolated as an impurity during a large scale preparation of ~ 60 g of (4S,8S,9R)-8-Cyclobutyl-14-(2,6-dimethylphenyl)-4-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen- 2-one. Early eluting fractions by flash chromatography on silica gel of the target product (5- 100% EtOAc in hexanes over 25 CV) containing the impurity were subjected to several rounds of reverse phase chromatography (typically 1-99% gradient of MeCN in aqueous 5 mM HCl) to provide (4S,8R,9R)-8-cyclobutyl-14-(2,6-dimethylphenyl)-4-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20-hexaen- 2-one (14 mg, <1% yield). ESI-MS m/z calc.785.2971, found 786.3(M+1)⁺; Retention time: 3.55 minutes. LC method N.1H NMR (500 MHz, acetone-d6) δ 5.85 (dd, J = 13.2 Hz, J = 4.5 Hz, 1H), 2.30 (m, 2H), 1.94 (m, 1H), 1.84 (m, 1H), 1.82 (m, 2H), 1.92 (m, 1H), 4.36 (dt, J = 12.3 Hz, J = 4.5 Hz, 1H), 6.01 (dd, J = 11.5 Hz, J = 5.1 Hz, 1H), 5.02 (t, J = 11.7, 1H), 6.87 (s, 1H), 7.68 (d, J = 7.8 Hz, 1H), 7.61 (d, J = 7.8 Hz, 1H), 7.87 (d, J = 7.8 Hz, 1H), 8.83 (br s, 1H), 7.14 (d, J = 7.6 Hz, 2H), 7.27 (d, J = 7.6 Hz, 1H), 2.19 (s, 6H), 2.47 (m, 1H), 1.71 (m, 1H), 1.07 (m, 1H), 1.61 (m, 1H), 1.47 (m, 1H), 6.80 (br s, 1H), 8.58 (s, 1H). Example 84: Preparation of Compound 134 Step 1: tert-Butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-3-vinyl- pyrrolidine-1-carboxylate [00708] A flame-dried flask was charged with copper iodide (9.2 g, 48.307 mmol) in anhydrous THF (100 mL). The heterogeneous mixture was cooled down to - 78 °C and a solution of vinyl magnesium bromide in THF (100 mL of 1 M, 100.00 mmol) was cannulated to the solution. The reaction was stirred at - 78 °C for 1 h after which time a solution of tert-butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-2H-pyrrole-1-carboxylate (10.05 g, 22.119 mmol) in anhydrous THF (20 mL) was added dropwise over 90 min keeping the internal temperature below - 78 °C. The dry-ice bath was then removed allowing the temperature to slowly reach room temperature over 30 minutes. The reaction mixture was put in an ice-cold bath and was quenched by a saturated aqueous solution of ammonium chloride (100 mL). The THF was then removed under reduced pressure and water (100 mL) was added to the mixture. The solution was extracted with DCM (6 x 100 mL) and combined organic layers were eluted onto a silica pad. The pad was washed with DCM (50 mL) and the filtrate was evaporated under reduced pressure. The product was purified by flash chromatography (gradient: 0 to 40 % EtOAc in heptanes, 11 CV). Fractions containing the desired product were evaporated under reduced pressure to afford tert-butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-3- vinyl-pyrrolidine-1-carboxylate (8.05 g, 74%) as a yellow oil 1H NMR (400 MHz, CDCl3 ) δ 7.71 - 7.56 (m, 4H), 7.52 - 7.34 (m, 6H), 5.97 - 5.79 (m, 1H), 5.10 (d, J = 5.4 Hz, 1H), 5.08 - 5.01 (m, 1H), 3.98 - 3.88 (m, 2H), 3.80 - 3.69 (m, 1H), 3.07 - 2.91 (m, 2H), 2.41 - 2.28 (m, 1H), 1.49 - 1.38 (m, 9H), 1.11 - 1.00 (m, 9H). ESI-MS m/z calc.479.2492, found 380.2 (M-99)+; Retention time: 2.31 minutes. LC method I. Step 2: tert-Butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-3- (3,3,3-trifluoropropyl)pyrrolidine-1-carboxylate [00709] In a dry-flamed flask under nitrogen atmosphere was charged silver nitrate (73 mg, 0.4297 mmol), iodobenzene diacetate (2.3 g, 7.1407 mmol) and sodium acetate (anhydrous) (660 mg, 8.0455 mmol). The flask was evacuated and backfilled with nitrogen three times. Tert- butyl (2R,3S)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5-oxo-3-vinyl-pyrrolidine-1-carboxylate (1.82 g, 3.7752 mmol) in solution in dry NMP (20 mL) was added to the mixture under nitrogen atmosphere. Then 1,3-dioxolane (2.7560 g, 2.6 mL, 37.204 mmol) and trifluoromethyltrimethylsilane (1.3440 g, 1.4 mL, 9.4518 mmol) were added and the reaction was stirred at room temperature overnight. The crude was quenched with water (70 mL) and EtOAc (50 mL) was added. The phases were separated, and the organic layer was washed with brine (3 x 40 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by reverse-phase chromatography (C18 column. Gradient: 65-100% MeCN in water with 0.1 % formic acid). The fractions containing the desired compound were concentrated under reduced pressure to afford tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5- oxo-3-(3,3,3-trifluoropropyl)pyrrolidine-1-carboxylate (1.13 g, 41%) as a light-yellow oil.1H NMR (400 MHz, Chloroform-d) δ 7.65 - 7.58 (m, 4H), 7.46 - 7.37 (m, 6H), 3.96 - 3.81 (m, 2H), 3.76 - 3.70 (m, 1H), 3.00 - 2.91 (m, 1H), 2.39 - 2.31 (m, 1H), 2.02 (s, 2H), 1.86 - 1.58 (m, overlapped with water, 3H), 1.46 - 1.43 (m, 9H), 1.07 - 1.05 (m, 9H).19F NMR (377 MHz, Chloroform-d) δ -66.09 (t, J = 10.9 Hz, 3F). ESI-MS m/z calc.549.2522, found 450.2 (M-99)+; Retention time: 2.53 minutes. LC method U. Step 3: tert-Butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-5,5,5- trifluoro-2-(2-hydroxyethyl)pentyl]carbamate [00710] To a 50 °C solution of tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5- oxo-3-(3,3,3-trifluoropropyl)pyrrolidine-1-carboxylate (1.13 g, 1.5623 mmol) in ethanol (20 mL), under nitrogen atmosphere, was added sodium borohydride (360 mg, 9.5156 mmol) portion-wise. The reaction was stirred for 2 hours at 50 °C. The flask was cooled to room temperature and then to 0 °C. A saturated aqueous solution of ammonium chloride (40 mL) was slowly introduced into the reaction and the resulting solution was stirred for 20 minutes at room temperature. The ethanol was evaporated under reduced pressure and ethyl acetate (40 mL) was added. The layers were separated and the aqueous one was extracted with ethyl acetate (3 x 30 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by reversed-phase chromatography on C18 column (gradient 60% to 100% acetonitrile in water). The fractions containing the desired product were combined, concentrated under reduced pressure to afford tert-butyl N-[(1R,2R)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-5,5,5-trifluoro-2-(2- hydroxyethyl)pentyl]carbamate (886 mg, 89%) as an orange oil.1H NMR (400 MHz, Chloroform-d) δ 7.70 - 7.62 (m, 4H), 7.48 - 7.37 (m, 6H), 4.82 (br. d, J = 8.6 Hz, 1H), 3.81 - 3.58 (m, 5H), 2.27 - 1.99 (m, overlapped with MeCN, 2H), 1.93 - 1.79 (m, 1H), 1.71 - 1.58 (m, 1H), 1.55 - 1.46 (m, overlapped with water peak, 4H), 1.44 (s, 9H), 1.07 (s, 9H).19F NMR (377 MHz, Chloroform-d) δ -66.42 (s, 3F). ESI-MS m/z calc.553.2835, found 454.2 (M-99)+; Retention time: 2.49 minutes. LC method U. Step 4: tert-Butyl N-[(1R,2R)-5,5,5-trifluoro-2-(2-hydroxyethyl)-1- (hydroxymethyl)pentyl]carbamate [00711] To a stirred solution of tert-butyl N-[(1R,2R)-1-[[tert- butyl(diphenyl)silyl]oxymethyl]-5,5,5-trifluoro-2-(2-hydroxyethyl)pentyl]carbamate (886 mg, 1.3953 mmol) in anhydrous THF (15 mL) at 0 °C was added a solution of TBAF solution in THF (4.5 mL of 1 M, 4.5000 mmol) dropwise. The reaction was stirred at 0 °C for 30 minutes, then allowed to reach room temperature and stirred for 2 hours. The reaction was quenched by the addition of a saturated aqueous solution of sodium bicarbonate (30 mL). EtOAc was added (30 mL) and the layers were separated. The aqueous layer was extracted with ethyl acetate (2 x 20 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by reversed-phase chromatography on C18 column (gradient 5% to 60% acetonitrile in water). The fractions containing the desired product were combined, concentrated under reduced pressure to afford tert-butyl N-[(1R,2R)-5,5,5-trifluoro-2-(2-hydroxyethyl)-1- (hydroxymethyl)pentyl]carbamate (333 mg, 72%) as a pale yellow oil.1H NMR (400 MHz, Chloroform-d) δ 5.01 (br. d, J = 7.8 Hz, 1H), 3.85 - 3.75 (m, 2H), 3.74 - 3.64 (m, 4H), 2.46 - 2.39 (m, 1H), 2.34 - 2.20 (m, 1H), 2.18 - 2.05 (m, 2H), 1.96 - 1.87 (m, 1H), 1.74 - 1.61 (m, overlapped with water, 3H), 1.46 (s, 9H).19F NMR (377 MHz, Chloroform-d) δ -66.40 (s, 3F). ESI-MS m/z calc.315.1657, found 216.2 (M-99)+; Retention time: 1.66 minutes. LC method U. Step 5: tert-Butyl (4R)-2,2-dimethyl-4-[(1R)-4,4,4-trifluoro-1-(2- hydroxyethyl)butyl]oxazolidine-3-carboxylate [00712] To a stirred solution of tert-butyl N-[(1R,2R)-5,5,5-trifluoro-2-(2-hydroxyethyl)-1- (hydroxymethyl)pentyl]carbamate (333 mg, 1.0032 mmol) in dry THF (10 mL), were added 2,2- dimethoxypropane (765.00 mg, 0.9 mL, 7.3453 mmol) and para toluenesulfonic acid monohydrate (195 mg, 1.0251 mmol). The reaction was stirred at room temperature for 4 hours. Acetone (8 mL) and isopropyl acetate (8 mL) were added, then a saturated aqueous solution of sodium bicarbonate (15 mL) was added, and the mixture was stirred at room temperature for 5 minutes. Organic solvents were evaporated under reduced pressure and co-evaporated with isopropyl acetate (3 x 8 mL). The aqueous layer was extracted with dichloromethane (2 x 15 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude mixture was purified by reversed- phase chromatography on a C18 cartridge, eluting with a gradient of 50 to 100 % of acetonitrile in water. The fractions containing the product were combined, evaporated to afford tert-butyl (4R)-2,2-dimethyl-4-[(1R)-4,4,4-trifluoro-1-(2-hydroxyethyl)butyl]oxazolidine-3-carboxylate (282.6 mg, 75%) as a colorless oil.1H NMR (400 MHz, Chloroform-d) δ 4.11 (br. s., 1H), 3.99 - 3.91 (m, 1H), 3.87 - 3.68 (m, 3H), 2.76 - 2.65 (m, 1H), 2.27 - 2.15 (m, 1H), 2.11 - 1.98 (m, 2H), 1.94 - 1.85 (m, 1H), 1.69 - 1.59 (m, overlapped with water, 3H), 1.53 - 1.39 (m, 15H).19F NMR (377 MHz, Chloroform-d) δ -66.51 (br. s., 3F). ESI-MS m/z calc.355.197, found 300.4 (M-55)+; Retention time: 1.98 minutes. LC method U. Step 6: tert-Butyl (4R)-2,2-dimethyl-4-[(1R)-4,4,4-trifluoro-1-(2- oxoethyl)butyl]oxazolidine-3-carboxylate [00713] To a solution of tert-butyl (4R)-2,2-dimethyl-4-[(1R)-4,4,4-trifluoro-1-(2- hydroxyethyl)butyl]oxazolidine-3-carboxylate (282 mg, 0.7538 mmol) in dry dichloromethane (8 mL) at 0 °C, were added sodium bromide (30 mg, 0.2916 mmol) and TEMPO (34 mg, 0.2176 mmol). A stirred mixture of sodium bicarbonate (308 mg, 3.6664 mmol) in sodium hypochlorite (in water) (3.75 mL of 0.53 M, 1.9875 mmol) was prepared at 0 °C, filtered and added dropwise to the reaction under vigorous stirring. The mixture was stirred at 0 °C for 3 hours. A saturated aqueous solution of sodium bicarbonate (12 mL) was added. The mixture was diluted with DCM (15 mL) and the layers were separated. The organic layer was washed with a saturated aqueous solution of sodium bicarbonate (15 mL), a 10% aqueous solution of Na2S2O3 (15 mL) and brine (20 mL), then dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford the crude mixture of tert-butyl (4R)-2,2-dimethyl-4-[(1R)-4,4,4-trifluoro-1-(2- oxoethyl)butyl]oxazolidine-3-carboxylate (276 mg, 98%) as an orange oil.1H NMR (400 MHz, Chloroform-d) δ 9.79 (br. t, J = 1.7 Hz, 1H), 3.99 - 3.94 (m, 1H), 3.81 (dd, J = 7.9, 4.8 Hz, 1H), 2.59 - 2.33 (m, 3H), 2.26 - 2.06 (m, 2H), 1.69 - 1.60 (m, 3H), 1.52 - 1.43 (m, 15H).19F NMR (377 MHz, Chloroform-d) δ -66.25 - -66.61 (m, 3F). ESI-MS m/z calc.353.1814, no iomization, Retention time: 2.09 minutes. LC method U. Step 7: tert-Butyl (4R)-4-[(E,1R)-5-[methoxy(methyl)amino]-5-oxo-1-(3,3,3- trifluoropropyl)pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00714] To a suspension of heptane washed sodium hydride in mineral oil (50 mg, 60 %w/w, 1.2501 mmol) in dry THF (3 mL), was added 2-diethoxyphosphoryl-N-methoxy-N-methyl- acetamide (232.60 mg, 0.2 mL, 0.9724 mmol) at room temperature. After the evolution of hydrogen stopped, this solution was transferred to a solution of tert-butyl (4R)-2,2-dimethyl-4- [(1R)-4,4,4-trifluoro-1-(2-oxoethyl)butyl]oxazolidine-3-carboxylate (215 mg, 0.6084 mmol) in dry THF (3 mL) under nitrogen atmosphere. The reaction was stirred at room temperature overnight. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (10 mL). The phases were separated, and the aqueous phase was extracted with ethyl acetate (3 x 10 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reverse-phase chromatography on C18 column (gradient 50 % to 100 % acetonitrile in water). The fractions containing the desired product were combined, concentrated under reduced pressure to afford tert-butyl (4R)-4-[(E,1R)-5-[methoxy(methyl)amino]-5-oxo-1-(3,3,3- trifluoropropyl)pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate (289 mg, 94%) as a colorless oil.1H NMR (400 MHz, Chloroform-d) δ 6.99 - 6.86 (m, 1H), 6.57 - 6.39 (m, 1H), 4.02 (br. s., 1H), 3.96 - 3.91 (m, 1H), 3.84 - 3.78 (m, 1H), 3.73 - 3.68 (m, 4H), 3.25 (s, 3H), 3.23 - 3.19 (m, 1H), 2.37 - 2.02 (m, 5H), 1.51 - 1.46 (m, 15H).19F NMR (377 MHz, Chloroform-d) δ -66.13 - -66.85 (m, 3F). ESI-MS m/z calc.438.2342, found 339.2 (M-99)+; Retention time: 2.08 minutes. LC method U. Step 8: tert-Butyl (4R)-4-[(1S)-5-[methoxy(methyl)amino]-5-oxo-1-(3,3,3- trifluoropropyl)pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00715] Nitrogen was bubbled to a stirred solution of tert-butyl (4R)-4-[(E,1R)-5- [methoxy(methyl)amino]-5-oxo-1-(3,3,3-trifluoropropyl)pent-3-enyl]-2,2-dimethyl-oxazolidine- 3-carboxylate (289 mg, 0.4877 mmol) in dry methanol (6 mL) for 10 minutes, then palladium on carbon (170 mg, 10 %w/w, 0.1597 mmol) was added. Nitrogen was bubbled to the mixture for 10 minutes, followed by hydrogen for 15 minutes. The reaction was then stirred for 1 hour, at room temperature, under hydrogen atmosphere. Nitrogen was bubbled to the mixture for 10 minutes. The mixture was filtered on a pad of Celite, washed with MeOH (20 mL) and concentrated in vacuo. The crude mixture was purified by reverse phase chromatography on a C18 cartridge, eluting with a gradient of 50 to 80% of acetonitrile in water. The fractions containing the desired product were combined, concentrated in vacuo to afford tert-butyl (4R)-4- [(1S)-5-[methoxy(methyl)amino]-5-oxo-1-(3,3,3-trifluoropropyl)pentyl]-2,2-dimethyl- oxazolidine-3-carboxylate (92 mg, 38%) as a light-yellow oil.1H NMR (400 MHz, Chloroform- d) δ 4.05 (br. s., 1H), 3.98 - 3.91 (m, 1H), 3.83 - 3.77 (m, 1H), 3.69 (s, 3H), 3.18 (s, 3H), 2.51 - 2.37 (m, 2H), 2.25 - 2.13 (m, 1H), 2.11 - 1.95 (m, 2H), 1.78 - 1.68 (m, 1H), 1.66 - 1.57 (m, overlapped with water, 