Patent Application
20240383898
The present invention relates to a pyridocyclic compound and its preparation method, intermediates, compositions and uses thereof.
Janus kinases (JAK) belong to a family of tyrosine kinases involved in inflammation, autoimmune diseases, proliferative diseases, graft rejection, diseases associated with impaired cartilage renewal, congenital cartilage malformations and/or diseases associated with excessive IL6 secretion.
JAK kinases are cytoplasmic tyrosine kinases that transduce cytokine signals from membrane receptors to STAT transcription factors. The prior art has described four JAK kinase family members: JAK1, JAK2, JAK3, and TYK2. When cytokines bind to their receptors, JAK family members autophosphorylate and/or transphosphorylate each other, followed by phosphorylation of STATs, which then migrate into the nucleus to regulate transcription. JAK-STAT intracellular signaling applies to interferons, most interleukins, and a variety of cytokines and endocrine factors, such as EPO, TPO, GH, OSM, LIF, CNTF, GM-CSF, and PRL.
Combinatorial studies using genetic models and small molecule JAK inhibitors have revealed the therapeutic potential of several JAKs. JAK3 was confirmed as an immunosuppressive target by mouse and human genetics. JAK3 inhibitors were successfully used in clinical development, initially for organ transplant rejection but were subsequently used for other immunoinflammatory indications such as rheumatoid arthritis (RA), psoriasis, and Crohn's disease. TYK2 is a potential target for immunoinflammatory diseases and has been confirmed by human genetics and gene knockout studies in mice. JAK2 is currently an effective target in the treatment of myeloproliferative diseases, with two drugs for the treatment of myelofibrosis already on the market. JAK1 is a novel target in the field of immunoinflammatory diseases as the heterodimerization of JAK1 with other JAKs is known to transduce cytokine-driven pro-inflammatory signaling. Thus, the inhibition of JAK1 and/or other JAKs is expected to be therapeutically beneficial for a range of inflammatory conditions and other diseases driven by JAK-mediated signaling.
Tofacitinib was developed by Pfizer Inc. and successfully launched in the United States on Nov. 7, 2012 for the treatment of rheumatoid arthritis under the trade name Xeljanz.
Ruxolitinib, jointly developed by incyte and Novartis, was launched in the United States in 2011 for the treatment of myelofibrosis under the trade name Jakafi.
Baricitinib, jointly developed by Incyte and Eli Lilly, was launched in the United States in 2018 for the treatment of rheumatoid arthritis under the trade name Olumiant.
Upadacitinib, a selective JAK1 inhibitor developed by AbbVie and marketed in 2019 for the treatment of rheumatoid arthritis, has been reported in the literature to have activity of JAK1 IC50=43 nM and JAK2 IC50=200 nM.
Fedratinib is a selective JAK2 inhibitor developed by Sanofi and launched in 2019 for the treatment of myelofibrosis, has been reported in the literature to have an activity of JAK2 IC50=3 nM.
The present invention provides a pyridocyclic compound and its preparation method, intermediates, compositions and uses.
On one hand, the present invention provides a compound shown in formula I.
or a tautomer, stereoisomer, racemate or isotopic derivative thereof, or a pharmaceutically acceptable salt of any of the foregoing (referring to the foregoing compound as shown in Formula I, its tautomer, stereoisomer, racemate or isotopic derivative), or a crystalline or solvated form of any of the foregoing (referring to the foregoing compound as shown in Formula I, its tautomer, stereoisomer, racemate, isotopic derivative or pharmaceutically acceptable salt as shown in Formula I).
Each R3 is independently a halogen (e.g., fluorine), a cyano group, a C1-4 alkyl group (e.g., methyl group), a CIA haloalkyl group (e.g., C1-4 fluoroalkyl group, e.g., —CF3), a —O—C1-4 alkyl group (e.g., —O—CH3), or a —O—C1-4 haloalkyl group (e.g. a —O—C1-4 fluoroalkyl group, e.g. —OCF3).
Each R4 is independently a halogen (e.g., fluorine), a hydroxyl group, a C1-4 alkyl group (e.g., methyl group), a C1-4 haloalkyl group (e.g., C1-4 fluoroalkyl group, e.g., —CF3), an —O—C1-4 alkyl group (e.g., —O—CH3), a —O—C1-4 haloalkyl group (e.g. —O—C1-4 fluoroalkyl group, e.g. —OCF3) or a C1-4 hydroxyalkyl group (e.g. —CH2OH).
Alternatively, two R4 located on the same carbon atom or on different carbon atoms are interconnected to form —CH2— or —(CH2)2— (e.g., to form
for example
R5b, R5c, R5d, R5e and R5k are each independently a C1-6 alkyl group, a C1-6 haloalkyl group, -L1-R5f, an unsubstituted or substituted 6-10-membered aryl group (said 6-10-membered aryl group e.g. phenyl or naphthyl group) or an unsubstituted or substituted 5-10 membered heteroaryl group (said 5-10-membered heteroaryl group e.g. 5-6-membered heteroaryl group), said substituted 6-10-membered aryl group and substituted 5-10-membered heteroaryl group are defined as structures where 1, 2, 3 or 4 hydrogen atoms in the 6-10-membered-aryl group and the 5-10-membered-heteroaryl group are independently substituted by R5h.
Each R5g and R5h is independently a halogen (e.g., fluorine), a cyano group, a C1-4 alkyl group (e.g., methyl group), a C1-4 haloalkyl group (e.g., a C1-4 fluoroalkyl group, e.g., —CF3), a —O—C1-4 alkvl group (e.g., —O—CH3), a —O—C1-4 haloalkyl group (e.g. —OCF3) or a phenyl group.
Each L1 is independently —[C(RaRb)]1-5— (e.g. —CH2—, —(CH2)2—, —(CH2)3—, —(CH2)4—, —(CH2)5— or
—[C(RaRb)]1-2—C(O)—[C(RaRb)]1-2—, —[C(RaRb)]1-2—C(O)NH—[C(RaRb)]1-2— (e.g. —C(RaRb)—C(O)NH—C(RaRb)—R5f), —[C(RaRb)]1-2—NHC(O)—[C(RaRb)]1-2—, —[C(RaRb)]1-2—S(O)2—[C(RaRb)]1-2—, —[C(RaRb))]1-2—NHS(O)2—[C(RaRb)]1-2— or —[C(RaRb)]1-2—S(O)2NH—[C(RaRb)]1-2—.
Each R5f is independently H, F, CHF2, CH2F, CF3 or CN.
Each Ra is independently H, a halogen (e.g., fluorine), or a C1-3 alkyl group (e.g., methyl or ethyl group).
Each Rb is independently H, a halogen (e.g., fluorine), or a C1-3 alkyl group (e.g., methyl or ethyl group).
Alternatively, Ra and Rb along with the carbon atom joining them, form a cyclopropyl group.
m is 0, 1, 2, 3 or 4.
n is 0, 1, 2, 3 or 4.
(ii) R1 is
(e.g. trans or cis).
Ring C is a benzene ring or a 5-6-membered heteroaromatic ring (e.g., imidazole, thiazole, furan, thiophene, or pyridine, e.g.,
Each R6 is independently a halogen (e.g., fluorine or chlorine, e.g., fluorine), a hydroxyl group, an amino group, a cyano group, a C1-4 alkyl group (e.g., methyl group), a C1-4 haloalkyl group (e.g., C1-4 fluoroalkyl group, e.g., CF3), an —O—C1-4 alkyl group (e.g., —O—CH3), an O—C1-4 haloalkyl group (e.g. —O—C1-4 fluoroalkyl group, e.g. —O—CF3), a —S—C1-4 alkyl group (e.g. —S—CH3), a —S(O)2—C1-4 alkyl group (e.g. —S(O)2—CH3) or a C1-4 hydroxyalkyl group (e.g. —CH2—OH).
Each R7 is independently R4 (i.e., R7 and R4 in the compound as shown in Formula I in any of the embodiments of the present invention have the same definition).
Alternatively, two R7 located on the same carbon atom or on different carbon atoms are interconnected to form —CH2— or —(CH2)2—.
R8 is R5 (i.e., R8 and R5 in the compound as shown in Formula I in any of the embodiments of the present invention have the same definition), —S(O)2R8a, —C(O)R8b, —C(O)NR8cR8d, —C(O)OR8e or —C(O)NHR8k.
R8a, R8b, R8c, R8d, R8e and R8k are each independently a methyl group, —CF3, a C2-6 alkenyl group (e.g. vinyl group) or a C3-6, cycloalkyl group (e.g. cyclopropyl, cyclobutyl or cyclopentyl group).
Each R9 is independently R4.
Alternatively, two R9 located on the same carbon atom or on different carbon atoms are interconnected to form —CH2— or —(CH2)2—.
z is 0, 1, 2, 3 or 4.
y is 0, 1, 2, 3 or 4.
t is 0, 1, 2, 3 or 4.
(iii) R1 is H, CF3 or
Ring D is a C3-6 cycloalkyl group (e.g., cyclopropyl or cyclobutyl group), a benzene or a 5-6-membered heteroaromatic ring (e.g., furan or thiophene, e.g.,
Ring E is a benzene ring or a 5-6-membered heteroaromatic ring (e.g., furan or thiophene, e.g.,
Each R10 and R11 is independently a halogen (e.g., fluorine or chlorine, e.g., fluorine), a hydroxyl group, an amino group, a cyano group, a C1-4 alkyl group (e.g., methyl group), a C1-4 haloalkyl group (e.g., C1-4 fluoroalkyl group, e.g., CF3), a —O—C1-4 alkyl group (e.g., —O—CH3), a —O—C1-4 haloalkyl group (e.g. —O—C1-4 fluoroalkyl group, e.g. —O—CF3), a —S—C1-4 alkyl group (e.g. —S—CH3), a —S(O)2—C1-4 alkyl group (e.g. —S(O)2—CH3) or a C1-4 hydroxyalkyl group (e.g. —CH2—OH).
R12 is a cyano group, -L3-R12f or
Each R14a is independently a halogen (e.g., fluorine), a cyano group, a C1-4 alkyl group (e.g., methyl group), a cyano-substituted C1-4 alkyl group (e.g., —CH2CN), a C1-4 haloalkyl group (e.g., C1-4 fluoroalkyl group, e.g., CF3), a —O—C1-4 alkyl group (e.g., —O—CH3), a —O—C1-4 haloalkyl group (e.g. —O—C1-4 fluoroalkyl group, e.g. —O—CF3), or a C1-4 hydroxyalkyl group (e.g. —CH2OH).
R14b is R5 (i.e., R14b and R5 in the compound in any of the embodiments of the present invention as shown in Formula I have the same definition) or —S(O)2—C1-4 alkyl (e.g., —S(O)2—CH2CH3, —S(O)2—CH2CH2CH3 or —S(O)2—CH2CH2CH2CH3).
L3 is —[C(ReRf)]1-5— (e.g. —CH2—, —(CH2)2—, —(CH2)3—, —(CH2)4— or —(CH2)5—), —C(O)—, —C(O)NH—, —NHC(O)—, —S(O)2—, —NHS(O)2—, —S(O)2NH—, —[C(ReRf)]1-2—C(O)—[C(ReRf)]1-2, —[C(ReRf)]1-2—C(O)NH—[C(ReRf)]1-2—, —[C(ReRf)]1-2—NHC(O)—[C(ReRf)]1-2—, —[C(ReRf)]1- 2—S(O)2[C(ReRf)]1-2—, —[C(ReRf)]1-2—NHS(O)2—[C(ReRf)]1-2— or —[C(ReRf)]1-2—S(O)2NH—[C(ReRf)]1-2—.
R12f is H, F, CHF2, CH2F, CF3 or CN.
Each Re and Rf is independently H, a halogen (e.g., fluorine), a C1-3 alkyl group (e.g., methyl or ethyl group), or a C3-6 cycloalkyl group (e.g., cyclopropyl, cyclobutyl, or cyclopentyl group).
