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WO2025012057A1 - Macrocycles pour le traitement d'une maladie auto-immune - Google Patents

Macrocycles pour le traitement d'une maladie auto-immune Download PDF

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Publication number
WO2025012057A1
WO2025012057A1 PCT/EP2024/068795 EP2024068795W WO2025012057A1 WO 2025012057 A1 WO2025012057 A1 WO 2025012057A1 EP 2024068795 W EP2024068795 W EP 2024068795W WO 2025012057 A1 WO2025012057 A1 WO 2025012057A1
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Prior art keywords
methyl
fluoro
pyrazolo
pyrimidin
oxa
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PCT/EP2024/068795
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English (en)
Inventor
Min Jiang
Buyu KOU
Haixia Liu
Hong Shen
Yao Wu
Zhiwei Zhang
Wei Zhu
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F. Hoffmann-La Roche Ag
Hoffmann-La Roche Inc.
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Publication of WO2025012057A1 publication Critical patent/WO2025012057A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B59/00—Introduction of isotopes of elements into organic compounds ; Labelled organic compounds per se
    • C07B59/002—Heterocyclic compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/06—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/05—Isotopically modified compounds, e.g. labelled

Definitions

  • Case 38652 Macrocycles for the treatment of autoimmune disease
  • the present invention relates to organic compounds useful for therapy and/or prophylaxis in a mammal, and in particular to antagonist of STING useful for treating autoimmune diseases.
  • FIELD OF THE INVENTION Autoimmune diseases, such as rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), and inflammatory bowel diseases (IBD), refer to a spectrum of conditions where the immune system mistakenly attacks one's own body, leading to unresolved and inappropriately activated inflammation that become pathogenic. Many of the autoimmune diseases are poorly managed by existing treatments that provide only symptomatic relief. Steroid and broad immunosuppressant drugs (e.g.
  • mycophenolate and cyclophosphamide constitute the stand of care, but are associated with significant treatment-related toxicity.
  • Pathway selective agents such as Adalizumab (anti-TNF antibody, for RA and IBD) occasionally resulting in infection or insufficient tumor surveillance.
  • Belimumab anti-BAFF antibody, the only FDA-approved new drug for SLE shows a slow onset of remission with modest efficacy in the clinic.
  • the heterogeneity of many autoimmune diseases with no-existing treatment illustrates the difficulty in finding efficacy through the blockade of one immune pathway.
  • Stimulator of interferon genes is an endoplasmic reticulum (ER)-located transmembrane protein that is pivotal in mediating the host's innate sensing of pathogen-/ damage-associated molecular patterns (PAMPs or DAMPs).
  • PAMPs or DAMPs pathogen-/ damage-associated molecular patterns
  • cGAS cyclic-GMP-AMP synthase
  • cGAS double-stranded DNA
  • dsDNA double-stranded DNA
  • cGAMP 2'3'- cyclic GMP-AMP
  • STING translocates from ER to Golgi and recruits TANK-binding kinase 1 (TBK1), which phosphorylates interferon regulatory factor 3 (IRF3) and nuclear factor-kappa B (NF- ⁇ B) to initiate the expression of type-I IFN and a myriad of pro-inflammatory cytokines (e.g., IL-6 and TNF ⁇ ), respectively.
  • TNK1 TANK-binding kinase 1
  • IRF3 interferon regulatory factor 3
  • NF- ⁇ B nuclear factor-kappa B
  • STING can be activated by other types of cyclic-di-nucleotides (CDNs), such as c-di-AMP, c-di- GMP, and 3’,3’-cGAMP from bacteria.
  • CDNs cyclic-di-nucleotides
  • STING is rapidly degraded to prevent it from constitutive signaling of the inflammatory responses. While eliciting robust host defense responses, aberrant STING signaling fuels dysregulated immune responses associated with many pathologies.
  • Gain-of-function (GoF) human STING mutations are the root cause of STING-associated vasculopathy with onset in infancy (SAVI), a monogenic disease characterized by the onset of auto-inflammation conditions called type I interferonopathies.
  • STING is implicated in DNA-driven inflammations, such as Aicardi-Gout Italian Syndrome (AGS) and genetic forms of lupus known as familial chilblain lupus (FCL).
  • AGS Aicardi-Gout Italian Syndrome
  • FCL familial chilblain lupus
  • STING mediated continuous innate immune activation in AGS is caused by deficiencies in self-DNA clearance and metabolisms due to mutations in endonuclease gene TREX1 and/or DNASE2.
  • genetic and pharmacological inhibition of STING ameliorates systemic inflammation and morbidity in the Trex1-/- mouse model.
  • a direct link between the cGAS-STING pathway and SLE was established by observing that PBMC from a subset of SLE patients has elevated cytosolic cGAMP than healthy controls.
  • membrane vesicles from apoptotic cells in SLE sera have high ISGs- stimulating activities dependent on cGAS-STING.
  • disrupting STING signaling ameliorated the development of lupus-like phenotypes in Fc ⁇ rIIb-/- mice.
  • multiple recent studies associate STING with distinct types of neurodegeneration.
  • cGAS- and cGAMP-independent mode of STING activation also affects neuropathology and provides a therapeutic target for the treatment of Niemann-Pick disease type C (NPC).
  • NPC Niemann-Pick disease type C
  • STING also mediates tumorigenic DNA responses caused by chromosomal instability during cancer metastasis, and that STING-deficiency confers protection against colorectal and skin cancer in the mouse.
  • the present invention relates to novel compounds of formula (I-2), (I-2), wherein R 1 is H, C1-6alkyl, or together with R 9 form an optionally substituted heterocyclic ring; R 2 is C1-6alkyl, or together with R 9 form an optionally substituted heterocyclic ring; M 1 is optionally substituted heterocyclylene which is further substituted by R 3 ; , R 4 is H, ((haloC 1-6 alkyl)azetidinyl)C 1-6 alkyl, 2,5-diazabicyclo[2.2.1]heptanyl substituted by C1-6alkyl, haloC1-6alkyl or oxetanyl, 2-oxo-3,6-diazabicyclo[3.1.1]heptanyl substituted by C 1-6 alkyl, C 3- 7cycloalkylC1-6alkyl or oxetanyl, 3,6-diazabicyclo[3.1.1]heptanyl substituted by C
  • Another object of the present invention is related to novel compounds of formula (I), (Ia), (I-1), (I-1a) or (I-2).
  • the compounds of formula (I), (Ia), (I-1), (I-1a) or (I-2) show superior STING antagonism activity.
  • the compounds of formula (I), (Ia), (I-1), (I-1a) or (I-2) also show good cytotoxicity, phototoxicity, solubility, hPBMC, metabolic stability, hERG and SDPK profiles, as well as low CYP inhibition.
  • BRIEF DESCRIPTION OF THE FIGURE Figure 1. X-ray crystallographic analysis of compound 46a-1.
  • Figure 2. X-ray crystallographic analysis of compound 67b.
  • DETAILED DESCRIPTION OF THE INVENTION DEFINITIONS The term “deuterio” or “deuterium” denotes the isotope of hydrogen that has a single neutron as well as a proton in the nucleus.
  • C1-6alkyl denotes a saturated, linear or branched chain alkyl group containing 1 to 6, particularly 1 to 4 carbon atoms, for example methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl and the like. Particular “C 1-6 alkyl” groups are methyl, ethyl and n-propyl.
  • C1-6alkylene denotes a divalent C1-6alkyl. Examples of C1-6alkylene groups include methylene, ethylene, propylene, 2-methylpropylene, butylene, 2-ethylbutylene, pentylene, hexylene.
  • C1-6alkoxy denotes C1-6alkyl-O-.
  • halogen and “halo” are used interchangeably herein and denote fluoro, chloro, bromo, or iodo.
  • haloC1-6alkyl denotes a C1-6alkyl group wherein at least one of the hydrogen atoms of the C1-6alkyl group has been replaced by same or different halogen atoms, particularly fluoro atoms.
  • haloalkyl examples include monofluoro-, difluoro- or trifluoro-methyl, -ethyl or -propyl, for example 3,3,3-trifluoropropyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, fluoromethyl, or trifluoromethyl.
  • haloC 1-6 alkoxy denotes haloC 1-6 alkyl-O-.
  • C3-7cycloalkyl denotes a monovalent saturated monocyclic or bicyclic hydrocarbon group of 3 to 7 ring carbon atoms. Bicyclic means consisting of two saturated carbocycles having one or more carbon atoms in common.
  • Examples for monocyclic cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl.
  • Examples for bicyclic cycloalkyl are bicyclo[1.1.0]butyl, bicyclo[2.2.1]heptanyl, bicyclo[1.1.1]pentanyl, or bicyclo[2.2.2]octanyl.
  • C 3-7 cycloalkylene denotes a divalent C 3-7 cycloalkyl.
  • oxetanylene denotes a divalent oxetanyl.
  • heterocyclic group refers to any mono-, bi-, tricyclic, spiro or bridged, saturated, partially saturated or unsaturated, non-aromatic ring system, having 3 to 20 ring atoms, where the ring atoms are carbon, and at least one atom in the ring or ring system is a heteroatom selected from nitrogen, sulfur, oxygen or silicon. If any ring atom of a cyclic system is a heteroatom, that system is a heterocycle, regardless of the point of attachment of the cyclic system to the rest of the molecule.
  • heterocyclyl includes 3-11 ring atoms (“members”) and includes monocycles, bicycles, tricycles, spiro, and bridged ring systems, wherein the ring atoms are carbon, where at least one atom in the ring or ring system is a heteroatom selected from nitrogen, sulfur, oxygen or silicon.
  • heterocyclyl includes 4-10 or 5-10 ring atoms.
  • heterocyclyl includes 1 to 4 heteroatoms.
  • heterocyclyl includes 1 to 3 heteroatoms.
  • heterocyclyl includes 3- to 7-membered monocycles having 1-2, 1-3 or 1-4 heteroatoms selected from nitrogen, sulfur, oxygen or silicon.
  • heterocyclyl includes 4- to 6-membered monocycles having 1-2, 1-3 or 1-4 heteroatoms selected from nitrogen, sulfur, oxygen or silicon.
  • heterocyclyl includes 3-membered monocycles.
  • heterocyclyl includes 4-membered monocycles.
  • heterocyclyl includes 5-6 membered monocycles.
  • a heterocycloalkyl includes at least one nitrogen.
  • the heterocyclyl group includes 0 to 3 double bonds. Any nitrogen or sulfur heteroatom may optionally be oxidized (e.g., NO, SO, SO2), and any nitrogen heteroatom may optionally be quaternized (e.g., [NR4] + Cl-, [NR4] + OH-).
  • heterocycles include oxiranyl, aziridinyl, thiiranyl, azetidinyl, oxetanyl, thietanyl, 1,2-dithietanyl, 1,3-dithietanyl, pyrrolidinyl, dihydro-1H-pyrrolyl, dihydrofuranyl, tetrahydrofuranyl, dihydrothienyl, tetrahydrothienyl, imidazolidinyl, piperidinyl, piperazinyl, isoquinolinyl, tetrahydroisoquinolinyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholinyl, dihydropyranyl, tetrahydropyranyl, hexahydrothiopyranyl, hexahydropyrimidinyl, oxazinanyl, thiazinanyl, thi
  • heterocyclylene denotes a divalent heterocyclyl group.
  • a heterocyclyl group or a heteroaryl group is attached at a carbon atom of the heterocyclyl group or the heteroaryl group.
  • carbon bonded heterocyclyl groups include bonding arrangements at position 2, 3, 4, 5, or 6 of a pyridine ring, position 3, 4, 5, or 6 of a pyridazine ring, position 2, 4, 5, or 6 of a pyrimidine ring, position 2, 3, 5, or 6 of a pyrazine ring, position 2, 3, 4, or 5 of a furan, tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole ring, position 2, 4, or 5 of an oxazole, imidazole or thiazole ring, position 3, 4, or 5 of an isoxazole, pyrazole, or isothiazole ring, position 2 or 3 of an aziridine ring, position 2, 3, or 4 of an azetidine ring, position 2, 3, 4, 5, 6, 7, or 8 of a quinoline ring or position 1, 3, 4, 5, 6, 7, or 8 of an isoquinoline ring.
  • the heterocyclyl group or heteroaryl group is N-attached.
  • nitrogen bonded heterocyclyl or heteroaryl groups include bonding arrangements at position 1 of an aziridine, azetidine, pyrrole, pyrrolidine, 2-pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline, 3-imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline, piperidine, piperazine, indole, indoline, 1H-indazole, position 2 of a isoindole, or isoindoline, position 4 of a morpholine, and position 9 of a carbazole, or ⁇ -carboline.
  • an optionally substituted group may be unsubstituted or substituted by one or more (e.g., 0, 1, 2, 3, 4, or 5 or more, or any range derivable therein) of the substituents listed for that group in which said substituents may be the same or different.
  • an optionally substituted group has 1 substituent.
  • an optionally substituted group has 2 substituents.
  • an optionally substituted group has 3 substituents.
  • an optionally substituted group has 4 substituents.
  • an optionally substituted group has 5 substituents.
  • Suitable substituents of an “optionally substituted” group may be selected from deuterio, halogen, cyano, carboxy, R, RO, RS, RNH, (R)2N, RCO, RSO2, RNHSO2, R2NSO2, RSO(NR), R3Si, ROC1-6alkyl, RSC1-6alkyl, RNHC1-6alkyl, (R)2NC1-6alkyl, RSO2C1-6alkyl, RNHSO2C1- 6 alkyl, R 2 NSO 2 C 1-6 alkyl, RSO(NR)C 1-6 alkyl and R 3 SiC 1-6 alkyl; wherein R is H, R a , R b or R c ; R a is C 1-6 alkyl optionally substituted by R b or R c ; R b is C 3-7 cycloalkyl, heterocyclyl, heteroaryl or aryl, said C3-7cycloalkyl, heterocyclyl, heteroaryl and aryl
  • PG denotes protecting groups.
  • cis and trans denote the relative stereochemistry of the molecule or moiety.
  • trans-cyclopentane-1,3-diamine used in Example 4 synthesis refers to a mixture of .