3H), 1.52 - 1.29 (m, 17H).19F NMR (377 MHz, Chloroform-d) δ -66.40 (d, J = 133.5 Hz, 3F). ESI-MS m/z calc.440.2498, found 442.2 (M+1)+; Retention time: 2.08 minutes. LC method U. Step 9: tert-Butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-5-oxo-1-(3,3,3-trifluoropropyl)pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate [00716] In a flame-dried flask under nitrogen atmosphere was dissolved 3-bromo-6-tert-butyl- 5-methyl-pyrrolo[2,3-b]pyrazine (92 mg, 0.3373 mmol) in dry THF (1 mL), at -78 °C. To this solution was added n-butyllithium (solution in hexanes) (0.15 mL of 2.5 M, 0.3750 mmol) dropwise. As soon as the addition of n-BuLi was over, a solution of tert-butyl (4R)-4-[(1S)-5- [methoxy(methyl)amino]-5-oxo-1-(3,3,3-trifluoropropyl)pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (92 mg, 0.1855 mmol) in dry THF (1 mL) was added dropwise. The reaction was left stirring 2.5 hours at -78 °C. A saturated aqueous solution of ammonium chloride (2 mL) and ethyl acetate (3 mL) were added. The mixture was slowly warmed up to room temperature and stirred for 10 minutes. The layers were separated, and the aqueous one was extracted with ethyl acetate (3 x 5 mL). The combined organic layers were washed with brine (3 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude was purified by reversed-phase chromatography on C18 column (gradient 50 to 100% acetonitrile in water). The desired fractions were concentrated under reduced pressure to afford tert-butyl (4R)- 4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-5-oxo-1-(3,3,3- trifluoropropyl)pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (50 mg, 45%) as a light-yellow semi-solid.1H NMR (400 MHz, Chloroform-d) δ 9.11 (s, 1H), 6.60 (s, 1H), 4.07 (s, 3H), 3.99 - 3.93 (m, 1H), 3.86 - 3.80 (m, 1H), 3.32 - 3.25 (m, 2H), 2.27 - 2.18 (m, 1H), 2.17 - 1.96 (m, 3H), 1.95 - 1.82 (m, 2H), 1.68 - 1.57 (m, 4H), 1.55 (s, 9H), 1.48 (br. s., 15H).19F NMR (377 MHz, Chloroform-d) δ -66.39 (d, J = 145.8 Hz, 3F). ESI-MS m/z calc.568.3236, found 569.4 (M+1)+; Retention time: 2.41 minutes. LC method U. Step 10: (5S)-5-[(1R)-1-Amino-2-hydroxy-ethyl]-1-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-8,8,8-trifluoro-octan-1-one [00717] To a solution of tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-5-oxo-1-(3,3,3-trifluoropropyl)pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (50 mg, 0.0878 mmol) in 1,4-dioxane (1 mL) and water (0.25 mL), was added hydrochloric acid (solution in 1,4-dioxane) (1.15 mL of 4 M, 4.6000 mmol). The reaction was stirred at room temperature for 3 hours. The solvents were removed under reduced pressure and the mixture was co-evaporated with 1,4-dioxane (5 x 10 mL). The material was dissolved in dichloromethane (10 mL) and the organic layer was washed with a saturated aqueous solution of sodium bicarbonate (10 mL) and brine (10 mL). The organic layer was dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure, to afford the crude (5S)-5-[(1R)-1-amino-2-hydroxy- ethyl]-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8,8,8-trifluoro-octan-1-one (40 mg, 101%) as a yellow oil.1H NMR (400 MHz, Chloroform-d) δ 8.70 (s, 1H), 6.50 (s, 1H), 4.08 (s, 1H), 4.03 (s, 3H), 3.90 - 3.88 (m, 2H), 3.80 - 3.77 (m, 1H), 3.68 - 3.63 (m, 2H), 2.23 - 2.14 (m, overlapped with acetone, 2H), 1.93 - 1.81 (m, 5H), 1.73 - 1.64 (m, 4H), 1.51 (s, 9H).19F NMR (377 MHz, Chloroform-d) δ -66.24 (s, 3F), -66.42. ESI-MS m/z calc.428.2399, found 429.2 (M+1)+; Retention time: 1.51 minutes. LC method U. Step 11: [(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (3,3,3-trifluoropropyl)azepan-2-yl]methanol and [(2R,3S,7R)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(3,3,3-trifluoropropyl)azepan-2- yl]methanol [00718] To a solution of (5S)-5-[(1R)-1-amino-2-hydroxy-ethyl]-1-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-8,8,8-trifluoro-octan-1-one (40 mg, 0.0887 mmol) in dry DCM (1 mL) at 0 °C, was added NaBH(OAc)3 (115 mg, 0.5318 mmol). The reaction mixture was stirred for 3 hours at room temperature and a saturated aqueous solution of sodium bicarbonate (5 mL) was added at 0 °C. The layers were separated, and the aqueous layer was extracted with DCM (3 x 8 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography on a C18 column (55 to 70 % acetonitrile in basic water 10 mM ammonium bicarbonate buffer: pH = 10). The fractions containing the first eluting product were combined and the volatiles were removed under reduced pressure to afford [(2R,3S,7S)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(3,3,3-trifluoropropyl)azepan-2-yl]methanol (13 mg, 31%) as a white solid with 99.0 % ee.1H NMR (400 MHz, Benzene-d6) δ 8.57 (s, 1H), 6.59 (s, 1H), 3.79 (dd, J = 10.0, 4.4 Hz, 1H), 3.57 (dd, J = 10.5, 3.7 Hz, 1H), 3.51 (s, 3H), 3.09 (br. t, J = 10.0 Hz, 1H), 2.36 (td, J = 9.0, 3.9 Hz, 1H), 1.99 - 1.91 (m, 1H), 1.84 - 1.65 (m, 2H), 1.58 - 1.44 (m, 3H), 1.39 - 1.18 (m, 4H), 1.12 (s, 9H), 1.11 - 0.97 (m, 3H).19F NMR (377 MHz, Benzene-d6) δ -66.16 (s, 3F). ESI-MS m/z calc.412.245, found 413.2 (M+1)+; Retention time: 3.56 minutes. The fractions containing the second eluting product were combined and the volatiles were removed under reduced pressure to afford [(2R,3S,7R)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-(3,3,3-trifluoropropyl)azepan-2-yl]methanol (3 mg, 8%) as a white solid.99.0 % ee.1H NMR (400 MHz, Benzene-d6) δ 8.60 (s, 1H), 6.60 (s, 1H), 3.77 (dd, J = 11.4, 2.6 Hz, 1H), 3.62 - 3.58 (m, 1H), 3.57 (s, 3H), 3.36 (t, J = 10.4 Hz, 1H), 2.40 (td, J = 9.7, 4.3 Hz, 1H), 2.13 - 2.06 (m, 1H), 1.73 - 1.40 (m, 5H), 1.38 - 1.28 (m, 2H), 1.18 - 1.11 (m, 1H), 1.10 (s, 9H), 1.07 - 0.95 (m, 4H).19F NMR (377 MHz, Benzene-d6) δ -66.16 (s, 3F). ESI-MS m/z calc. 412.245, found 413.3 (M+1)+; Retention time: 3.65 minutes. LC method W. Step 12: 3-[[4-[[(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-(3,3,3-trifluoropropyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin- 2-yl]sulfamoyl]benzoic acid
[00719] To a solution of [(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (3,3,3-trifluoropropyl)azepan-2-yl]methanol (12 mg, 0.0254 mmol) and 3-[[4-chloro-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (13 mg, 0.0311 mmol) in dry THF (0.4 mL) was added portion-wise sodium tert-butoxide (14 mg, 0.1457 mmol). The reaction was stirred at room temperature for 4 h. The mixture was cooled down to 0 °C with an ice-cold bath then slowly acidified with an aqueous solution of HCl 1 N (3 mL). The phases were separated, and the aqueous layer was extracted with Me-THF (3 x 3 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Purification by reverse-phase chromatography (Column: C18. Gradient: 50-75 % acetonitrile in acidic water (0.1% w/w of HCl)) afforded 3-[[4-[[(2R,3S,7S)-7-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-3-(3,3,3-trifluoropropyl)azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (18 mg, 77%) as a yellow solid.1H NMR (400 MHz, methanol-d4) δ 8.69 - 8.63 (m, 1H), 8.60 - 8.36 (m, 1H), 8.25 - 8.20 (m, 1H), 8.13 - 8.07 (m, 1H), 7.67 - 7.61 (m, 1H), 7.30 - 7.25 (m, 1H), 7.14 (d, J = 7.6 Hz, 2H), 6.62 - 6.56 (m, 1H), 6.34 - 6.31 (m, 1H), 4.63 - 4.56 (m, 1H), 4.12 - 4.06 (m, 3H), 3.89 - 3.81 (m, 1H), 2.42 - 2.21 (m, 6H), 2.12 - 1.94 (m, 12H), 1.83 - 1.75 (m, 2H), 1.55 - 1.52 (m, 6H), three labile protons are missing, 2 protons are obscured by water peak.19F NMR (377 MHz, methanol-d4) δ -67.57 (s, 3F). ESI-MS m/z calc.793.3233, found 794.2 (M+1)+; Retention time: 1.61 minutes. LC method U. Step 13: (4S,8S,9R)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6- dimethylphenyl)-18,18-dioxo-8-(3,3,3-trifluoropropyl)-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (Compound 134) [00720] In a flame-dried flask under nitrogen atmosphere was dissolved 3-[[4-[[(2R,3S,7S)-7- (6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(3,3,3-trifluoropropyl)azepan-2- yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (18 mg, 0.0204 mmol) in dry NMP (0.4 mL) and acetone (0.8 mL). TsCl (12 mg, 0.0629 mmol) was added followed by a rapid dropwise addition of 4-methylmorpholine (13.800 mg, 0.015 mL, 0.1364 mmol). The reaction was stirred at 55 °C for 3 hours. The reaction was cooled to room temperature, the acetone was evaporated, and the crude product was purified by reverse phase chromatography (acetonitrile in water with 0.1 % formic acid, 60 to 100%). The fraction was concentrated and freeze-dried in a mixture of MeCN/distilled water to afford (4S,8S,9R)-4-(6- tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-18,18-dioxo-8-(3,3,3- trifluoropropyl)-11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (7.2 mg, 45%) as a light-yellow solid.1H NMR (400 MHz, DMSO-d6) δ 8.62 (br. s, 1H), 8.48 (s, 1H), 7.92 - 7.82 (m, 1H), 7.73 - 7.54 (m, 2H), 7.30 - 7.22 (m, 1H), 7.17 - 7.10 (m, 2H), 6.45 (s, 1H), 6.34 (br. s., 1H), 5.91 - 5.84 (m, 1H), 5.76 (dd, J = 11.6, 5.7 Hz, 1H), 4.96 (br. t, J = 11.4 Hz, 1H), 4.34 - 4.27 (m, 1H), 4.08 (s, 3H), 3.29 (s, overlapped with water, 6H), 2.19 - 1.96 (m, 10H), 1.83 - 1.75 (m, 2H), 1.47 (s, 9H). ESI-MS m/z calc.775.3128, found 776.2 (M+1)+; Retention time: 3.94 minutes. LC method 1B. Example 85: Preparation of Compound 135 Step 1: tert-Butyl (4R)-2,2-dimethyl-4-[(1S)-5-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-1-(3,3,3- trifluoropropyl)pentyl]oxazolidine-3-carboxylate [00721] To a solution of 3-bromo-5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazine (73 mg, 0.2274 mmol) in THF (2 mL), at -78 °C, was added n-butyllithium (solution in hexanes) (0.1 mL of 2.5 M, 0.2500 mmol) dropwise. As soon as the addition of n-BuLi was over, a solution of tert-butyl (4R)-4-[(1S)-5-[methoxy(methyl)amino]-5-oxo-1-(3,3,3- trifluoropropyl)pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (70 mg, 0.1398 mmol) in THF (0.5 mL) was added dropwise. The reaction was left stirring 1 hour at -78 °C. A saturated aqueous solution of ammonium chloride (1 mL) and ethyl acetate (2 mL) were added. The mixture was slowly warmed up to room temperature and stirred for 15 minutes. The layers were separated, and the aqueous one was extracted with ethyl acetate (3 x 5 mL). The combined organic layers were washed with brine (2 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude was purified by reversed-phase chromatography on C18 column (gradient 40% to 80% acetonitrile in water). The desired fractions were concentrated under reduced pressure, then co-evaporated with acetonitrile (4 x 5 mL), to afford tert-butyl (4R)-2,2-dimethyl-4-[(1S)-5-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazin-3-yl]-5-oxo-1-(3,3,3-trifluoropropyl)pentyl]oxazolidine-3-carboxylate (38 mg, 40%) as a yellow semi-solid.1H NMR (400 MHz, CDCl3) δ 9.32 - 9.13 (m, 1H), 6.85 (s, 1H), 4.16 - 3.90 (m, 5H), 3.85 - 3.75 (m, 1H), 3.42 - 3.15 (m, 2H), 2.28 - 1.98 (m, 3H), 1.96 - 1.71 (m, 3H), 1.70 - 1.53 (m, overlapped with water, 4H), 1.52 - 1.25 (m, 18H).19F NMR (377 MHz, CDCl3) δ -66.11 - -66.66 (m, 3F), -68.54 (s, 3F). ESI-MS m/z calc.620.2797, found 621.2 (M+1)+; Retention time: 2.43 minutes. LC method U. Step 2: (5S)-5-[(1R)-1-Amino-2-hydroxy-ethyl]-8,8,8-trifluoro-1-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]octan-1-one [00722] To a solution of tert-butyl (4R)-2,2-dimethyl-4-[(1S)-5-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-5-oxo-1-(3,3,3- trifluoropropyl)pentyl]oxazolidine-3-carboxylate (38 mg, 0.0557 mmol) in 1,4-dioxane (1 mL) and water (0.25 mL), was added hydrochloric acid (solution in 1,4-dioxane) (0.7 mL of 4 M, 2.8000 mmol). The reaction was stirred at room temperature for 20 hours, under nitrogen. The solvents were removed under reduced pressure and the mixture was co-evaporated with 1,4- dioxane (6 x 3 mL). The material was dissolved in dichloromethane (10 mL) and the organic layer was washed with a saturated aqueous solution of sodium bicarbonate (2 mL) and brine (1 mL). The organic layer was dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure, to afford (5S)-5-[(1R)-1-amino-2-hydroxy-ethyl]-8,8,8-trifluoro-1-[5-methyl- 6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]octan-1-one (26 mg, 93%) as a yellow semi-solid.19F NMR (377 MHz, CDCl3) δ -66.17 - -66.56 (m, 3F), -68.43 - -68.92 (m, 3F). ESI-MS m/z calc.480.196, found 481.2 (M+1)+; Retention time: 1.5 minutes. LC method U. Step 3: [(2R,3S,7S)-7-[5-Methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazin-3-yl]-3-(3,3,3-trifluoropropyl)azepan-2-yl]methanol [00723] To a solution of (5S)-5-[(1R)-1-amino-2-hydroxy-ethyl]-8,8,8-trifluoro-1-[5-methyl- 6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]octan-1-one (26 mg, 0.0541 mmol) in DCM (1 mL), at 0 °C, was added NaBH(OAc)3 (70 mg, 0.3237 mmol). The reaction mixture was stirred for 6 hours at room temperature and a saturated aqueous solution of sodium bicarbonate (2 mL) was added at 0 °C. The layers were separated, and the aqueous layer was extracted with DCM (3 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography on a C18 column (isocratic 50% acetonitrile in 10 mM ammonium bicarbonate buffer: pH = 10). The fractions containing the first eluting product were combined and the volatiles were removed under reduced pressure. The residue was co- evaporated with acetonitrile (6 x 10 mL). The resulting product was dried under high vacuum to afford [(2R,3S,7S)-7-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3- (3,3,3-trifluoropropyl)azepan-2-yl]methanol (12 mg, 47%) as a yellow oil.99.9% ee, 99.9% de. 1H NMR (400 MHz, benzene-d6) δ 8.55 (s, 1H), 6.70 (s, 1H), 3.77 (dd, J = 10.1, 4.3 Hz, 1H), 3.61 - 3.53 (m, 1H), 3.42 (s, 3H), 3.15 - 3.04 (m, 1H), 2.35 (td, J = 9.0, 3.8 Hz, 1H), 1.92 (ddt, J = 13.7, 6.8, 3.5 Hz, 1H), 1.82 - 1.65 (m, 2H), 1.59 - 1.42 (m, overlapped with impurities, 3H), 1.41 - 0.88 (m, overlapped with impurities, 9H), 0.58 - 0.49 (m, 2H).19F NMR (377 MHz, benzene-d6) δ -66.14 - -66.25 (m, overlapped with impurities, 3F), -68.65 (s, 3F). ESI-MS m/z calc.464.2011, found 465.2 (M+1)+; Retention time: 3.53 minutes. The fractions containing the second eluting product were combined and the volatiles were removed under reduced pressure. The residue was co-evaporated with acetonitrile (6 x 10 mL). The resulting product was dried under high vacuum to afford [(2R,3S,7R)-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3-(3,3,3-trifluoropropyl)azepan-2- yl]methanol (4 mg, 16%) as a yellow oil.99.9% ee, 99.9% de.1H NMR (400 MHz, benzene-d6) δ 8.59 (s, 1H), 6.72 (s, 1H), 3.74 (dd, J = 11.5, 2.4 Hz, 1H), 3.58 (dd, J = 10.5, 4.2 Hz, 1H), 3.49 (s, 3H), 3.31 (t, J = 10.4 Hz, 1H), 2.38 (td, J = 9.7, 4.3 Hz, 1H), 2.11 - 2.00 (m, 1H), 1.79 - 1.26 (m, 6H), 1.21 - 0.75 (m, 7H), 0.51 - 0.43 (m, 2H). (1H missing, labile proton).19F NMR (377 MHz, BENZENE-d6) δ -66.16 (t, J = 10.9 Hz, 3F), -68.67 (s, 3F). ESI-MS m/z calc. 464.2011, found 465.2 (M+1)+; Retention time: 3.6 minutes. LC method W. Step 4: 3-[[4-(2,6-Dimethylphenyl)-6-[[(2R,3S,7S)-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3-(3,3,3- trifluoropropyl)azepan-2-yl]methoxy]pyrimidin-2-yl]sulfamoyl]benzoic acid