Alternatively, Re and Rf along with their connecting carbon atom, form a cyclopropyl group.
p is 0, 1, 2, 3 or 4.
q is 0, 1, 2, 3 or 4.
v is 0, 1, 2 or 3.
u is 0, 1 or 2.
The heteroatom in the above heteroaryl group is independently N, O or S, and the number of heteroatoms in the above heteroaryl group is independently 1, 2, 3 or 4.
In some embodiments, said compound shown in formula I has the structure shown in formula I-1 as follows.
wherein R1a, R4, R5, X and n are defined as described in the compound shown in Formula I as defined in any of the embodiments of the present invention.
In some embodiments, a compound as shown in Formula I or I-1 has a structure as shown in Formula I-1C, I-1D, I-1E or I-1F as follows.
wherein R1a, R4, R5, X and n are defined as described in the compound shown in Formula I as defined in any of the embodiments of the present invention.
In some embodiments, said compound shown in formula I or I-1 has the structure shown in formula I-1a, I1b, I-1c or I-1d as follows, preferably having the structure shown in I-1d.
wherein R1a, R4, R5g, X, n, L1 and R5f are defined as described in the compound shown in Formula I as defined in any of the embodiments of the present invention.
Each f is independently 0, 1, 2, 3 or 4, e.g. 0 or 1.
In some embodiments, said compound shown in formula I has the structure shown in formula I-2, I-2A or I-2B as follows
wherein R1a, R3, X and m are defined as described in the compound shown in Formula I as defined in any of the embodiments of the present invention.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, I-1F, 1-1a, I-1b, 1-1c, 1-1d, I-2, I-2A, or I-2B, X is CH.
In some embodiments, as in the compounds shown in Formula I, I-1, 1-1C, 1-1D, I-1E, I-1F, 1-1a, I-1b, I-1c, I-1d, I-2, I-2A, or I-2B, X is N.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, I-1F, I-1a, I-1b, I-1c, I-1d, I-2, I-2A, or I-2B, R1a is a methyl group.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1a, I-1b, I-1c, I-1d, or I-2,
preferably
(i) In some embodiments, as in the compounds shown in Formula I, I-2, I-2A or I-2B, m is 1.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1a, I-1b, I-1c, or I-1d,
In some embodiments, is
In some embodiments,
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, I-1F, I-1a, I-1b, I-1c or I-1d, each R4 is independently a halogen (e.g., fluorine) or a C1-4 hydroxyalkyl group (e.g., —CH2OH), or, alternatively, two R4 located at the same carbon atom or at different carbon atoms are interconnected to form —CH2— or —(CH2)2—.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, I-1F, I-1a, I-1b, I-1c, I-1d, I-2, I-2A, or I-2B, n is 0, 1, or 2.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, I-1F, I-1a, I-1b, I-1c, or I-1d,
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, I-1F, I-1a, I-1b, I-1c, or I-1d,
In some embodiments, the ring B is a benzene ring in the compound as shown in Formula 1.
In some embodiments, m is 1 in the compound as shown in Formula I.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E or I-1F, R5a is a C2-6 alkyl group (e.g., ethyl, n-propyl or n-butyl group), an unsubstituted or substituted 6-10-membered aryl group (said 6-10-membered aryl group e.g., phenyl or naphthyl group) or an unsubstituted or substituted 5-10-membered heteroaryl group (said 5-10-membered heteroaryl group e.g. 5-6-membered heteroaryl group).
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, or I-1F, the “unsubstituted or substituted 6-10-membered aryl group” in the definition of R5a may be “unsubstituted or substituted phenyl group”, or “unsubstituted or substituted naphthyl group”, wherein said unsubstituted or substituted phenyl group may be, for example, an unsubstituted phenyl group, or a phenyl group substituted with 1, 2, 3 and 4 (e.g., 1) of the functional groups independently selected from the following: halogens (e.g., fluorine), C1-4 alkyl groups (e.g., methyl), —O—C1-4 haloalkyl groups (e.g., —OCF3) and phenyl groups, such as
Wherein, said unsubstituted or substituted naphthyl group may be, for example, an unsubstituted naphthyl group, such as
In some embodiments, the “unsubstituted or substituted 6-10-membered-aryl group” in R5a is preferably a —OCF3 substituted phenyl group
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E or I-1F, “unsubstituted or substituted 5-10-membered-heteroaryl group” in the definition of R5a may be “unsubstituted or substituted 5-6-membered heteroaryl group”, such as an unsubstituted or substituted thiophenyl group, preferably
In some embodiments, as in the compound shown in Formula I, I-1, I-1C, I-1D, I-1E, I-1F, I-1a, I-1b, or I-1c, each R5g is independently a halogen (e.g., fluorine), a C1-4 alkyl group (e.g., methyl group), a —O—C1-4 haloalkyl group (e.g. —OCF3) or a phenyl group, preferably —OCF3.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E or I-1F, —S(O)2R5f in the definition of R5 is —S(O)2(CH2)2—CF3, —S(O)2(CH2)2—CH3, —S(O)2CH2—CH3, —S(O)2(CH2)3—CH3, —S(O)2(CH2)2—CF3,
preferably —S(O)2(CH2)2—CF3, —S(O)2(CH2)2—CH3, —S(O)2CH2—CH3, —S(O)2(CH2)3—CH3, —S(O)2(CH2)2—CF3, or
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, or I-1F, R5b is -L1-R5f, or an unsubstituted or substituted phenyl group.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, or I-1F, -L1-R5f, in the definition of R5b is —C(RaRb)—R5f, for example
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E or I-1F, the “unsubstituted or substituted 6-10-membered aryl group” in the definition of R5b may be “unsubstituted or substituted phenyl group”, or “unsubstituted or substituted naphthyl group”; wherein unsubstituted or substituted phenyl group may be, for example, an unsubstituted phenyl group, or a phenyl group substituted with 1, 2, 3 and 4 (e.g., 1) of the functional groups independently selected from the following: halogens (e.g., fluorine) and cyano-substituted phenyl groups such as
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, or I-1F, the —C(O)R5b in the definition of R5 is
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, or I-1F, R5e is a C1-6 alkyl group, such as tert-butyl group.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, or I-1F, —C(O)OR5e in the definition of R5 is —C(O)OC(CH3)2—CH3.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, or I-1F, R5c, R5d, and R5k are each independently -L1-R5f.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E or I-1F, -L1-R5f in the definitions of R5c, R5d and R5k are —C(RaRb)—R5f, e.g. —CH2—R5f, e.g. —CH2—CF3.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, or I-1F, —C(O)NHR5k in the definition of R5 is —C(O)NHC(RaRb)—R5f, e.g. —C(O)NHCH2-R5f, e.g. —C(O)NHCH2—CF3.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E or I-1F, -L1-R5f in the definition of R5 is —[C(RaRb)]1-5—R5f or —[C(RaRb)]1-2—C(O)NH—[C(RaRb)]1-2—, preferably —C(RaRb)—R5f, —[C(RaRb)]2—R5f, —[C(RaRb)]3—R5f, —[C(RaRb)]4-R5f or —[C(RaRb)]5-R5f, further preferably —[C(RaRb)]2—R5f, —[C(RaRb)]3-R5f.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, or I-1F, -L1-R5f in the definition of R5 is —C(RaRb)—R5f, —[C(RaRb)]2—R5f, —[C(RaRb)]3—R5f, —[C(RaRb)]4—R5f, —[C(RaRb)]5-R5f or —C(RaRb)—C(O)NH—C(RaRb)—R5f, preferably —[C(RaRb)]2—R5f or —[C(RaRb)3-R5f.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, or I-1F, each R5f is independently F, CF3, or CN. In some embodiments, each R5f is independently F. In some embodiments, each R5f is independently CF3. In some embodiments, each R5f is independently CN.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, or I-1F, -L1-R5f in the definition of R5 is —C(RaRb)—CN, —[C(RaRb)]2—CN, —[C(RaRb)]3—CN, —[C(RaRb)]4—CN, —[C(RaRb)]—CF3, —[C(RaRb)]2—CF3, —[C(RaRb)]3—CF3, —C(RaRb)C(O)NH—C(RaRb)CF3 or —[C(RaRb)]5—F, preferably —C(RaRb)—CN, —[C(RaRb)]2—CN, —[C(RaRb)]3—CN, —[C(RaRb)]4—CN, —[C(RaRb)]—CF3, —[C(RaRb)]2—CF3, —[C(RaRb)]3—CF3, or —[C(RaRb)]5—F.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E or I-1F, each Ra is independently H, fluorine, a methyl or ethyl group; each Rb is independently H, fluorine, a methyl or ethyl group; or, Ra and Rb along with the carbon atom connecting them, form a cyclopropyl group together.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E or I-1F, each Rb is independently H, fluorine, a methyl group or an ethyl group.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E or I-1F, each Rb is independently H or fluorine.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E or I-1F, each Rb is independently H, fluorine, a methyl group or an ethyl group.
In some embodiments, as in the compounds such as those shown in Formula I, I-1, I-1C, I-1D, I-1E or I-1F, each Rb is independently H, fluorine, a methyl group or an ethyl group.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E or I-1F, -L1-R5f in the definition of R5 is —CH2—CN, —(CH2)2—CN, —(CH2)3—CN, —(CH2)4—CN, —(CH2)2—CF3, —CH2(CF2)2—CF3, —(CF2)3—CF3, —(CH2)2CF2—CF3, —(CH2)3—CF3, —CH(CH2CH3)C(O)NH—CH2CF3, —CH2C(O)NHCH2—CF3, —CH(CH3)C(O)NHCH2—CF3 or —(CH2)5—F, preferably —(CH2)2—CN, —(CH2)3—CN, —(CH2)2—CF3, —CH2(CF2)2—CF3, —(CF2)3—CF3, —(CH2)2CF2—CF3 or —(CH2)3—CF3.
In some embodiments, as in the compounds shown in Formula I, I-1, I-1C, I-1D, I-1E, or I-1F, R5 is —CH2—CN, —(CH2)2—CN, —(CH2)3—CN, —(CH2)4—CN, —(CH2)2—CF3, —CH2(CF2)2—CF3, —(CF2)3—CF3, —(CH2)CF2—CF3, —(CH2)3—CF3, —CH(CH2CH3)C(O)NH—CH2CF3, —CH2C(O)NHCH2—CF3, —CH(CH3)C(O)NHCH2—CF3, —(CH2)5—F,
—S(O)2(CH2)2—CF3, —C(O)NHCH2—CF3, —S(O)2(CH2)2—CH3, —S(O)2CH2—CH3, —S(O)2(CH2)3—CH3, —S(O)2(CH2)2—CF3,
C(O)OC(CH3)2—CH3, or
preferably —CH2—CN, —(CH2)2—CN, —(CH2)3—CN, —(CH2)4—CN, —(CH2)2—CF3, —CH2(CF2)2—CF3, —(CF2)3—CF3, —(CH2)2CF2—CF3, —(CH2)3—CF3, —CH(CH2CH3)C(O)NH—CH2CF3, —CH2C(O)NHCH2—CF3, —CH(CH3)C(O)NHCH2—CF3, —(CH2)5—F, —S(O)2(CH2)2—CF3, —S(O)2CH2—CH3, —S(O)2(CH2)3—CH3, —S(O)2(CH2)2—CF3,
In some embodiments, said compound shown in formula I has the structure shown in formula I-3, I-3A or I-3B as follows.
wherein X, Y, , ring C, R6, R7, R9, z and y are defined as described in the
compound shown in Formula I as defined in any of the embodiments of the present invention.
In some embodiments, said compound shown in formula I has the structure shown in formula I-4 as follows.
wherein X, Y, , ring C, R6, R9, z and t are defined as described in the compound shown in Formula I as defined in any of the embodiments of the present invention.
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, or I-4, is a double bond and Y is N.
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, or I-4, is a single bond and Y is NH.