  • the way of showing relative stereochemistry also applies to the final compounds.
  • pharmaceutically acceptable salts denotes salts which are not biologically or otherwise undesirable. Pharmaceutically acceptable salts include both acid and base addition salts.
  • pharmaceutically acceptable acid addition salt denotes those pharmaceutically acceptable salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, and organic acids selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic, and sulfonic classes of organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, maloneic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, embonic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluene
  • pharmaceutically acceptable base addition salt denotes those pharmaceutically acceptable salts formed with an organic or inorganic base.
  • acceptable inorganic bases include sodium, potassium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts.
  • Salts derived from pharmaceutically acceptable organic nontoxic bases includes salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, trimethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperizine, piperidine, N-ethylpiperidine, and polyamine resins.
  • substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, trieth
  • a pharmaceutically active metabolite denotes a pharmacologically active product produced through metabolism in the body of a specified compound or salt thereof. After entry into the body, most drugs are substrates for chemical reactions that may change their physical properties and biologic effects. These metabolic conversions, which usually affect the polarity of the compounds of the invention, alter the way in which drugs are distributed in and excreted from the body. However, in some cases, metabolism of a drug is required for therapeutic effect.
  • therapeutically effective amount denotes an amount of a compound or molecule of the present invention that, when administered to a subject, (i) treats or prevents the particular disease, condition or disorder, (ii) attenuates, ameliorates or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, condition or disorder described herein.
  • the therapeutically effective amount will vary depending on the compound, the disease state being treated, the severity of the disease treated, the age and relative health of the subject, the route and form of administration, the judgement of the attending medical or veterinary practitioner, and other factors.
  • pharmaceutical composition denotes a mixture or solution comprising a therapeutically effective amount of an active pharmaceutical ingredient together with pharmaceutically acceptable excipients to be administered to a mammal, e.g., a human in need thereof.
  • pharmaceutically acceptable excipient can be used interchangeably and denote any pharmaceutically acceptable ingredient in a pharmaceutical composition having no therapeutic activity and being non-toxic to the subject administered, such as disintegrators, binders, fillers, solvents, buffers, tonicity agents, stabilizers, antioxidants, surfactants, carriers, diluents or lubricants used in formulating pharmaceutical products.
  • the present invention relates to (i) a compound of formula (I-2), (I-2), wherein R 1 is H, C1-6alkyl, or together with R 9 form an optionally substituted heterocyclic ring; R 2 is C1-6alkyl, or together with R 9 form an optionally substituted heterocyclic ring; M 1 is optionally substituted heterocyclylene which is further substituted by R 3 ; , ((haloC1-6alkyl)azetidinyl)C1-6alkyl, 2,5-diazabicyclo[2.2.1]heptanyl substituted by C1-6alkyl, haloC1-6alkyl or oxetanyl, 2-oxo-3,6-diazabicyclo[3.1.1]heptanyl substituted by C1-6alkyl, C3- 7cycloalkylC1-6alkyl or oxetanyl, 3,6-diazabicyclo[3.1.1]heptanyl substituted by C 1-6 alkyl
  • Another embodiment of present invention is (ii) a compound of formula (I), wherein R 1 is H, C1-6alkyl, or together with R 9 form a heterocyclic ring; R 2 is C 1-6 alkyl, or together with R 9 form a heterocyclic ring; R 3 is ; wherein R 4 is H, ((haloC 1-6 alkyl)azetidinyl)C 1-6 alkyl, 2,5-diazabicyclo[2.2.1]heptanyl substituted by C1-6alkyl, haloC1-6alkyl or oxetanyl, 2-oxo-3,6-diazabicyclo[3.1.1]heptanyl substituted by C 1-6 alkyl, C 3-7 cycloalkyl, C 3-7 cycloalkylC 1-6 alkyl or oxetanyl, 3,6-diazabicyclo[3.1.1]heptanyl substituted by C1-6alkyl, C3- 7cycloalkylcarbonyl
  • Another embodiment of present invention is (iii) a compound of formula (Ia) according to (i) or (ii), wherein R 1 is H, C1-6alkyl, or together with R 9 form a heterocyclic ring; R 2 is C 1-6 alkyl, or together with R 9 form a heterocyclic ring; 3 R is ; wherein R 4 is H, ((haloC 1-6 alkyl)azetidinyl)C 1-6 alkyl, 2,5-diazabicyclo[2.2.1]heptanyl substituted by C1-6alkyl, haloC1-6alkyl or oxetanyl, 2-oxo-3,6-diazabicyclo[3.1.1]heptanyl substituted by C 1-6 alkyl, C 3-7 cycloalkyl, C3-7cycloalkylC1-6alkyl or oxetanyl, 3,6-diazabicyclo[3.1.1]heptanyl substituted by C1-6alkyl, C3
  • Another embodiment of present invention is (iv) a compound of formula (I-1) according to any one of (i) to (iii), wherein R 1 is H, C 1-6 alkyl, or together with R 9 form a heterocyclic ring; R 2 is C 1-6 alkyl, or together with R 9 form a heterocyclic ring; ; wherein R 4 is H, ((haloC 1-6 alkyl)azetidinyl)C 1-6 alkyl, 2,5-diazabicyclo[2.2.1]heptanyl substituted by C1-6alkyl, haloC1-6alkyl or oxetanyl, 2-oxo-3,6-diazabicyclo[3.1.1]heptanyl substituted by C1-6alkyl, C3-7cycloalkyl, C 3-7 cycloalkylC 1-6 alkyl or oxetanyl, 3,6-diazabicyclo[3.1.1]heptanyl substituted by C1-6alkyl, C
  • a further embodiment of present invention is (v) a compound of formula (I-1a), wherein R 1 is H, C 1-6 alkyl, or together with R 9 form a heterocyclic ring; R 2 is C1-6alkyl, or together with R 9 form a heterocyclic ring; ; wherein R 4 is H, ((haloC1-6alkyl)azetidinyl)C1-6alkyl, 2,5-diazabicyclo[2.2.1]heptanyl substituted by C 1-6 alkyl, haloC 1-6 alkyl or oxetanyl, 2-oxo-3,6-diazabicyclo[3.1.1]heptanyl substituted by C1-6alkyl, C3-7cycloalkyl, C3-7cycloalkylC1-6alkyl or oxetanyl, 3,6-diazabicyclo[3.1.1]heptanyl substituted by C 1-6 alkyl, C 3- 7cycloalkylcarbonyl, C3-7cycl
  • a further embodiment of present invention is (vi) a compound of formula (I), (Ia), (I-1), (I- 1a) or (I-2), according to any one of (i) to (v), or a pharmaceutically acceptable salt thereof, wherein R 1 is C 1-6 alkyl, or together with R 9 form a heterocyclic ring, wherein R 1 is C 1-6 alkylene and R 9 is -C1-6alkylene-O-.
  • a further embodiment of present invention is (vii) a compound of formula (I), (Ia), (I-1), (I-1a) or (I-2) according to any one of (i) to (vi), wherein R 1 is methyl, or together with R 9 form a heterocyclic ring, wherein R 1 is methylene and R 9 is -methylene-O-.
  • a further embodiment of present invention is (viii) a compound of formula (I), (Ia), (I-1), (I-1a) or (I-2), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (vii), wherein R 2 is C 1-6 alkyl, or together with R 9 form a heterocyclic ring, wherein R 2 is C 1-6 alkylene and R 9 is a bond, O or -C1-6alkylene-O-.
  • a further embodiment of present invention is (ix) a compound of formula (I), (Ia), (I-1), (I-1a) or (I-2), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (viii), wherein R 2 is methyl, or together with R 9 form a heterocyclic ring, wherein R 2 is ethylene or propylene, and R 9 is a bond, O or -methylene-O-.
  • a further embodiment of present invention is (x) a compound of formula (I), (Ia), (I-1), (I- 1a) or (I-2), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (ix), wherein R 4 is H, ((haloC1-6alkyl)azetidinyl)C1-6alkyl, 2,5-diazabicyclo[2.2.1]heptanyl substituted by haloC1-6alkyl or oxetanyl, 2-oxo-3,6-diazabicyclo[3.1.1]heptanyl substituted by C 1-6 alkyl, C 3-7 cycloalkyl, C 3- 7cycloalkylC1-6alkyl or oxetanyl, 3,6-diazabicyclo[3.1.1]heptanyl substituted by haloC1-6alkyl or oxetanyl, 3-oxa-6-azabicyclo[3.1.1]heptanyl, C1-6alkoxyC1-6al
  • a further embodiment of present invention is (xi) a compound of formula (I), (Ia), (I-1), (I- 1a) or (I-2), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (x), wherein R 4 is H, [1-(2,2,2-trifluoroethyl)azetidin-3-yl]methyl, 3-(2,2-difluoroethyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl, 3-(2-fluoroethyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl, 3- (cyclopropylmethyl)-2-oxo-3,6-diazabicyclo[3.1.1]heptan-6-yl, 3-(oxetan-3-yl)-2-oxo-3,6- diazabicyclo[3.1.1]heptan-6-yl, 3-(oxetan-3-yl)-3,6- diaza
  • a further embodiment of present invention is (xii) a compound of formula (I), (Ia), (I-1), (I-1a) or (I-2), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (xi), wherein R 5 is phenyl which is twice substituted by substituents independently selected from halogen, C 1-6 alkoxy and trideuterioC 1-6 alkoxy.
  • a further embodiment of present invention is (xiii) a compound of formula (I), (Ia), (I-1), (I-1a) or (I-2), or a pharmaceutically acceptable salt thereof, according to any one of (i) to (xii), wherein R 5 is phenyl which is twice substituted by substituents independently selected from fluoro, methoxy and trideuteriomethoxy.
  • a further embodiment of present invention is (xiv) a compound of formula (I), (Ia), (I-1), (I-1a) or (I-2), according to any one of (i) to (xiii), wherein R 5 is 2,4-difluorophenyl, 4-fluoro-2- methoxy-phenyl or 4-fluoro-2-(trideuteriomethoxy)phenyl.
  • a further embodiment of present invention is (xv) a compound of formula (I), (Ia), (I-1), (I-1a) or (I-2), according to any one of (i) to (xiv), wherein R 7 is H or fluoro.
  • a further embodiment of present invention is (xvi) a compound of formula (I), (Ia), (I-1), (I-1a) or (I-2), according to any one of (i) to (xv), wherein Q 1 is CR 8 R 9 .
  • a further embodiment of present invention is (xvii) a compound of formula (I), (Ia), (I-1), (I-1a) or (I-2), according to any one of (i) to (xvi), wherein R 8 is H.
  • a further embodiment of present invention is (xviii) a compound of formula (I), (Ia), (I-1), (I-1a) or (I-2), according to any one of (i) to (xvii), wherein m is 0, 1 or 2.
  • a further embodiment of present invention is (xix) a compound of formula (I), (Ia), (I-1), (I-1a) or (I-2), according to any one of (i) to (v), wherein R 1 is C1-6alkyl, or together with R 9 form a heterocyclic ring, wherein R 1 is C1-6alkylene and R 9 is -C 1-6 alkylene-O-; R 2 is C 1-6 alkyl, or together with R 9 form a heterocyclic ring, wherein R 2 is C 1-6 alkylene and R 9 is a bond, O or -C1-6alkylene-O-; R 4 is H, ((haloC1-6alkyl)azetidinyl)C1-6alkyl, 2,5-diazabicyclo[2.2.1]heptanyl substituted by haloC 1-6 alkyl or oxetanyl, 2-oxo-3,6-diazabicyclo[3.1.1]heptanyl substituted by C 1-6 alkyl
  • a further embodiment of present invention is (xx) a compound of formula (I), (Ia), (I-1), (I-1a) or (I-2), according to (xix), wherein R 1 is methyl, or together with R 9 form a heterocyclic ring, wherein R 1 is methylene and R 9 is -methylene-O-; R 2 is methyl, or together with R 9 form a heterocyclic ring, wherein R 2 is ethylene or propylene, and R 9 is a bond, O or -methylene-O-; 3 R is ; wherein R 4 is H, [1-(2,2,2-trifluoroethyl)azetidin-3-yl]methyl, 3-(2,2-difluoroethyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl, 3-(2-fluoroethyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl, 3-(cyclopropylmethyl)-2-ox
  • Another embodiment of present invention is a compound of formula (I), (Ia), (I-1), (I- 1a) or (I-2) selected from the following: (8S,11S,18S)-10-[1-(2,4-difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-16-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one; (8S,11S,18R)-10-[1-(2,4-difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-16-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .
  • Another embodiment of present invention is a process for the preparation of a compound according to any one of (i) to (xxi) comprising the following step: a) the formation of compound of formula (I) via nucleophilic substitution between compound of formula (VI), (VI), and R 3 X, in the presence of a base; or via condensation reaction between compound of formula (VI) and R 3 OH in the presense of coupling reagent; b) the formation of compound of formula (IX), via nucleophilic substitution between compound of (Ic), when R 4 contains reactive primary or secondary amino group; or via photoredox coupling between compound of formula (Ic) and halide R 4 X or XR c BoC in the presence of a catalyst; c) the formation of compound of formula (IX) via reaction between compound of formula (VIII), anhydride or acid; wherein X is halogen; R e is a divalent substituent bearing reactive primary or secondary amino groups; the base in step a) is DIEA; the
  • Another embodiment of present invention is related to a compound or pharmaceutically acceptable salt according to any one of (i) to (xxi) for use as therapeutically active substance.
  • Another embodiment of present invention is related to a pharmaceutical composition comprising a compound in accordance with any one of (i) to (xxi) and a pharmaceutically acceptable excipient.
  • Another embodiment of present invention is related to the use of a compound according to any one of (i) to (xxi) for the treatment or prophylaxis of autoimmune diseases, inflammatory diseases, neurological disorders diseases, metabolic diseases, cardiovascular diseases, ocular diseases, or selective types of cancers where overexpression or activation of STING is implicated.