[00724] To a solution of [(2R,3S,7S)-7-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-3-(3,3,3-trifluoropropyl)azepan-2- yl]methanol (12 mg, 0.0255 mmol) and 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (14 mg, 0.0332 mmol) in dry THF (0.5 mL) was added portion-wise sodium tert-butoxide (15 mg, 0.1514 mmol). The reaction was stirred at room temperature for 4 h. More sodium tert-butoxide (15 mg, 0.1514 mmol) was added and the mixture was stirred for 1 hour at room temperature, 1 hour at 30 °C and 16 hours at room temperature. More sodium tert-butoxide (15 mg, 0.1514 mmol) was added and the mixture was stirred for 1 hour at room temperature and 6 hours at 50 °C. The mixture was cooled down to 0 °C with an ice-cold bath, then was slowly acidified with an aqueous solution of HCl (1 M, 400 μL) until pH = 2. The layers were separated, and the aqueous layer was extracted with Me-THF (3 x 3 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by reversed-phase chromatography on C18 column (gradient 5% to 44% acetonitrile in acidic water (0.1% w/w of HCl)). The fractions containing the desired product were combined and concentrated under reduced pressure, co- evaporated with acetonitrile (5 x 5 mL), co-evaporated with CD3OD (0.5 mL), co-evaporated with acetonitrile (0.5 mL) and co-evaporated with acetone (0.5 mL), to afford 3-[[4-(2,6- dimethylphenyl)-6-[[(2R,3S,7S)-7-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazin-3-yl]-3-(3,3,3-trifluoropropyl)azepan-2-yl]methoxy]pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (12 mg, 53%) as a yellow solid.1H NMR (400 MHz, methanol-d4) δ 8.74 - 8.58 (m, 1H), 8.29 - 8.00 (m, 2H), 7.74 - 7.51 (m, 1H), 7.31 - 7.21 (m, 1H), 7.20 - 7.06 (m, 2H), 6.90 (br. s, 1H), 6.40 - 6.28 (m, 1H), 5.05 - 4.70 (m, overlapped with water, 3H), 4.65 - 4.45 (m, 1H), 4.08 - 3.75 (m, 3H), 2.51 - 2.14 (m, 5H), 2.14 - 2.01 (m, 9H), 2.01 - 1.72 (m, 4H), 1.69 - 1.55 (m, 2H), 1.46 - 1.30 (m, 2H). (3H missing, labile protons).19F NMR (377 MHz, methanol-d4) δ -67.53 (t, J = 10.2 Hz, 3F), -70.05 (s, 3F). ESI-MS m/z calc.845.2794, found 846.2 (M+1)+; Retention time: 1.64 minutes. LC method U. Step 5: (4S,8S,9R)-14-(2,6-dimethylphenyl)-4-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-8-(3,3,3- trifluoropropyl)-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one (Compound 135) [00725] In a heat-dried flask, under nitrogen atmosphere, was dissolved 3-[[4-(2,6- dimethylphenyl)-6-[[(2R,3S,7S)-7-[5-methyl-6-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3- b]pyrazin-3-yl]-3-(3,3,3-trifluoropropyl)azepan-2-yl]methoxy]pyrimidin-2-yl]sulfamoyl]benzoic acid (12 mg, 0.0142 mmol) in dry NMP (0.25 mL) and acetone (0.6 mL). TsCl (8 mg, 0.0415 mmol) was added, followed by a dropwise addition of 4-methylmorpholine (9.1080 mg, 10 μL, 0.0891 mmol) over 15 seconds. The reaction was stirred at 55 °C for 4 hours. The reaction was cooled to room temperature and the acetone was evaporated. The crude material was purified by reversed-phase chromatography on C18 column (gradient 5% to 75% acetonitrile in acidic water (0.1% v/v of formic acid in water). The fractions containing the desired product were combined, concentrated under reduced pressure, co-evaporated with acetonitrile (6 x 5 mL), co-evaporated with CD3OD (0.5 mL), co-evaporated with acetonitrile (3 x 1 mL) and dried under high vacuum, to afford (4S,8S,9R)-14-(2,6-dimethylphenyl)-4-[5-methyl-6-[1- (trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazin-3-yl]-18,18-dioxo-8-(3,3,3-trifluoropropyl)- 11-oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (8.2 mg, 69%) as an off-white solid.1H NMR (400 MHz, methanol-d4) δ 8.74 (s, 1H), 8.55 (br. s, 1H), 8.05 - 7.93 (m, 1H), 7.75 - 7.59 (m, 2H), 7.34 - 7.23 (m, 1H), 7.15 (d, J = 7.6 Hz, 2H), 6.75 (s, 1H), 6.26 (s, 1H), 6.09 (dd, J = 11.5, 4.9 Hz, 1H), 5.87 (dd, J = 12.6, 4.5 Hz, 1H), 5.07 (t, J = 11.7 Hz, 1H), 4.48 - 4.35 (m, 1H), 4.10 (s, 3H), 2.40 - 1.85 (m, 15H), 1.83 - 1.72 (m, 1H), 1.66 - 1.51 (m, 3H), 1.45 - 1.28 (m, overlapped with impurities, 3H).19F NMR (377 MHz, methanol-d4) δ -67.84 (t, J = 10.9 Hz, 3F), -70.18 (s, 3F). ESI-MS m/z calc.827.2688, found 828.2 (M+1)+; Retention time: 4.04 minutes. LC method 1B. Example 86: Preparation of Compound 136 Step 1: tert-Butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-(1,1- dibromo-2,2,2-trifluoro-ethyl)-5-oxo-pyrrolidine-1-carboxylate [00726] To a stirred solution of tert-butyl (2R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-5- oxo-2H-pyrrole-1-carboxylate (5 g, 10.163 mmol) and 2,2-dibromo-1,1,1-trifluoro-ethane (3.4303 g, 1.5 mL, 14.043 mmol) in dry THF (90 mL), at -78°C, was added LHMDS in THF (30 mL of 1 M, 30.000 mmol) dropwise over 1 hour. The mixture was stirred at -78°C for 4 hours, then a saturated aqueous solution of ammonium chloride (75 mL) was added at -78°C, the solution was allowed to warm to room temperature and stirred for 30 minutes at room temperature. The THF was removed under reduced pressure. The mixture was extracted with ethyl acetate (3 x 100 mL), the combined organic layers were washed with brine (25 mL) dried over anhydrous sodium sulfate, filtrated and concentrated under reduced pressure. The crude material was purified by reversed-phase chromatography on C18 column (isocratic 95% methanol in water). The fractions containing the desired product were combined, concentrated under reduced pressure and co-evaporated with acetonitrile (5 x 20 mL), to afford tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3-(1,1-dibromo-2,2,2-trifluoro-ethyl)-5-oxo- pyrrolidine-1-carboxylate (4.25 g, 43%) as an orange semi-solid. ESI-MS m/z calc.691.0576, found 592.0 (M-99)+; Retention time: 2.61 minutes. LC method U. Step 2: tert-Butyl N-[(1R,2S)-1-[[tert-butyl(diphenyl)silyl]oxymethyl]-4,4,4- trifluoro-2-(2-hydroxyethyl)butyl]carbamate [00727] To a 50 °C solution of tert-butyl (2R,3R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-3- (1,1-dibromo-2,2,2-trifluoro-ethyl)-5-oxo-pyrrolidine-1-carboxylate (4.25 g, 4.3391 mmol) in ethanol (100 mL), under nitrogen, was added sodium borohydride (1.3 g, 34.362 mmol) portion- wise. The reaction was stirred for 2 hours at 50 °C. A saturated aqueous solution of ammonium chloride (150 mL) was slowly introduced into the reaction and the resulting solution was stirred for 30 minutes at room temperature. The ethanol was evaporated under reduced pressure and ethyl acetate (200 mL) was added. The layers were separated, and the aqueous one was extracted with ethyl acetate (3 x 100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by reversed-phase chromatography on C18 column (gradient 60% to 90% acetonitrile in water). The fractions containing the desired product were combined, concentrated under reduced pressure and co-evaporated with acetonitrile (4 x 25 mL), to afford tert-butyl N-[(1R,2S)-1- [[tert-butyl(diphenyl)silyl]oxymethyl]-4,4,4-trifluoro-2-(2-hydroxyethyl)butyl]carbamate (2.19 g, 64%) as an orange oil.99.9% ee.1H NMR (400 MHz, CDCl3) δ 7.72 - 7.55 (m, 4H), 7.51 - 7.32 (m, 6H), 5.02 - 4.81 (m, 1H), 3.93 - 3.78 (m, 1H), 3.78 - 3.52 (m, 4H), 2.40 - 2.21 (m, 1H), 2.16 - 2.01 (m, 1H), 1.76 - 1.54 (m, overlapped with water, 2H), 1.46 - 1.38 (m, 9H), 1.07 (s, 9H). (2H missing, labile protons).19F NMR (377 MHz, CDCl3) δ -63.84 (t, J = 10.2 Hz, 3F). ESI-MS m/z calc.539.2679, found 440.2 (M-99)+; Retention time: 4.39 minutes. LC method 1B. Step 3: tert-Butyl N-[(1R,2S)-4,4,4-trifluoro-2-(2-hydroxyethyl)-1- (hydroxymethyl)butyl]carbamate [00728] To a stirred solution of crude tert-butyl N-[(1R,2S)-1-[[tert- butyl(diphenyl)silyl]oxymethyl]-4,4,4-trifluoro-2-(2-hydroxyethyl)butyl]carbamate (2.16 g, 2.7535 mmol) in anhydrous THF (50 mL) at 0 °C was added a solution of TBAF in THF (5.5 mL of 1 M, 5.5000 mmol) dropwise. The reaction was stirred at 0 °C for 30 minutes, then allowed to reach room temperature and stirred for 16 hours at room temperature. The reaction was quenched by the addition of a saturated aqueous solution of sodium bicarbonate (20 mL) and the THF was removed under reduced pressure. Brine (10 mL) was added, and the aqueous mixture was extracted with ethyl acetate (4 x 50 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude material was purified by reversed-phase chromatography on C18 column (gradient 5% to 40% acetonitrile in water). The fractions containing the desired product were combined, concentrated under reduced pressure and co-evaporated with acetonitrile (5 x 10 mL), to afford tert-butyl N-[(1R,2S)-4,4,4-trifluoro-2-(2-hydroxyethyl)-1- (hydroxymethyl)butyl]carbamate (788 mg, 90%) as a pale yellow oil.1H NMR (400 MHz, CDCl3) δ 5.28 - 5.06 (m, 1H), 3.92 - 3.55 (m, 5H), 2.96 - 2.69 (m, 1H), 2.59 - 2.23 (m, 3H), 2.22 - 2.03 (m, 1H), 1.86 - 1.55 (m, overlapped with water, 2H), 1.53 - 1.34 (m, 9H).19F NMR (377 MHz, CDCl3) δ -63.78 (t, J = 10.9 Hz, 3F). ESI-MS m/z calc.301.1501, found 202.2 (M-99)+; Retention time: 1.58 minutes. LC method U. Step 4: tert-Butyl (4R)-2,2-dimethyl-4-[(1S)-3,3,3-trifluoro-1-(2- hydroxyethyl)propyl]oxazolidine-3-carboxylate [00729] To a stirred solution of tert-butyl N-[(1R,2S)-4,4,4-trifluoro-2-(2-hydroxyethyl)-1- (hydroxymethyl)butyl]carbamate (688 mg, 2.1692 mmol) in dry DCM (20 mL), were added 2,2- dimethoxypropane (935.00 mg, 1.1 mL, 8.9776 mmol) and para toluenesulfonic acid monohydrate (41 mg, 0.2155 mmol). The mixture was stirred at room temperature for 2 hours. Acetone (10 mL) and isopropyl acetate (10 mL) were added, then a saturated aqueous solution of sodium bicarbonate (10 mL) was added, and the mixture was stirred at room temperature for 5 minutes. Organic solvents were evaporated under reduced pressure and co-evaporated with isopropyl acetate (3 x 5 mL). The aqueous layer was extracted with dichloromethane (2 x 50 mL) and with isopropyl acetate (50 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude mixture was purified by reversed-phase chromatography on a C18 cartridge, eluting with a gradient of 5 to 65% of acetonitrile in water. The fractions containing the product were combined, evaporated and co-evaporated with acetonitrile (3 x 5 mL), to afford tert-butyl (4R)- 2,2-dimethyl-4-[(1S)-3,3,3-trifluoro-1-(2-hydroxyethyl)propyl]oxazolidine-3-carboxylate (620 mg, 80%) as a colorless oil.1H NMR (400 MHz, methanol-d4) δ 4.11 - 3.95 (m, 2H), 3.94 - 3.83 (m, 1H), 3.72 - 3.56 (m, 2H), 2.59 - 2.40 (m, 2H), 2.18 - 1.99 (m, 1H), 1.86 - 1.67 (m, 1H), 1.60 (s, 3H), 1.57 - 1.51 (m, 1H), 1.48 (s, 9H), 1.45 (s, 3H). (1H missing, labile proton).19F NMR (377 MHz, methanol-d4) δ -65.74 (t, J = 10.9 Hz, 1.75F), -65.91 (t, J = 9.5 Hz, 1.25F). ESI-MS m/z calc.341.1814, found 286.1 (M-55)+; Retention time: 1.92 minutes. LC method U. Step 5: tert-Butyl (4R)-2,2-dimethyl-4-[(1S)-3,3,3-trifluoro-1-(2- oxoethyl)propyl]oxazolidine-3-carboxylate [00730] To a solution of tert-butyl (4R)-2,2-dimethyl-4-[(1S)-3,3,3-trifluoro-1-(2- hydroxyethyl)propyl]oxazolidine-3-carboxylate (620 mg, 1.7254 mmol) in dichloromethane (25 mL) at 0 °C, were added sodium bromide (59 mg, 0.5734 mmol) and TEMPO (73 mg, 0.4672 mmol). A stirred mixture of sodium bicarbonate (660 mg, 7.8565 mmol) in sodium hypochlorite (in water) (2.4 mL of 1.6 M, 3.8400 mmol) was prepared at 0 °C, filtered and added portion-wise to the reaction (in a way that the red coloration disappeared between each portion) over 60 minutes under vigorous stirring. A saturated aqueous solution of sodium bicarbonate (25 mL) was added. The mixture was diluted with DCM (25 mL) and the layers were separated. The aqueous layer was extracted with DCM (2 x 25 mL). The combined organic layers were washed with a saturated aqueous solution of sodium bicarbonate (25 mL), a 10% aqueous solution of Na2S2O3 (25 mL) and brine (10 mL), then dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford the crude mixture of rotamers of tert-butyl (4R)- 2,2-dimethyl-4-[(1S)-3,3,3-trifluoro-1-(2-oxoethyl)propyl]oxazolidine-3-carboxylate (619 mg, 91%) as an orange oil.1H NMR (400 MHz, methanol-d4) δ 9.68 (s, 1H), 4.59 (s, 1H), 4.23 - 3.75 (m, 3H), 2.80 - 2.34 (m, 2H), 2.34 - 1.96 (m, 1H), 1.93 - 1.68 (m, 1H), 1.68 - 1.36 (m, 15H).19F NMR (377 MHz, methanol-d4) δ -65.15 - -66.08 (m, 3F). ESI-MS m/z calc.339.1657, found 362.2 (M+23)+; Retention time: 2.03 minutes. LC method U. Step 6: tert-Butyl (4R)-4-[(E,1S)-5-[methoxy(methyl)amino]-5-oxo-1-(2,2,2- trifluoroethyl)pent-3-enyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00731] To a suspension of heptane washed sodium hydride in mineral oil (1.8 mg, 60 %w/w, 0.0450 mmol) in THF (0.25 mL), was added 2-diethoxyphosphoryl-N-methoxy-N-methyl- acetamide (11 mg, 0.0460 mmol) at room temperature. After the evolution of hydrogen stopped, this solution was transferred to a solution of tert-butyl (4R)-2,2-dimethyl-4-[(1S)-3,3,3-trifluoro- 1-(2-oxoethyl)propyl]oxazolidine-3-carboxylate (12 mg, 0.0285 mmol) in THF (0.25 mL) and left to react at room temperature for 2 hours. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (0.5 mL) and extracted with ethyl acetate (3 x 5 mL). The combined organic layers were washed with brine (1 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reversed- phase chromatography on C18 column (gradient 40% to 75% acetonitrile in water). The fractions containing the desired product were combined, concentrated under reduced pressure and co- evaporated with acetonitrile (3 x 1 mL), to afford tert-butyl (4R)-4-[(E,1S)-5- [methoxy(methyl)amino]-5-oxo-1-(2,2,2-trifluoroethyl)pent-3-enyl]-2,2-dimethyl-oxazolidine- 3-carboxylate (10 mg, 83%) as a colorless oil.1H NMR (400 MHz, methanol-d4) δ 7.04 - 6.78 (m, 1H), 6.73 - 6.44 (m, 1H), 4.07 - 3.80 (m, 3H), 3.74 (s, 3H), 3.24 (s, 3H), 2.74 - 2.23 (m, 4H), 2.22 - 2.04 (m, 1H), 1.60 (s, 3H), 1.54 - 1.41 (m, 12H).19F NMR (377 MHz, methanol-d4) δ - 65.71 - -65.91 (m, 3F). ESI-MS m/z calc.424.2185, found 425.4 (M+1)+; Retention time: 2.05 minutes. LC method U. Step 7: tert-Butyl (4R)-4-[(1S)-5-[methoxy(methyl)amino]-5-oxo-1-(2,2,2- trifluoroethyl)pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00732] Nitrogen was bubbled to a stirred solution of tert-butyl (4R)-4-[(E,1S)-5- [methoxy(methyl)amino]-5-oxo-1-(2,2,2-trifluoroethyl)pent-3-enyl]-2,2-dimethyl-oxazolidine- 3-carboxylate (890 mg, 1.5118 mmol) in dry methanol (50 mL) for 10 minutes, then 10% palladium on carbon (50% wet) (420 mg, 5 %w/w, 0.1973 mmol) was added. Nitrogen was bubbled to the mixture for 10 minutes, followed by hydrogen for 5 minutes. The reaction was then stirred for 1.5 hours, at room temperature, under hydrogen atmosphere. Nitrogen was bubbled to the mixture for 10 minutes. The mixture was filtered on a pad of Celite, washed with MeOH (20 mL) and concentrated in vacuo. The crude mixture was purified by reversed-phase chromatography on a C18 cartridge, eluting with a gradient of 40 to 70% of acetonitrile in water. The fractions containing the desired product were combined, concentrated in vacuo, then co- evaporated with acetonitrile (3 x 2 mL) and THF (3 x 1 mL), to afford tert-butyl (4R)-4-[(1S)-5- [methoxy(methyl)amino]-5-oxo-1-(2,2,2-trifluoroethyl)pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (537 mg, 82%) as a pale yellow oil.1H NMR (400 MHz, methanol-d4) δ 4.11 - 3.95 (m, 2H), 3.92 - 3.79 (m, 1H), 3.72 (s, 3H), 3.18 (s, 3H), 2.63 - 2.24 (m, 4H), 2.16 - 1.92 (m, 1H), 1.82 - 1.62 (m, 2H), 1.59 (s, 3H), 1.57 - 1.51 (m, 1H), 1.48 (s, 9H), 1.45 (s, 3H), 1.41 - 1.27 (m, 1H).19F NMR (377 MHz, methanol-d4) δ -65.64 - -66.03 (m, 3F). ESI-MS m/z calc.426.2342, found 427.1 (M+1)+; Retention time: 3.72 minutes. LC method W. Step 8: tert-Butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-5-oxo-1-(2,2,2-trifluoroethyl)pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00733] To a solution of 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine (62 mg, 0.2273 mmol) in THF (1.5 mL), at -78 °C, was added n-butyllithium (solution in hexanes) (0.1 mL of 2.5 M, 0.2500 mmol) dropwise. As soon as the addition of n-BuLi was over, a solution of tert-butyl (4R)-4-[(1S)-5-[methoxy(methyl)amino]-5-oxo-1-(2,2,2-trifluoroethyl)pentyl]-2,2- dimethyl-oxazolidine-3-carboxylate (60 mg, 0.1397 mmol) in THF (0.4 mL) was added dropwise. The reaction was left stirring 1 hour at -78 °C. More n-butyllithium (solution in hexanes) (0.1 mL of 1.75 M, 0.1750 mmol) was added and the reaction was left stirring 1 hour at -78 °C. A saturated aqueous solution of ammonium chloride (0.5 mL) and ethyl acetate (1 mL) were added. The mixture was slowly warmed up to room temperature and stirred for 30 minutes. The layers were separated, and the aqueous one was extracted with ethyl acetate (3 x 5 mL). The combined organic layers were washed with brine (2 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude was purified by reversed-phase chromatography on C18 column (gradient 50 to 80% acetonitrile in water). The desired fractions were concentrated under reduced pressure, then co-evaporated with acetonitrile (3 x 5 mL), to afford tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-5-oxo-1-(2,2,2-trifluoroethyl)pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (40 mg, 52%) as a white solid.1H NMR (400 MHz, CDCl3) δ 9.13 (s, 1H), 6.62 (s, 1H), 4.21 - 3.98 (m, 5H), 3.82 (d, J = 9.3 Hz, 1H), 3.40 - 3.25 (m, 2H), 2.64 - 2.36 (m, 2H), 2.05 - 1.77 (m, 3H), 1.74 - 1.61 (m, 4H), 1.57 (s, overlapped with water, 9H), 1.50 (s, 12H), 1.45 - 1.27 (m, 1H).19F NMR (377 MHz, CDCl3) δ -63.90 - -64.30 (m, 3F). ESI-MS m/z calc.554.308, found 555.2 (M+1)+; Retention time: 2.4 minutes. LC method U. Step 9: (5S)-5-[(1R)-1-Amino-2-hydroxy-ethyl]-1-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-7,7,7-trifluoro-heptan-1-one [00734] To a solution of tert-butyl (4R)-4-[(1S)-5-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-5-oxo-1-(2,2,2-trifluoroethyl)pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (432 mg, 0.7664 mmol) in 1,4-dioxane (12 mL) and water (3 mL), was added hydrochloric acid (solution in 1,4-dioxane) (10 mL of 4 M, 40.000 mmol). The reaction was stirred at room temperature for 20 hours, under nitrogen. The solvents were removed under reduced pressure and the mixture was co-evaporated with 1,4-dioxane (5 x 10 mL). The material was dissolved in dichloromethane (100 mL) and the organic layer was washed with a saturated aqueous solution of sodium bicarbonate (20 mL) and brine (5 mL). The organic layer was dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure, to afford (5S)-5-[(1R)-1-amino- 2-hydroxy-ethyl]-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-7,7,7-trifluoro-heptan-1- one (327 mg, 100%) as a yellow oil.1H NMR (400 MHz, CDCl3) δ 8.68 (s, 1H), 6.49 (s, 1H), 4.02 (s, 3H), 3.89 (d, J = 6.4 Hz, 1H), 3.83 - 3.74 (m, 3H), 3.70 (s, 1H), 3.68 - 3.62 (m, 1H), 2.68 - 2.50 (m, 1H), 2.36 - 2.14 (m, 2H), 2.14 - 1.96 (m, 3H), 1.91 - 1.78 (m, 1H), 1.74 - 1.62 (m, 2H), 1.61 - 1.46 (m, overlapped with water 9H).19F NMR (377 MHz, CDCl3) δ -63.54 (t, J = 11.6 Hz, 3F). ESI-MS m/z calc.414.2243, found 415.4 (M+1)+; Retention time: 1.44 minutes. LC method U. Step 10: [(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3- (2,2,2-trifluoroethyl)azepan-2-yl]methanol and [(2R,3S,7R)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(2,2,2-trifluoroethyl)azepan-2-yl]methanol [00735] To a solution of (5S)-5-[(1R)-1-amino-2-hydroxy-ethyl]-1-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-7,7,7-trifluoro-heptan-1-one (327 mg, 0.7692 mmol) in DCM (12 mL), at 0 °C, was added NaBH(OAc)3 (1.2 g, 5.5487 mmol). The reaction mixture was stirred for 2 hours at room temperature and a saturated aqueous solution of sodium bicarbonate (20 mL) was added at 0 °C. The layers were separated, and the aqueous layer was extracted with DCM (3 x 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography on a C18 column (isocratic 55% acetonitrile in 10 mM ammonium bicarbonate buffer: pH = 10). The fractions containing the first eluting product were combined and the volatiles were removed under reduced pressure. The residue was co-evaporated with acetonitrile (5 x 10 mL). The resulting product was dried under high vacuum to afford [(2R,3S,7S)-7-(6-tert- butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(2,2,2-trifluoroethyl)azepan-2-yl]methanol (204 mg, 66%) as a white solid.99.9% ee, 99.9% de.1H NMR (400 MHz, Benzene-d6) δ 8.56 (s, 1H), 6.59 (s, 1H), 3.71 (dd, J = 10.8, 4.2 Hz, 1H), 3.54 - 3.44 (m, 4H), 3.01 (t, J = 10.0 Hz, 1H), 2.29 (td, J = 9.0, 3.9 Hz, 1H), 2.00 - 1.88 (m, 1H), 1.80 - 1.43 (m, 7H), 1.29 - 1.14 (m, 1H), 1.14 - 1.07 (m, 9H). (2H missing, labile protons).19F NMR (377 MHz, Benzene-d6) δ -63.50 (t, J = 11.6 Hz, 3F). ESI-MS m/z calc.398.2293, found 399.2 (M+1)+; Retention time: 3.53 minutes. The fractions containing the second eluting product were combined and the volatiles were removed under reduced pressure. The residue was co-evaporated with acetonitrile (5 x 10 mL). The resulting product was dried under high vacuum to afford [(2R,3S,7R)-7-(6-tert-butyl-5- methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(2,2,2-trifluoroethyl)azepan-2-yl]methanol (37 mg, 12%) as a white solid.99.9% ee, 97.4% de.1H NMR (400 MHz, Benzene-d6) δ 8.54 (s, 1H), 6.59 (s, 1H), 3.68 (dd, J = 11.2, 2.7 Hz, 1H), 3.60 - 3.47 (m, 4H), 3.36 (t, J = 10.3 Hz, 1H), 2.34 (td, J = 9.7, 4.3 Hz, 1H), 2.06 - 1.95 (m, 1H), 1.74 - 1.57 (m, 3H), 1.57 - 1.28 (m, 3H), 1.27 - 1.00 (m, 11H). (2H missing, labile protons).19F NMR (377 MHz, Benzene-d6) δ -63.50 (t, J = 11.6 Hz, 3F). ESI-MS m/z calc.398.2293, found 399.2 (M+1)+; Retention time: 3.63 minutes. LC method W. Step 11: 3-[[4-[[(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 3-(2,2,2-trifluoroethyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00736] 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (101 mg, 0.2417 mmol) and [(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(2,2,2- trifluoroethyl)azepan-2-yl]methanol (96 mg, 0.2409 mmol) were combined in anhydrous THF (1.5 mL) under nitrogen atmosphere. After stirring a few minutes until all solids dissolved, sodium tert-butoxide (119 mg, 1.238 mmol) was added and the reaction mixture was stirred at room temperature for 4 hours, then stirred at 40 °C for 2 hours. An additional amount of sodium tert-butoxide (45 mg, 0.4682 mmol) was added and the reaction was stirred at 40 °C for 7 h. The reaction mixture was cooled to room temperature and diluted with EtOAc, brine and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (3 x). The organic layers were combined, washed with brine, dried over sodium sulfate and concentrated under reduced pressure. The residue was triturated in ethyl acetate, filtered and dried to give as a yellow solid 3-[[4-[[(2R,3S,7S)-7-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(2,2,2- trifluoroethyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (91 mg, 43%). ESI-MS m/z calc.779.3077, found 780.86 (M+1)⁺; Retention time: 1.53 minutes. LC method A. Step 12: (4S,8S,9R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-14- (2,6-dimethylphenyl)-18,18-dioxo-8-(2,2,2-trifluoroethyl)-11-oxa-18λ6-thia- 3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (Compound 136) [00737] 3-[[4-[[(2R,3S,7S)-7-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-(2,2,2- trifluoroethyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (91 mg, 0.1115 mmol) was dissolved under nitrogen in anhydrous NMP (2 mL) and acetone (4 mL). TsCl (67 mg, 0.3514 mmol) was added followed by NMM (77 µL, 0.7004 mmol) and the reaction was stirred at 55 °C for 24 h. The reaction was poured into a mixture of ethyl acetate and 1 M HCl and the layers were separated. The aqueous layer was further extracted with ethyl acetate (2 x). The organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was taken up in DMSO/MeOH and purified by reverse phase HPLC (10-99% acetonitrile/5 mM aqueous HCl over 25 min) to give after evaporation (4S,8S,9R)-4-(6-tert-butyl-5-methyl- pyrrolo[2,3-b]pyrazin-3-yl)-14-(2,6-dimethylphenyl)-18,18-dioxo-8-(2,2,2-trifluoroethyl)-11- oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (45 mg, 53%) as a yellow solid.1H NMR (400 MHz, CDCl3) δ 8.82 (s, 1H), 8.45 (s, 1H), 7.79 (d, J = 7.7 Hz, 2H), 7.58 (t, J = 7.7 Hz, 1H), 7.25 - 7.21 (m, 1H), 7.10 (d, J = 7.6 Hz, 2H), 6.52 (s, 1H), 6.31 (s, 1H), 5.97 (dd, J = 11.9, 5.3 Hz, 1H), 5.91 (dd, J = 13.3, 4.7 Hz, 1H), 4.89 (t, J = 11.9 Hz, 1H), 4.57 (dt, J = 11.1, 4.9 Hz, 1H), 4.15 (s, 3H), 2.45 - 2.32 (m, 2H), 2.26 - 2.12 (m, 2H), 2.07 (s, 6H), 2.05 - 1.95 (m, 2H), 1.92 - 1.74 (m, 3H), 1.50 (s, 9H).19F NMR (376 MHz, CDCl3) δ -64.06 (t, J = 10.9 Hz). ESI- MS m/z calc.761.2971, found 762.4 (M+1)⁺; Retention time: 2.06 minutes. LC method A. Example 87: Preparation of Compound 137 Step 1: (3S,7aR)-3-Phenyl-3,6,7,7a-tetrahydro-1H-pyrrolo[1,2-c]oxazol-5-one [00738] To a solution of (5R)-5-(hydroxymethyl)pyrrolidin-2-one (25 g, 217.15 mmol) in toluene (100 mL) was added benzaldehyde (22.880 g, 22 mL, 215.60 mmol) and PTSA hydrate (2.06 g, 10.830 mmol). The reaction was stirred at reflux overnight. The reaction was allowed to cool down to room temperature and the solvent was removed under reduced pressure. The crude residue was purified by silica gel chromatography eluting from 30-80 % EtOAc in heptanes) to give (3S,7aR)-3-phenyl-3,6,7,7a-tetrahydro-1H-pyrrolo[1,2-c]oxazol-5-one (2.71 g, 6%) as a dark orange oil.1H NMR (400 MHz, DMSO-d6) δ 7.98 - 7.88 (m, 1H), 7.65 - 7.58 (m, 1H), 7.41 - 7.36 (m, 3H), 6.07 (s, 1H), 4.21 - 4.09 (m, 2H), 3.49 - 3.40 (m, 1H), 2.79 - 2.66 (m, 1H), 2.47 - 2.36 (m, 1H), 2.34 - 2.22 (m, 1H), 1.97 - 1.85 (m, 1H). ESI-MS m/z calc.203.0946, found 204.2 (M+1)+; Retention time: 1.56 minutes. LC method U. Step 2: (3S,7aR)-3-Phenyl-6-phenylselanyl-3,6,7,7a-tetrahydro-1H-pyrrolo[1,2- c]oxazol-5-one [00739] To a solution of NaHMDS (in THF) (55 mL of 1 M, 55.000 mmol) in anhydrous THF (120 mL) at -78 °C, was added a solution of (3S,7aR)-3-phenyl-3,6,7,7a-tetrahydro-1H- pyrrolo[1,2-c]oxazol-5-one (11.6 g, 52.853 mmol) in anhydrous THF (20 mL) dropwise over 10 minutes. The reaction was stirred at -78 °C for 30 minutes then a solution of TMSCl (8.0464 g, 9.4 mL, 74.063 mmol) in anhydrous THF (15 mL) was added dropwise over 10 minutes. The reaction was allowed to warm to 0 °C in an ice-cold bath and the reaction was stirred at that temperature for 3 hours. To the reaction mixture at 0 °C was added a solution of PhSeCl (12.0 g, 62.658 mmol) in anhydrous THF (15 mL) dropwise over 5 minutes and the reaction was stirred at room temperature for 1 hour. To the reaction mixture was added an aqueous saturated solution of sodium bicarbonate (50 mL) and EtOAc (50 mL). The phases were separated, and the aqueous layer was extracted with EtOAc (2 x 50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography (Column: C18. Gradient: 5-80 % MeOH in water). The fractions containing the product were collected and the volatiles were removed under reduced pressure. The aqueous solution was diluted with brine (100 mL) and extracted with DCM (3 x 100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford (3S,7aR)-3-phenyl-6-phenylselanyl-3,6,7,7a-tetrahydro-1H-pyrrolo[1,2- c]oxazol-5-one (13 g, 65%) as an orange oil as a mixture of diastereomers in an approximate 60/40 ratio.1H NMR (400 MHz, CDCl3) δ 7.72 - 7.62 (m, 2H), 7.48 - 7.42 (m, 0.6H), 7.40 - 7.30 (m, 5H), 7.29 - 7.24 (overlapped with solvent signal, m, 1H), 7.23 - 7.15 (m, 1.4H), 6.28 (s, 0.6H), 6.27 (s, 0.4H), 4.37 (t, J = 9.3 Hz, 0.4H), 4.16 - 4.09 (m, 1H), 4.09 - 4.01 (m, 0.4H), 3.99 (dd, J = 6.8, 