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, or I-4, the 5-6-member e r aromatic ring in the definition of ring C is an imidazole
thiazol
furan
thiophene
or pyridine
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, I-4, I-4A, I-4, I-4C or I-4D, when R2 is
ring C is a benzene ring, an imidazole
a furan
or a pyridine
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, I-4, I-4A, I-4B, I-4C or I4D, when R2 is
ring C is a benzene ring, a thiazole
a furan
or a thiophene
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, I-4 I-4A, I-4B, I-4C, or I-4D
In some embodiments, said compound shown in formula I has the structure shown in formula I-3C, I-3D, I-3E, I-3F or I-3G as follows.
wherein X, R6, R7, R8, z and y are defined as described in any of the embodiments of the present invention.
In some embodiments, as in the compounds shown in Formula I, I-3, I-3C, I-3D, I-3E, I-3F, or I-3G,
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, I-3C, I-3D, I-3E, I-3F or I-3G, R7 is R4, which means that R7 and R4 in the compound as shown in Formula I in any of the embodiments of the present invention have the same definition.
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, I-3C, I-3D, I-3E, I-3F or I-3G, R8 is R5, which means that R8 and R5 in the compound as shown in Formula I in any of the embodiments of the present invention have the same definition.
In some embodiments, as in the compound shown in Formula I, I-3, I-3A, I-3B, I-3C, I-3D, I-3E, I-3F, or I-3G, y is 0.
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, I-3C, I-3D, I-3E, I-3F or I-3G, R8 is R5, —S(O)2R8a or —C(O)R8b, wherein R5 is defined in the same way as R5 as defined in the compound shown in Formula I in any of the embodiments of the present invention.
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, I-3C, I-3D, I-3E, I-3F, or I-3G, R8 is a —S(O)2C(RaRb)—C1-6 alkyl group, —S(O)2—CF3, —S(O)2C(RaRb)—R5f, —S(O)2[C(RaRb)2]—R5f, —S(O)2[C(RaRb)3]—R5f, —C(RaRb)—R5f, —[C(RaRb)2]—R5f, —[C(RaRb)3]—R5f,
—C(O)CH═CH2 or —C(O)NHC(RaRb)—R5f; wherein R5f is defined in the same way as R5f as defined in the compound shown in Formula I in any of the embodiments of the present invention, e.g. R5f is CF3 or CN.
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3D, I-3C, I-3D, I-3E, I-3F or I-3G, R8 is —S(O)2C(RaRb)—CH3, —S(O)2—CF3, —C(O)NHC(RaRb)—CF3, —C(O)CH═CH2,
—C(RaRb)—CN, —[C(RaRb)2]—CN or —[C(RaRb)3]—CF3, preferably —S(O)2C(RaRb)—CH3, —S(O)2—CF3,
—[C(RaRb)2]—CN or —[C(RaRb)3]—CF3.
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, I-3C, I-3D, I-3E, I-3F, or I-3G, R8 is —S(O)2(CH2)2—CH3, —S(O)2—CF3, —C(O)NHCH2—CF3, —C(O)CH═CH2,
—CH2—CN, —(CH2) —CN, or —(CH2)3—CF3, preferably —S(O)2(CH2)2—CH3, —S(O)2—CF3,
—(CH2)2—CN or —(CH2)3—CF3.
In some embodiments, said compound shown in Formula I has the structure shown in Formula I-4E, I-4F, I-4J, I-4L or I-4M as follows.
wherein X, R6, R9, z and t are defined as described in the compound shown in Formula I as defined in any of the embodiments of the present invention.
In some embodiments, compounds such as those shown in Formula I, I-4, I-4E, I-4F, I-4J, I-4L, or I-4M,
are in the cis or trans configuration, e.g., trans.
In some embodiments, as in the compounds shown in Formula I, I-4, I-4E, I-4F, I-4J, I-4L, or I-4M, t is 0.
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, I-3C, I-3D, I-3E, I-3F, I-3G, I-4, I-4E, I-4F, I-4J, I-4L, or I-4M, X is CH.
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, I-3C, I-3D, I-3E, I-3F, I-3G, I-4, I-4E, I-4F, I-4J, I-4L, or I-4M, X is N.
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3D, I-3C, I-3D, I-3E, I-3F, I-3G, I-4, I-4E, I-4F, I-4J, I-4L, or I-4M, z is 0, 1, or 2.
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, I-3C, I-3D, I-3E, I-3F, I-3G, I-4, I-4E, I-4F, I-4J, I-4L, or I-4M, each R6 is independently a halogen (e.g., fluorine), a hydroxyl group, an amino group, a cyano group, a C1-4 alkyl group (e.g., methyl group), a C1-4 haloalkyl group (e.g. C1-4 fluoroalkyl group, e.g., CF3), a —O—C1-4 alkyl group (e.g., —O—CH3), a —S—C1-4 alkyl group (e.g., —S—CH3), a —S(O)2—C1-4 alkyl group (e.g. —S(O)2—CH3), or a C1-4 hydroxyalkyl group (e.g. —CH2—OH).
In some embodiments, as in the compounds shown in Formula I, I-3, I-3A, I-3B, I-3C, I-3D, I-3E, I-3F, I-3G, I-4, I-4E, I-4F, I-4J, I-4L or I-4M, each R6 is independently fluorine, a hydroxyl group, an amino group, a cyano group, a methyl group, CF3, —O—CH3, —S—CH3, —S(O)2—CH3 or —CH2—OH.
In some embodiments, z is (in the compound as shown in Formulas I-3C.
In some embodiments, as in the compounds shown in Formula I-3D or I-4F,
is independently
wherein R6 is defined as described in the compounds shown in Formula I as defined in any of the embodiments of the present invention, such as a C1-4 alkyl group (e.g., methyl group) or a C1-4 hydroxyalkyl group (e.g., —CH2—OH). For example, in some embodiments,
can be
for example, in some embodiments,
can be
In some embodiments, as in the compounds shown in Formula I-3E, I-3G, I-4L or I-4M,
is independently
wherein each R6 is defined independently as described in the compounds shown in Formula I as defined in any of the embodiments of the present invention, e.g. halogen (e.g. fluorine), hydroxyl group, C1-4 haloalkyl group (e.g. C1-4 fluoroalkyl group, e.g. CF3), —O—C1-4 alkyl group (e.g. —O—CH3), —S—C1-4 alkyl group (e.g. —S—CH3) or —S(O)2—C1-4 alkyl group (e.g. —S(O)2—CH3).
In some embodiments, as in the compound shown in Formula I-3F, is
wherein R6 is defined as described in the compounds shown in Formula I as defined in any of the embodiments of the present invention, e.g., amino group.
In some embodiments, as in the compound shown in Formula I4E,
wherein R6 is defined as described in the compounds shown in Formula I as defined in any of the embodiments of the present invention, such as an amino group or a C1-4 alkyl group (e.g., methyl group). For example, in some embodiments,
For example, in some embodiments,
In some embodiments, as in the compound shown in Formula I-4J,
wherein R6 is defined as described in the compounds shown in Formula I as defined in any of the embodiments of the present invention, such as a C1-4 alkyl group (e.g., methyl group). For example, in some embodiments,
can be
In some embodiments, said compound shown in formula I has the structure shown in formula I-5 as follows.
wherein X, R11, R12 and q are defined as described in the compound shown in Formula I as defined in any of the embodiments of the present invention.
In some embodiments, said compound shown in formula I has the structure shown in formula I-6 as follows.
wherein X, R11, R12 and q are defined as described in the compound shown in Formula I as defined in any of the embodiments of the present invention.
In some embodiments, as in the compound shown in Formula I-5 or I-6, R12 is independently —C(ReRf)—CN, —[C(ReRf)2]—CN or —[C(ReRf)3]—CN, such as —CH2—CN, —(CH2)2—CN, —(CH2)3—CN, or
In some embodiments, as in the compound shown in Formula I-5, R12 is preferably —[C(ReRf)3]—CN, for example —(CH2)3—CN.
In some embodiments, as in the compound shown in Formula I-6, R12 is preferably —CH2—CN or
In some embodiments, said compound shown in formula I has the structure shown in formula I-7 as follows.
wherein X, Y, , ring D, ring E, R10, R11, R12, p and q are defined as described in the compound shown in Formula I as defined in any of the embodiments of the present invention.
In some embodiments, as in the compound shown in Formula I or I-7, is a double bond and Y is N.
In some embodiments, as in the compound shown in formula I or I-7 as described, is a single bond and Y is NH.
In some embodiments, as in the compound shown in Formula I or I-7, “the 5-6-membered heteroaromatic ring” in the definition of ring D is a furan
or a thiophene
In some embodiments, as in the compound shown in Formula I or I-7, ring D is a cyclopropyl group, a cyclobutyl group, a benzene ring,
In some embodiments, as in the compound shown in Formula I or I-7, is
is independently
In some embodiments, said compound shown in Formula I has the structure shown in Formula I-7A, I-7B, I-7C, I-7D, I-7E or I-7F as follows.
wherein X, R10, R11, R12, p and q are defined as described in any of the embodiments of the present invention.
In some embodiments, as in the compounds in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E or I-7F, R12 is a cyano group, —C(ReRf)—R12f, —[C(ReRf)2]—R12f, —[C(ReRf)3]—R12f or
In some embodiments, as in the compounds shown in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E, or I-7F, v is 0.
In some embodiments, as in the compounds shown in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E, or I-7F,
for example
In some embodiments, as in the compounds shown in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E, or I-7F, R14a is a cyano-substituted C1-4 alkyl group, such as —CH2CN.
In some embodiments, as in the compounds shown in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E, or I-7F, R12 is —CN, —C(ReRf)—CN, —[C(RR)2]—CN, —[C(ReRf)3]—CN or
In some embodiments, as in the compounds shown in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E or I-7F, each Re is independently H or a C1-3 alkyl group.
In some embodiments, as in the compounds shown in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E or I-7F, each Rf is independently H or a C1-3 alkyl group.
In some embodiments, as in the compounds shown in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E or I-7F, R12 is —CN, —CH2—CN, —(CH2)2—CN, —(CH2)3—CN,
preferably —CH2—CN, —(CH2)2—CN, —(CH2)3—CN, further preferably —(CH2)2—CN.
In some embodiments, as in the compounds shown in formula I-7A, I-7B or I-7C, R12 is a cyano group.
In some embodiments, as in the compounds shown in Formula I-7E or I-7F, R12 is —C(ReRf)—R12f, —[C(ReRf)2]—R12f, —C(ReRf)3]—R12f, or
preferably —C(ReRf)—R12f, —[C(ReRf)2]—R12f or —[C(ReRf)3]—R12f, further preferably —[C(ReRf)2]—R12f.
In some embodiments, said compound shown in Formula I, I-7E or I-7F has the structure shown in Formula I-7G or I-7H as follows.
wherein X, R10, R11, R14b, p and q are defined as described in any of the embodiments of the present invention.
In some embodiments, as in the compounds shown in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E, I-7F, I-7G or I-7H, R14b is R5, which means that R14b and R5 in the compound as shown in Formula I in any of the embodiments of the present invention have the same definition
In some embodiments, as in the compounds shown in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E, I-7F, I-7G or I-7H, R14b is a —S(O)2—C1-4 alkyl group, such as —S(O)2—CH2CH3, —S(O)2—CH2CH2CH3 or —S(O)2—CH2CH2CH2CH3.
In some embodiments, as in the compounds shown in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E, I-7F, I-7G, or I-7H, X is N.
In some embodiments, as in the compounds shown in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E, I-7F, 1-7G, or I-7H, X is CH.
In some embodiments, as in the compounds shown in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E, I-7F, I-7G, or I-7H, q is 0.
In some embodiments, as in the compounds shown in Formula I, I-5, 1-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E, I-7F, I-7G, or I-7H, p is 0, 1, or 2.