  • Another embodiment of present invention is related to a compound or pharmaceutically acceptable salt according to any one of (i) to (xxi) for the treatment or prophylaxis of autoimmune diseases, inflammatory diseases, neurological disorders diseases, metabolic diseases, cardiovascular diseases, ocular diseases, or selective types of cancers where overexpression or activation of STING is implicated.
  • Another embodiment of present invention is related to the use of a compound according to any one of (i) to (xxi) for the treatment to subjects suffered from an inteferonopathy or auto-inflammatory diseases in which the STING activation are the root-cause of disease pathologies.
  • Another embodiment of present invention is related to the use of a compound according to any one of (i) to (xxi) for the treatment or prophylaxis of systemic lupus erythematosus (SLE), dermatomyositis, diabetic kidney disease (DKD), diabetic retinopathy (DR), age-related macular degeneration (AMD), Anti-Neutrophilic Cytoplasmic Autoantibodies (ANCA) vasculitis, STING-associated vasculopathy with onset in infancy (SAVI), familial chilblain lupus (FCL), Niemann-Pick disease type C (NPC), Aicardi-Goutines Syndrome (AGS), COPA syndrome or Wiskott-Aldrich syndrome.
  • SLE systemic lupus erythematosus
  • DKD diabetic kidney disease
  • DR diabetic retinopathy
  • AMD age-related macular degeneration
  • ANCA Anti-Neutrophilic Cytoplasmic Autoantibodies
  • Another embodiment of present invention is related to the use of a compound according to any one of (i) to (xxi) for the preparation of a medicament for the treatment or prophylaxis of systemic lupus erythematosus (SLE), dermatomyositis, diabetic kidney disease (DKD), diabetic retinopathy (DR), age-related macular degeneration (AMD), Anti-Neutrophilic Cytoplasmic Autoantibodies (ANCA) vasculitis, STING-associated vasculopathy with onset in infancy (SAVI), familial chilblain lupus (FCL), Niemann-Pick disease type C (NPC), Aicardi- Goutines Syndrome (AGS), COPA syndrome or Wiskott-Aldrich syndrome.
  • SLE systemic lupus erythematosus
  • DKD diabetic kidney disease
  • DR diabetic retinopathy
  • AMD age-related macular degeneration
  • ANCA Anti-Neutrophilic Cyto
  • Another embodiment of present invention is related to the use of a compound according to any one of (i) to (xxi) for the inhibition of STING.
  • Another embodiment of present invention is related to the use of a compound according to any one of (i) to (xxi) for the preparation of a medicament for the inhibition of STING.
  • Another embodiment of present invention is related to a compound or pharmaceutically acceptable salt according to any one of (i) to (xxi), when manufactured according to a process of (xxii).
  • Another embodiment of present invention is related to a method for the treatment or prophylaxis of autoimmune diseases, which method comprises administering a therapeutically effective amount of a compound as defined in any one of (i) to (xxi).
  • PHARMACEUTICAL COMPOSITIONS AND ADMINISTRATION Another embodiment provides pharmaceutical compositions or medicaments containing the compounds of the invention and a therapeutically inert carrier, diluent or excipient, as well as methods of using the compounds of the invention to prepare such compositions and medicaments.
  • compounds of formula (I) may be formulated by mixing at ambient temperature at the appropriate pH, and at the desired degree of purity, with physiologically acceptable carriers, i.e., carriers that are non-toxic to recipients at the dosages and concentrations employed into a galenical administration form.
  • physiologically acceptable carriers i.e., carriers that are non-toxic to recipients at the dosages and concentrations employed into a galenical administration form.
  • the pH of the formulation depends mainly on the particular use and the concentration of compound, but preferably ranges anywhere from about 3 to about 8.
  • a compound of formula (I) is formulated in an acetate buffer, at pH 5.
  • the compounds of formula (I) are sterile.
  • the compound may be stored, for example, as a solid or amorphous composition, as a lyophilized formulation or as an aqueous solution.
  • compositions are formulated, dosed, and administered in a fashion consistent with good medical practice.
  • Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners.
  • the “effective amount” of the compound to be administered will be governed by such considerations, and is the minimum amount necessary to inhibit STING interaction with IRF3, NF-kB, NLRP3 etc., for blocking downstream type I IFN and pro-inflammatory cytokine (e.g. IL-6, TNFa, ISGs) production, cellular responses/conditions (e.g.
  • the pharmaceutically effective amount of the compound of the invention administered parenterally per dose will be in the range of about 0.1 to 1000 mg/kg, alternatively about 0.1 to 1000 mg/kg of patient body weight per day, with the typical initial range of compound used being 0.1 to 1000 mg/kg/day.
  • oral unit dosage forms such as tablets and capsules, preferably contain from about 0.1 to about 1000 mg of the compound of the invention.
  • the compounds of the invention may be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural and intranasal, and, if desired for local treatment, intralesional administration.
  • Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.
  • the compounds of the present invention may be administered in any convenient administrative form, e.g., tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc.
  • compositions may contain components conventional in pharmaceutical preparations, e.g., diluents, carriers, pH modifiers, sweeteners, bulking agents, and further active agents.
  • a typical formulation is prepared by mixing a compound of the present invention and a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, e.g., Ansel, Howard C., et al., Ansel’s Pharmaceutical Dosage Forms and Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R., et al. Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C.
  • the formulations may also include one or more buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present invention or pharmaceutical composition thereof) or aid in the manufacturing of the pharmaceutical product (i.e., medicament).
  • buffers stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present invention or pharmaceutical composition thereof) or aid in the manufacturing
  • An example of a suitable oral dosage form is a tablet containing about 0.1 to 1000 mg of the compound of the invention compounded with about 0.1 to 1000 mg anhydrous lactose, about 0.1 to 1000 mg sodium croscarmellose, about 0.1 to 1000 mg polyvinylpyrrolidone (PVP) K30, and about 0.1 to 1000 mg magnesium stearate.
  • the powdered ingredients are first mixed together and then mixed with a solution of the PVP.
  • the resulting composition can be dried, granulated, mixed with the magnesium stearate and compressed to tablet form using conventional equipment.
  • An example of an aerosol formulation can be prepared by dissolving the compound, for example 0.1 to 1000 mg, of the invention in a suitable buffer solution, e.g.
  • An embodiment includes a pharmaceutical composition comprising a compound of formula (I), or a stereoisomer or pharmaceutically acceptable salt thereof.
  • a pharmaceutical composition comprising a compound of formula (I), or a stereoisomer or pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier or excipient.
  • Another embodiment includes a pharmaceutical composition comprising a compound of formula (I) for use in the treatment of interferonopathies, autoimmune and inflammatory diseases.
  • composition A A compound of the present invention can be used in a manner known per se as the active ingredient for the production of tablets of the following composition: Per tablet Active ingredient 200 mg Microcrystalline cellulose 155 mg Corn starch 25 mg Talc 25 mg Hydroxypropylmethylcellulose 20 mg 425 mg.
  • Composition B A compound of the present invention can be used in a manner known per se as the active ingredient for the production of capsules of the following composition: Per capsule Active ingredient 100.0 mg Corn starch 20.0 mg Lactose 95.0 mg Talc 4.5 mg Magnesium stearate 0.5 mg 220.0 mg INDICATIONS AND METHODS OF TREATMENT The compound of the invention inhibit the binding of cGAMP to STING and its downstream signaling.
  • the compound of the invention are useful for blocking STING activation, signaling, downstream cytokine, chemokine production and cellular processes such as apoptosis and autophagy.
  • Compounds of the invention are useful for inhibition of STING.
  • compounds of invention are useful for the treatment or prophylaxis of systemic lupus erythematosus (SLE), dermatomyositis, diabetic kidney disease (DKD), diabetic retinopathy (DR), age-related macular degeneration (AMD), Anti-Neutrophilic Cytoplasmic Autoantibodies (ANCA) vasculitis, STING-associated vasculopathy with onset in infancy (SAVI), familial chilblain lupus (FCL), Niemann-Pick disease type C (NPC), Aicardi- Goutines Syndrome (AGS), COPA syndrome or Wiskott-Aldrich syndrome.
  • SLE systemic lupus erythematosus
  • DKD diabetic kidney
  • the compound can be useful for the treatment or prophylaxis of autoimmune diseases, inflammatory diseases, neurological disorders diseases, metabolic diseases, cardiovascular diseases, ocular diseases, or selective types of cancers where overexpression or activation of STING is implicated.
  • compounds of the invention are useful for the treatment or prophylaxis of autoimmune diseases.
  • compounds of the invention are useful for the treatment or prophylaxis of inflammatory diseases.
  • compounds of the invention are useful for the treatment or prophylaxis of neurological disorders diseases.
  • compounds of the invention are useful for the treatment or prophylaxis of cardiovascular diseases.
  • compounds of the invention are useful for the treatment or prophylaxis of ocular diseases.
  • compounds of the invention are useful for the treatment or prophylaxis of selective types of cancers where overexpression or activation of STING is implicated.
  • compounds of the invention are useful for the treatment of subjects suffered from an inteferonopathy or auto-inflammatory diseases in which the STING activation are the root-cause of disease pathologies. More broadly, the compounds can be used for the treatment of all pathological cellular processes which are STING dependent.
  • Another embodiment includes a method of treating or preventing cancer in a mammal in need of such treatment, wherein the method comprises administering to said mammal a therapeutically effective amount of a compound of formula (I), a stereoisomer, tautomer, prodrug or pharmaceutically acceptable salt thereof.
  • Rf is H or PG, wherein PG can be, for example, Boc or Cbz; X is halogen.
  • Scheme 1 the synthesis of compounds of the present invention started from boronic ester compound of formula (II). Suzuki coupling between compound of formula (II) and compound of formula (IIb) with a catalyst, such as Pd(dppf)Cl2, and a base, such as K2CO3, provides compound of formula (III).
  • Compound of formula (III) is hydrolyzed in the presence of LiOH directly and followed by appropriate deprotection of Rf to give compound of formula (IV) (Boc deprotection: HCl in dioxane or TFA in DCM; Cbz deprotection: Pd/C or Pd(OH)2/C under H 2 ).
  • Compound of formula (IV) can be cyclized to give compound of formula (V) in the presence of a coupling reagent, such as HATU, and a base, such as DIPEA.
  • This invention also relates to a process for the preparation of a compound of formula (I) comprising the following step: a) the formation of compound of formula (I) via nucleophilic substitution between compound of formula (VI), (VI), and R 3 X, in the presence of a base; or via condensation reaction between compound of formula (VI) and R 3 OH in the presence of coupling reagent; b) the formation compound of formula (IX), via nucleophilic substitution between compound of (Ic), when R 4 contains reactive primary or secondary amino group; or via photoredox coupling between compound of formula (Ic) and halide R 4 X or XR c BoC in the presence of a catalyst; c) the formation of compound of formula (IX) via reaction between compound of formula (VIII), anhydride or acid; wherein the base in step a) can be, for example, DIEA; the coupling reagent in step a) can be, for example, PyBOP; the catalyst in step b
  • a compound of formula (I), (Ia), (I-1), (I-1a) or (I-2) when manufactured according to the above process is also an object of the invention.
  • EXAMPLES The invention will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of the invention.
  • ABBREVIATIONS The invention will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of the invention.
  • Waters AutoP purification System (Sample Manager 2767, Pump 2525, Detector: Micromass ZQ and UV 2487, solvent system: acetonitrile and 0.1% ammonium hydroxide in water; acetonitrile and 0.1% FA in water or acetonitrile and 0.1% TFA in water).
  • Or Gilson-281 purification System (Pump 322, Detector: UV 156, solvent system: acetonitrile and 0.05% ammonium hydroxide in water; acetonitrile and 0.225% FA in water; acetonitrile and 0.05% HCl in water; acetonitrile and 0.075% TFA in water; or acetonitrile and water).
  • LC/MS spectra of compounds were obtained using a LC/MS (Waters TM Alliance 2795- Micromass ZQ, Shimadzu Alliance 2020-Micromass ZQ or Agilent Alliance 6110-Micromass ZQ), LC/MS conditions were as follows (running time 3 or 1.5 mins): Acidic condition I: A: 0.1% TFA in H2O; B: 0.1% TFA in acetonitrile; Acidic condition II: A: 0.0375% TFA in H 2 O; B: 0.01875% TFA in acetonitrile; Basic condition I: A: 0.1% NH 3 ⁇ H 2 O in H 2 O; B: acetonitrile; Basic condition II: A: 0.025% NH3 ⁇ H2O in H2O; B: acetonitrile; Neutral condition: A: H2O; B: acetonitrile.
  • Mass spectra generally only ions which indicate the parent mass are reported, and unless otherwise stated the mass ion quoted is the positive mass ion (MH) + .
  • NMR Spectra were obtained using Bruker Avance 400 MHz, 500 MHz. The microwave assisted reactions were carried out in a Biotage Initiator Sixty microwave synthesizer. All reactions involving air-sensitive reagents were performed under an argon or nitrogen atmosphere. Reagents were used as received from commercial suppliers without further purification unless otherwise noted.
  • Step 2 preparation of tert-butyl 6-[(2-amino-6-bromo-anilino)methyl]-1,4-oxazepane- 4-carboxylate (intermediate A1-b) To a flask was added nickel (273 mg, 4.65 mmol), MeOH (20 mL) and tert-butyl 6-[(2- bromo-6-nitro-anilino)methyl]-1,4-oxazepane-4-carboxylate (intermediate A1-a, 2 g, 4.65 mmol), the suspension was stirred vigorously and hydrazine (2.98 g, 2.9 mL, 46.48 mmol) was then added dropwise under ice-water bath.
  • Step 3 preparation of tert-butyl 6-[(7-bromobenzimidazol-1-yl)methyl]-1,4- oxazepane-4-carboxylate (intermediate A1-c)
  • Step 4 preparation of tert-butyl 6-[[7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2- yl)benzimidazol-1-yl]methyl]-1,4-oxazepane-4-carboxylate (intermediate A1)
  • tert-butyl 6-[(7-bromobenzimidazol-1-yl)methyl]-1,4-oxazepane-4- carboxylate (intermediate A1-c, 1.1 g, 2.68 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2- dioxaborolane) (1.36 g, 5.36 mmol), potassium acetate (526 mg,
  • Step 1 preparation of N-[(4,6-dichloropyrimidin-5-yl)methyleneamino]-2,4-difluoro- aniline (compound C2-b)
  • a mixture of 4,6-dichloro-5-pyrimidinecarbaldehyde (compound C2-a, 20.0 g, 113 mmol), 2,4-difluorophenylhydrazine hydrochloride (24.9 g, 137.9 mmol) and potassium carbonate (21.2 g, 153.2 mmol) in DCM (400 mL) was stirred at 20 °C for 12 h. Then the mixture was washed with water and brine, the organic layer was dried and concentrated to give the crude compound C2-b (34.5 g).