4.6 Hz, 0.6H), 3.67 - 3.58 (m, 0.6H), 3.42 (t, J = 8.1 Hz, 0.6H), 3.10 (t, J = 7.9 Hz, 0.4H), 2.84 (ddd, J = 13.9, 9.5, 7.6 Hz, 0.4H), 2.55 - 2.48 (m, 1.2H), 2.03 (ddd, J = 14.1, 8.9, 5.4 Hz, 0.4H). ESI-MS m/z calc.359.0425, found 360.2 (M+1)+; Retention time: 1.93 minutes. LC method U. Step 3: (3S,7aR)-3-Phenyl-3,7a-dihydro-1H-pyrrolo[1,2-c]oxazol-5-one [00740] To a solution of (3S,7aR)-3-phenyl-6-phenylselanyl-3,6,7,7a-tetrahydro-1H- pyrrolo[1,2-c]oxazol-5-one (13.0 g, 34.396 mmol) in THF (200 mL) at 0 °C was added an aqueous solution of hydrogen peroxide (in water) (1.1 g, 0.9910 mL of 35 %w/w, 11.319 mmol) dropwise. The reaction mixture was stirred at this temperature for 1 hour, then was stirred at room temperature for 3 hours. The reaction mixture was cooled in an ice-cold bath and a solution of hydrogen peroxide (in water) (5.6 g, 5.0450 mL of 35 %w/w, 57.622 mmol) was again added when the internal temperature was 15 °C. The bath was removed, and the reaction was stirred at room temperature for 15 minutes. To the reaction mixture was added a saturated aqueous solution of sodium bicarbonate (100 mL) and EtOAc (100 mL). The phases were separated, and the aqueous layer was extracted with EtOAc (2 x 100 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography (Column: C18. Gradient: 5-40 % MeOH in water). The fractions containing the product were collected and the volatiles were removed under reduced pressure. The aqueous solution was diluted with brine (100 mL) and extracted with DCM (3 x 100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford (3S,7aR)-3-phenyl-3,7a-dihydro-1H-pyrrolo[1,2-c]oxazol-5-one (6.47 g, 93%) as a beige solid.1H NMR (400 MHz, CDCl3) δ 7.58 - 7.51 (m, 2H), 7.44 - 7.33 (m, 3H), 7.28 (dd, J = 5.6, 2.0 Hz, 1H), 6.21 (s, 1H), 6.18 (dd, J = 5.6, 1.5 Hz, 1H), 4.64 (ddt, J = 8.5, 6.9, 1.6 Hz, 1H), 4.31 - 4.26 (m, 1H), 3.45 (t, J = 8.3 Hz, 1H). ESI-MS m/z calc.201.079, found 202.2 (M+1)+; Retention time: 1.57 minutes. LC method U. Step 4: (1R,2S,4R,7S)-3,3-Dimethyl-7-phenyl-8-oxa-6- azatricyclo[4.3.0.02,4]nonan-5-one [00741] To a solution of isopropyl(triphenyl)phosphonium iodide (75 mg, 0.1735 mmol) in anhydrous THF (0.5 mL) at -30 °C was added a solution of n-BuLi (in hexanes) (0.06 mL of 2.5 M, 0.1500 mmol). The reaction was stirred for 30 minutes at 0 °C then cooled down to -30 °C. To the reaction mixture at that temperature was added a solution of (3S,7aR)-3-phenyl-3,7a- dihydro-1H-pyrrolo[1,2-c]oxazol-5-one (25 mg, 0.1241 mmol) in anhydrous THF (0.3 mL) dropwise. The reaction was stirred in the cooling bath for 45 minutes allowing the temperature to slowly reach 0 °C then the bath was removed, and the reaction was stirred at room temperature for 18 hours. To the reaction mixture was added water (5 mL) and the aqueous solution was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography (Column: C18. Gradient: 50-100 % acetonitrile in water). The fractions containing the product were collected and the volatiles were removed under reduced pressure. The aqueous solution was diluted with brine (20 mL) and extracted with DCM (3 x 10 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (Gradient: 0-30 % EtOAc in heptane) to afford (1R,2S,4R,7S)-3,3-dimethyl-7-phenyl-8-oxa-6-azatricyclo[4.3.0.02,4]nonan-5-one (9 mg, 30%) as a white solid.1H NMR (400 MHz, CDCl3) δ 7.43 - 7.28 (m, 5H), 6.31 (s, 1H), 4.22 (dd, J = 7.7, 6.2 Hz, 1H), 3.73 (dd, J = 9.3, 6.4 Hz, 1H), 3.53 (dd, J = 9.3, 7.8 Hz, 1H), 1.97 - 1.86 (m, 2H), 1.33 (s, 3H), 1.19 (s, 3H). ESI-MS m/z calc.243.1259, found 244.2 (M+1)+; Retention time: 1.74 minutes. LC method I. Step 5: (1R,4R,5S)-4-(Hydroxymethyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexan-2- one [00742] Trifluoroacetic acid (18.500 g, 12.5 mL, 162.25 mmol) and then water (12.000 g, 12 mL, 666.10 mmol) were added to a solution of (1R,2S,4R,7S)-3,3-dimethyl-7-phenyl-8-oxa-6- azatricyclo[4.3.0.02,4]nonan-5-one (2.2 g, 8.9157 mmol) in DCM (125 mL) at room temperature and the solution was stirred overnight at this temperature. The mixture was concentrated under reduced pressure. The resulting residue was partitioned between concentrated aqueous sodium bicarbonate (50 mL) and ethyl acetate (100 mL). The aqueous phase was separated and extracted with ethyl acetate (4 x 100 mL). The combined ethyl acetate extracts were dried over magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by reverse phase chromatography on a C18 column using a 5 to 100% gradient of acetonitrile in pure water to provide (1R,4R,5S)-4-(hydroxymethyl)-6,6- dimethyl-3-azabicyclo[3.1.0]hexan-2-one (1.12 g, 77%) as a white solid.1H NMR (400 MHz, CDCl3) δ 6.51 (br. s, 1H), 3.73 - 3.64 (m, 1H), 3.63 - 3.54 (m, 1H), 3.50 - 3.44 (m, 1H), 2.99 (br. s, 1H), 1.73 (d, J = 6.4 Hz, 1H), 1.53 (d, J = 5.6 Hz, 1H), 1.14 (s, 3H), 1.13 (s, 3H). ESI-MS m/z calc.155.0946, found 156.1 (M+1)+; Retention time: 0.81 minutes. LC method 1E. Step 6: (1R,4R,5S)-4-[[tert-Butyl(diphenyl)silyl]oxymethyl]-6,6-dimethyl-3- azabicyclo[3.1.0]hexan-2-one [00743] TBDPSCl (7 g, 25.467 mmol) was added to a solution of (1R,4R,5S)-4- (hydroxymethyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexan-2-one (1.98 g, 12.120 mmol) and imidazole (3.6 g, 52.881 mmol) in DCM (40 mL) at 0 °C then left to warm up to room temperature and stirred for 3 h. The reaction mixture was quenched with water (50 mL) and extracted with DCM (3 x 50 mL). The combined organics were dried over magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by reverse phase chromatography on a C18 column using a 5 to 100% gradient of acetonitrile in pure water to provide (1R,4R,5S)-4-[[tert-butyl(diphenyl)silyl]oxymethyl]-6,6-dimethyl-3- azabicyclo[3.1.0]hexan-2-one (4.17 g, 86%) as a thick oil 1H NMR (400 MHz, CDCl3) δ 7.69 - 7.63 (m, 4H), 7.49 - 7.38 (m, 6H), 5.56 (br. s, 1H), 3.63 (d, J = 6.1 Hz, 2H), 3.51 - 3.41 (m, 1H), 1.68 (dt, J = 6.3, 1.4 Hz, 1H), 1.44 (dd, J = 6.1, 1.0 Hz, 1H), 1.16 (s, 3H), 1.10 (s, 3H), 1.07 (s, 9H). ESI-MS m/z calc.393.2124, found 394.2 (M+1)+; Retention time: 2.97 minutes. LC method 1E. Step 7: tert-butyl (1S,2R,5R)-2-[[tert-Butyl(diphenyl)silyl]oxymethyl]-6,6- dimethyl-4-oxo-3-azabicyclo[3.1.0]hexane-3-carboxylate [00744] To (1R,4R,5S)-4-[[tert-butyl(diphenyl)silyl]oxymethyl]-6,6-dimethyl-3- azabicyclo[3.1.0]hexan-2-one (25 mg, 0.0632 mmol) and DMAP (16 mg, 0.1310 mmol) in acetonitrile (500 μL) was added di-tert-butyl dicarbonate (27 mg, 0.1237 mmol) at room temperature and the reaction mixture was left stirring overnight. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by reverse phase chromatography on a C18 column using a 5 to 100% gradient of acetonitrile in pure water to provide tert-butyl (1S,2R,5R)-2-[[tert-butyl(diphenyl)silyl]oxymethyl]-6,6-dimethyl-4-oxo-3- azabicyclo[3.1.0]hexane-3-carboxylate (31.9 mg, 100%) as a clear film 1H NMR (400 MHz, CDCl3) δ 7.71 - 7.59 (m, 4H), 7.49 - 7.33 (m, 6H), 4.00 - 3.78 (m, 3H), 1.88 (dd, J = 6.2, 1.3 Hz, 1H), 1.69 (d, J = 6.4 Hz, 1H), 1.40 (s, 9H), 1.15 (s, 3H), 1.11 (s, 3H), 1.06 (s, 9H). ESI-MS m/z calc.493.2648, found 394.0 (M-99)+; Retention time: 3.39 minutes. LC method 1C. Step 8: tert-Butyl N-[(1R)-2-[tert-butyl(diphenyl)silyl]oxy-1-[(1S,3R)-3- (hydroxymethyl)-2,2-dimethyl-cyclopropyl]ethyl]carbamate [00745] NaBH4 (7.6 g, 200.89 mmol) was added to a solution of tert-butyl (1S,2R,5R)-2-[[tert- butyl(diphenyl)silyl]oxymethyl]-6,6-dimethyl-4-oxo-3-azabicyclo[3.1.0]hexane-3-carboxylate (5 g, 9.9957 mmol) in ethanol (160 mL) maintained at 80 °C with an oil bath. The reaction mixture was stirred at this temperature for 1 h then concentrated under reduced pressure and the resulting residue was partitioned between distilled water (500 mL) and ethyl acetate containing 20% heptanes (250 mL). The aqueous phase was separated and extracted with ethyl acetate containing 20% heptanes (2 x 250 mL). The combined organics were dried over magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by reverse phase chromatography on a C18 column using a 5 to 100% gradient of acetonitrile in pure water to provide tert-butyl N-[(1R)-2-[tert-butyl(diphenyl)silyl]oxy-1-[(1S,3R)-3- (hydroxymethyl)-2,2-dimethyl-cyclopropyl]ethyl]carbamate (4.3 g, 79%) as a clear oil that solidified under high vacuum 1H NMR (400 MHz, Chloroform-d) δ 7.67 (ddd, J = 7.2, 5.7, 1.5 Hz, 4H), 7.52 - 7.35 (m, 6H), 5.02 (br s, 1H), 3.87 - 3.70 (m, 2H), 3.68 - 3.50 (m, 3H), 3.16 (br d, J = 2.0 Hz, 1H), 1.46 (s, 9H), 1.08 (s, 9H), 1.05 - 0.96 (m, 4H), 0.90 - 0.79 (m, 4H). ESI-MS m/z calc.497.2961, found 498.2 (M+1)+; Retention time: 3.33 minutes. LC method 1C. Step 9: tert-Butyl N-[(1R)-2-hydroxy-1-[(1S,3R)-3-(hydroxymethyl)-2,2-dimethyl- cyclopropyl]ethyl]carbamate [00746] TBAF (in THF) (10 mL of 1 M, 10.000 mmol) was added to a solution of tert-butyl N-[(1R)-2-[tert-butyl(diphenyl)silyl]oxy-1-[(1S,3R)-3-(hydroxymethyl)-2,2-dimethyl- cyclopropyl]ethyl]carbamate (4.3 g, 7.9306 mmol) in THF (46 mL) maintained at 0 °C and the reaction was left stirring at this temperature for 1 h. More TBAF (in THF) (25 mL of 1 M, 25.000 mmol) was added, and the reaction was left stirring for another 1 h. The reaction mixture was quenched with a saturated aqueous solution of sodium bicarbonate (100 mL). The mixture was extracted with ethyl acetate (3 x 100 mL). The combined organics were dried over magnesium sulfate, filtered and concentrated under reduced pressure to provide the crude tert- butyl N-[(1R)-2-hydroxy-1-[(1S,3R)-3-(hydroxymethyl)-2,2-dimethyl- cyclopropyl]ethyl]carbamate (11.9 g, 100%) as a clear yellow oil. It was used directly in the next reaction. Step 10: tert-Butyl (4R)-4-[(1S,3R)-3-(hydroxymethyl)-2,2-dimethyl- cyclopropyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00747] PTSA hydrate (300 mg, 0.2804 mL, 1.5771 mmol) was added to a solution of tert- butyl N-[(1R)-2-hydroxy-1-[(1S,3R)-3-(hydroxymethyl)-2,2-dimethyl- cyclopropyl]ethyl]carbamate (11.8 g, 7.8715 mmol) in 2,2-dimethoxypropane (40 g, 384.07 mmol) and DCM (200 mL) and the reaction was left stirring for 2 h at room temperature. The reaction mixture was concentrated under reduced pressure. and the resulting residue quenched with aqueous sodium bicarbonate (250 mL) and extracted with DCM (3 x 200 mL). The combined organics were dried over magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by reverse phase chromatography on a C18 column using a 5 to 100% gradient of acetonitrile in pure water to provide tert-butyl (4R)-4-[(1S,3R)-3- (hydroxymethyl)-2,2-dimethyl-cyclopropyl]-2,2-dimethyl-oxazolidine-3-carboxylate (2.44 g, 98%) as a clear oil. ESI-MS m/z calc.299.2097, found 300.2 (M+1)+; Retention time: 2.42 minutes. LC method 1C. Step 11: tert-Butyl (4R)-4-[(1S,3R)-3-[(E)-3-[methoxy(methyl)amino]-3-oxo- prop-1-enyl]-2,2-dimethyl-cyclopropyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00748] A cold solution 0 °C of sodium hypochlorite (in water) (0.432 mL of 1.7 M, 0.7344 mmol) with sodium bicarbonate (73 mg, 0.8690 mmol) was added dropwise to a stirring suspension of tert-butyl (4R)-4-[(1S,3R)-3-(hydroxymethyl)-2,2-dimethyl-cyclopropyl]-2,2- dimethyl-oxazolidine-3-carboxylate (200 mg, 0.6346 mmol), NaBr (3.4 mg, 0.0330 mmol), and TEMPO (2.1 mg, 0.0134 mmol) in DCM (10 mL), water (2.5 mL) maintained at 0 °C. The reaction was stirred at this temperature for 1 h then isopropanol was added and the reaction was left stirring for another 10 min. The aqueous phase was separated and extracted with DCM (10 mL). The combined organics were dried over magnesium sulfate, filtered and concentrated under reduced pressure to provide crude tert-butyl (4R)-4-[(1S,3R)-3-formyl-2,2-dimethyl- cyclopropyl]-2,2-dimethyl-oxazolidine-3-carboxylate (190 mg) as a clear oil. Diethyl (2- (methoxy(methyl)amino)-2-oxoethyl)phosphonate (340 mg, 1.3426 mmol) in THF (10 mL) was added to heptane washed NaH (in oil) (27 mg, 60 %w/w, 0.6751 mmol). The mixture was left stirring at room temperature for 10 min. The crude product tert-butyl (4R)-4-[(1S,3R)-3-formyl- 2,2-dimethyl-cyclopropyl]-2,2-dimethyl-oxazolidine-3-carboxylate from the step above was dissolved in THF (1 mL) and added to the reaction mixture maintained at room temperature. This mixture was left stirring for 1 h then quenched with water (10 mL) and DCM (20 mL). The aqueous phase was separated and washed with DCM (3 x 10 mL). The organics were combined and dried over magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by reverse phase chromatography on a C18 column using a 5 to 65 % gradient followed by 65% isocratic acetonitrile in pure water to provide tert-butyl (4R)-4- [(1S,3R)-3-[(E)-3-[methoxy(methyl)amino]-3-oxo-prop-1-enyl]-2,2-dimethyl-cyclopropyl]-2,2- dimethyl-oxazolidine-3-carboxylate (172 mg, 64%) as a clear oil. ESI-MS m/z calc.382.2468, found 383.2 (M+1)+; Retention time: 2.48 minutes. LC method 1C. Step 12: tert-Butyl (4R)-4-[(1S,3R)-3-[3-[methoxy(methyl)amino]-3-oxo-propyl]- 2,2-dimethyl-cyclopropyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00749] Nitrogen was bubbled to a stirred solution of tert-butyl (4R)-4-[(1S,3R)-3-[(E)-3- [methoxy(methyl)amino]-3-oxo-prop-1-enyl]-2,2-dimethyl-cyclopropyl]-2,2-dimethyl- oxazolidine-3-carboxylate (1.3 g, 3.3953 mmol) in dry methanol (55 mL), at 0 °C, for 10 minutes, then 10% palladium on carbon (50% wet) (140 mg, 5 %w/w, 0.0658 mmol) was added. Nitrogen was bubbled to the mixture for 10 minutes, followed by hydrogen for 5 minutes. The reaction was then stirred for 80 minutes, at 0 °C, under hydrogen atmosphere. Nitrogen was bubbled to the mixture for 10 minutes. The mixture was filtered on a pad of Celite, washed with MeOH (20 mL) and concentrated in vacuo. The crude mixture was purified by reversed-phase chromatography on a C18 cartridge, eluting with a gradient of 50 to 70% of acetonitrile in water. The fractions containing the desired product were combined, concentrated in vacuo, then co- evaporated with acetonitrile (5 x 10 mL), to afford tert-butyl (4R)-4-[(1S,3R)-3-[3- [methoxy(methyl)amino]-3-oxo-propyl]-2,2-dimethyl-cyclopropyl]-2,2-dimethyl-oxazolidine-3- carboxylate (1.067 g, 82%) as a colorless oil.1H NMR (400 MHz, methanol-d4) δ 4.02 (dd, J = 8.4, 5.5 Hz, 1H), 3.83 - 3.62 (m, 5H), 3.19 (br. s, 3H), 2.71 - 2.34 (m, 2H), 2.12 - 1.96 (m, 1H), 1.77 - 1.55 (m, 4H), 1.50 (s, 3H), 1.48 (s, 9H), 1.07 (s, 3H), 1.00 (s, 3H), 0.84 (t, J = 9.5 Hz, 1H), 0.67 (td, J = 8.9, 5.5 Hz, 1H). ESI-MS m/z calc.384.2624, found 385.2 (M+1)+; Retention time: 2.1 minutes. LC method X. Step 13: tert-Butyl (4R)-4-[(1S,3R)-3-[3-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-oxo-propyl]-2,2-dimethyl-cyclopropyl]-2,2-dimethyl- oxazolidine-3-carboxylate [00750] To a solution of 3-bromo-6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazine (569 mg, 2.0859 mmol) in THF (10 mL), at -78 °C, was added n-butyllithium (solution in hexanes) (1.1 mL of 1.75 M, 1.9250 mmol) dropwise. As soon as the addition of n-BuLi was over, a solution of tert-butyl (4R)-4-[(1S,3R)-3-[3-[methoxy(methyl)amino]-3-oxo-propyl]-2,2-dimethyl- cyclopropyl]-2,2-dimethyl-oxazolidine-3-carboxylate (500 mg, 1.2991 mmol) in THF (4 mL) was added dropwise. The reaction was left stirring 1 hour at -78 °C. A saturated aqueous solution of ammonium chloride (20 mL) and ethyl acetate (10 mL) were added. The mixture was slowly warmed up to room temperature and stirred for 30 minutes. The layers were separated, and the aqueous one was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude was purified by reversed-phase chromatography on C18 column (gradient 60 to 85% acetonitrile in water). The desired fractions were concentrated under reduced pressure, then co-evaporated with acetonitrile (5 x 10 mL), to afford tert-butyl (4R)-4-[(1S,3R)-3-[3-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-3-oxo-propyl]-2,2- dimethyl-cyclopropyl]-2,2-dimethyl-oxazolidine-3-carboxylate (440 mg, 66%) as a pale yellow solid.1H NMR (400 MHz, methanol-d4) δ 8.97 (s, 1H), 6.61 (s, 1H), 4.13 (s, 3H), 4.01 (dd, J = 8.1, 5.1 Hz, 1H), 3.80 - 3.69 (m, 2H), 3.45 - 3.25 (m, overlapped with MeOH, 2H), 2.28 - 2.12 (m, 1H), 1.88 - 1.67 (m, 1H), 1.59 (s, 3H), 1.57 (s, 9H), 1.48 (s, overlapped with impurity, 3H), 1.37 (s, 9H), 1.07 - 1.00 (m, 6H), 0.91 - 0.83 (m, 1H), 0.82 - 0.71 (m, 1H). ESI-MS m/z calc. 512.3363, found 513.4 (M+1)+; Retention time: 2.47 minutes. LC method U. Step 14: 3-[(1R,3S)-3-[(1R)-1-Amino-2-hydroxy-ethyl]-2,2-dimethyl- cyclopropyl]-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)propan-1-one [00751] To a solution of tert-butyl (4R)-4-[(1S,3R)-3-[3-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-3-oxo-propyl]-2,2-dimethyl-cyclopropyl]-2,2-dimethyl-oxazolidine-3- carboxylate (440 mg, 0.8574 mmol) in 1,4-dioxane (12 mL) and water (3 mL), was added hydrochloric acid (solution in 1,4-dioxane) (11 mL of 4 M, 44.000 mmol). The mixture was stirred at room temperature for 2 hours, under nitrogen. The solvents were removed under reduced pressure and the mixture was co-evaporated with 1,4-dioxane (5 x 10 mL). The material was dissolved in dichloromethane (50 mL) and the organic layer was washed with a saturated aqueous solution of sodium bicarbonate (6 mL) and brine (2 mL). The aqueous layer was extracted with dichloromethane (3 x 50 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered and evaporated under reduced pressure, to afford 3-[(1R,3S)-3-[(1R)-1-amino-2-hydroxy-ethyl]-2,2-dimethyl-cyclopropyl]-1- (6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)propan-1-one (317 mg, 99%) as an orange semi-solid.1H NMR (400 MHz, CDCl3) δ 9.14 (s, 1H), 6.52 (s, 1H), 4.03 (s, 3H), 3.87 - 3.72 (m, 3H), 3.70 (s, 2H), 3.68 - 3.61 (m, 1H), 3.55 - 3.43 (m, 1H), 3.03 - 2.92 (m, 1H), 2.05 - 1.88 (m, 2H), 1.52 (s, 9H), 1.21 (s, 3H), 1.06 (s, 3H), 0.98 - 0.87 (m, 1H), 0.83 - 0.73 (m, 1H). ESI- MS m/z calc.372.2525, found 373.4 (M+1)+; Retention time: 1.46 minutes. LC method U. Step 15: [(1S,2R,4S,7R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)- 8,8-dimethyl-3-azabicyclo[5.1.0]octan-2-yl]methanol [00752] To a solution of 3-[(1R,3S)-3-[(1R)-1-amino-2-hydroxy-ethyl]-2,2-dimethyl- cyclopropyl]-1-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)propan-1-one (319 mg, 0.8555 mmol) in DCM (10 mL), at 0 °C, was added NaBH(OAc)3 (1.3 g, 6.0111 mmol). The reaction mixture was stirred for 2 hours at room temperature and a saturated aqueous solution of sodium bicarbonate (20 mL) was added at 0 °C. The layers were separated, and the aqueous layer was extracted with DCM (4 x 20 mL). The combined organic layers were washed with brine (2 x 2 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The crude residue was purified by reversed-phase chromatography on a C18 column (gradient 40 to 50% acetonitrile in 10 mM ammonium bicarbonate buffer : pH = 10). The fractions containing the product were combined and the volatiles were removed under reduced pressure. The residue was co-evaporated with acetonitrile (5 x 10 mL). The resulting product was dried under high vacuum to afford [(1S,2R,4S,7R)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-8,8-dimethyl-3-azabicyclo[5.1.0]octan-2-yl]methanol (200 mg, 66%) as a white solid.1H NMR (400 MHz, Benzene-d6) δ 8.59 (s, 1H), 6.61 (s, 1H), 3.79 - 3.67 (m, 1H), 3.64 (dd, J = 10.5, 4.4 Hz, 1H), 3.51 (s, 3H), 3.23 (t, J = 10.1 Hz, 1H), 2.58 (td, J = 10.1, 4.4 Hz, 1H), 2.04 - 1.83 (m, 3H), 1.30 - 1.16 (m, 1H), 1.13 (s, 9H), 1.01 (s, 3H), 0.89 (s, 3H), 0.67 - 0.56 (m, 1H), 0.26 - 0.14 (m, 1H). (2H missing, labile protons). ESI-MS m/z calc.356.2576, found 357.2 (M+1)+; Retention time: 3.48 minutes. LC method W. Step 16: 3-[[4-[[(1S,2R,4S,7R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3- yl)-8,8-dimethyl-3-azabicyclo[5.1.0]octan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid [00753] 3-[[4-Chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (102 mg, 0.2441 mmol) and [(1S,2R,4S,7R)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8,8- dimethyl-3-azabicyclo[5.1.0]octan-2-yl]methanol (87.9 mg, 0.2466 mmol) were combined in anhydrous THF (1.5 mL) under nitrogen atmosphere. After stirring a few minutes until all solids dissolved, sodium tert-butoxide (157 mg, 1.634 mmol) was added and the reaction mixture was stirred at room temperature for 2 hours, then stirred at 40 °C for 3 hours. An additional amount of sodium tert-butoxide (52 mg, 0.5411 mmol) was added and the reaction was stirred at 40 °C for 19 h. The reaction mixture was cooled to room temperature and diluted with EtOAc, brine and 1 M HCl. The organic phase was separated, and the aqueous layer was extracted with EtOAc (3 x). The organic layers were combined, washed with brine, dried over sodium sulfate and concentrated under reduced pressure. The residue was triturated in ethyl acetate, filtered and dried to give as a yellow solid 3-[[4-[[(1S,2R,4S,7R)-4-(6-tert-butyl-5-methyl-pyrrolo[2,3- b]pyrazin-3-yl)-8,8-dimethyl-3-azabicyclo[5.1.0]octan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (68 mg, 36%). ESI- MS m/z calc.737.33594, found 738.44 (M+1)⁺; Retention time: 1.54 minutes. LC method A. Step 17: (4S,7R,9S,10R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-15- (2,6-dimethylphenyl)-8,8-dimethyl-19,19-dioxo-12-oxa-19λ6-thia-3,16,18,25- tetrazapentacyclo[18.3.1.113,17.03,10.07,9]pentacosa- 1(23),13,15,17(25),20(24),21-hexaen-2-one (Compound 137)
[00754] 3-[[4-[[(1S,2R,4S,7R)-4-(6-tert-Butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-8,8- dimethyl-3-azabicyclo[5.1.0]octan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid (hydrochloride salt) (68 mg, 0.08781 mmol) was dissolved under nitrogen in anhydrous NMP (1.5 mL) and acetone (3 mL). TsCl (53 mg, 0.2780 mmol) was added followed by NMM (64 µL, 0.5821 mmol) and the reaction was stirred at 55 °C for 3 days. The reaction was poured into a mixture of ethyl acetate and 1 M HCl and the layers were separated. The aqueous layer was further extracted with ethyl acetate (2 x). The organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was taken up in DMSO/MeOH and purified by reverse phase HPLC (10-99% acetonitrile/5 mM aqueous HCl over 25 min) to give after evaporation (4S,7R,9S,10R)- 4-(6-tert-butyl-5-methyl-pyrrolo[2,3-b]pyrazin-3-yl)-15-(2,6-dimethylphenyl)-8,8-dimethyl- 19,19-dioxo-12-oxa-19λ6-thia-3,16,18,25-tetrazapentacyclo[18.3.1.113,17.03,10.07,9]pentacosa- 1(23),13,15,17(25),20(24),21-hexaen-2-one (15.9 mg, 24%) as a tan solid.1H NMR (400 MHz, CDCl3) δ 9.46 (broad s, 1H), 8.91 (s, 1H), 8.53 (s, 1H), 8.20 (d, J = 7.8 Hz, 1H), 7.85 (d, J = 7.6 Hz, 1H), 7.69 (t, J = 7.7 Hz, 1H), 7.16 (t, J = 7.6 Hz, 1H), 6.99 (d, J = 7.6 Hz, 2H), 6.50 (s, 1H), 6.36 (t, J = 8.3 Hz, 1H), 6.04 (s, 1H), 5.09 (dd, J = 10.8, 4.5 Hz, 1H), 4.51 - 4.38 (m, 1H), 4.06 (s, 3H), 3.58 (t, J = 11.4 Hz, 1H), 2.78 - 2.64 (m, 1H), 2.63 - 2.46 (m, 1H), 2.35 - 2.18 (m, 1H), 1.94 (s, 6H), 1.53 - 1.38 (m, 10H), 1.22 (q, J = 9.2 Hz, 1H), 0.96 - 0.77 (m, 6H), 0.49 (dd, J = 9.2, 2.1 Hz, 1H). ESI-MS m/z calc.719.3254, found 720.44 (M+1)⁺; Retention time: 2.15 minutes. LC method A. Example 88: Preparation of Compound 139 Step 1: tert-Butyl (4R)-4-[(1S)-5-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-1- (cyclopropylmethyl)-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate [00755] To a solution of 2-bromo-6-tert-butyl-furo[2,3-b]pyrazine (1.74 g, 6.5955 mmol) in anhydrous THF (25 mL) at -94 °C under nitrogen atmosphere was added n-BuLi (solution in hexane) (2.8 mL of 2.4 M, 6.7200 mmol) at a rate which maintained the internal temperature below -87 °C. Ten minutes after the end of the addition, a solution of tert-butyl (4R)-4-[(1S)-1- (cyclopropylmethyl)-5-[methoxy(methyl)amino]-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3- carboxylate (2.26 g, 5.6651 mmol) in anhydrous THF (12 mL) was added at a rate which maintained the internal temperature below -87 °C. Ten minutes after the end of the addition a 2M aqueous solution of ammonium chloride (10 mL) was added at a rate which maintained the internal temperature below -40 °C. The reaction mixture was allowed to warm up to room temperature. The resulting reaction mixture of tert-butyl (4R)-4-[(1S)-5-(6-tert-butylfuro[2,3- b]pyrazin-3-yl)-1-(cyclopropylmethyl)-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (2.91 g, 100%) was combined with other batches for work up and purification. To the heterogeneous mixture of several batches of tert-butyl (4R)-4-[(1S)-5-(6-tert-butylfuro[2,3- b]pyrazin-2-yl)-1-(cyclopropylmethyl)-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (2.3 g, 4.4776 mmol), (2.3 g, 4.4776 mmol) and (2.91 g, 5.6651 mmol) in THF (105 mL) and ammonium chloride (2M aqueous solution) (30 mL) was added EtOAc (50 mL). Phases were separated and the aqueous solution was extracted once with EtOAc (50 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. Purification by reverse-phase chromatography (Column: C18. Gradient: 5-100% MeCN in water) afforded tert-butyl (4R)-4-[(1S)-5-(6-tert-butylfuro[2,3- b]pyrazin-2-yl)-1-(cyclopropylmethyl)-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (5.5 g, 73%) as an orange oil.1H NMR (400 MHz, CDCl3) δ 8.91 (s, 1H), 6.68 (s, 1H), 4.14 - 3.97 (m, 1H), 3.96 - 3.88 (m, 1H), 3.82 (dd, J = 9.0, 2.0 Hz, 1H), 3.30 - 3.20 (m, 2H), 2.22 - 2.03 (m, 1H), 1.92 - 1.77 (m, 2H), 1.72 - 1.54 (overlapped with water, m, 4H), 1.48 - 1.45 (m, 21H), 1.38 - 1.12 (overlapped with grease, m, 3H), 0.76 - 0.66 (m, 1H), 0.53 - 0.38 (m, 2H), 0.10 - -0.08 (m, 2H). ESI-MS m/z calc.513.3203, found 414.2 (M-99)+; Retention time: 2.76 minutes. LC method 1D. Step 2: (5S,6R)-6-Amino-1-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-5- (cyclopropylmethyl)-7-hydroxy-heptan-1-one [00756] To a solution of tert-butyl (4R)-4-[(1S)-5-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-1- (cyclopropylmethyl)-5-oxo-pentyl]-2,2-dimethyl-oxazolidine-3-carboxylate (5.5 g, 10.697 mmol) in dioxane (40 mL) and water (0.65 mL) was added HCl (solution in dioxane) (21.5 mL of 4 M, 86.000 mmol). The solution was stirred for 3 h then solvent was removed under reduced pressure. Dioxane (40 mL) was added and evaporated under reduced pressure (operation repeated twice). The residue was then dissolved in acetonitrile (40 mL) and concentrated to afford (5S,6R)-6-amino-1-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-5-(cyclopropylmethyl)-7- hydroxy-heptan-1-one (hydrochloride salt) (4.54 g, 98%) as a yellow solid which was used without further purification 1H NMR (400 MHz, MeCN-d3) δ 8.82 - 8.75 (m, 1H), 7.77 (br. s., 3H), 6.79 (s, 1H), 4.34 (br. s., 1H), 3.85 - 3.67 (m, 2H), 3.44 (br. s, 1H), 3.26 (t, J = 7.1 Hz, 2H), 1.81 - 1.72 (m, 2H), 1.66 - 1.57 (m, 2H), 1.46 - 1.34 (m, 12H), 0.80 - 0.69 (m, 1H), 0.50 - 0.37 (m, 2H), 0.17 - 0.02 (m, 2H). ESI-MS m/z calc.373.2365, found 374.4 (M+1)+; Retention time: 1.53 minutes. LC method U Step 3: [(2R,3S,7S)-7-(6-tert-Butylfuro[2,3-b]pyrazin-2-yl)-3- (cyclopropylmethyl)azepan-2-yl]methanol [00757] A suspension of (5S,6R)-6-amino-1-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-5- (cyclopropylmethyl)-7-hydroxy-heptan-1-one (hydrochloride salt) (4.54 g, 10.521 mmol) in anhydrous MeCN (65 mL) was sparged with nitrogen for 5 min. Then RuCl(p-cymene) [(R,R)- Ts-DPEN] (80 mg, 0.1257 mmol) was added followed by triethylamine (3.6300 g, 5 mL, 35.873 mmol) and formic acid (1.7080 g, 1.4 mL, 37.110 mmol). The mixture was sparged with nitrogen for a further 5 minutes and it was stirred for 16 h at 45 °C. Then the flask was cooled down to 0 °C and stirred for 45 min. The solution was then filtered, and the solid was washed with ice-cold MeCN (20 mL). The mother liquors were kept aside. The solid was then dissolved in DCM (50 mL) and washed with water (50 mL), 1 M aqueous solution of NaOH (50 mL) then brine (50 mL). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford 1.94 g of pure product. The previous mother liquors were then concentrated under reduced pressure. The residue was taken up with EtOAc (50 mL), then filtered through a silica pad eluted with EtOAc (500 mL). The mother liquors were concentrated under reduced pressure. The residue was triturated in ice-cold MeCN (15 mL) and the solids were washed with ice-cold acetonitrile (15 mL) to afford 600 mg of pure product. The two batches of solid were diluted in DCM (20 mL), combined to afford after evaporation of solvent under reduced pressure [(2R,3S,7S)-7-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-3- (cyclopropylmethyl)azepan-2-yl]methanol (2.54 g, 67%) as a pinkish solid.1H NMR (400 MHz, DMSO-d6) δ 8.29 (s, 1H), 6.86 (s, 1H), 4.56 (br. s, 1H), 3.91 - 3.79 (m, 1H), 3.53 (dd, J = 10.6, 3.3 Hz, 1H), 3.10 (dd, J = 10.3, 9.0 Hz, 1H), 2.44 (td, J = 8.6, 3.3 Hz, 1H), 2.03 - 1.84 (m, 2H), 1.79 - 1.70 (m, 1H), 1.70 - 1.53 (m, 3H), 1.52 - 1.41 (m, 1H), 1.40 - 1.31 (m, 10H), 1.11 - 1.02 (m, 1H), 0.73 - 0.65 (m, 1H), 0.49 - 0.34 (m, 2H), 0.10 - -0.06 (m, 2H), (1H missing, labile proton). ESI-MS m/z calc.357.2416, found 358.2 (M+1)+; Retention time: 3.78 minutes. LC method W. Step 4: 3-[[4-[[(2R,3S,7S)-7-(6-tert-Butylfuro[2,3-b]pyrazin-2-yl)-3- (cyclopropylmethyl)azepan-2-yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2- yl]sulfamoyl]benzoic acid [00758] In a 100 mL flask equipped with an internal temperature probe, [(2R,3S,7S)-7-(6-tert- butylfuro[2,3-b]pyrazin-2-yl)-3-(cyclopropylmethyl)azepan-2-yl]methanol (2.14 g, 5.986 mmol) and 3-[[4-chloro-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (2.75 g, 6.581 mmol) were combined under nitrogen in a mixture of anhydrous THF (20 mL) and NMP (4 mL). After stirring at room temperature until all solids dissolved, the mixture was cooled down in an ice-water bath until internal temperature reached 3 °C. NaOtBu (9 mL of 2 M, 18.00 mmol) (2M in THF) was added through syringe at a rate which maintained internal temperature below 10 °C (over 6 min). The ice bath was removed, and the mixture was stirred without external cooling for 3.5 h. The reaction mixture was added via syringe in an ice-cold mixture of EtOAc (50 mL) and aqueous HCl (60 mL of 1 M, 60.00 mmol) under stirring at a rate that maintained internal temperature below 9°C (over 10 min). After warming up to room temperature, the two phases were separated. The aqueous phase was extracted with EtOAc (50 mL). The combined organic extracts were dried quickly over sodium sulfate and filtered. After evaporation to a resin, EtOAc (100 mL) was added, and the solvent was evaporated. The residue was triturated and sonicated in EtOAc (200 mL) and the solid was filtered and rinsed with a bit of EtOAc. Drying under vacuum provided as a beige solid 3-[[4-[[(2R,3S,7S)-7-(6-tert- butylfuro[2,3-b]pyrazin-2-yl)-3-(cyclopropylmethyl)azepan-2-yl]methoxy]-6-(2,6- dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (2.82 g, 60%). ESI- MS m/z calc.738.31995, found 739.09(M+1)⁺; Retention time: 1.49 minutes. LC method A. Step 5: (4S,8S,9R)-4-(6-tert-Butylfuro[2,3-b]pyrazin-2-yl)-8-(cyclopropylmethyl)- 14-(2,6-dimethylphenyl)-18,18-dioxo-11-oxa-18λ6-thia-3,15,17,24- tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa-1(22),12,14,16(24),19(23),20- hexaen-2-one [00759] Under nitrogen, in a 250 mL flask equipped with a temperature probe, 3-[[4- [[(2R,3S,7S)-7-(6-tert-butylfuro[2,3-b]pyrazin-2-yl)-3-(cyclopropylmethyl)azepan-2- yl]methoxy]-6-(2,6-dimethylphenyl)pyrimidin-2-yl]sulfamoyl]benzoic acid (hydrochloride salt) (2.72 g, 3.460 mmol) was dissolved in ethyl acetate (70 mL) and anhydrous NMP (13 mL). Methanesulfonylchloride (0.80 mL, 10.34 mmol) was added, followed by NMM (2.3 mL, 20.92 mmol), then the reaction mixture was stirred at 50 °C (internal temperature) for 22 h. After cooling down to room temperature, the reaction was treated with EtOAc (100 mL), water (50 mL) and brine (50 mL). The layers were separated and the organic phase was washed with a 1:1 (v:v) mixture of water and brine (3 x 50 mL), dried over sodium sulfate and evaporated. The residue was dissolved in DCM and purified by flash chromatography on silica gel (120 g column) using a gradient of ethyl acetate (15 to 100% over 25 min) in heptane. After evaporation, the resulting solid was dissolved in 20 mL of EtOAc. Precipitation was induced by pouring the solution in heptane under stirring (150 mL). The solid was filtered and washed with heptane. Drying under vacuum provided as an off-white solid (4S,8S,9R)-4-(6-tert- butylfuro[2,3-b]pyrazin-2-yl)-8-(cyclopropylmethyl)-14-(2,6-dimethylphenyl)-18,18-dioxo-11- oxa-18λ6-thia-3,15,17,24-tetrazatetracyclo[17.3.1.112,16.03,9]tetracosa- 1(22),12,14,16(24),19(23),20-hexaen-2-one (957 mg, 38%).1H NMR (400 MHz, CDCl3) δ 10.20 (s, 1H), 8.63 (s, 1H), 8.29 (s, 1H), 7.95 (d, J = 7.9 Hz, 1H), 7.78 - 7.67 (m, 1H), 7.56 (t, J = 7.8 Hz, 1H), 7.21 (t, J = 7.6 Hz, 1H), 7.03 (d, J = 7.6 Hz, 2H), 6.66 (s, 1H), 6.18 (s, 1H), 5.82 (dd, J = 13.0, 4.5 Hz, 1H), 5.72 (dd, J = 11.8, 5.2 Hz, 1H), 4.97 (t, J = 11.9 Hz, 1H), 4.40 (dt, J = 12.3, 4.8 Hz, 1H), 2.37 - 2.22 (m, 1H), 2.21 - 2.11 (m, 1H), 2.08 - 1.75 (m, 11H), 1.41 (s, 9H), 1.24 - 1.12 (m, 1H), 1.01 - 0.92 (m, 1H), 0.45 - 0.34 (m, 1H), 0.32 - 0.22 (m, 2H), -0.11 - -0.24 (m, 2H). ESI-MS m/z calc.720.3094, found 721.09(M+1)⁺; Retention time: 2.25 minutes. LC method A. X. Characterization of Compounds A. Separation methods for stereoisomers of CFTR correctors [00760] The table below describes the separation methods utilized for separating the individual stereoisomers of the CFTR correctors. Table 4. Separation methods for stereoisomers of CFTR correctors
B. LCMS Data [00761] The compounds in the table(s) below were prepared by procedures analogous to those disclosed in the specification, and the analytical data were consistent with the reported structure. Table 5.
C. NMR Data Table 6.
D. Bioactivity Data 1. HBE assay procedure (a) Ussing Chamber Assay of CFTR-mediated short-circuit currents [00762] Ussing chamber experiments were performed using human bronchial epithelial (HBE) cells derived from CF subjects heterozygous for F508del and a minimal function (MF) CFTR mutation (F508del/MF-HBE) and cultured as previously described (Neuberger T, Burton B, Clark H, Van Goor F Methods Mol Biol 2011:741:39-54). After four days, the apical media was removed and the cells were grown at an air liquid interface for >14 days prior to use. This resulted in a monolayer of fully differentiated columnar cells that were ciliated, features that are characteristic of human bronchial airway epithelia. [00763] To isolate the CFTR-mediated short-circuit (ISC) current, F508del/MF-HBE grown on Costar® Snapwell™ cell culture inserts were mounted in an Ussing chamber and the transepithelial ISC was measured under voltage-clamp recording conditions (Vhold= 0 mV) at 37 oC. The basolateral solution contained (in mM) 145 NaCl, 0.83 K2HPO4, 3.3 KH2PO4, 1.2 MgCl2, 1.2 CaCl2, 10 Glucose, 10 HEPES (pH adjusted to 7.4 with NaOH) and the apical solution contained (in mM) 145 NaGluconate, 1.2 MgCl2, 1.2 CaCl2, 10 glucose, 10 HEPES (pH adjusted to 7.4 with NaOH) and 30 µM amiloride to block the epithelial sodium channel. Forskolin (20 µM) was added to the apical surface to activate CFTR, followed by apical addition of a CFTR inhibitor cocktail consisting of BPO, GlyH-101, and CFTR inhibitor 172 (each at 20 µM final assay concentration) to specifically isolate CFTR currents. The CFTR-mediated ISC (µA/cm2) for each condition was determined from the peak forskolin response to the steady-state current following inhibition. (b) Identification of Corrector Compounds [00764] The activity of the CFTR corrector compounds on the CFTR-mediated ISC was determined in Ussing chamber studies as described above. The F508del/MF-HBE cell cultures were either incubated with the corrector compounds at a range of concentrations in combination with 33 nM (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol and 18 µM Tezacaftor for 18 – 24 hours at 37 oC and in the presence of 20% human serum. The concentration of corrector compounds with 33 nM (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa- 3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol and 18 µM Tezacaftor during the 18-24 hours incubations was kept constant throughout the Ussing chamber measurement of the CFTR-mediated ISC to ensure compounds were present throughout the entire experiment. The efficacy and potency of the putative F508del correctors was compared to that of the known Vertex corrector, (14S)-8-[3-(2-{Dispiro[2.0.2.1]heptan-7-yl}ethoxy)-1H-pyrazol- 1-yl]-12,12-dimethyl-2λ6-thia-3,9,11,18,23-pentaazatetracyclo[17.3.1.111,14.05,10]tetracosa- 1(22),5,7,9,19(23),20-hexaene-2,2,4-trione, in combination with 18 µM Tezacaftor and 1 µM Ivacaftor. Ranges of concentrations to measure potency and efficacy Assay A: 0.00002 µM to 30 µM, 12 concentrations Assay B 0.03 µM to 30 µM, 6 concentrations [00765] The following assays C and D were run in identical conditions as stated above using a human bronchial epithelial (HBE) cell line derived from CF subjects heterozygous for F508del and a minimal function (MF) CFTR mutation (F508del/MF-HBE), however, this subject harbored a different MF CFTR mutation than in the cell line used in assay A and B: Assay C: 0.00002 µM to 30 µM, 12 concentrations Assay D: 0.03 µM to 30 µM, 6 concentrations 3. Biological Activity Data [00766] The following table represents CFTR modulating activity for representative compounds of the invention generated using one or more of the assays disclosed herein (EC50: +++ is < 1 µM; ++ is 1-3 µM; + is 3-30 µM; % Activity: +++ is > 60%; ++ is 30-60%; + is < 30%). Table 8. Biological Activity Data for Compounds 1-107 Table 9. Biological Activity Data for Compounds 108-139
Other Embodiments [00767] The foregoing discussion discloses and describes merely exemplary embodiments of this disclosure. One skilled in the art will readily recognize from such discussion and from the accompanying drawings and claims, that various changes, modifications and variations can be made therein without departing from the spirit and scope of this disclosure as defined in the following claims.

Claims

CLAIMS 1. A compound selected from compounds of Formula I: tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein: Ring A is selected from 7- and 8-membered heterocycles containing 0-2 additional heteroatoms selected from O and N, wherein Ring A is substituted with 0-4 groups selected from: C3-C6 cycloalkyl (further substituted with 0-4 groups selected from C1-C4 alkyl); C1-C5 alkyl (substituted with 0-4 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, oxo, and 4- to 6-membered heterocycle); C1-C4 alkoxy (substituted with 0-4 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, and oxo); 5- and 6-membered heterocycle; OH; NH2; and oxo; wherein two substituents on Ring A taken together with the atoms to which they are bonded may form a ring selected from C3-C6 cycloalkyl (further substituted with 0-2 groups selected from C1-C4 alkyl and C1-C4 alkoxy); Ring B is a 9-12 membered heteroaryl group containing 1-4 heteroatoms selected from N, O, and S, wherein Ring B is substituted with 1-4 groups selected from: C1-C6 alkyl (substituted with 0-3 groups selected from halogen, C1-C3 alkoxy, and OH); C3-C6 cycloalkyl, substituted with 0-3 groups selected from: C1-C6 alkyl (substituted with 0-3 groups selected from C1-C3 alkoxy and halogen), OH, and halogen; halogen; and 5- and 6-membered heterocycle; each X is independently selected from CH and N; Z is selected from CH and N; R1 is selected from H, C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy (substituted with 0-5 halogens), and halogen; each R2 is independently selected from C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy (substituted with 0-5 halogens), OH, and halogen; each R3 is independently selected from C1-C3 alkyl, C1-C3 alkoxy, OH, and halogen; n is an integer selected from 2 to 5; and p is an integer selected from zero to 3.
2. A compound selected from compounds of Formula I′: tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein: Ring A is selected from 7- and 8-membered heterocycles containing 0-2 additional heteroatoms selected from O and N, wherein Ring A is substituted with 0-2 groups selected from: C3-C6 cycloalkyl (substituted with 0-2 groups selected from C1-C4 alkyl); C1-C5 alkyl (substituted with 0-2 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, oxo, and C4-C6 heterocycle); C1-C4 alkoxy (substituted with 0-2 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, and oxo); 5- and 6-membered heterocycle; OH; NH2; and oxo; Ring B is a 9-12 membered heteroaryl group containing 1-4 heteroatoms selected from N, O, and S, wherein Ring B is substituted with 1-4 groups selected from: C1-C6 alkyl (substituted with 0-3 groups selected from halogen, C1-C3 alkoxy, and OH); C3-C6 cycloalkyl, substituted with 0-3 groups selected from: C1-C6 alkyl (substituted with 0-3 groups selected from: C1-C3 alkoxy and halogen), OH, and halogen; halogen; and 5- and 6-membered heterocycle; each X is independently selected from CH and N; Z is selected from CH and N; R1 is selected from H, C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy (substituted with 0-5 halogens), and halogen; each R2 is independently selected from C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy (further substituted with 0-5 halogens), OH, and halogen; each R3 is independently selected from C1-C3 alkyl, C1-C3 alkoxy, OH, and halogen; n is an integer selected from 2 to 5; and p is zero to 3.
3. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to claim 1 or claim 2, wherein the compound is selected from compounds of Formula Ia: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
4. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to claim 1 or claim 2, wherein the compound is selected from compounds of Formula Ib: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
5. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to claim 1 or claim 2, wherein the compound is selected from compounds of Formula Ic: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
6. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to claim 1 or claim 2, wherein the compound is selected from compounds of Formula II: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein each R4 is independently selected from: C3-C6 cycloalkyl (substituted with 0-3 groups selected from halogen and C1-C4 alkyl); C1-C5 alkyl (substituted with 0-3 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, oxo, and 4- to 6-membered heterocycle); C1-C4 alkoxy (substituted with 0-3 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, and oxo); 5- and 6-membered heterocycle; OH; NH2; and oxo.