In some embodiments, as in the compounds shown in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E, I-7F, I-7G, or I-7H, each R10 is independently a halogen (e.g., fluorine or chlorine), a hydroxyl group, an amino group, a cyano group, a C1-4 alkyl group (e.g., methyl group), a C1-4 haloalkyl group (e.g., C1-4 fluoroalkyl group, e.g., CF3), a —O—C1-4 alkyl group (e.g. —O—CH3), a —S(O)2—C1-4 alkyl group (e.g. —S(O)2—CH3), or a C1-4 hydroxyalkyl group (e.g. —CH2—OH).
In some embodiments, as in the compounds shown in Formula I, I-5, I-6, I-7, I-7A, I-7B, I-7C, I-7D, I-7E, I-7F, I-7G or I-7H, each R10 is independently fluorine, chlorine, a hydroxy group, a methyl group, —S(O)2—CH3 or —CH2—OH.
In some embodiments, as in the compounds shown in Formula I-7A, I-7F, or I-7H,
is independently,
wherein each R10 is defined as described in any of the embodiments of the present invention, such as a C1-4 alkyl group (e.g., methyl group) or a C1-4 hydroxyalkyl group (e.g., —CH2OH). For example, in some embodiments,
can be
For example, in some embodiments,
may be
In some embodiments, as in the compounds shown in Formula I-7E or I-7G,
is independently
wherein each R10 is defined as described in any of the embodiments of the present invention, e.g., a C1-4 alkyl group (e.g., methyl group).
In some embodiments, as in the compounds shown in Formula I-7B, I-7C or I-7D,
wherein each R10 is defined as described in any of the embodiments of the present invention, such as a hydroxyl group, a halogen (e.g., fluorine or chlorine) or a —S(O)2—C1-4 alkyl group (e.g. —S(O)2—CH3).
In some embodiments, said compound as shown in Formula I has the following structure.
On the other hand, the present invention also provides a method for the preparation of a compound as shown in formula I, which includes the following steps: reacting a compound as shown in formula 1I with a base (e.g. sodium hydroxide) in an organic solvent (e.g. a mixture of tetrahydrofuran and methanol) to obtain said compound as shown in formula I.
wherein X, Y, , R1 and R2 are defined as described in any of the embodiments of the present invention.
On the other hand, the present invention also provides a compound as shown in Formula 1I.
wherein X, Y, , R1 and R2 are defined as described in any of the embodiments of the present invention.
On the other hand, the present invention also provides a pharmaceutical composition comprising:
On the other hand, the present invention also provides a compound as described in Formula I, or a tautomer, stereoisomer, racemate or isotopic derivative thereof, or a pharmaceutically acceptable salt of any of the foregoing, or a crystalline or solvated form of any of the foregoing, or an application of said pharmaceutical composition as a drug.
On the other hand, the present invention also provides a compound as described in Formula I, or a tautomer, stereoisomer, racemate or isotopic derivative thereof, or a pharmaceutically acceptable salt of any of the foregoing, or a crystalline or solvated form of any of the foregoing, or an application of said pharmaceutical composition which acts as an inhibitor of Janus kinase (e.g., JAK1 and/or JAK2).
On the other hand, the present invention also provides a method of inhibiting Janus kinase (e.g. JAK1 and/or JAK2) in vivo, in vitro or ex vivo, which includes the contact of said compound shown in Formula I, or a tautomer, stereoisomer, racemate or isotopic derivative thereof, or a pharmaceutically acceptable salt of any of the foregoing, or a crystalline or solvated form of any of the foregoing, or said pharmaceutical composition with said Janus kinase.
On the other hand, the present invention also provides a compound as described in Formula I, or a tautomer, stereoisomer, racemate or isotopic derivative thereof, or a pharmaceutically acceptable salt of any of the foregoing, or a crystalline or solvated form of any of the foregoing, or a pharmaceutical composition as described in the preparation of drugs for the treatment of diseases associated with Janus kinases (e.g. JAK1 and/or JAK2).
On the other hand, the present invention also provides a compound as shown in Formula I, or a tautomer, stereoisomer, racemate or isotopic derivative thereof, or a pharmaceutically acceptable salt of any of the foregoing, or a crystalline or solvated form of any of the foregoing, or an application of said pharmaceutical composition in the preparation of a drug for the treatment of an autoimmune disease or cancer.
On the other hand, the present invention also provides a method of treating a disease associated with a Janus kinase (e.g., JAK1 and/or JAK2) which includes administering a therapeutically effective amount of said compound shown in Formula I, or a tautomer, stereoisomer, racemate or isotopic derivative thereof, or a pharmaceutically acceptable salt of any of the foregoing, or a crystalline or solvated form of any of the foregoing, or a pharmaceutical composition as described to a subject in need of such treatment.
On the other hand, the present invention also provides a method of treating an autoimmune disease or cancer which includes administering a therapeutically effective amount of said compound shown in Formula I, or a tautomer, stereoisomer, racemate or isotopic derivative thereof, or a pharmaceutically acceptable salt of any of the foregoing, or a crystalline or solvated form of any of the foregoing, or a pharmaceutical composition as described to a subject in need of such treatment.
The diseases associated with Janus kinases (e.g., JAK1 and/or JAK2) described in the present invention can be autoimmune diseases or cancers.
The autoimmune diseases described herein or autoimmune diseases associated with Janus kinase can be, for example, psoriasis, rheumatoid arthritis, inflammatory bowel disease, Sjogren's syndrome, leukoaraiosis, multiple sclerosis, or systemic lupus erythematosus.
The cancer or cancers associated with Janus kinase described in the present invention may be, for example, Kapozi's sarcoma, giant lymph node hyperplasia, lymphoma, leukemia multiple myeloma, or myeloproliferative disorders. Myeloproliferative disorders described may be, for example, polycythemia vera (PV), essential thrombocytosis (ET), primary myelofibrosis (PMF), chronic myeloid leukemia (CML), chronic monocytic leukemia (CMML), eosinophilic leukocytosis syndrome (HES), idiopathic myelofibrosis (IMF), or systemic mastocytosis (SMCD).
Unless otherwise stated, terms used in the present invention have the following definitions, and terms not covered below are defined as common knowledge understood by those skilled in the art of the present invention.
The term “tautomer” refers to a functional group isomer that results from the rapid movement of an atom in two positions in a molecule. For example, acetone and 1-propen-2-ol can transform into each other through the rapid movement of the hydrogen atom between the oxygen atom and a -carbon.
The term “stereoisomers” refers to isomers of molecules in which atoms or groups of atoms are attached to each other in the same order but are in different spatial arrangements, such as cis-trans isomers (e.g., Z-isomer, E-isomer), optical isomers (e.g., enantiomeric isomers, diastereomeric isomers), and atropisomers. These stereoisomers can be separated, purified and enriched by asymmetric synthesis methods or chiral separation methods (including but not limited to thin-layer chromatography, rotational chromatography, column chromatography, gas chromatography, high-pressure liquid chromatography, etc.). Stereoisomers can also be obtained by bonding (chemical bonding, etc.) or salt formation (physical bonding, etc.) with other chiral compounds via chiral resolution. Optical isomers include enantiomers and diastereoisomers. All of these isomers, as well as their mixtures, are included in the scope of the present invention.
i. The term “isotopic derivative” refers to the substitution of one or more atoms in a compound by one or more atoms having a specific atomic mass or mass number. Examples of isotopes that can be incorporated into a compound include, but are not limited to, isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, sulfur, and chlorine (e.g., 2H, 3H, 13C, 14C, 15N, 18O, 17O, 18F, 35S, and 36Cl). Isotopic compounds can typically be prepared by replacing non-isotopically labeled reagents with isotopically labeled reagents according to the methods described herein. Typical examples of isotopic derivatives include deuterated compounds.
The term “pharmaceutically acceptable salt” refers to the salt of a compound prepared with a relatively non-toxic, pharmaceutically acceptable acid or base. When a compound contains a relatively acidic functional group, a base addition salt may be obtained via the exposure of the neutral form of such compound to a sufficient amount of a pharmaceutically acceptable base in a pure solution or a suitable inert solvent. The pharmaceutically acceptable base addition salts include, but are not limited to: lithium, sodium, potassium, calcium, aluminum, magnesium, zinc, bismuth, ammonium, and diethanolamine salts. When the compounds of the present invention contain relatively basic functional groups, the corresponding acid addition salts can be obtained by exposing the neutral form of such compounds to a sufficient amount of a pharmaceutically acceptable acid in a pure solution or a suitable inert solvent. Said pharmaceutically acceptable acids include inorganic acids, said inorganic acids including but not limited to: hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, phosphoric acid, phosphorous acid, sulfuric acid, etc. Said pharmaceutically acceptable acids include organic acids, said organic acids including, but not limited to: acetic acid, propionic acid, oxalic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanedioic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, salicylic acid, tartaric acid, methanesulfonic acid, isonicotinic acid, acid citric acid, oleic acid, tannic acid, pantothenic acid, tartaric acid hydrogen, ascorbic acid, gentianic acid, fumaric acid, gluconic acid, glycolic acid, formic acid, ethanesulfonic acid, dihydroxynaphthalic acid (i.e., 4,4′-methylene-bis(3-hydroxy-2-naphthoic acid)), amino acids (e.g., glutamic acid, arginine), etc. When the compounds contain relatively acidic and relatively basic functional groups, they can be converted into base addition salts or acid addition salts, respectively. For details, see Berge et al. “Pharmaceutical Salts”, Journal of Pharmaceutical Science 66: 1-19 (1977), or, Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl and Camille G. Wermuth, ed., Wiley-VCH, 2002).
In some embodiments, a pharmaceutically acceptable salt of a compound with the structure shown in general formula (I) as described herein may be an acid addition salt formed by a compound of general formula (I) with a pharmaceutically acceptable acid, said acid including, but not limited to: hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid p-toluenesulfonic acid or ferulic acid. A pharmaceutically acceptable salt of said compound with the structure shown in general formula (I) may be prepared by reaction with an equivalent or excess amount of an acid (inorganic or organic) in a suitable solvent or solvent mixture. Said acids include, but are not limited to, hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, or ferulic acid. Said solvents include, but are not limited to, methanol, ethanol, methylene chloride, acetone, ethyl acetate, toluene, or tetrahydrofuran, or a mixture of any of these acids.
The term “crystalline” is defined as a structure where the ions or molecules are arranged in a defined way, in a three-dimensional space, and in a strictly periodic manner, and have a regular pattern of periodic recurrence at a certain distance apart, because of the periodic arrangement mentioned above, there can be a variety of crystalline forms, that is, the phenomenon of polycrystalline forms, also known as polymorphism.
The term “solvate” refers to a substance formed by combining molecules with a stoichiometric or non-stoichiometric solvent. The solvent molecules in a solvate may exist in an ordered or unordered arrangement. Said solvents include, but are not limited to, water, methanol, ethanol, etc.
The term “halogen” refers to fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine.
The term “amino group” refers to the —NH2 group.
The term “hydroxyl group” refers to the —OH group.
The term “cyano group” refers to the —CN group.
The term “alkyl group” refers to a saturated straight or branched monovalent hydrocarbon group having a certain number of carbon atoms. A C1-4 alkyl group refers to an alkyl group having 1-4 carbon atoms, including C1 alkyl group, C2 alkyl group, C3 alkyl group, and C4 alkyl group, such as methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, tert-butyl group butyl grouptert-butyl group. Examples of alkyl groups include, but are not limited to, methyl group, ethyl group, n-propyl group, isopropyl group, n-butylgroup, isobutyl group, sec-butyl group, tert-butyl group, and pentyl group.
The term “haloalkyl group” refers to an alkyl group substituted with one or more (e.g., 1, 2, 3 or 4)halogens (e.g., fluorine, chlorine, bromine or iodine, preferably fluorine), including but not limited to —CHF2, —CH2F, —CF3, —CHF—CH2Cl.