  • Step 2 preparation of (4-fluoro-2-methoxy-phenyl)hydrazine hydrochloride (compound C11-d) A mixture of compound C11-c (11.0 g, 34.34 mmol) in ethanol (100.0 mL) and con. HCl (10.0 mL, 120 mmol) was stirred for 12 h at 60 °C. The mixture was concentrated, and then EtOAc (100 mL) was added.
  • Step 3 preparation of N-[(4,6-dichloropyrimidin-5-yl)methyleneamino]-4-fluoro-2- methoxy-aniline (compound C11-f)
  • a mixture of compound C11-d (4.3 g, 22.32 mmol), 4,6-dichloro-5- pyrimidinecarbaldehyde (compound C11-e, 4.3 g, 24.3 mmol) and triethylamine (3.5 g, 34.59 mmol) in anhydrous THF (100.0 mL) was stirred for 12 h at 20 °C.
  • the reaction solution was diluted with EtOAc (100.0 mL), washed with water and brine.
  • Step 4 preparation of 4-chloro-1-(4-fluoro-2-methoxy-phenyl)pyrazolo[3,4- d]pyrimidine (Intermediate C11)
  • Step 2 preparation of (5-amino-1-(4-fluoro-2-methoxy-phenyl)pyrazole-4- carbonitrile (compound C12-c)
  • compound C12-c To a solution of (4-fluoro-2-methoxy-phenyl)hydrazine;hydrochloride (compound C12-b, 11.5 g, 59.7 mmol) in ethanol (10 mL) was added ethoxymethylenemalononitrile (7.29 g, 59.7 mmol) and DIEA (32 mL, 180 mmol). The mixture was stirred at 80 °C for 12 h. The reaction mixture was concentrated and the residue was diluted with EA.
  • Step 3 preparation of (N-[4-cyano-2-(4-fluoro-2-methoxy-phenyl)pyrazol-3- yl]acetamide (compound C12-d)
  • a solution of 5-amino-1-(4-fluoro-2-methoxy-phenyl)pyrazole-4-carbonitrile (compound C12-c, 5.2 g, 22.39 mmol) in acetic anhydride (2.29 g, 22.39 mmol) was stirred at 100 °C for 12 h.
  • Step 4 preparation of 1-(4-fluoro-2-methoxy-phenyl)-6-methyl-pyrazolo[3,4- d]pyrimidin-4-ol (compound C12-e)
  • acetic acid 13 mL
  • phosphorus oxychloride 1.3 mL, 13.95 mmol
  • Step 5 preparation of 4-chloro-1-(4-fluoro-2-methoxy-phenyl)-6-methyl- pyrazolo[3,4-d]pyrimidine (Intermediate C12)
  • Step 1 preparation of 2-(cyclopropoxy)-4-fluoro-1-nitro-benzene (compound C17-a) A mixture of 2,4-difluoro-1-nitro-benzene (10.0 g, 62.86 mmol), cyclopropanol (5.0 g, 86.09 mmol) and sodium tert-butoxide (12.1 g, 0.12 mol) in THF (0.2 L) was stirred at 50 °C for 1 h.
  • Step 2 ⁇ 6 preparation of 4-chloro-1-[2-(cyclopropoxy)-4-fluoro-phenyl]pyrazolo[3,4- d]pyrimidine (Intermediate C17)
  • the title compound was prepared in analogy to the preparation of intermediate C18 by using compound C17-a instead of compound C18-a.
  • Step 1 preparation of 2-ethoxy-4-fluoro-1-nitro-benzene (compound C18-a)
  • a mixture of 5-fluoro-2-nitro-phenol (1.0 g, 6.4 mmol), potassium carbonate (1.7 g, 12.7 mmol) and iodoethane (0.6 mL, 7.64 mmol, 1.2 eq) in DMF (10.0 mL) was stirred at 30 °C for 2 h. The mixture was poured into water and extracted with EtOAc.
  • Step 2 preparation of 2-ethoxy-4-fluoro-aniline (compound C18-b) To the mixture of 2-ethoxy-4-fluoro-1-nitro-benzene (compound C18-a, 1.0 g, 5.4 mmol) in methanol (10.0 mL) was added Pd/C (0.1 g) and the mixture was stirred at 20 °C for 4 h under H2 atmosphere. The mixture was poured into water (20.0 mL) and extracted with EtOAc (20.0 mL twice). The organic layer was concentrated, the residue was purified by flash chromatography column to give compound C18-b, (0.8 g).
  • Step 3 preparation of 4-chloro-6-(2-ethoxy-4-fluoro-anilino)pyrimidine-5- carbaldehyde (compound C18-c) To a mixture of 4,6-dichloropyrimidine-5-carbaldehyde (0.8 g, 4.51 mmol) and TEA (0.9 g, 9.0 mmol) in chloroform (20.0 mL) was added 2-ethoxy-4-fluoro-aniline (compound C18-b, 0.7 g, 4.5 mmol) dropwise and the mixture was stirred at 0 °C for 1 h.
  • Step 4 preparation of [[4-chloro-6-(2-ethoxy-4-fluoro-anilino)pyrimidin-5- yl]methyleneamino] hydrogen sulfate (compound C18-d)
  • Step 5 preparation of 1-(2-ethoxy-4-fluoro-phenyl)pyrazolo[3,4-d]pyrimidin-4-ol (compound C18-e) To the mixture of compound C18-d (0.1 g, 0.3 mmol) in ACN (5.0 mL) was added NaOH a.q (2.0 mL, 1M) and the reaction mixture was stirred at 25 °C for 2 h under N2. The mixture was purified by prep-HPLC to give compound C18-e (30.0 mg). LCMS (M+H + ): 275.
  • Step 6 preparation of 4-chloro-1-(2-ethoxy-4-fluoro-phenyl)pyrazolo[3,4- d]pyrimidine (compound C18-f)
  • a solution of 1-(2-ethoxy-4-fluoro-phenyl)pyrazolo[3,4-d]pyrimidin-4-ol (compound C18- e, 20.0 mg, 0.07 mmol) in POCl 3 (1.0 mL) was stirred at 100 °C for 1 h. The mixture was concentrated to give intermediate C18 (20.0 mg).
  • Step 1 preparation of 2-(difluoromethoxy)-4-fluoro-1-nitro-benzene (C19-a) To a solution of 5-fluoro-2-nitro-phenol (5.0 g, 31.83 mmol) and sodium chlorodifluoroacetate (5.8 g, 38.2mmol) in DMF (60.0 mL) was added Na 2 CO 3 (6.75 g, 63.65 mmol). The mixture was stirred at 100 °C for 2 h, and then poured into water (200.0 mL).
  • Step 2-6 preparation of 4-chloro-1-[2-(difluoromethoxy)-4-fluoro- phenyl]pyrazolo[3,4-d]pyrimidine (Intermediate C19)
  • the title compound was prepared in analogy to the preparation of intermediate C18 by using compound C19-a instead of compound C18-a.
  • LCMS (M+H) + 315.
  • Step 2 preparation of 1-(2,4-difluorophenyl)-6-(methoxymethyl)-5H-pyrazolo[3,4- d]pyrimidin-4-one (compound C28-b)
  • a mixture of compound C28-a (700 mg, 2.62 mmol), 2-methoxyacetonitrile (372 mg, 5.24 mmol) in 4 M HCl and dioxane (8 mL) was stirred at 100 °C for 14 hours, then the solvent was removed in vacuo and the residue was partitioned between DCM and water.
  • Step 3 preparation of [4-chloro-1-(4-fluoro-2-methoxy-phenyl)pyrazolo[3,4- d]pyrimidin-6-yl]amine (Intermediate C28) A suspension of compound C28-c (200 mg, 0.68 mmol)) in phosphorus oxychloride (2 mL) was stirred at 90 °C for 0.5 hours.
  • Step 2 preparation of 2,4-difluoro-6-methoxy-aniline (compound C61-c)
  • compound C61-c 1,5-difluoro-3-methoxy-2-nitro-benzene (compound C61-b, 10.0 g, 52.9 mmol) in anhydrous EtOAc (200 mL)
  • Pd/C 1.0 g
  • the reaction mixture was stirred at room temperature for 12 h under hydrogen.
  • the mixture was filtered, the filtrate was concentrated to give compound C61-c (7.5 g).
  • LCMS (M+H) + 160.
  • Step 3 ⁇ 5 preparation of 1-(2,4-difluoro-6-methoxy-phenyl)pyrazolo[3,4-d]pyrimidin- 4-ol (compound C61-f)
  • Compound C61-f was prepared in analogy to the preparation of intermediate C43 by using 2,4-difluoro-6-methoxy-aniline instead of 2-bromo-4-fluoro-aniline.
  • LCMS (M+H) + 279.
  • Step 6 preparation of 4-chloro-1-(2,4-difluoro-6-methoxy-phenyl)pyrazolo[3,4- d]pyrimidine (Intermediate C61)
  • a mixture of 1-(2,4-difluoro-6-methoxy-phenyl)pyrazolo[3,4-d]pyrimidin-4-ol (compound C61-f, 4.6 g, 16.5 mmol) in POCl3 (18 mL) was stirred at 100 °C for 2 h. The mixture was concentrated, the residue was taken in EtOAc (100 mL) and poured into aq.NaHCO 3 (100 mL).
  • Step 2 preparation of tert-butyl 6-[4-benzyloxy-1-(2,4-difluorophenyl)pyrazolo[3,4- d]pyrimidin-6-yl]-3,6-diazabicyclo[3.1.1]heptane-3-carboxylate (compound C80-b)
  • compound C80-a 1.25 g, 3.35 mmol
  • tert-butyl 3,6- diazabicyclo[3.1.1]heptane-3-carboxylate 731.32 mg, 3.69 mmol
  • N,N-diisopropylethylamine (1.75 mL, 10 mmol
  • Step 4 preparation of 1-(2,4-difluorophenyl)-6-[3-(2-fluoroethyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl]pyrazolo[3,4-d]pyrimidin-4-ol (intermediate C80)
  • 1-fluoro-2-iodoethane (1.90 g, 10.91 mmol) in ACN (20 mL) was added triethylamine (1.5 mL, 10.91 mmol) and then the mixture was stirred at 80 °C for 12 hrs.
  • Step 2 preparation of 6-[(1R,4R)-2,5-diazabicyclo[2.2.1]heptan-2-yl]-1-(2,4- difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-ol (compound C88-b)
  • TFA 3.04 mL, 12.16 mmol
  • the reaction mixture was concentrated to give compound c88-b (1500 mg) as a yellow oil.
  • Step 3 preparation of 1-(2,4-difluorophenyl)-6-[(1R,4R)-5-(oxetan-3-yl)-2,5- diazabicyclo[2.2.1]heptan-2-yl]pyrazolo[3,4-d]pyrimidin-4-ol (Intermediate C88)
  • 3-oxetanone 720.0 mg, 9.99 mmol
  • sodium triacetoxyborohydride (1.04 g, 4.91 mmol) at 0 °C and then stirred at 25 °C for 1 h.
  • Step 2 preparation of 3-methyl-3,6-diazabicyclo[3.1.1]heptan-2-one (compound C90- b)
  • a mixture of compound C90-a (1.30 g, 5.75 mmol) in 4 M HCl in dioxane (15.0 mL,) was stirred at 25 °C for 2 h. Then the reaction mixture was concentrated and the residue was lyophilized to give compound C90-b (800 mg) as a yellow oil.
  • Step 3 preparation of 6-[4-benzyloxy-1-(2,4-difluorophenyl)pyrazolo[3,4- d]pyrimidin-6-yl]-3-methyl-3,6-diazabicyclo[3.1.1]heptan-2-one (compound C90-c)
  • Example 1 and 2 (8S,11S,18S)-10-[1-(2,4-difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-16-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one and (8S,11S,18R)-10-[1-(2,4- difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-16-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(26),2(30),3,5,21,23(27),24-heptaen-
  • Step 1 preparation of N-methyl-3-[2-methyl-7-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)benzimidazol-1-yl]propan-1-amine
  • (1a) A mixture of tert-butyl 6-[[7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzimidazol-1- yl]methyl]-1,4-oxazepane-4-carboxylate (intermediate A1, 1 g, 2.19 mmol, as the “BORONIC REAGENT” in table 2) in DCM (10 mL) and HCl in Dioxane (4 M, 10 mL, ) was stirred at rt for 4 hours, then the reaction was filtered to give compound 1a (800 mg).
  • Step 3 preparation of ditert-butyl (8S,11S)-12-oxo-16-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(26),2(30),3,5,21,23(27),24-heptaene- 7,10-dicarboxylate (compound 1c) To a mixture of HATU (478 mg, 1.26 mmol ) and DIPEA (406 mg, 3.14 mmol ) in acetonitrile (66 mL) was added (2S,4S)-1-tert-butoxycarbonyl-4-[tert-butoxycarbonyl-[6-[3-(1,4- oxazepan-6-ylmethyl)benzimidazol-4-yl]-2-pyridyl]amino]pyrrolidine-2-carboxylic acid (compound 1b, 400
  • Step 4 preparation of (8S,11S)-16-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(26),2(30),3,5,21,23(27),24-heptaen- 12-one (compound 1d)
  • Step 5 preparation of (8S,11S,18S)-10-[1-(2,4-difluorophenyl)pyrazolo[3,4- d]pyrimidin-4-yl]-16-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(26),2(30),3,5,21,23(27),24-heptaen- 12-one and (8S,11S,18R)-10-[1-(2,4-difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-16-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]
  • Example 1 11 mg, faster eluted
  • Example 2 (19 mg, slower eluted).