7. A compound selected from compounds of Formula II: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein each R4 is independently selected from: C3-C6 cycloalkyl (substituted with 0-2 groups selected from halogen and C1-C4 alkyl); C1-C5 alkyl (substituted with 0-2 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, oxo, and C3-C6 heterocycle); C1-C4 alkoxy (substituted with 0-2 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, and oxo); 5- and 6-membered heterocycle; OH; NH2; and oxo. Ring B is a 9-12 membered heteroaryl group containing 1-4 heteroatoms selected from N, O, and S, wherein Ring B is substituted with 1-4 groups selected from: C1-C6 alkyl (substituted with 0-3 groups selected from halogen, C1-C3 alkoxy, and OH); C3-C6 cycloalkyl, substituted with 0-3 groups selected from: C1-C6 alkyl (substituted with 0-3 groups selected from C1-C3 alkoxy and halogen), OH, and halogen; halogen; and 5- and 6-membered heterocycle; each X is independently selected from CH and N; Z is selected from CH and N; R1 is selected from H, C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy (substituted with 0-5 halogens), and halogen; each R2 is independently selected from C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy (further substituted with 0-5 halogens), OH, and halogen; each R3 is independently selected from C1-C3 alkyl, C1-C3 alkoxy, OH, and halogen; n is an integer selected from 2 to 5; and p is zero to 3.
8. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to claim 6 or claim 7, wherein the compound is selected from compounds of Formula IIa: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
9. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to claim 6 or claim 7, wherein the compound is selected from compounds of Formula IIb: (IIb), tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
10. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to claim 6 or claim 7, wherein the compound is selected from compounds of Formula IIc: tautomers of those compounds, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
11. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to claim 1, wherein the compound is selected from compounds of Formula IIIa: (IIIa), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein: Y1 is selected from O and -CH2-; Y2 is selected from NR9 and CH; Y3 is selected from O and CH; R4 is selected from: C1-C4 alkyl (substituted with 0-3 groups selected from halogen and cyclopropyl); C1-C4 alkoxy; and C3-C4 cycloalkyl; and R8 is selected from C1-C4 alkyl and C3-C4 cycloalkyl, each of which is substituted with 0-1 trifluoromethyl groups and, if R9 is present, R9 is methyl; or R8 and R9 are taken together to form a five-membered heterocyclic ring, substituted with 0-2 methyl groups.
12. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to claim 1, wherein the compound is selected from compounds of Formula IIIb: (IIIb), tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein: R4 is selected from: C1-C4 alkyl (substituted with 0-3 groups selected from halogen and cyclopropyl); C1-C4 alkoxy; and C3-C4 cycloalkyl; and R8 is selected from C3 alkyl and C3-C4 cycloalkyl, each of which is substituted with 0-1 trifluoromethyl groups and R9 is methyl; or R8 and R9 are taken together to form a five-membered heterocyclic ring, substituted with 0-2 methyl groups.
13. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to any one of claims 1 to 5, wherein: Ring A is selected from azepane and azocane, wherein one CH2 group is optionally replaced by an O atom, and wherein Ring A is substituted with 0-2 groups selected from: C3 and C4 cycloalkyl (substituted with 0-1 methyl groups), C1-C5 alkyl (substituted with 0-1 groups selected from C3 and C4 cycloalkyl); and C1-C3 alkoxy (substituted with 0-1 groups selected from C3 and C4 cycloalkyl); Ring B is a 9-12 membered heteroaryl group containing 2-4 heteroatoms independently selected from N and O, wherein Ring B is substituted with 1-3 groups selected from: C1-C4 alkyl (substituted with 0-3 groups selected from OMe, halogen, and OH); C3 and C4 cycloalkyl, substituted with 0-1 groups selected from: C1 and C2 alkyl (substituted with 0-3 halogen atoms) and CH2OCH3; halogen; and 6-membered heterocycle; X is N; Z is selected from CH and N; R1 is selected from H, CH3, OCH3, and F; each R2 is CH3; R3 is absent; n is 2; and p is zero.
14. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to any one of claims 1 to 5, wherein Ring A is selected from:
15. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to any one of claims 1 to 10, wherein Ring B is selected from: Q is selected from C and N; each W is independently selected from CH and N; W1 is selected from N, CH, and CF; Y is selected from N, O, S, and CR; Ring B contains 1 to 4 nitrogen atoms; R is selected from H and C1-C3 alkyl (substituted with 0-1 groups selected from C1-C3 alkoxy); R5 is selected from H, C1-C4 alkyl, and C3 and C4 cycloalkyl; R6 is selected from: H; C1-C5 alkyl; C1-C5 haloalkyl; C3 and C4 cycloalkyl (substituted with 0-1 groups selected from C1-C4 alkyl, CH2OCH3, and C1-C3 haloalkyl); and 5- and 6-membered heterocycle; R7 is selected from C1-C3 alkyl; and R8 is selected from C1-C4 alkyl; and wherein R5 and R6 are not both hydrogen.
16. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to any one of claims 1 to 10, wherein Ring B is selected from: R is selected from H and C1-C3 alkyl (substituted with 0-1 groups selected from C1-C3 alkoxy); R5 is selected from H, C1-C4 alkyl, and C3 and C4 cycloalkyl; R6 is selected from: H; C1-C5 alkyl; C1-C5 haloalkyl; C3 and C4 cycloalkyl (substituted with 0-1 groups selected from C1-C4 alkyl, CH2OCH3, and C1-C3 haloalkyl); and 5- and 6-membered heterocycle; R7 is selected from C1-C3 alkyl; R8 is selected from C1-C4 alkyl; and G is selected from halogens; and wherein R, R5 and R6 are not all simultaneously hydrogen, R5 and R6 are not simultaneously hydrogen when R is not present, and R6 is not hydrogen when R and R5 are not present.
17. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to any one of claims 1 to 10, wherein Ring B is selected from: , , , , ,
18. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to claim 1, selected from: tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
19. The compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to claim 1, selected from tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing.
20. A compound selected from compounds of Formula I: tautomers thereof, deuterated derivatives of those compounds and tautomers, and pharmaceutically acceptable salts of any of the foregoing, wherein: Ring A is selected from 7- and 8-membered heterocycles containing 0-2 additional heteroatoms selected from O and N, wherein Ring A is substituted with 0-4 groups selected from: C3-C6 cycloalkyl (further substituted with 0-4 groups selected from C1-C4 alkyl); C1-C5 alkyl (substituted with 0-4 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, oxo, and 4- to 6-membered heterocycle); C1-C4 alkoxy (substituted with 0-4 groups selected from C3-C6 cycloalkyl, halogen, OH, NH2, and oxo); 5- and 6-membered heterocycle; OH; NH2; and oxo; wherein two substituents on Ring A taken together with the atoms to which they are bonded may form a ring selected from C3-C6 cycloalkyl (further substituted with 0-2 groups selected from C1-C4 alkyl and C1-C4 alkoxy); Ring B is a 9-12 membered heteroaryl group containing 1-4 heteroatoms selected from N, O, and S, wherein Ring B is substituted with 1-4 groups selected from: C1-C6 alkyl (substituted with 0-3 groups selected from halogen, C1-C3 alkoxy, and OH); C3-C6 cycloalkyl, substituted with 0-3 groups selected from: C1-C6 alkyl (substituted with 0-3 groups selected from C1-C3 alkoxy and halogen), OH, and halogen; halogen; and 5- and 6-membered heterocycle; each X is independently selected from CH and N; Z is selected from CH and N; R1 is selected from H, C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy (substituted with 0-5 halogens), and halogen; each R2 is independently selected from C1-C5 alkyl, C1-C5 haloalkyl, C1-C5 alkoxy (substituted with 0-5 halogens), OH, and halogen; each R3 is independently selected from C1-C3 alkyl, C1-C3 alkoxy, OH, and halogen; n is an integer selected from 2 to 5; p is an integer selected from zero to 3; and wherein one or more carbon atoms are replaced by silicon and/or germanium.
21. A pharmaceutical composition comprising a compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to any one of claims 1 to 20 and at least one pharmaceutically acceptable carrier.
22. The pharmaceutical composition according to claim 21, further comprising at least one additional active therapeutic agent.
23. The pharmaceutical composition according to claim 22, wherein at least one additional active therapeutic agents are (a) at least one compound selected from tezacaftor, lumacaftor, and deuterated derivatives and pharmaceutically acceptable salts thereof; and (b) at least one compound selected from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing.
24. The pharmaceutical composition according to claim 22, wherein the at least one additional active therapeutic agents are (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)- 13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol, and optionally tezacaftor.
25. A method of treating cystic fibrosis comprising the step of administering the compound, tautomer, deuterated derivative, or pharmaceutically acceptable salt according to any one of claims 1 to 20 or the pharmaceutical composition according to any one of claims 21 to 24.
26. The method according to claim 25, wherein the method further comprises administering at least one additional active therapeutic agent.
27. The method according to claim 26, wherein the at least one additional active therapeutic agent to be administered is (a) at least one compound selected from tezacaftor, lumacaftor, and deuterated derivatives and pharmaceutically acceptable salts thereof; and (b) at least one compound selected from ivacaftor, deutivacaftor, (6R,12R)-17-amino-12-methyl-6,15- bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16- pentaen-6-ol, (6R)-17-amino-12,12-dimethyl-6,15-bis(trifluoromethyl)-19-oxa-3,4,13,18- tetrazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol, and deuterated derivatives and pharmaceutically acceptable salts of any of the foregoing.
28. The method according to claim 26, wherein the method further comprises the step of administering (6R,12R)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18- triazatricyclo[12.3.1.12,5] nonadeca-1(18),2,4,14,16-pentaen-6-ol, and optionally administering tezacaftor.
EP24794995.1A 2023-10-04 2024-10-03 Modulators of cystic fibrosis transmembrane conductance regulator Pending EP4788429A1 (en)

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Family Cites Families (41)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2881078C (en) 2004-06-24 2018-10-02 Vertex Pharmaceuticals Incorporated Modulators of atp-binding cassette transporters
PT2404919E (en) 2005-11-08 2013-10-22 Vertex Pharma A HETEROCYCLIC USEFUL COMPOSITE AS AN ATP CONNECTOR CASSETTE CONVERTER MODULATOR
LT1993360T (en) 2005-12-28 2017-06-12 Vertex Pharmaceuticals Incorporated Solid forms of n-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide
US7645789B2 (en) 2006-04-07 2010-01-12 Vertex Pharmaceuticals Incorporated Indole derivatives as CFTR modulators
ATE534383T1 (en) 2006-05-12 2011-12-15 Vertex Pharma COMPOSITIONS OF N-Ä2,4-BIS(1,1-DIMETHYLETHYL)-5-HYDROXYPHENYLÜ-1,4-DIHYDRO-4-OXOCINOLIN-3-CARBOXAMIDE
CA2989620C (en) 2007-12-07 2022-05-03 Vertex Pharmaceuticals Incorporated Processes for producing cycloalkylcarboxamido-pyridine benzoic acids
HUE049642T2 (en) 2007-12-07 2020-09-28 Vertex Pharma Solid forms of 3- (6- (1- (2,2-Difluorobenzo [d] [1,3] dioxol-5-yl) cyclopropanecarboxamido) -3-methylpyridin-2-yl) benzoic acid
US20100074949A1 (en) 2008-08-13 2010-03-25 William Rowe Pharmaceutical composition and administration thereof
RS56894B1 (en) 2008-08-13 2018-04-30 Vertex Pharma Pharmaceutical composition of n-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4- oxoquinoline-3-carboxamide and administration thereof
WO2010037066A2 (en) 2008-09-29 2010-04-01 Vertex Pharmaceuticals Incorporated Dosage units of 3-(6-(1-(2,2-difluorobenzo [d] [1,3] dioxol-5-yl) cyclopropanecarboxamido)-3-methylpyridin-2-yl)benzoic acid
KR20160013251A (en) 2008-11-06 2016-02-03 버텍스 파마슈티칼스 인코포레이티드 Modulators of atp-binding cassette transporters
SI3330255T1 (en) 2009-03-20 2021-03-31 Vertex Pharmaceuticals Incorporated Process for making modulators of cystic fibrosis transmembrane conductance regulator
EP2563778B1 (en) 2010-03-25 2016-08-24 Vertex Pharmaceuticals Incorporated Crystalline form of (r)-1(2,2 -difluorobenzo[d][1,3]dioxol-5yl)-n-(1-(2,3-dihyderoxypropyl)-6-fluoro-2-(1-hydroxy-2-methylpropan-2yl)-1h-indol-5yl)cyclopropanecarboxamide
CN102811938B (en) 2010-04-09 2016-10-12 艾克索仿生技术公司 Exoskeleton load handling system and method of use
RU2569678C2 (en) 2010-04-22 2015-11-27 Вертекс Фармасьютикалз Инкорпорейтед Method for producing cycloalkylcarboxamido-indole compounds
NZ603042A (en) 2010-04-22 2015-02-27 Vertex Pharma Pharmaceutical compositions comprising cftr modulators and administrations thereof
CA2809263A1 (en) 2010-08-27 2012-03-01 Eleni Dokou Pharmaceutical composition and administrations thereof
HUE047354T2 (en) 2011-05-18 2020-04-28 Vertex Pharmaceuticals Europe Ltd Deuterated derivatives of ivacaftor
ME03652B (en) 2011-05-18 2020-07-20 Vertex Pharmaceuticals Europe Ltd Deuterated derivatives of ivacaftor
EP2819670B1 (en) 2012-02-27 2026-05-20 Vertex Pharmaceuticals Incorporated Pharmaceutical preparations and their administration
AU2013290444B2 (en) 2012-07-16 2018-04-26 Vertex Pharmaceuticals Incorporated Pharmaceutical compositions of (R)-1-(2,2-diflurorbenzo[d][1,3]dioxol-5-yl)-N-(1-(2,3-dihydroxypropyl)-6-fluoro-2-(1-hydroxy-2-methylpropan-2-yl)-1H-indol-5-yl) cyclopropanecarboxamide and administration thereof
KR102396897B1 (en) 2012-11-02 2022-05-13 버텍스 파마슈티칼스 인코포레이티드 Pharmaceutical compositions for the treatment of cftr mediated diseases
WO2014078842A1 (en) 2012-11-19 2014-05-22 Concert Pharmaceuticals, Inc. Deuterated cftr potentiators
PL3925607T5 (en) 2014-04-15 2026-06-01 Vertex Pharmaceuticals Incorporated Pharmaceutical compositions for the treatment of cystic fibrosis transmembrane conductance regulator mediated diseases
US9840513B2 (en) 2015-07-16 2017-12-12 Abbvie S.Á.R.L. Substituted tricyclics and method of use
HK1258472A1 (en) 2015-09-21 2019-11-15 Vertex Pharmaceuticals (Europe) Limited Administration of deuterated cftr potentiators
AU2017352206B2 (en) 2016-10-27 2022-03-03 Vertex Pharmaceuticals (Europe) Limited Methods of treatment with deuterated CFTR potentiators
CA3158057A1 (en) 2019-11-12 2021-05-20 Genzyme Corporation 5-membered heteroarylaminosulfonamides for treating conditions mediated by deficient cftr activity
MX2022005810A (en) 2019-11-12 2022-06-08 Genzyme Corp 6-MEMBRED HETEROARYLAMINOSULPHONAMIDES FOR TREATING DISEASES AND CONDITIONS MEDIATED BY DEFICIENT CYSTIC FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR (CFTR) ACTIVITY.
WO2021113809A1 (en) 2019-12-05 2021-06-10 Genzyme Corporation Arylamides and methods of use thereof
WO2021113808A1 (en) 2019-12-05 2021-06-10 Genzyme Corporation Cftr-modulating arylamides
WO2021113806A1 (en) 2019-12-05 2021-06-10 Genzyme Corporation Arylamides and methods of use thereof
CR20230120A (en) 2020-08-07 2023-09-01 Vertex Pharma Modulators of cystic fibrosis transmembrane conductance regulator
BR112023006470A2 (en) * 2020-10-07 2023-09-26 Vertex Pharma Cystic Fibrosis Transmembrane Conductance Regulator Modulators
PE20231380A1 (en) 2020-11-18 2023-09-07 Vertex Pharma MACROCYCLE CONTAINING A 1,3,4-OXADIAZOLE RING FOR USE AS MODULATORS OF THE REGULATOR OF TRANSMEMBRANE CONDUCTANCE IN CYSTIC FIBROSIS
WO2023012230A2 (en) 2021-08-06 2023-02-09 Telefonaktiebolaget Lm Ericsson (Publ) Generative adversarial-based attack in federated learning
IL311038A (en) 2021-09-03 2024-04-01 Genzyme Corp Indole compounds and methods of use
WO2023034946A1 (en) 2021-09-03 2023-03-09 Genzyme Corporation Indole compounds and uses thereof in the treatement of cystic fibrosis
US20250320226A1 (en) 2022-02-03 2025-10-16 Vertex Pharmaceuticals Incorporated Methods of preparing and crystalline forms of (6a,12a)-17-amino-12-methyl-6,15-bis(trifluoromethyl)-13,19-dioxa-3,4,18-triazatricyclo[12.3.1.12,5]nonadeca-1(18),2,4,14,16-pentaen-6-ol
AU2023273614A1 (en) 2022-05-16 2024-11-07 Vertex Pharmaceuticals Incorporated Methods of treatment for cystic fibrosis
US20250340568A1 (en) 2022-05-16 2025-11-06 Vertex Pharmaceuticals Incorporated Solid forms of a macrocyclic compounds as cftr modulators and their preparation

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