The term “hydroxyalkyl group” refers to an alkyl group substituted with one or more (e.g., 1, 2, 3 or 4)hydroxyl groups, including but not limited to —CH2OH, —CHOH—CH2OH. C1-4 hydroxyalkyl groups include but are not limited to —CH2OH, —CHOH—CH2OH, —CH(CH2OH)—CH2OH.
The term “alkenyl group” refers to a straight or branched monovalent hydrocarbon group with a certain number of carbon atoms and contains at least one carbon-carbon double bond, where the carbon-carbon double bond can be located anywhere within the alkenyl group
A C2-C6 alkenyl group refers to an alkenyl group with 2-6 carbon atoms. These include the C2 alkenyl group, the C3 alkenyl group, C4 alkenyl group, C5 alkenyl group, and C6 alkenyl group. Examples of alkenyl groups include, but are not limited to, vinyl group, propenyl group, butenyl group, pentenyl group, hexenyl group, butadienyl group, pentadienyl group, hexadienyl group.
The term “alkoxy group” refers to —O—RX, where RX is an alkyl group as defined above. In some embodiments, a C1-4 alkoxy group can be a methoxy group, ethoxy group, n-propoxy group, isopropoxy group, n-butoxy group, isobutoxy group, sec-butoxy group, or tert-butoxy group.
The term “cycloalkyl group” refers to a saturated monocyclic or polycyclic (e.g., concatenated, spiro, or bridged) hydrocarbon group formed by carbon atoms. In some embodiments, the cycloalkyl group is a monocyclic group. In some embodiments, a C3-6 cycloalkyl group can be a cyclopropyl group, cyclobutyl group, cyclopentyl group, or cyclohexyl group. In some embodiments, a C3-8 cycloalkyl group can be a C3-6 cycloalkyl group, such as a cyclopropyl group, cyclobutyl group, cyclopentyl group, or cyclohexyl group.
i. The term “heteroaryl group” or “heteroaromatic ring” refers to an aromatic cyclic group formed by a carbon atom and at least one heteroatom, wherein the heteroatom may be N, O and S. A 5-membered heteroaryl group or heteroaromatic ring is, for example, a furan, thiophene, pyrrole, pyrazole, oxazole, thiazole, imidazole or triazole. 6-membered heteroaryl group or heteroaromatic ring or, for example, pyrazine, pyridazine, pyridine or pyrimidine.
The “x-y membered” in the cyclic group described in the present invitation means that the number of atoms in the ring is x-y. For example, cyclopropyl group is a 3-membered group, tetrahydropyrrolyl group is a 5-membered group, and piperidinyl group is a 6-membered group.
The term “substitution” or “substituent” refers to the substitution of one or more hydrogen atoms by a specified group. When no substitution position is specified, the substitution may be at any position, but only the formation of a stable or chemically viable chemical is permitted.
When any variable (e.g. R) appears more than once in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2 R's, said group may optionally be substituted with up to two R's, and there are independent options for R in each case. Furthermore, combinations of substituents and/or variables are only allowed if such combinations would yield stable compounds. For example.
where w is 0, 1 or 2 and each R is independently a methyl group or fluorine, then
includes
etc.
(i) The term “treatment” refers to curative therapy. In the context of a specific disease or condition, treatment means (1) alleviating one or more biological manifestations of the disease or condition, (2) interfering with (a) one or more points in the biological cascade causing or contributing to the disease or condition or (b) one or more biological manifestations of the disease or condition, (3) ameliorating one or more symptoms, effects or side effects associated with the disease or condition, or one or more symptoms, effects or side effects associated with the disease, condition or its treatment, or (4) slowing the progression of the disease, condition or one or more biological manifestations of the disease or condition.
The term “therapeutically effective amount” means an amount of a compound that, when given to a patient, is sufficient to effectively treat or prevent the disease or condition described herein. The “therapeutically effective amount” will vary depending on the compound, the disease or condition, and its severity, and the age of the patient to be treated, but may be adjusted by those skilled in the art as needed. In general, the compounds of the present invention are used in the therapeutic dose range of 1-1000 mg/day for human use. Doses beyond this range may also be used depending on the dosage form and the severity of the disease or condition.
The pharmaceutical compositions can be made into various types of dosing unit dosage forms depending on the therapeutic purpose, such as tablets, pills, powders, liquids, suspensions, emulsions, granules, pellets, capsules and injections (solutions or suspensions), preferably tablets, capsules, liquids, suspensions and injections (solutions or suspensions).
The compounds described in the present invention can be administered clinically by means of oral administration, injection, etc.
The term “subject” means any animal, preferably mammalian, preferably human, that is about to receive or has received the administration of a compound or composition. The term “mammal” includes any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., with humans being preferred.
All patents and published publications covered herein are incorporated herein by reference in their entirety.
Each of the above preferred conditions can be combined in any way to obtain a better example of the invention without violating the common knowledge in the art.
The reagents and raw materials used in this invention are commercially available.
The positive progressive effect of the present invention is that the present invention provides a novel pyridocyclic compound that exhibits JAK inhibitory activity.
In order to further illustrate the invention, a series of embodiments are given below, which are entirely illustrative and are used only to describe the invention specifically and should not be construed as a limitation of the invention.
Step 1: Dissolve 4-chloro-7-azaindole (25.0 g, 163.8 mmol) in 250 mL of dichloromethane, add DMAP (2.0 g, 16.5 mmol), triethylamine (34.0 mL, 245.8 mmol) and stir at room temperature for 30 min. Dissolve benzenesulfonyl chloride (23.3 mL, 180.3 mmol) in 50 mL of dichloromethane and slowly add dropwise to the above reaction solution, stir for about 4 h at room temperature and then filter, collect the filtrate and concentrate under vacuum to obtain a brown solid. 4-chloro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine is obtained as an off-white solid (43.0 g, 90%) by triturating and curing with an appropriate amount of methanol. The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C13H10ClNO22S calculated value 293.0146, measured value 293.0139.
Step 2: Dissolve 4-chloro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (30.0 g, 102.7 mmol) in 300 mL of dichloromethane, add tetramethylammonium nitrate (28.0 g, 205.5 mmol) to the above solution at 25° C. and stir. Add trifluoroacetic anhydride (57.3 mL, 410.8 mmol) slowly dropwise while keeping the temperature of the reaction solution below 30° C. The mixture is stirred at room temperature for 5 h. The completion of the reaction is monitored by TLC. Saturated sodium bicarbonate is then added until the reaction solution was weakly basic, and the organic phase is separated. The aqueous phase is then extracted twice using dichloromethane. Combine the organic phases and wash with saturated NaCl solution, followed by drying over anhydrous sodium sulfate and concentrating under vacuum to yield a yellow solid. Triturating and curing with an appropriate amount of methanol yields 4-chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine as a light yellow solid (25.9 g, 75%). The product can be used directly in the next step without further purification. 1HNMR (300 MHz, DMSO-d6): δ=9.09 (s, 1H), 8.29 (d, J=3.0 Hz, 1H), 8.17 (t, J=4.5 Hz, 2H), 7.79 (t, J=7.5 Hz, 1H), 7.68 (t, J=7.5 Hz, 2H), 7.11 (d, J=3.0 Hz, 1H) ppm: HRMS (ESI): m/z [M+Na]+. C13H8ClN3NaO4S calculated value 359.9816, measured value 359.9801.
Step 3: Dissolve 4-chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (10.0 g, 29.7 mmol) in 100 mL of tetrahydrofuran, add DIPEA (7.7 g, 59.3 mmol) and 1-Boc-3-aminopyrrolidine (8.3 g, 44.5 mmol). Stir the mixture under reflux for 4 h at an elevated temperature. The reaction was monitored for completion by TLC. The reaction is then concentrated under vacuum to yield a yellow oily liquid. Tert-butyl 3-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate is obtained as a yellow solid (10.1 g, 70%) by triturating and curing with an appropriate amount of methanol. The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C22H26N5O6S calculated value 488.1598, measured value 488.1607.
Step 4: Dissolve tert-butyl 3-(((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (10.0 g, 20.5 mmol) in 100 mL of methanol, add palladium carbon (1 g, 10%) and keep the reaction under hydrogen atmosphere after replacing the air in the reaction flask with hydrogen gas more than three times. The reaction is carried out under hydrogen gas atmosphere at room temperature with stirring for 12 h. The completion of reaction is monitored by TLC. After extraction, the filtrate is then collected and concentrated under vacuum to yield 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidin-1-tert-butyl carboxylate as a light pink foamy solid (9.2 g, 98%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C22H28N5O4S calculated value 458.1857, measured value 458.1862.
Step 5: Dissolve triethyloxonium tetrafluoroborate (11.2 g, 59.1 mmol) and (R)-lactamide (5.3 g, 59.1 mmol) in 100 mL of tetrahydrofuran, stir at room temperature for 3 h and concentrate under vacuum to obtain an oily mixture, followed by the addition of 100 mL of ethanol to dissolve and the addition of tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (9.0 g, 19.7 mmol) The reaction is then stirred while maintaining reflux for 3 h. The completion of reaction is monitored by TLC. Saturated sodium bicarbonate is then added until the reaction solution is weakly basic, and the organic phase is separated. The aqueous phase is then extracted twice using dichloromethane. Combine the organic phases and wash with saturated NaCl solution, followed by drying over anhydrous sodium sulfate and concentrating under vacuum. The residue is then purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield tert-butyl 3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate as a light green oil (6.0 g, 60%). HRMS (ESI): m/z [M+H]+. C25H30N5O5S calculated value 512.1962, measured value 512.1983.
Step 6: Dissolve tert-butyl 3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate (6.0 g, 11.7 mmol) in 60 mL dichloromethane, slowly add trifluoroacetic acid (13.4 g, 117.4 mmol) and stir at room temperature for 12 h. The product is then concentrated under vacuum to yield Intermediate-1: (1R)-1-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol as a light brown oil (4.8 g, 100%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C20H22N5O3S calculated value 412.1438, measured value 412.1448.
Step 1: A solution of 2-diethoxyphosphoryl acetonitrile (5.7 g, 32.1 mmol) in THF (50 mL) is added to a solution of NaH (1.2 g, 30.7 mmol) in THF (50 mL) at 0° C. The mixture is then stirred at room temperature for 1 h. Then cool the mixture to 0° C., and add a solution of tert-butyl 3-oxazetidine-1-carboxylate (5.0 g, 29.2 mmol) in THF (50 mL) within 1 h. Stir the above reaction solution at room temperature for 16 hours. After the reaction is finished, add an appropriate amount of water and extract the aqueous phase twice with EA. Combine the organic layers and wash with saturated NaCl solution, dry over sodium sulfate and concentrate under vacuum to yield tert-butyl 3-(cyanomethyl)azetidine-1-carboxylate as a yellow solid (5.2 g, 78%). The product can be used directly in the next step without further purification. HRMS (EST): m/z [M+H]+. C10H15N2O2 calculated value 195.1128, measured value 195.1121.
Step 2: Add DBU (11.8 g, 77.4 mmol) to a solution of tert-butyl 3-(cyanomethyl)azetidine-1-carboxylate (5.0 g, 25.8 mmol) and 4-nitropyrazole (3.2 g, 28.7 mmol) in acetonitrile (30 mL). Stir the mixture at room temperature for 16 h. At the end of the reaction, add an appropriate amount of water and extract the aqueous phase twice with EA. Combine the organic layers and wash with saturated NaCl solution, dry over sodium sulfate and concentrate under vacuum. The residue is then purified using a silica gel column (petroleum ether:ethyl acetate=4:1) to yield tert-butyl 3-(cyanomethyl)-3-(4-nitro-1H-pyrazol-1-yl)azetidine-1-carboxylate as a white oil (4.0 g, 50%). HRMS (EST): m/z [M+H]+. C13H18N5O4 calculated value 308.1353, measured value 308.1346.