  • Example 1 LCMS (M+H + ): 649.
  • 1 H NMR (500 MHz, METHANOL-d4) ⁇ 8.60 - 8.34 (m, 1H), 8.30 - 8.20 (m, 2H), 7.88 - 7.79 (m, 1H), 7.76 - 7.69 (m, 1H), 7.69 - 7.58 (m, 1H), 7.51 - 7.41 (m, 1H), 7.37 - 7.26 (m, 2H), 7.23 - 7.15 (m, 1H), 7.08 - 7.01 (m, 1H), 6.83 - 6.73 (m, 1H), 5.68 - 5.49 (m, 2H), 4.80 - 4.74 (m, 1H), 4.65 - 4.56 (m, 1H), 4.52 - 4.43 (m, 1H), 4.38 - 4.30 (m, 2H),
  • HALIDE 1 1 .0 ]triaconta- 7.25 (m, 1H), 7.13 - 7.01 (m, Intermediate B9 1(26),2(30),3,5,21,23(27),24- 2H), 6.94 - 6.88 (m, 1H), HALIDE 2: heptaen-12-one and 6.84 - 6.71 (m, 1H), 6.00 - Intermediate C11 (8S,11S,18R)-10-[1-(4-fluoro-2- 5.75 (m, 1H), 5.64 - 5.49 (m, methoxy-phenyl)pyrazolo[3,4- 1H), 4.82 - 4.74 (m, 1H), d]pyrimidin-4-yl]-21-methyl-16- 4.64 - 4.56 (m, 1H), 4.42 - oxa-7,10,13,20,22,30- 4.25 (m, 4H), 4.13 -
  • Example 3 (8S,11S,16R)-10-[1-(2,4-difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-7,10,13,18,20,28- 2,6 8,11 13,16 21,25 hexazahexacyclo[16.6.1.1 .1 .1 .0 ]octacosa-1(24),2(28),3,5,19,21(25),22-heptaen- 12-one
  • Example 3 was prepared in analogy to the preparation of Example 1 by using intermediate A2 instead of intermediate A1 and intermediate B8 instead of intermediate B9.
  • Step 1 preparation of tert-butyl N-[trans-3-(2-bromo-6-nitro- anilino)cyclopentyl]carbamate (compound 4a)
  • trans-cyclopentane-1,3-diamine;dihydrochloride (2.99 g, 17.27 mmol)
  • K 2 CO 3 5.97 g, 43.18 mmol
  • methanol 50 mL
  • acetonitrile 50 mL
  • the suspension was heated to 70 o C and a solution of 1-bromo-2-fluoro-3-nitro-benzene (1.9 g, 8.64 mmol) in acetonitrile (20 mL) was added dropwise in about 40 mins.
  • the mixture was kept stirring for about 1 hr.
  • the mixture was cooled and filtered through celite, the collected solid was washed with ACN, the filtrate was concentrated to give a brown oil.
  • the crude oil was dissolved in about 100 mL EA and washed with 300 mL sat. NH4Cl.
  • the mixture was separated, the aqueous layer was basified with 2 N aq. solution of NaOH (pH>10), then extracted with 50 mL DCM/MeOH (10:1) five times.
  • the DCM layer was dried over Na 2 SO 4 and concentrated, an orange oil was obtained.
  • Step 2 preparation of tert-butyl N-[trans-3-(2-amino-6-bromo- anilino)cyclopentyl]carbamate (compound 4b)
  • compound 4a 1.8 g, 4.5 mmol, 1 eq
  • aluminum-nickel alloy 577.92 mg, 6.75 mmol,
  • MeOH 50 mL
  • hydrazine;hydrate 689 mg, 13.49 mmol
  • Step 5 preparation of tert-butyl N-methyl-N-[ trans-3-[7-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)benzimidazol-1-yl]cyclopentyl]carbamate (compound 4e)
  • compound 4d 1.2 g, 3 mmol
  • 4,4,4',4',5,5,5',5'- octamethyl-2,2'-bi(1,3,2-dioxaborolane) (2 g, 7.88 mmol
  • bis(1-adamantyl)-butyl-phosphine (218 mg, 0.609 mmol
  • potassium acetate 597 mg, 6.09 mmol
  • DMSO 20 mL
  • Step 6 preparation of O1-benzyl O2-methyl (2S,4S)-4-[[6-[3-[(1S)-3-[tert- butoxycarbonyl(methyl)amino]cyclopentyl]benzimidazol-4-yl]-2- pyridyl]amino]pyrrolidine-1,2-dicarboxylate (compound 4f)
  • compound 4e 1.1 g, 2.49 mmol
  • K 2 CO 3 (689 mg, 4.98 mmol
  • 1,4-dioxane 8 mL
  • water (0.500 mL).
  • Step 7 preparation of (2S,4S)-1-benzyloxycarbonyl-4-[[6-[3-[ trans -3- (methylamino)cyclopentyl]benzimidazol-4-yl]-2-pyridyl]amino]pyrrolidine-2-carboxylic acid (compound 4g)
  • compound 4f 1.1 g, 1.64 mmol
  • tetrahydrofuran 4 mL
  • methanol 4 mL
  • 2 M LiOH (2N aq.) 4 mL, 8 mmol
  • Step 8 preparation of benzyl (8S,11S)-6-methyl-7-oxo-1,6,9,12,23,26- 2,5 8,11 13,17 22,25 hexazahexacyclo[16.6.1.1 .1 .1 .0 ]octacosa-13,15,17(26),18,20,22(25),23-heptaene- 9-carboxylate (compound 4h)
  • HATU 760 mg, 2 mmol
  • DIEA 620 mg, 4.8 mmol
  • N,N- dimethylformamide (2 mL) and acetonitrile (150 mL) was added dropwise another suspension of compound 4g (535 mg, 0.800 mmol) in N,N-dimethylformamide (2 mL) and acetonitrile (150 mL), it took about 2 hrs.
  • Step 11 preparation of (2R,5R,8S,11S)-9-[1-(2,4-difluorophenyl)pyrazolo[3,4- 2,5 8,11 13,17 22,25 d]pyrimidin-4-yl]-6-methyl-1,6,9,12,23,26-hexazahexacyclo[16.6.1.1 .1 .1 .0 ] octacosa-13,15,17(26),18,20,22(25),23-heptaen-7-one and (2S,5S,8S,11S)-9-[1-(2,4- difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-6-methyl-1,6,9,12,23,26-hexazahexacyclo 2,5 8,11 13,17 22,25 [16.6.1.1 .1 .1 .0 ]octacosa-13,15,17(26),18,20,22(25),23-hepta
  • Example 8 (8S,11S)-10-[1-(4-fluoro-2-methoxy-phenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-13,18-dimethyl- 2,6 8,11 20,24 spiro[7,10,13,17,19,26-hexazapentacyclo[15.6.1.1 .1 .0 ]hexacosa- 1(23),2(26),3,5,18,20(24),21-heptaene-15,3'-oxetane]-12-one
  • the title compound was prepared according to the following scheme: Step 1 ⁇ 6: preparation of O1-benzyl O2-methyl (2S,4S)-4-[[6-[3-[[3-[[tert- butoxycarbonyl(methyl)amino]methyl]oxetan-3-yl]methyl]-2-methyl-benzimidazol-4-yl]-2- pyridyl]amino]pyrrolidine-1
  • Step 8 preparation of benzyl (8S,11S)-13,18-dimethyl-12-oxo-spiro[7,10,13,17,19,26- 2,6 8,11 20,24 hexazapentacyclo[15.6.1.1 .1 .0 ]hexacosa-1(23),2(26),3,5,18,20(24),21-heptaene-15,3'- oxetane]-10-carboxylate (8h) To a solution of HATU (408 mg, 1.07 mmol) and DIEA (333 mg, 2.58 mmol) in acetonitrile (75 mL) was added dropwise a solution of compound 8g (300 mg, 0.429 mmol) in acetonitrile (75 mL) in about 2 hrs.
  • Step 9 preparation of (8S,11S)-13,18-dimethylspiro[7,10,13,17,19,26- 2,6 8,11 20,24 hexazapentacyclo[15.6.1.1 .1 .0 ]hexacosa-1(23),2(26),3,5,18,20(24),21-heptaene-15,3'- oxetane]-12-one (8i) To the flask containing compound 8h (120 mg, 0.211 mmol) was added methanol (50 mL) and palladium hydroxide (20% on carbon with 50% H2O) (30 mg, 0.214 mmol).
  • Step 10 preparation of (8S,11S)-10-[1-(4-fluoro-2-methoxy-phenyl)pyrazolo[3,4- d]pyrimidin-4-yl]-13,18-dimethyl-spiro[7,10,13,17,19,26- 2,6 8,11 20,24 hexazapentacyclo[15.6.1.1 .1 .0 ]hexacosa-1(23),2(26),3,5,18,20(24),21-heptaene-15,3'- oxetane]-12-one (Example 8) To a microwave tube was added compound 8i (54 mg, 0.125 mmol), DIEA (48 mg, 0.375 mmol), acetonitrile (2 mL) and 4-chloro-1-(4-fluoro-2-methoxy-phenyl)pyrazolo[3,4- d]pyrimidin-4-yl]-13,18-dimethyl-spiro[7,10
  • Example 8 as a white powder.
  • Example 21 and 22 (8S,11S,18S)-10-[1-[2-(difluoromethoxy)-4-fluoro-phenyl]pyrazolo[3,4-d]pyrimidin-4-yl]- 25-fluoro-21-methyl-17-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(26),2(30),3,5,21,23(27),24-heptaen- 12-one (Example 21) and (8S,11S,18R)-10-[1-[2-(difluoromethoxy)-4-fluoro- phenyl]pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6
  • Step 1 ⁇ 3 preparation of benzyl (8S,11S)-25-fluoro-21-methyl-12-oxo-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaene-10-carboxylate (compound 21c)
  • Compound 21c was prepared in analogy to the preparation of compound 1c by using intermediate A6 instead of intermediate A1 and intermediate B10 instead of intermediate B9.
  • Step 4 preparation of (8S,11S)-25-fluoro-21-methyl-17-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(26),2(30),3,5,21,23(27),24-heptaen- 12-one (compound 21d)
  • Example 23 and 24 (8S,11S,18S)-10-[1-[2-(cyclopropoxy)-4-fluoro-phenyl]pyrazolo[3,4-d]pyrimidin-4-yl]-25- fluoro-21-methyl-17-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(26),2(30),3,5,21,23(27),24-heptaen- 12-one (Example 23) and (8S,11S,18R)-10-[1-[2-(cyclopropoxy)-4-fluoro- phenyl]pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .
  • Example 24 slower eluted, LCMS (M+H + ): 719.
  • 1 H NMR (500 MHz, METHANOL-d 4 ) ⁇ 8.51 - 8.24 (m, 1H), 8.20 - 7.74 (m, 1H), 7.70 - 7.38 (m, 2H), 7.37 - 7.26 (m, 2H), 7.09 - 6.97 (m, 1H), 6.96 - 6.40 (m, 3H), 5.42 - 5.20 (m, 1H), 4.65 - 4.36 (m, 2H), 4.29 - 4.05 (m, 2H), 4.00 - 3.76 (m, 5H), 3.74 - 3.54 (m, 2H), 3.19 - 2.83 (m, 3H), 2.73 - 2.42 (m, 4H), 2.15 - 1.93 (m, 1H), 1.90 - 1.74 (m, 1H), 0.84 - 0.66 (m, 2H), 0.66 - 0.39 (m, 2H).
  • Example 25 and 26 (8S,11S,18S)-10-[1-(2-ethoxy-4-fluoro-phenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21- 2,6 8,11 13,18 23,27 methyl-17-oxa-7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one (Example 25) and (8S,11S,18R)-10-[1-(2- ethoxy-4-fluoro-phenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]
  • Example 26 slower eluted, LCMS (M+H + ): 707.
  • Example 27 and 28 (8S,11S,18S)-25-fluoro-10-[1-(4-fluoro-2-methoxy-phenyl)pyrazolo[3,4-d]pyrimidin-4-yl]- 2,6 8,11 13,18 23,27 21-methyl-17-oxa-7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one (Example 27) and (8S,11S,18R)-25-fluoro-10- [1-(4-fluoro-2-methoxy-phenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0
  • Example 28 slower eluted, LCMS (M+H + ): 693.
  • 1 H NMR (500 MHz, METHANOL-d4) ⁇ 8.56 - 8.40 (m, 1H), 8.24 - 8.11 (m, 1H), 7.56 - 7.37 (m, 2H), 7.35 - 7.22 (m, 1H), 7.12 - 6.81 (m, 4H), 6.64 - 6.50 (m, 2H), 5.41 - 5.24 (m, 1H), 4.65 - 4.41 (m, 3H), 4.28 - 4.08 (m, 2H), 4.04 - 3.81 (m, 5H), 3.71 - 3.59 (m, 2H), 3.16 - 2.90 (m, 3H), 2.66 - 2.42 (m, 5H), 2.16 - 1.98 (m, 1H), 1.92 - 1.78 (m, 1H).
  • Example 29 and 30 (8S,11S,18S)-25-fluoro-10-[1-(4-fluoro-2-methoxy-phenyl)-6-methyl-pyrazolo[3,4- d]pyrimidin-4-yl]-21-methyl-17-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(26),2(30),3,5,21,23(27),24-heptaen- 12-one (Example 29) and (8S,11S,18R) -25-fluoro-10-[1-(4-fluoro-2-methoxy-phenyl)-6- methyl-pyrazolo[3,4-d]pyrimidin-4-yl]-21-methyl-17-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.
  • Example 30 slower eluted, LCMS (M+H + ): 707.
  • 1 H NMR (500 MHz, METHANOL-d 4 ) ⁇ 8.46 - 8.30 (m, 1H), 7.52 - 7.35 (m, 2H), 7.33 - 7.25 (m, 1H), 7.09 - 6.99 (m, 2H), 6.92 - 6.52 (m, 3H), 5.37 - 5.15 (m, 1H), 4.63 - 4.53 (m, 1H), 4.52 - 4.33 (m, 2H), 4.25 - 4.03 (m, 2H), 4.03 - 3.80 (m, 4H), 3.79 - 3.59 (m, 5H), 3.13 - 2.88 (m, 2H), 2.62 - 2.33 (m, 7H), 2.12 - 1.97 (m, 1H), 1.92 - 1.79 (m, 1H).
  • Example 31 and 32 (8S,11S,18R)-25-fluoro-10-[1-(4-fluoro-2-methoxy-phenyl)pyrazolo[3,4-d]pyrimidin-4-yl]- 2,6 8,11 13,18 23,27 21-methyl-17-oxa-5,7,10,13,20,22,30-heptazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one (Example 31) and (8S,11S,18S)-25-fluoro-10-[1- (4-fluoro-2-methoxy-phenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 5,7,10,13,20,22,30-heptazahexacyclo[18.6.1.1 .1 .1
  • Example 31 and 32 were obtained via SFC separation.
  • Example 32 slower eluted, LCMS (M+H + ): 694.
  • 1 H NMR (500 MHz, METHANOL-d4) ⁇ 8.58 - 8.26 (m, 2H), 8.24 - 7.77 (m, 1H), 7.54 - 7.33 (m, 2H), 7.30 - 7.19 (m, 1H), 7.13 - 7.03 (m, 1H), 7.03 - 6.97 (m, 1H), 6.96 - 6.85 (m, 1H), 5.92 - 5.73 (m, 1H), 5.64 - 5.44 (m, 1H), 4.83 - 4.66 (m, 1H), 4.58 - 4.49 (m, 1H), 4.42 - 4.11 (m, 4H), 4.10 - 4.01 (m, 1H), 3.91 - 3.71 (m, 4H), 3.53 - 3.37 (m, 3H), 3.21 - 3.11 (m, 1H), 3.10 - 2.90 (m, 1H), 2.76 - 2.50 (m
  • Example 37 and 38 (8S,11S,18R)-10-[1-(2,4-difluorophenyl)-6-(methoxymethyl)pyrazolo[3,4-d]pyrimidin-4-yl]- 25-fluoro-21-methyl-17-oxa-5,7,10,13,20,22,30- 2,6 8,11 13,18 23,27 heptazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(26),2(30),3,5,21,23(27),24-heptaen- 12-one (Example 37) and (8S,11S,18S)-10-[1-(2,4-difluorophenyl)-6- (methoxymethyl)pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 5,7,10,13,20,22,30-heptazahexacyclo[18
  • Example 37 faster eluted, LCMS (M+H + ): 726.
  • 1 H NMR (500 MHz, METHANOL-d4) ⁇ 8.56 - 7.82 (m, 1H), 7.70 - 7.53 (m, 2H), 7.46 - 7.36 (m, 1H), 7.34 - 7.26 (m, 1H), 7.24 - 7.16 (m, 1H), 7.03 - 6.85 (m, 1H), 6.76 - 6.56 (m, 1H), 5.41 - 5.19 (m, 1H), 4.64 - 4.33 (m, 5H), 4.30 - 3.81 (m, 5H), 3.79 - 3.57 (m, 1H), 3.56 - 3.44 (m, 2H), 3.43 - 3.36 (m, 2H), 3.24 - 2.97 (m, 3H), 2.91 - 2.77 (m, 3H), 2.69 - 2.45 (m, 1H), 2.20 - 2.07 (m, 1H), 2.03 - 1.85 (m
  • Example 38 slower eluted, LCMS (M+H + ): 726.
  • Example 39 (8S,11S)-10-[1-(2,4-difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-16- 2,6 8,11 13,17 23,27 oxa-7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ] triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one
  • Example 39 was prepared in analogy to the preparation of Example 21 by using intermediate A9 instead of intermediate A6 in step 1 and using intermediate C2 instead of intermediate C19 in step 5.
  • Example 40 and 41 (8S,11S,18R)-10-[1-(2,4-difluoro-6-methoxy-phenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-25- fluoro-21-methyl-17-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(26),2(30),3,5,21,23(27),24-heptaen- 12-one (Example 40) and (8S,11S,18S)-10-[1-(2,4-difluoro-6-methoxy-phenyl)pyrazolo[3,4- d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1
  • Example 40 faster eluted, LCMS (M+H + ): 711.
  • 1 H NMR (500 MHz, METHANOL-d4) ⁇ 8.63 - 8.41 (m, 1H), 8.23 - 8.09 (m, 1H), 7.64 - 7.48 (m, 2H), 7.43 - 7.30 (m, 1H), 7.04 - 6.52 (m, 4H), 5.40 - 5.24 (m, 1H), 4.63 - 4.16 (m, 4H), 4.12 - 3.87 (m, 4H), 3.85 - 3.74 (m, 3H), 3.71 - 3.57 (m, 2H), 3.22 - 2.96 (m, 3H), 2.89 - 2.77 (m, 3H), 2.72 - 2.45 (m, 1H), 2.17 - 2.03 (m, 1H), 1.92 - 1.78 (m, 1H).
  • Example 41 slower eluted, LCMS (M+H + ): 711.
  • 1 H NMR (500 MHz, METHANOL-d4) ⁇ 8.63 - 8.54 (m, 1H), 8.25 - 8.17 (m, 1H), 7.90 - 7.82 (m, 1H), 7.59 - 7.48 (m, 2H), 7.11 - 7.03 (m, 1H), 7.00 - 6.94 (m, 1H), 6.91 - 6.84 (m, 1H), 6.83 - 6.78 (m, 1H), 6.45 - 6.27 (m, 1H), 5.66 - 5.48 (m, 1H), 4.80 - 4.75 (m, 1H), 4.67 - 4.58 (m, 1H), 4.38 - 4.29 (m, 3H), 4.08 - 3.97 (m, 2H), 3.85 - 3.74 (m, 4H), 3.56 - 3.47 (m, 2H), 3.21 - 3.09 (m, 1H), 2.97 - 2.78 (m,
  • Example 42 and 43 (8S,11S,18R)-10-[1-(2,4-difluorophenyl)-6-[3-methyl-3,6-diazabicyclo[3.1.1]heptan-6- yl]pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(26),2(30),3,5,21,23(27),24-heptaen- 12-one (Example 42) and (8S,11S,18S)-10-[1-(2,4-difluorophenyl)-6-[3-methyl-3,6- diazabicyclo[3.1.1]heptan-6-yl]pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl
  • Example 44 (8S,11S,18S)-10-[1-(2,4-difluorophenyl)-6-[(1R,4R)-5-methyl-2,5-diazabicyclo[2.2.1]heptan- 2-yl]pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(26),2(30),3,5,21,23(27),24-heptaen- 12-one
  • Example 44 was prepared in analogy to the preparation of Example 43 by using (1R,4R)-2- methyl-2,5-diazabicyclo[2.2.1]heptane (dihydrochloride salt) instead of 3-methyl-3,6- diazabicyclo[3.1.1]heptane (di-2,2,2-tri
  • Example 45 (8S,11S)-10-[1-(2,4-difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-22-fluoro-13,18- 2,6 8,11 20,24 dimethyl-spiro[7,10,13,17,19,26-hexazapentacyclo[15.6.1.1 .1 .0 ]hexacosa- 1(23),2(26),3,5,18,20(24),21-heptaene-15,1'-cyclopropane]-12-one
  • the title compound was prepared according to the following scheme:
  • Step 1 preparation of [1-[(2-bromo-4-fluoro-6-nitro- anilino)methyl]cyclopropyl]methanol (compound 45a)
  • DMF 60 mL
  • DIEA 8.02 g, 75.63 mmol
  • the reaction mixture was stirred at room temperature for 1 hr.
  • the reaction mixture was poured into water and extracted with EtOAc for twice.
  • the combined organic layer was washed with brine, dried and concentrated to give some residue.
  • Step 2 preparation of [1-[(2-amino-6-bromo-4-fluoro- anilino)methyl]cyclopropyl]methanol (compound 45b) To a solution of [1-[(2-bromo-4-fluoro-6-nitro-anilino)methyl]cyclopropyl]methanol (compound 45a, 7.0 g, 21.93 mmol) in ethyl acetate (100 mL) was added Pt/C (1.0 g), the reaction mixture was stirred at room temperature for 1 hr under hydrogen.
  • Step 4 preparation of 7-bromo-1-[[1-(chloromethyl)cyclopropyl]methyl]-5-fluoro-2- methyl-benzimidazole (compound 45d)
  • a solution of [1-[(7-bromo-5-fluoro-2-methyl-benzimidazol-1- yl)methyl]cyclopropyl]methanol (compound 45c, 4.0 g, 12.77 mmol) in SOCl2 (20.0 mL) was stirred at room temperature for 1 hr.
  • the reaction mixture was concentrated under reduced pressure to give compound 45d (4.2 g) as a white solid, which was used in next step without further purification.
  • Step 5 preparation of tert-butyl N-[[1-[(7-bromo-5-fluoro-2-methyl-benzimidazol-1- yl)methyl]cyclopropyl]methyl]-N-methyl-carbamate (compound 45e)
  • tert-butyl N-methylcarbamate 3.32 g, 25.33 mmol
  • 7-bromo-1-[[1- (chloromethyl)cyclopropyl]methyl]-5-fluoro-2-methyl-benzimidazole compound 45d, 4.2 g, 12.67 mmol
  • DMF 40 mL
  • Step 6 preparation of tert-butyl N-[[1-[[7-(6-chloro-2-pyridyl)-5-fluoro-2-methyl- benzimidazol-1-yl]methyl]cyclopropyl]methyl]-N-methyl-carbamate (compound 45f)
  • compound 45f To a mixture of tert-butyl N-[[1-[(7-bromo-5-fluoro-2-methyl-benzimidazol-1- yl)methyl]cyclopropyl]methyl]-N-methyl-carbamate (compound 45e, 600.0 mg, 1.41 mmol), 6- chloropyridine-2-boronic acid pinacol ester (337 mg, 1.41 mmol) and potassium phosphate (500 mg, 4.22 mmol) in 1,4-dioxane (6 mL) and water (1 mL) was added Pd-Ad 2 nBuP (93.9 mg, 0.14 mmol).
  • Step 7 preparation of O1-benzyl O2-methyl (2S,4S)-4-[[6-[3-[[1-[[tert- butoxycarbonyl(methyl)amino]methyl]cyclopropyl]methyl]-6-fluoro-2-methyl- benzimidazol-4-yl]-2-pyridyl]amino]pyrrolidine-1,2-dicarboxylate (compound 45g)
  • Step 8 preparation of (2S,4S)-1-benzyloxycarbonyl-4-[[6-[3-[[1-[[tert- butoxycarbonyl(methyl)amino]methyl]cyclopropyl]methyl]-6-fluoro-2-methyl- benzimidazol-4-yl]-2-pyridyl]amino]pyrrolidine-2-carboxylic acid (compound 45h)
  • Step 9 preparation of (2S,4S)-1-benzyloxycarbonyl-4-[[6-[6-fluoro-2-methyl-3-[[1- (methylaminomethyl)cyclopropyl]methyl]benzimidazol-4-yl]-2-pyridyl]amino]pyrrolidine- 2-carboxylic acid (compound 45i)
  • Step 11 preparation of (8S,11S)-22-fluoro-13,18-dimethyl-spiro[7,10,13,17,19,26- 2,6 8,11 20,24 hexazapentacyclo[15.6.1.1 .1 .0 ]hexacosa-1(23),2(26),3,5,18,20(24),21-heptaene-15,1'- cyclopropane]-12-one (compound 45k) A mixture of (8S,11S)-22-fluoro-13,18-dimethyl-12-oxo-spiro[7,10,13,17,19,26- 2,6 8,11 20,24 hexazapentacyclo[15.6.1.1 .1 .0 ]hexacosa-1(23),2(26),3,5,18,20(24),21-heptaene-15,1'- cyclopropane]-10-carboxylate (compound 45j, 5.0 mg, 0.01 mmol) and Pd
  • Step 12 preparation of (8S,11S)-10-[1-(2,4-difluorophenyl)pyrazolo[3,4-d]pyrimidin- 4-yl]-22-fluoro-13,18-dimethyl-spiro[7,10,13,17,19,26- 2,6 8,11 20,24 hexazapentacyclo[15.6.1.1 .1 .0 ]hexacosa-1(23),2(26),3,5,18,20(24),21-heptaene-15,1'- cyclopropane]-12-one (Example 45) A mixture of (8S,11S)-22-fluoro-13,18-dimethyl-spiro[7,10,13,17,19,26- 2,6 8,11 20,24
  • Example 45 (1.1 mg) as a white solid.
  • Example 46 (8S,11S,18S)-25-fluoro-10-[1-[4-fluoro-2-(trideuteriomethoxy)phenyl]-6-(3-oxa-6- azabicyclo[3.1.1]heptan-6-yl)pyrazolo[3,4-d]pyrimidin-4-yl]-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one
  • Step 1 preparation of tert-butyl (8S,11S,18S)-25-fluor
  • Example 47 (8S,11S,18S)-10-[1-(2,4-difluorophenyl)-6-(3-oxa-6-azabicyclo[3.1.1]heptan-6- yl)pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(26),2(30),3,5,21,23(27),24-heptaen- 12-one
  • Example 47 was prepared in analogy to the preparation of Example 43 by using 3-oxa-6- azabicyclo[3.1.1] heptane instead of 3-methyl-3,6-diazabicyclo[3.1.1]heptane (di-2,2,2- trifluoroacetic acid salt).
  • Example 48 and 49 (13R,18S,21S)-19-[1-(2,4-difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-16-methyl-11-oxa- 6,9 18,21 2,7 8,13 8,16,19,22,27,29-hexazahexacyclo[21.3.1.1 .1 .0 .0 ]nonacosa- 1(26),2(7),3,5,9(29),23(27),24-heptaen-17-one and (13S,18S,21S)-19-[1-(2,4- difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-16-methyl-11-oxa-8,16,19,22,27,29- 6,9 18,21 2,7 8,13 hexazahexacyclo[21.3.1.1 .1 .0 .0 ]nonacosa-1(26),2(7),3,5,9(29),23
  • Example 48 & 49 Step 1 preparation of tert-butyl N-[3-(2-bromo-6-nitro-anilino)propyl]-N-methyl- carbamate (compound 48b)
  • a mixture of 3-bromo-2-fluoronitrobenzene (5.0 g, 22.73 mmol), compound 48a (5.5 g, 29.21 mmol) and triethylamine (3.0 g, 29.65 mmol) in THF (100 mL) was stirred for 12 hrs at 20 °C.