Step 3: Dissolve tert-butyl 3-(cyanomethyl)-3-(4-nitro-1H-pyrazol-1-yl)azetidine-1-carboxylate (4.0 g, 13.0 mmol) in 50 mL of methanol, add palladium carbon (0.4 g, 10%) and then use hydrogen gas to replace the air in the reaction flask more than three times. Keep the reaction under hydrogen gas atmosphere and stir for 12 h at room temperature. The completion of the reaction is monitored by TLC. The filtrate is then collected and concentrated under vacuum to yield tert-butyl 3-(4-amino-1H-pyrazol-1-yl)-3-(cyanomethyl)azetidine-1-carboxylate as a light brown foam solid (3.5 g, 98%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C13H20N5O2 calculated value 278.1612, measured value 278.1610.
Step 4: Dissolve 4-chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (3.5 g, 10.5 mmol) in 100 mL of tetrahydrofuran. Add DIPEA (4.1 g, 31.5 mmol) and tert-butyl 3-(4-amino-1H-pyrazol-1-yl)-3-(cyanomethyl)azetidine-1-carboxylate (3.5 g, 12.6 mmol) and then stir under reflux at an elevated temperature for 4 h. The completion of the reaction is monitored by TLC. The reaction is then concentrated under vacuum to yield a yellow oily liquid. Tert-butyl 3-(cyanomethyl)-3-(4-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)azetidine-1-carboxylate is obtained as a yellow solid (4.3 g, 70%) by triturating and curing using an appropriate amount of methanol. The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C26H27N5O6S calculated value 579.1769, measured value 579.1782.
Step 5: Dissolve tert-butyl 3-(cyanomethyl)-3-(4-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)azetidine-1-carboxylate ester (4.3 g, 7.4 mmol) in 50 mL of methanol, add palladium carbon (0.4 g, 10%) and replace the air in the reaction flask with hydrogen gas more than three times. The reaction is then carried out under hydrogen gas atmosphere with stirring at room temperature for 12 h. The completion of the reaction is monitored by TLC. After extraction, collect the filtrate and concentrate under vacuum to yield tert-butyl 3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)-3-(cyanomethyl)azetidine-1-carboxylate as a light brown foamy solid (4.0 g, 98%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C26H29N5O4S calculated value 549.2027, measured value 549.2043.
Step 6: Dissolve tert-butyl 3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)-3-(cyanomethyl)azetidine-1-carboxylate (4.0 g, 7.3 mmol) in 50 mL of DMF. Then add Na2S2O5 (6.9 g, 36.5 mmol) and 5-hydroxymethyl furfural (1.8 g, 14.6 mmol), and the reaction is stirred for 12 h at 90° C. The completion of the reaction is monitored by TLC. Saturated sodium bicarbonate is then added until the reaction solution is weakly basic, and the organic phase is separated. Extract the aqueous phase twice using dichloromethane. Combine the organic phases and wash with saturated NaCl solution, dry over anhydrous sodium sulfate and concentrate under vacuum. The residue is then purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield 3-(cyanomethyl)-3-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidine-1-carboxylate as a yellow solid (3.3 g, 69%). HRMS (ESI): m/z [M+H]+. C32H31N8O6S calculated value 655.2082, measured value 655.2063.
Step 7: Dissolve 3-(cyanomethyl)-3-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidine-1-carboxylate (3.0 g, 4.6 mmol) in 60 mL of dichloromethane, slowly add trifluoroacetic acid (5.2 g, 46.0 mmol) and stir at room temperature for 12 h. Concentrate under vacuum to yield Intermediate 2: 2-(3-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl) acetonitrile as a light yellow oil (2.3 g, 91%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C27H23N5O4S calculated value 555.1557, measured value 555.1555.
Step 1: Dissolve tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (3.0 g, 6.6 mmol) in 50 mL of DMF and add Na2S2O5 (6.2 g, 32.8 mmol) and 5-hydroxymethylfurfural (1.7 g, 13.2 mmol). The mixture is then stirred at 90° C. for 12 h. the completion of the reaction is monitored by TLC. Saturated sodium bicarbonate is then added until the reaction solution is weakly basic, and the organic phase is separated. Extract the aqueous phase twice with dichloromethane. Combine the organic phases and wash with saturated NaCl solution, dry over anhydrous sodium sulfate and concentrate under vacuum. The residue is then purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield tert-butyl 3-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl pyrrolidine-1-carboxylate as a yellow solid (2.4 g, 65%). HRMS (ESI): m/z [M+H]+. C28H30N5O6S calculated value 564.1911, measured value 564.1921.
Step 2: Dissolve tert-butyl 3-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl pyrrolidine-1-carboxylate (2.0 g, 3.6 mmol) in 50 mL dichloromethane, slowly add trifluoroacetic acid (4.1 g, 36.0 mmol) at room temperature and then stir for 12 h. The product is then concentrated under vacuum to yield
Intermediate-3:(5-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)furan-2-methanol as a light yellow oil (1.5 g, 91%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C23H22N5O4S calculated value 464.1387, measured value 464.1385.
Step 3: Dissolve tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (3.0 g, 6.6 mmol) in 50 mL of DMF and add Na2S2O5 (6.2 g, 32.8 mmol) and 2-imidazolecarboxaldehyde (1.3 g, 13.2 mmol). The mixture is then stirred at 90° C. for 12 h. The completion of the reaction is monitored by TLC. Saturated sodium bicarbonate is then added until the reaction solution is weakly basic, and the organic phase is separated. Extract the aqueous phase twice using dichloromethane. Combine the organic phases and wash with saturated NaCl solution, dry over anhydrous sodium sulfate and concentrate under vacuum. The residue is then purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield tert-butyl 3-(2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate as a yellow solid (2.2 g, 63%). HRMS (ESI): m/z [M+H]+. C26H28N7O4S calculated value 534.1918, measured value 534.1910.
Step 4: Dissolve tert-butyl 3-(2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate (2.0 g, 3.8 mmol) in 50 mL of dichloromethane, slowly add trifluoroacetic acid (4.3 g, 38.0 mmol) and stir at room temperature for 12 h. The product is then concentrated under vacuum to yield Intermediate 4: 2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine as a light yellow oil (1.2 g, 74%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C21H20N7O2S calculated value 434.1394, measured value 434.1390.
Step 1: Dissolve 4-chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (5.0 g, 14.8 mmol) in 50 mL of tetrahydrofuran, add DIPEA (3.8 g, 29.7 mmol) and (R)-(+)-1-Boc-3-aminopyrrolidine (4.1 g, 22.3 mmol). The mixture is then stirred while maintaining reflux at an elevated temperature for 4 hours. The completion of the reaction is monitored by TLC. Concentrate under vacuum to yield a yellow oily liquid. Triturating and curing with an appropriate amount of methanol yields tert-butyl 3-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (R) as a yellow solid (4.8 g, 66%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C22H26N5O6S calculated value 488.1598, measured value 488.1605.
Step 2: Dissolve tert-butyl 3-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (R) (4.8 g, 9.9 mmol) in 50 mL of methanol, add palladium carbon (0.5 g, 10%) and replace the air in the reaction flask more than three times using hydrogen gas. The reaction is then carried out under hydrogen gas atmosphere with stirring at room temperature for 12 h. The completion of the reaction is monitored by TLC. The filtrate is then collected and concentrated under vacuum to yield (R) tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate as a light pink foamy solid (4.2 g, 93%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z. [M+H]+. C22H28N5O4S calculated value 458.1857, measured value 458.1850.
Step 3: Dissolve triethyloxonium tetrafluoroborate (5.0 g, 26.3 mmol) and (R)-lactamide (2.3 g, 26.3 mmol) in 80 mL of tetrahydrofuran, stir for 3 h at room temperature and then concentrate the mixture under vacuum to obtain an oil-like substance, followed by adding 80 mL of ethanol to dissolve and adding (R)tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (4.0 g, 8.8 mmol). The reaction is then stirred while maintaining reflux for 3 h and the completion of the reaction is monitored by TLC. Saturated sodium bicarbonate is then added until the reaction solution is weakly basic, and the organic phase is separated. Extract the aqueous phase twice using dichloromethane. Combine the organic phases and wash with saturated NaCl solution, dry over anhydrous sodium sulfate and concentrate under vacuum. The residue is then purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield (R)-tert-butyl 3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-pyrrolidine-1-carboxylate as a light green oil (3.2 g, 72%). HRMS (ESI): m/z [M+H]+. C25H30N5O5S calculated value 512.1962, measured value 512.1974.
Step 4: Dissolve (R)-tert-butyl 3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-pyrrolidine-1-carboxylate (3.0 g, 5.9 mmol) in 50 mL of dichloromethane, slowly add trifluoroacetic acid (6.7 g, 58.6 mmol) and then stir for 12 h at room temperature. Concentrate under vacuum to yield Intermediate 5: (R)-1-(6-(phenylsulfonyl)-1-((R-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol as a light brown oil (2.2 g, 91%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C20H22N5O3S calculated value 412.1438, measured value 412.1433.
Step 5: Dissolve 4-chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (8.0 g, 23.7 mmol) in 80 mL of tetrahydrofuran. Add DIPEA (6.1 g, 47.5 mmol) and (S)-(−)-1-Boc-3-aminopyrrolidine (6.6 g, 35.6 mmol). Then, stir the reaction while maintaining reflux at an elevated temperature for 4 hours and monitor the reaction for completion by TLC. Concentrated under vacuum to yield a yellow oily liquid. Triturate and cure with an appropriate amount of methanol to yield tert-butyl 3-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (S) as a yellow solid (8.1 g, 70%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C22H26N5O6S calculated value 488.1598, measured value 488.1603.
Step 6: Dissolve tert-butyl 3-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (S) (8.0 g, 16.4 mmol) in 80 mL of methanol, add palladium carbon (0.8 g, 10%) and replace the air in the reaction vial more than three times using hydrogen gas. The reaction is then carried out under hydrogen gas atmosphere and stirred at room temperature for 12 h. The completion of the reaction is monitored by TLC. The filtrate is then collected and concentrated under vacuum to yield (S) tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate as a light pink foamy solid (7.4 g, 99%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C22H28N5O4S calculated value 458.1857, measured value 458.1850.
Step 7: Dissolve triethyloxonium tetrafluoroborate (8.7 g, 46.0 mmol) and (R)-lactamide (4.1 g, 46.0 mmol) in 100 mL of tetrahydrofuran, stir for 3 h at room temperature and then concentrate the mixture in vacuum to obtain an oily substance, followed by the addition of 100 mL of ethanol to dissolve, and the addition of (S) tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (7.0 g, 15.3 mmol) The mixture is then stirred while maintaining reflux at an elevated temperature for 3 h and the completion of the reaction is monitored by TLC. Saturated sodium bicarbonate is then added until the reaction solution is weakly basic, and the organic phase is separated. Extract the aqueous phase twice using dichloromethane. Combine the organic phases and wash with saturated NaCl solution, dry over anhydrous sodium sulfate and concentrate under vacuum. The residue is then purified using a silica gel column (petroleum ether:ethyl acetate =1:1) to yield (S)-tert-butyl 3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-pyrrolidine-1-carboxylate as a light green oil (5.3 g, 68%). HRMS (ESI): m/z [M+H]+. C25H30N5O5S calculated value 512.1962, measured value 512.1969.
Step 8: Dissolve (S)-tert-butyl 3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-pyrrolidine-1-carboxylate (5.0 g, 9.8 mmol) in 50 mL of dichloromethane, slowly add trifluoroacetic acid (11.2 g, 97.8 mmol) and then stir for 12 h at room temperature. Concentrate under vacuum to yield Intermediate 6: (R)-1-(6-(phenylsulfonyl)-1-((S-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol as a light brown oil (3.8 g, 94%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C20H22N5O3S calculated value 412.1438, measured value 412.1433.