  • Step 2 preparation of tert-butyl N-[3-(2-amino-6-bromo-anilino)-4-hydroxy-butyl]-N- methyl-carbamate(compound 48c)
  • Raney Ni 2.25 g, 38.25 mmol
  • hydrazine hydrate 3.2 g, 63.92 mmol
  • Step 4 preparation of tert-butyl N-[3-[7-bromo-2-(hydroxymethyl)benzimidazol-1- yl]-4-hydroxy-butyl]-N-methyl-carbamate (compound 48e)
  • compound 48d 3.0 g, 5.53 mmol
  • methanol 30 mL
  • potassium fluoride 1.61 g, 27.65 mmol
  • the reaction mixture was concentrated under reduced pressure and the residue was purified by prep-HPLC to give compound 48e (1.7 g) as a brown oil.
  • LCMS M+2+H + ): 430.
  • Step 5 preparation of tert-butyl N-[2-(6-bromo-3,4-dihydro-1H-[1,4]oxazino[4,3- a]benzimidazol-4-yl)ethyl]-N-methyl-carbamate(compound 48f)
  • compound 48e 400 mg, 0.79 mmol
  • tributylphosphane 482 mg, 2.38 mmol
  • toluene 40 mL
  • Step 7 preparation of N-methyl-2-[6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)- 3,4-dihydro-1H-[1,4]oxazino[4,3-a]benzimidazol-4-yl]ethanamine(compound 48h)
  • DCM dimethyl-2-[6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)- 3,4-dihydro-1H-[1,4]oxazino[4,3-a]benzimidazol-4-yl]ethanamine(compound 48h)
  • DCM 10 mL
  • HCl/dioxane 4 mL
  • the reaction mixture was concentrated to give compound 48h (300 mg) as a brown solid.
  • Step 8 preparation of O1-tert-butyl O2-methyl (2S,4S)-4-[[6-[4-[2- (methylamino)ethyl]-3,4-dihydro-1H-[1,4]oxazino[4,3-a]benzimidazol-6-yl]-2- pyridyl]amino]pyrrolidine-1,2-dicarboxylate (compound 48i)
  • a mixture of compound 48h 300 mg, 0.84 mmol
  • O1-tert-butyl O2-methyl (2S,4S)-4- [(6-bromo-2-pyridyl)amino] pyrrolidine-1,2-dicarboxylate (336 mg, 0.84 mmol), 1,1'- bis(diphenylphosphino)ferrocene-palladium(II)dichloride dichloromethane complex (137 mg, 0.17 mmol) and potassium carbonate (232 mg, 1.68 mmol) in 1,4-
  • Step 10 preparation of tert-butyl (13R,18S,21S)-16-methyl-17-oxo-11-oxa- 6,9 18,21 2,7 8,13 8,16,19,22,27,29-hexazahexacyclo[21.3.1.1 .1 .0 .0 ]nonacosa- 1(26),2(7),3,5,9(29),23(27),24-heptaene-19-carboxylate(compound 48k-1) and tert-butyl (13S,18S,21S)-16-methyl-17-oxo-11-oxa-8,16,19,22,27,29- 6,9 18,21 2,7 8,13 hexazahexacyclo[21.3.1.1 .1 .0 .0 ]nonacosa-1
  • Step 11 preparation of (13R,18S,21S)-16-methyl-11-oxa-8,16,19,22,27,29- 6,9 18,21 2,7 8,13 hexazahexacyclo [21.3.1.1 .1 .0 .0 ]nonacosa-1(26),2(7),3,5,9(29),23(27),24-heptaen- 17-one (compound 48l-1) and (13S,18S,21S)-16-methyl-11-oxa-8,16,19,22,27,29- 6,9 18,21 2,7 8,13 hexazahexacyclo[21.3.1.1 .1 .0 .0 ]nonacosa-1(26),2(7),3,5,9(29),23(27),24-heptaen- 17-one (compound 48l-1) and (13S,18S,21S)-16-methyl-11-oxa-8,16,19,22,27,29- 6,9 18,21 2,7 8,13 hexaza
  • Step 12 preparation of (13R,18S,21S)-19-[1-(2,4-difluorophenyl)pyrazolo[3,4- d]pyrimidin-4-yl]-16-methyl-11-oxa-8,16,19,22,27,29- 6,9 18,21 2,7 8,13 hexazahexacyclo[21.3.1.1 .1 .0 .0 ]nonacosa-1(26),2(7),3,5,9(29),23(27),24-heptaen- 17-one (Example 48) and (13S,18S,21S)-19-[1-(2,4-difluorophenyl)pyrazolo[3,4- d]pyrimidin-4-yl]-16-methyl-11-oxa-8,16,19,22,27,29- 6,9 18,21 2,7 8,13 hexazahexacyclo[21.3.1.1 .1 .0 .0 ]nonacosa-1(26),
  • Step 1 preparation of benzyl N-(3-hydroxypropyl)-N-methyl-carbamate (compound 50b)
  • Step 2 preparation of benzyl N-methyl-N-(3-oxopropyl)carbamate (compound 50c) To a solution of compound 50b (2.0 g, 8.96 mmol) in DCM (20 mL) was added DMP (4.56 g, 10.75 mmol) at 0 °C, then the mixture was stirred at 20 °C for 2 hrs.
  • Step 3 preparation of benzyl N-[2-(10-bromo-3-oxo-2,4-dihydro-1H-pyrazino[1,2- b]indazol-1-yl)ethyl]-N-methyl-carbamate (compound 50d)
  • 2-methyl- 2-propanesulfinamide 822 mg, 6.78 mmol
  • MgSO 4 814 mg, 6.78 mmol
  • pyridinium tosylate 1.7 g, 6.78 mmol.
  • Step 4 preparation of 2-[(4-bromoindazol-2-yl) methoxy] ethyl-trimethyl-silane (compound 50f)
  • compound 50e 3-(chloromethoxy)ethyl)trimethylsilane (3.05 g, 18.27 mmol)
  • dicyclohexylmethylamine 3.6 g, 18.27 mmol
  • Step 6 preparation of benzyl N-[3-amino-3-(4-bromo-2H-indazol-3-yl)propyl]-N- methyl-carbamate (compound 50h)
  • a solution of compound 50g (2.0 g, 3.07 mmol) in methanol (10 mL) was added aq.HCl (10.0 mL, 60.0 mmol). After being stirred at 70 °C for 2 h, the mixture was concentrated to give compound 50h (1.3 g) as light yellow semisolid.
  • Step 7 preparation of benzyl N-[3-(4-bromo-2H-indazol-3-yl)-3-[(2- chloroacetyl)amino]propyl]-N-methyl-carbamate (compound 50i)
  • compound 50h 1.3 g, 3.12 mmol
  • NaHCO3 524 mg, 6.23 mmol
  • chloroacetyl chloride 352 mg, 3.12 mmol
  • DCM 20 mL
  • Step 8 preparation of benzyl N-[3-amino-3-(4-bromo-2H-indazol-3-yl)propyl]-N- methyl-carbamate (compound 50j) To a suspension of cesium carbonate (198 mg, 0.61 mmol) in ACN (50 mL) was added compound 50i (300 mg, 0.61 mmol) in ACN (100 mL) slowly at 80 °C.
  • Step 9 preparation of benzyl N-methyl-N-[2-[3-oxo-10-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)-2,4-dihydro-1H-pyrazino[1,2-b]indazol-1-yl]ethyl]carbamate (compound 50k)
  • compound 50j 200 mg, 0.44 mmol
  • potassium acetate 86 mg, 0.87 mmol
  • Pd(dppf)Cl2 18 mg, 0.02 mmol
  • bis(pinacolato)diboron 167 mg, 0.66 mmol
  • Example 51 (8S,11S)-10-[1-(2,4-difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-7,10,13,16,22,24,31- 2,6 8,11 13,18 25,29 heptazahexacyclo[20.6.1.1 .1 .0 .0 ]hentriaconta-1(29),2,4,6(31),23,25,27-heptaen- 12-one
  • the title compound was prepared according to the following scheme:
  • Step 1 preparation of O4-benzyl O1-tert-butyl O2-methyl piperazine-1,2,4- tricarboxylate (compound 51b)
  • compound 51a 10 g, 40.93 mmol
  • triethylamine 8 mL, 61.4 mmol
  • benzyl carbonochloridate 7 g, 45.03 mmol
  • the mixture was stirred at 25 °C for 3 hs, and then the reaction was quenched with water (100 mL). After separation, the organic layer was washed with 5% citric acid, sat. NaHCO3 and brine.
  • Step 2 preparation of O4-benzyl O1-tert-butyl 2-(hydroxymethyl)piperazine-1,4- dicarboxylate (compound 51c) To a solution of compound 51b (22 g, 58.14 mmol) and CaCl 2 (9 g, 81.39 mmol) in Ethanol (200 mL) and THF (200 mL) was added sodium borohydride (4 g, 116.27 mmol) at 0 °C and stirred at 20 °C for 16 hrs.
  • Step 3 preparation of O4-benzyl O1-tert-butyl 2-formylpiperazine-1,4-dicarboxylate (compound 51d)
  • DCM 120 mL
  • Dess- Martin Peridinane 29 g, 68.49 mmol
  • the reaction mixture was poured into a solution (saturated sodium dicarbonate solution 120 mL, and saturated sodium hydrogen sulfite solution 120 mL) and extrated with DCM (20ml ⁇ 3), the organic layer was dried over Na2SO4, filtered and concentrated.
  • Step 5 preparation of O4-benzyl O1-tert-butyl 2-(3-ethoxy-3-oxo-propyl)piperazine- 1,4-dicarboxylate (compound 51f)
  • a mixture of compound 51e (2 g, 5.02 mmol) and PtO2 (200 mg, 5.02 mmol) in anhydrous THF (20 mL) was stirred for 2 hrs at 20 °C under hydrogen balloon. The mixture was filtered, and concentrated to give compound 51f (2 g), which was used in next step directly.
  • Step 8 preparation of O4-benzyl O1-tert-butyl 2-(3-aminopropyl)piperazine-1,4- dicarboxylate (compound 51i)
  • a mixture of compound 51h (3 g, 5.71 mmol) and monomethylamine in water (5.0 mL, 5.71 mmol) in ethanol (10 mL) was stirred for 12 hrs at 70 °C.
  • the mixture was diluted with DCM (100 mL), washed with water and brine, the organic layer was dried and concentrated to give compound 51i (2 g).
  • Step 9 ⁇ 12 preparation of O4-benzyl O1-tert-butyl 2-[3-[7-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)benzimidazol-1-yl]propyl]piperazine-1,4-dicarboxylate (compound 51m)
  • Compound 51m was prepared in analogy to the reparation of intermediate A1 by using compound 51i instead of compound tert-butyl 6-(aminomethyl)-1,4-oxazepane-4-carboxylate.
  • Step 13 ⁇ 17 preparation of benzyl (8S,11S)-10-[1-(2,4-difluorophenyl)pyrazolo[4,3- c]pyridin-4-yl]-12-oxo-7,10,13,16,22,24,31- 2,6 8,11 13,18 25,29 heptazahexacyclo[20.6.1.1 .1 .0 .0 ]hentriaconta-1(29),2,4,6(31),23,25,27-heptaene- 16-carboxylate (compound 51s)
  • Compound 51s was prepared in analogy to the reparation of Example 1 by using compound 51m instead of intermediate A1.
  • Step 18 preparation of (8S,11S)-10-[1-(2,4-difluorophenyl)pyrazolo[3,4-d]pyrimidin- 2,6 8,11 13,18 25,29 4-yl]-7,10,13,16,22,24,31-heptazahexacyclo[20.6.1.1 .1 .0 .0 ]hentriaconta- 1(29),2,4,6(31),23,25,27-heptaen-12-one (Example 51) A mixture of compound 51s (20 mg, 0.03 mmol) and trifluoroacetic acid (2 mL, 25.96 mmol) was stirred for 1 h at 70 °C.
  • Example 52 (8S,11S,18S)-25-fluoro-10-[1-[4-fluoro-2-(trideuteriomethoxy)phenyl]-6-[[1-(2,2,2- trifluoroethyl)azetidin-3-yl]methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one
  • Example 52 was prepared according to the following scheme: Step 1: preparation of tert-butyl 3-[[4-[(8S,11S,18S)-25-fluoro-21-methyl-12-oxo-17- oxa-7,10,13,20,22,30-hexazahexacyclo
  • Step 3 preparation of (8S,11S,18S)-25-fluoro-10-[1-[4-fluoro-2-(trideuteriomethoxy) phenyl]-6-[[1-(2,2,2-trifluoroethyl)azetidin-3-yl]methyl]pyrazolo[3,4-d]pyrimidin-4-yl]-21- methyl-17-oxa-7,10,13,20,22,30-hexazahexacyclo [18.6.1.1 2,6 .1 8,11 .1 13,18 .0 23,27 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one (Example 52) To a solution of compound 52b (20 mg
  • Example 52 (6.8 mg) as a white solid.
  • Example 53 (8S,11S,18S)-10-[1-(2,4-difluorophenyl)-6-[3-(2-fluoroethyl)-3,6-diazabicyclo[3.1.1]heptan- 6-yl]pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(26),2(30),3,5,21,23(27),24-heptaen- 12-one
  • Example 53 was prepared in analogy to the preparation of Example 58 by using intermediate C80 instead of compound 58f.
  • Example 54 (8S,11S,18S)-10-[1-(2,4-difluorophenyl)-6-[(1R,4R)-5-(2-fluoroethyl)-2,5- diazabicyclo[2.2.1]heptan-2-yl]pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one
  • Example 54 was prepared in analogy to the preparation of Example 58 by using intermediate C83 instead of compound 58f.
  • Example 55 (8S,11S,18S)-10-[1-(2,4-difluorophenyl)-6-[3-(oxetan-3-yl)-3,6-diazabicyclo[3.1.1]heptan-6- yl]pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa-7,10,13,20,22,30- 2,6 8,11 13,18 23,27 hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta-1(27),2,4,6(30),21,23,25-heptaen-12- one
  • Example 55 was prepared in analogy to the preparation of Example 58 by using intermediate C89 instead of compound 58f.
  • Example 56 (8S,11S,18S)-10-[6-[3-(2,2-difluoroethyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl]-1-(2,4- difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(27),2,4,6(30),21,23,25-heptaen-12-one
  • Example 56 was prepared in analogy to the preparation of Example 58 by using intermediate C81 instead of compound 58f.
  • Example 57 (8S,11S,18S)-10-[1-(2,4-difluorophenyl)-6-[(1R,4R)-5-(oxetan-3-yl)-2,5- diazabicyclo[2.2.1]heptan-2-yl]pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one
  • Example 57 was prepared in analogy to the preparation of Example 58 by using intermediate C88 instead of compound 58f.
  • Example 58 1-[1-(2,4-difluorophenyl)-4-[(8S,11S,18S)-25-fluoro-21-methyl-12-oxo-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-10-yl]pyrazolo[3,4-d]pyrimidin-6-yl]azetidine-3- carbonitrile
  • the title compound was prepared according to the following scheme: Step 1: preparation of methyl 1-[4-benzyloxy-1-(2,4-difluorophenyl)pyrazolo[3,4- d]pyrimidin-6-yl]azetidine-3-carboxylate (compound 58b) A mixture of compound C80-a (400 mg, 1.07 mmol),
  • Step 2 preparation of 1-[4-benzyloxy-1-(2,4-difluorophenyl)pyrazolo[3,4- d]pyrimidin-6-yl]azetidine-3-carboxylic acid (compound 58c)
  • a mixture of compound 58b (400 mg, 0.88 mmol) and lithium hydroxide (2.22 mL, 2 M) in methanol (8 mL) was stirred at rt for 4 hours, then the pH was adjusted to 6, and the mixture was diluted with water.
  • Step 3 preparation of 1-[4-benzyloxy-1-(2,4-difluorophenyl)pyrazolo[3,4- d]pyrimidin-6-yl]azetidine-3-carboxamide (compound 58d)
  • a mixture of compound 58c (380 mg, 0.61 mmol), ammonium chloride (325 mg, 6.08 mmol), DIPEA (786 mg, 6.08 mmol ) and HATU (462 mg, 1.22 mmol) in tetrahydrofuran (10 mL) was stirred at 40 o C for 2 hours, then the reaction mixture was concentrated and the residue was purified by silica gel column chromatography to give compound 58d (300 mg).
  • Step 6 preparation of 1-[1-(2,4-difluorophenyl)-4-[(8S,11S,18S)-25-fluoro-21-methyl- 2,6 8,11 13,18 23,27 12-oxo-17-oxa-7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-10-yl]pyrazolo[3,4-d]pyrimidin-6-yl]azetidine-3- carbonitrile (Example 58) A mixture of compound 58f (35 mg, 0.1 mmol), PyBOP (70 mg, 0.13 mmol) and DIPEA (57 mg, 0.44 mmol) in DMF (2 mL) was stirred at 50 o C for 3 hours.
  • Example 59 (8S,11S,18S)-10-[6-[3-(cyclopropanecarbonyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl]-1-(2,4- difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one
  • Step 1 preparation of tert-butyl 6-[1-(2,4-difluorophenyl)-4-[(8S,11S,18S)-25-fluoro- 21-methyl-12-oxo-17-oxa-7,10,13,20,22,
  • Example 59 (18 mg) as white powder, LCMS (M+H + ): 845.
  • 1 H NMR (400 MHz, METHANOL-d 4 ) ⁇ 8.29 - 8.05 (m, 1H), 7.73 - 7.60 (m, 1H), 7.56 - 7.42 (m, 1H), 7.21 - 6.98 (m, 4H), 6.92 - 6.78 (m, 1H), 6.64 - 6.48 (m, 1H), 5.99 - 5.79 (m, 1H), 5.39 - 5.32 (m, 1H), 4.61 - 4.33 (m, 8H), 4.32 - 4.02 (m, 5H), 4.02 - 3.78 (m, 2H), 3.78 - 3.57 (m, 2H), 3.39 - 3.30 (m, 2H), 3.15 - 2.83 (m, 2H), 2.67 - 2.33 (m, 6H), 1.79 - 1.63 (m, 1H
  • Example 60 (8S,11S,18S)-10-[1-(2,4-difluorophenyl)-6-[3-(1-hydroxycyclobutanecarbonyl)-3,6- diazabicyclo[3.1.1]heptan-6-yl]pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one
  • the title compound was prepared according to the following scheme: A mixture of compound 59b (30 mg, 0.039 mmol ), 1-hydroxycyclobutanecarboxylic acid (22 mg, 0.19 mmol), DIPEA (50 mg, 0.39 mmol) and HATU (29 mg, 0.078 m
  • Example 61 (8S,11S,18S)-10-[6-(3-cyclopropylsulfonyl-3,6-diazabicyclo[3.1.1]heptan-6-yl)-1-(2,4- difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one
  • Example 61 was prepared in analogy to the preparation of Example 59 by using cyclopropanesulfonyl chloride instead of cyclopropanecarbonyl chloride.
  • Example 62 (8S,11S,18S)-10-[6-[3-(2,2-difluoroacetyl)-3,6-diazabicyclo[3.1.1]heptan-6-yl]-1-(2,4- difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one
  • Example 62 was prepared in analogy to the preparation of Example 59 by using (2,2- difluoroacetyl) 2,2-difluoroacetate instead of cyclopropanecarbonyl chloride.
  • Examples 63 and 64 (8S,11S,18S)-10-[1-(2,4-difluorophenyl)-6-[ (1R,5S)-3-methyl-2-oxo-3,6- diazabicyclo[3.1.1]heptan-6-yl]pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(27),2,4,6(30),21,23,25-heptaen-12-one and (8S,11S,18S)-10-[1-(2,4-difluorophenyl)-6- [(1S,5R)-3-methyl-2-oxo-3,6-diazabicyclo[3.1.1]heptan-6-yl]pyrazolo[3,4-d]pyr
  • Examples 65 and 66 (8S,11S,18S)-10-[6-[(1R,5S)-3-(cyclopropylmethyl)-2-oxo-3,6-diazabicyclo[3.1.1]heptan-6- yl]-1-(2,4-difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(27),2,4,6(30),21,23,25-heptaen-12-one and (8S,11S,18S)-10-[6-[(1S,5R)-3- (cyclopropylmethyl)-2-oxo-3,6-diazabicyclo[3.1.1]heptan-6-yl]-1-(2,4- difluor
  • Example 66 (slower eluted), 16 mg, white powder.LCMS (M+H + ): 845.
  • Step 3 preparation of benzyl (1R,5S)-3-(oxetan-3-yl)-2-oxo-3,6- diazabicyclo[3.1.1]heptane-6-carboxylate (compound 67d)
  • DMA diazabicyclo[3.1.1]heptane-6-carboxylate
  • Step 4 preparation of (1S,5R)-3-(oxetan-3-yl)-3,6-diazabicyclo[3.1.1]heptan-2-one (compound 67e) To a solution of compound 67d (160 mg, 0.53 mmol) in methanol (1 mL) was added wet Pd/C (40 mg) under N 2 at 25 °C. The mixture was stirred under H 2 (balloon) at 50 °C for 2 h.
  • Step 5 preparation of (8S,11S,18S)-10-[1-(2,4-difluorophenyl)-6-[(1S,5R)-3-(oxetan-3- yl)-2-oxo-3,6-diazabicyclo[3.1.1]heptan-6-yl]pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21- methyl-17-oxa-7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 2,6 .1 8,1 1.1 13,18 .0 23,27 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one(Example 67) A mixture of compound 42a-2 (40.0 mg, 0.06 mmol), compound 67
  • Example 68 (8S,11S,18S)-10-[6-[(1S,5R)-3-cyclopropyl-2-oxo-3,6-diazabicyclo[3.1.1]heptan-6-yl]-1-(2,4- difluorophenyl)pyrazolo[3,4-d]pyrimidin-4-yl]-25-fluoro-21-methyl-17-oxa- 2,6 8,11 13,18 23,27 7,10,13,20,22,30-hexazahexacyclo[18.6.1.1 .1 .1 .0 ]triaconta- 1(26),2(30),3,5,21,23(27),24-heptaen-12-one
  • Step 1 preparation of tert-butyl (1S,5R)-3-cyclopropyl-2-oxo-3,6-diazabicyclo [3.1.1]heptane-6-carboxylate (compound 68a)
  • Step 1 preparation of tert-buty
  • Step 2 preparation of (1S,5R)-3-cyclopropyl-3,6-diazabicyclo[3.1.1]heptan-2-one (compound 68b) To a solution of compound 68a (180.0 mg, 0.71 mmol) in DCM (2 mL) was added TFA (1.0 mL) at 0 °C.
  • Example 69 Microliter plate-based TR-FRET assay for binders of STING This is the competition-binding assay to test the compounds’ potency to the C-terminal Domain (CTD) and ligand-binding domain of human stimulator of interferon genes (STING).
  • CCD C-terminal Domain
  • STING ligand-binding domain of human stimulator of interferon genes
  • Alexa-488 labeled active site probe (refer to patent WO2017/175156 A1) bounds to STING (139-379), it accepts the 485 nm emission from Tb-M2-Flag-STING and results in an increase in fluorescence at 520 nm. Compounds that compete for the probe-binding site will reduce 520 nm signal.
  • the assay was run in proxiplate- 384 plus (PerkinElmer, cat: 60150300) containing of 2.5 nM STING, 2.5 nM M2-Tb (Cisbio, 61FG2TLA, Lot: 17A) and 250 nM Alexa488 probe.
  • THP1-Dual Lucia Reporter Gene Assay This is the cellular reporter assay to evaluate compounds’ antagonism to interferon regulatory factor (IRF) pathway in THP1-DualTM cells (InvivoGen, cat.: thpd-nfis) .
  • IRF interferon regulatory factor
  • THP1- DualTM cells were derived from the human THP-1 monocyte cell line by stable integration lucia luciferase gene, a new secreted luciferase reporter gene, under the control of an ISG54 (interferon-stimulated gene) minimal promoter in conjunction with five interferon (IFN)- stimulated response elements.
  • ISG54 interferon-stimulated gene
  • THP1 ⁇ DualTM cells allow the study of the IRF pathway, by assessing the activity of Lucia luciferase. Lucia luciferase protein is readily measurable in the cell culture supernatant when using QUANTI ⁇ LucTM (InvivoGen, cat.
  • test medium RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum
  • test medium RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum
  • stimulator final concentration is 20 ⁇ M of 2’3’cGAMP, or final concentration is 10 MOI baculovirus virus,
  • compound solution per well final 1% DMSO
  • Example THP1_IC50 Example THP1_IC50
  • Example THP1_IC50 Example THP1_IC50 NO. (nM) NO. (nM) NO. (nM) 1 450 34 156 55 14 5 790 36 200 56 5 10 132 38 122 57 22 12 110 41 259 58 43 14 220 43 99 59 2 16 174 44 83 60 20 18 286 45 319 61 33 20 284 46 239 62 3 21 171 47 87 63 234 23 84 48 125 64 26 2 5 62 50 277 65 168 27 100 52 145 66 8 29 94 53 13 67 13 3 2 205 54 16 68 2

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Abstract

La présente invention concerne des composés de formule (I-2), dans laquelle R1 à R3, Q1, Q2, A1 à A7 et M1 sont tels que décrits dans la description, et leur sel pharmaceutiquement acceptable, et des compositions comprenant les composés et des procédés d'utilisation des composés.
PCT/EP2024/068795 2023-07-07 2024-07-04 Macrocycles pour le traitement d'une maladie auto-immune WO2025012057A1 (fr)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017175156A1 (fr) 2016-04-07 2017-10-12 Glaxosmithkline Intellectual Property Development Limited Amides hétérocycliques utiles en tant que modulateurs de protéine
EP3770161A1 (fr) * 2018-04-16 2021-01-27 Shenzhen TargetRx, Inc. Composé macrocyclique di(hétéro)aryle pour inhiber l'activité de la protéine kinase
WO2021014365A1 (fr) * 2019-07-22 2021-01-28 Lupin Limited Composés macrocycliques utilisés en tant qu'agonistes sting et procédés et utilisations de ceux-ci
WO2023237457A1 (fr) * 2022-06-06 2023-12-14 F. Hoffmann-La Roche Ag Macrocycles d'indazole pour le traitement d'une maladie auto-immune

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017175156A1 (fr) 2016-04-07 2017-10-12 Glaxosmithkline Intellectual Property Development Limited Amides hétérocycliques utiles en tant que modulateurs de protéine
EP3770161A1 (fr) * 2018-04-16 2021-01-27 Shenzhen TargetRx, Inc. Composé macrocyclique di(hétéro)aryle pour inhiber l'activité de la protéine kinase
WO2021014365A1 (fr) * 2019-07-22 2021-01-28 Lupin Limited Composés macrocycliques utilisés en tant qu'agonistes sting et procédés et utilisations de ceux-ci
WO2023237457A1 (fr) * 2022-06-06 2023-12-14 F. Hoffmann-La Roche Ag Macrocycles d'indazole pour le traitement d'une maladie auto-immune

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
ANSELHOWARD C. ET AL.: "Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems", 2004, WILLIAMS & WILKINS
GENNARO, ALFONSO R. ET AL.: "Remington: The Science and Practice of Pharmacy", 2000, WILLIAMS & WILKINS
ROWE, RAYMOND C: "Handbook of Pharmaceutical Excipients", 2005, PHARMACEUTICAL PRESS

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