Step 9: Dissolve (R) tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (3.0 g, 6.6 mmol) in 50 mL of DMF. Then, add NaS2O5 (6.2 g, 32.8 mmol) and 5-hydroxymethylfurfural (1.7 g, 13.2 mmol). Stir the mixture at 90° C. for 12 h. The completion of the reaction is monitored by TLC. Saturated sodium bicarbonate is then added until the reaction solution is weakly basic, and the organic phase is separated. Extract the aqueous phase twice using dichloromethane. Combine the organic phases and wash with saturated NaCl solution, dry over anhydrous sodium sulfate and concentrate under vacuum. The residue is then purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield (R)-tert-butyl 3-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate as a yellow solid (2.0 g, 54%). HRMS (EST): m/z [M+H]+. C28H30N5O6S calculated value 564.1911, measured value 564.1922.
Step 10: Dissolve (R)-tert-butyl 3-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate (2.0 g, 3.6 mmol) in 30 mL of dichloromethane, slowly add trifluoroacetic acid (4.1 g, 36.0 mmol) and stir for 12 h at room temperature. Intermediate 7, which is (R)-(5-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) furan-2-yl methanol is then obtained as a light yellow oil (1.6 g, 97%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C23H22N5O4S calculated value 464.1387, measured value 464.1380.
Step 11: Dissolve (R) tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (3.0 g, 6.6 mmol) in 50 mL of DMF. Then, add Na2S2O5 (6.2 g, 32.8 mmol) and 2-imidazolecarboxaldehyde (1.3 g, 13.2 mmol) and stir the mixture at 90° C. for 12 h. The completion of the reaction is monitored by TLC. Saturated sodium bicarbonate is then added until the reaction solution is weakly basic, and the organic phase is separated. Extract the aqueous phase twice using dichloromethane. Combine the organic phases and wash with saturated NaCl solution, dry over anhydrous sodium sulfate and concentrate under vacuum. The residue is then purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield (R)-tert-butyl 3-(2-(l H-imidazol-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-pyrrolidine-1-carboxylate as a yellow solid (2.5 g, 71%). HRMS (ESI): m/z [M+H]+. C26H28N7O4S calculated value 534.1918, measured value 534.1910.
Step 12: Dissolve (R)-tert-butyl 3-(2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-pyrrolidine-1-carboxylate (2.5 g, 4.7 mmol) in 30 mL of dichloromethane, slowly add trifluoroacetic acid (5.3 g, 46.9 mmol) and then stir at room temperature for 12 h. Concentrate under vacuum to obtain Intermediate 8: (R)-2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine as a light yellow oil (1.7 g, 84%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C21H20N7O2S calculated value 434.1394, measured value 434.1390.
Step 1: Dissolve 4-((tert-butoxycarbonyl)amino)cyclohexanecarboxylic acid (10.0 g, 41.1 mmol) in 100 mL of tetrahydrofuran and cool down to −15° C. Add isobutyl chloroformate (11.3 g, 82.2 mmol) at low temperature and stir at low temperature for 1 h. Then, slowly add 100 mL of ammonia and stir at room temperature for 4 h. After the reaction is complete, filter the product and tert-butyl (4-carbamoylcyclohexyl)carbamate is obtained as a white solid (8.0 g, 80%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C12H23N2O3 calculated value 243.1703, measured value 243.1698.
Step 2: Dissolve tert-butyl (4-carbamoylcyclohexyl)carbamate (8.0 g, 33.1 mmol) in 50 mL of pyridine and cool down to −15° C. Add phosphorus oxychloride (10 g, 66.2 mmol) slowly dropwise at low temperature and stir at low temperature for 3 h. After the reaction is complete, pour the reaction solution slowly into 200 mL of ice-water mixture. Extract the aqueous phase three times with EA. Combine the organic phases and wash with saturated NaCl solution, dry with anhydrous sodium sulfate and concentrate under vacuum to yield tert-butyl (4-cyanocyclohexyl)carbamate as a milky-white solid (5.5 g, 74%). HRMS (ESI): m/z [M+H]+. C12H21N2O2 calculated value 225.1598, measured value 225.1603.
Step 3: Dissolve tert-butyl (4-cyanocyclohexyl)carbamate (5.5 g, 24.6 mmol) in 50 mL of dichloromethane and slowly add trifluoroacetic acid (28.0 g, 245.5 mmol). Stir the mixture at room temperature for 12 h. The product is then concentrated under vacuum to yield trans-4-aminocyclohexanecarbonitrile as a light white solid (2.5 g, 82%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C7H13N2 calculated value 125.1073, measured value 125.1082.
Step 4: Dissolve 4-chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (4.5 g, 13.4 mmol) in 50 mL of tetrahydrofuran, add DIPEA (3.5 g, 26.9 mmol) and trans-4-aminocyclohexanecarbonitrile (2.5 g, 20.2 mmol) and stir for 4 h at an elevated temperature while maintaining reflux. The completion of the reaction is monitored by TLC. Concentrate under vacuum to obtain a yellow oily liquid. Triturate and cure with an appropriate amount of methanol to yield trans-4-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile as a yellow solid (4.2 g, 74%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C20H20N5O4S calculated value 426.1231, measured value 426.1237.
Step 5: Dissolve trans-4-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (4.2 g, 9.9 mmol) in 50 mL of methanol, add palladium carbon (0.4 g, 10%) and replace the air in the reaction flask with hydrogen gas more than three times. The reaction is then carried out under hydrogen gas atmosphere and stirred at room temperature for 12 h. The completion of the reaction is monitored by TLC. After filtration, collect the filtrate and concentrate under vacuum to yield Intermediate 9: trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile as a light brown foamy solid (3.5 g, 90%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C20H22N5O2S calculated value 396.1489, measured value 396.1492.
Step 1: Dissolve 4-nitropyrazole (5 g, 44.2 mmol) in 50 mL of DMF, add potassium carbonate (12.2 g, 88.4 mmol) and 3-bromopropionitrile (8.9 g, 66.3 mmol), Then, stir the mixture at room temperature for 12 h. The completion of the reaction is monitored by TLC. Saturated sodium bicarbonate is then added until the reaction solution is weakly basic, and the organic phase is separated. Extract the aqueous phase three times using dichloromethane. Combine the organic phases and wash with saturated NaCl solution, dry over anhydrous sodium sulfate and concentrate under vacuum to yield 3-(4-nitro-1H-pyrazol-1-yl)propanenitrile as a pale yellow solid (4.8 g, 65%) The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C6H7N4O2 calculated value 167.0564, measured value 167.0569.
Step 2. Dissolve 3-(4-nitro-1H-pyrazol-1-yl)propanenitrile (4.8 g, 28.9 mmol) in 50 mL of methanol, add palladium carbon (0.5 g, 10%) and use hydrogen gas to replace the air in the reaction flask more than three times The reaction is then carried out under hydrogen atmosphere and stirred at room temperature for 12 h. The completion of the reaction is monitored by TLC. After filtration, the filtrate is then collected and concentrated under vacuum to yield 3-(4-amino-1H-pyrazol-1-yl)propanenitrile as a light brown foamy solid (3.5 g, 89%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C6H9N4 calculated value 137.0822, measured value 137.0833.
Step 3: Dissolve 4-chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (5.8 g, 17.2 mmol) in 60 mL of tetrahydrofuran. Add DIPEA (4.4 g, 34.4 mmol) and 3-(4-amino-1H-pyrazol-1-yl)propanenitrile (3.5 g, 25.7 mmol). The mixture is stirred while maintaining reflux for 4 hours at an elevated temperature and the completion of the reaction is monitored by TLC. Subsequent concentration under vacuum yields a yellow oily liquid. Triturating and curing with an appropriate amount of methanol yields 3-(4-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)propanenitrile as a yellow solid (5.1 g, 68%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C19H16N7O4S calculated value 438.0979, measured value 438.0985.
Step 4: Dissolve 3-(4-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)propanenitrile (5.0 g, 11.4 mmol) in 50 mL of methanol, add palladium carbon (0.5 g, 10%) and replace the air in the reaction flask more than three times using hydrogen gas. The reaction is then carried out under hydrogen gas atmosphere and stirred at room temperature for 12 h. The completion of the reaction is monitored by TLC. After filtration, collect the filtrate and concentrate under vacuum to yield Intermediate 10: 3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)propanenitrile as a light yellow foamy solid (4.5 g, 97%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C19H18N7O2S calculated value 408.1237, measured value 408.1241.
Step 5: Dissolve 4-nitropyrazole (2.5 g, 22.1 mmol) in 50 mL of DMF, add potassium carbonate (6.1 g, 44.2 mmol) and 2-bromoacetonitrile (4.0 g, 33.2 mmol) and stir at room temperature for 12 h. The completion of the reaction is monitored by TLC. Saturated sodium bicarbonate is then added until the reaction solution is weakly basic, and the organic phase is separated. Extract the aqueous phase three times using dichloromethane. Combine the organic phases and wash with saturated NaCl solution, dry over anhydrous sodium sulfate and concentrate under vacuum to yield 3-(4-nitro-1H-pyrazol-1-yl)acetonitrile as a pale yellow solid (2.2 g, 65%) The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C5H5N4O2 calculated value 153.0407, measured value 153.0411.
Step 6: Dissolve 3-(4-nitro-1H-pyrazol-1-yl)acetonitrile (2.2 g, 14.5 mmol) in 30 mL of methanol, add palladium carbon (0.3 g, 10%) and then use hydrogen gas to replace the air in the reaction flask more than three times. The reaction is then carried out under hydrogen gas atmosphere and stirred at room temperature for 12 h. The completion of the reaction is monitored by TLC. After filtration, collect the filtrate and concentrate under vacuum to yield 3-(4-amino-1H-pyrazol-1-yl)acetonitrile as a light brown foamy solid (1.6 g, 91%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C5H7N4 calculated value 123.0665, measured value 123.0671.
Step 7: Dissolve 4-chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (2.8 g, 8.2 mmol) in 60 mL of tetrahydrofuran. Add DIPEA (2.1 g, 16.4 mmol) and 3-(4-amino-1H-pyrazol-1-yl)acetonitrile (1.5 g, 12.3 mmol) and then stir the mixture while maintaining reflux at an elevated temperature for 4 h. The completion of the reaction is monitored by TLC. Concentrate under vacuum to yield a yellow oily liquid. Then triturate and cure with an appropriate amount of methanol to yield 3-(4-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)acetonitrile as a yellow solid (2.5 g, 72%). The product can be used directly in the next step without further purification. HRMS (EST): m/z [M+H]+. C18H14N7O4S calculated value 424.0822, measured value 424.0833.
Step 8: Dissolve 3-(4-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)acetonitrile (2.5 g, 5.9 mmol) in 30 mL of methanol, add palladium carbon (0.3 g, 10%) and replace the air in the reaction flask more than three times using hydrogen gas. The reaction is then carried out under hydrogen gas atmosphere at room temperature with stirring for 12 h. The completion of the reaction is monitored by TLC. Collect the filtrate and concentrate under vacuum to yield Intermediate 11: 3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)acetonitrile as alight yellow foamy solid (2.0 g, 86%). The product can be used directly in the next step without further purification. HRMS (EST): m/z [M+H]+. C18H16N7O2S calculated value 394.1081, measured value 394.1084.
Step 1: Dissolve 4-chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (2.0 g, 5.9 mmol) in 50 mL of toluene. Then, add tris(dibenzalacetone)dipalladium (0.2 g, 10%), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.2 g, 10%) and p-cyanoaniline (1.4 g, 11.9 mmol). The reaction is then carried out under nitrogen gas protection and stirred while maintaining reflux at an elevated temperature for 5 h. The completion of the reaction is monitored by TLC. Saturated sodium bicarbonate is then added until the reaction solution is weakly basic, and the organic phase is separated. Extract the aqueous phase three times using dichloromethane. Combine the organic phases and wash with saturated NaCl solution, dry over anhydrous sodium sulfate and concentrate under vacuum. The residue is then purified using a silica gel column (petroleum ether:ethyl acetate=2:1) to yield 4-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)benzonitrile as a yellow solid (1.2 g, 48%). hRMS (ESI): m/z [M+H]+. C20H14N5O4S calculated value 420.0761, measured value 420.0771.
Step 2: Dissolve 4-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)benzonitrile (1.2 g, 2.9 mmol) in 30 mL of methanol, add palladium carbon (0.2 g, 10%) and then use hydrogen gas to replace the air in the reaction flask more than three times. The reaction is then carried out under hydrogen gas atmosphere and stirred at room temperature for 12 h. The completion of the reaction is monitored by TLC. Collect the filtrate and concentrate under vacuum to yield Intermediate 12: 4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2.3-b]pyridin-4-yl)amino)benzonitrile as a yellow foamy solid (1.1 g, 99%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C20H16N5O2S calculated value 390.1019, measured value 390.1023.
Step 1; Dissolve 4-chloro-1H-pyrazolo[3,4-b]pyridine (10.0 g, 65.4 mmol) in 100 mL of dichloromethane. Add DMAP (0.8 g, 6.5 mmol), triethylamine (18.0 mL, 130.7 mmol) and stir at room temperature for 30 min. Dissolve benzenesulfonyl chloride (10.1 mL, 78.5 mmol) in 50 mL of dichloromethane and slowly add the solution dropwise to the above reaction solution, stir for about 4 h at room temperature and then filter, collect the filtrate and concentrate under vacuum to obtain a brown solid. 4-chloro-1-(phenylsulfonyl)-1H-pyrazolo[3,4-b]pyridine is then obtained as an off-white solid (17.0 g, 89%) by triturating and curing with an appropriate amount of methanol. The product can be used directly in the next step without further purification. HRMS (EST): m/z [M+H]+. C12H9ClN3O2S calculated value 294.0099, measured value 293.0089.
Step 2: Dissolve 4-chloro-1-(phenylsulfonyl)-1H-pyrazolo[3,4-b]pyridine (15.0 g, 51.2 mmol) in 150 mL of dichloromethane, add tetramethylammonium nitrate (13.9 g, 102.3 mmol) to the above solution at 25° C. and stir. Trifluoroacetic anhydride (28.6 mL, 204.8 mmol) is then added slowly dropwise while maintaining the temperature of the reaction solution below 30° C. Stir the mixture at room temperature for 5 h. The completion of the reaction is monitored by TLC. Saturated sodium bicarbonate is then added until the reaction solution is weakly basic, and the organic phase is separated. Extract the aqueous phase twice using dichloromethane. Combine the organic phases and wash with saturated NaCl solution, dry over anhydrous sodium sulfate and concentrate under vacuum to yield a yellow solid. Triturate and cure with an appropriate amount of methanol to yield 4-chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrazolo[2,3-b]pyridine as alight yellow solid (12.3 g, 71%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C12H8ClN4O4S calculated value 338.9949, measured value 338.9930.
Step 3: Dissolve 4-chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrazolo[2,3-b]pyridine (5.0 g, 14.8 mmol) in 50 mL of tetrahydrofuran, followed by the addition of DIPEA (3.8 g, 29.6 mmol) and (S)-(−)-1-Boc-3-aminopyrrolidine (4.2 g, 22.2 mmol). The mixture is then stirred while maintaining reflux at an elevated temperature for 4 hours and the completion of the reaction is monitored by TLC. Concentrate under vacuum to yield a yellow oily liquid. Triturating and curing with an appropriate amount of methanol yields (S)tert-butyl 3-(((5-nitro-1-(phenylsulfonyl)-1H-pyrazolo[3,4-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate as a yellow solid (5.1 g, 71%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C21H25N6O6S calculated value 489.1551, measured value 488.1559.
Step 4: Dissolve (S)tert-butyl 3-(((5-nitro-1-(phenylsulfonyl)-1H-pyrazolo[3,4-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (5.0 g, 10.2 mmol) in 50 mL of methanol, add palladium carbon (0.5 g, 10%) and replace the air in the reaction flask with hydrogen gas more than three times. The reaction is then carried out under hydrogen gas atmosphere at room temperature with stirring for 12 h. The completion of the reaction is monitored by TLC. The filtrate is then collected and concentrated under vacuum to yield (S)tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrazolo[3,4-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate as a pink foamy solid (4.5 g, 96%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C21H27N6O4S calculated value 459.1809, measured value 459.1813.
Step 5: Dissolve triethyloxonium tetrafluoroborate (5.0 g, 26.1 mmol) and (R)-lactamide (2.3 g, 26.1 mmol) in 50 mL of tetrahydrofuran, stir at room temperature for 3 h and then concentrate the mixture under vacuum to obtain an oily mixture, followed by the addition of 50 mL of ethanol to dissolve and the addition of (S)tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrazolo[3,4-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (4.0 g, 8.7 mmol). The reaction is then stirred while maintaining reflux for 3 h at an elevated temperature. The completion of the reaction is monitored by TLC. Saturated sodium bicarbonate is then added until the reaction solution is weakly basic, and the organic phase is separated. Extract the aqueous phase twice using dichloromethane. Combine the organic phases and wash with saturated NaCl solution, dry over anhydrous sodium sulfate and concentrate under vacuum. The residue is then purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield (S)-tert-butyl 3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrazolo[3,4-b]pyridin-1(6H)-pyrrolidine-1-carboxylate as a green oil (2.2 g, 49%). HRMS (ESI): m/z [M+H]+. C24H29N6O5S calculated value 513.1915, measured value 513.1923.
Step 6: Dissolve (S)-tert-butyl 3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrazolo[3,4-b]pyridin-1(6H)-pyrrolidine-1-carboxylate (2.0 g, 3.9 mmol) in 20 mL of dichloromethane, slowly add trifluoroacetic acid (4.4 g, 39.0 mmol) and then stir for 12 h at room temperature. Concentrate under vacuum to yield Intermediate 13: (R)-1-(6-(phenylsulfonyl)-1-((S)-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrazolo[3,4-b]pyridin-2-yl) ethanol as a light brown oil (1.5 g, 91%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C19H21N6O3S calculated value 413.1390, measured value 413.1399.
Tert-butyl 3-(2-((R)-1-hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate
Step 1: Tert-butyl 3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate (0.4 g, 0.8 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, and stirred at room temperature for 5 h after adding 5 mL of 1 M sodium hydroxide. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS01: tert-butyl 3-(2-((R)-1-hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate (0.2 g, 69%), 69%). 1 HNMR (300 MHz, DMSO-d6): δ=11.95 (s, 1H), 8.59 (s, 1H), 7.51 (s, 1H), 6.35 (s, 1H), 5.75 (t, J=3.0 Hz, 2H), 5.17-5.21 (m, 1H), 3.82-3.93 (m, 2H), 3.76 (t, J=9.0 Hz, 1H), 3.40-3.47 (m, 1H), 2.58-2.74 (m, 1H), 2.28-2.41 (m, 1H), 1.65 (d, J 9.0 Hz, 3H), 1.45 (d, J=24.0 Hz, 9H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 153.23, 148.61, 148.53, 142.1, 129.04, 127.15, 120.71, 115.63, 99.37, 79.85, 63.60, 58.31, 53.58, 48.13, 28.47, 27.05, 22.81 ppm: HRMS (ESI): m/z [M+H]+. C19H26N5O3 calculated value 372.2030, measured value 372.2018.
Step 1: Dissolve (1R)-1-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) and then slowly add p-cyanobenzoyl chloride (0.2 g, 1.4 mmol) dropwise. Raise the temperature and stir under reflux for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phase was then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 4-(3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-carbonyl)benzonitrile as a brown oil (0.3 g, 76%). The product can be used directly in the next step without further purification. HRMS (ESI). m/z [M+H]+. C28H25N6O4S calculated value 541.1653, measured value 541.1643.
Step 2: 4-(3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-carbonyl)benzonitrile (0.3 g, 0.6 mmol) was dissolved in a solvent mixture of 5 mL of tetrahydrofuran and 5 mL of methanol and stirred at room temperature for 5 h after the addition of 5 mL of 1 M sodium hydroxide. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS02: 4-(3-(2-((R)-1-hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-carbonyl)benzonitrile (0.1 g, 45%). 1 HNMR (300 MHz, DMSO-d6): δ=11.96 (s, 1H), 8.87 (s, 1H), 8.21 (d, J=9.0 Hz, 2H), 8.10 (d, J=9.0 Hz, 2H), 7.66 (s, 1H), 6.88 (s, 1H), 4.28-4.69 (m, 1H), 3.81-3.92 (m, 2H), 3.74-3.79 (m, 1H), 3.30-3.45 (m, 2H), 1.93-2.15 (m, 2H), 1.49 (d, J=9.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO6) δ 172.53, 148.69, 148.51, 142.16, 139.53, 132.09, 129.04, 127.91, 127.16, 120.75, 118.63, 115.61, 113.69, 99.39, 63.60, 58.45, 52.57, 47.13, 27.11, 22.82 ppm; HRMS (ESI): m/z [M+H]+. C2H21N6O2 calculated value 401.1721, measured value 401.1723.
Step 1: Dissolve (1R)-1-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by slow dropwise addition of p-fluorobenzoyl chloride (0.2 g, 1.1 mmol). The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phase was then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (4-fluorophenyl)(3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)methanone as a brown oil (0.3 g, 77%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C27H25FN5O4S calculated value 534.1606, measured value 534.1619.
Step 2: (4-fluorophenyl)(3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)methanone (0.3 g, 0.6 mmol) was dissolved in a solvent mixture of 5 mL of tetrahydrofuran and 5 mL of methanol. After the addition of 5 mL of 1 M sodium hydroxide, the mixture was stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate =1:1) to yield LXS03: (4-Fluorophenyl)(3-(2-((R)-1-Hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)methanone (0.1 g, 45%). 1 HNMR (3M) MHz, DMSO-d6): δ=11.97 (s, 1H), 8.60 (d, J=6.0 Hz, 1H), 7.74-7-76 (m, 1H), 7.55 (d, J=15.0 Hz, 2H), 7.36 (t, J=9.0 Hz, 1H), 7.22 (t, J=9.0 Hz, 1H) Hz, 1H), 6.45 (d, J=48.0 Hz, 1H), 5.77 (s, 2H), 5.15-5.23 (m, 1H), 3.97-4.20 (m, 2H), 3.71-3.93 (m, 2H), 2.66-2.73 (m, 1H), 1.67 (d, J=6.0 Hz, 3H), 1.38 (t, J=37.5 Hz, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 172.58, 163.91, 148.83, 148.29, 142.54, 130.84, 129.03, 128.83, 127.29, 120.85, 115.61, 115.33, 99.57, 63.67, 58.49, 52.59, 47.83, 27.29, 22.89 ppm, HRMS (ESI): m/z [M+H]+. C21H21FNSO2 calculated value 394.1674, measured value 394.1682.
Step 1: Dissolve (1R)-1-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by the slow dropwise addition of ethyl sulfonyl chloride (0.2 g, 1.1 mmol). Then stir under reflux by heating for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phase was then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (1R)-1-(1-(1-(1-(ethylsulfonyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol as a yellow oil (0.3 g, 82%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C22H26N5O5S2 calculated value 504.1370, measured value 504.1389.
Step 2: (1R)-1-(1-(1-(1-(ethylsulfonyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.3 g, 0.6 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, and 5 mL of 1M sodium hydroxide was added and then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS04: (1R)-1-(1-(1-(ethylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-ol (0.2 g, 93%). 1 HNMR (300 MHz, DMSO-d6): δ=12.03 (s, 1H), 8.97 (s, 1H), 7.69 (s, 1H), 6.78 (s, 1H), 4.184.77 (m, 1H), 3.60-3.79 (m, 1H), 3.40-3.51 (m, 2H), 3.11-3.32 (m, 2H), 2.74-2.89 (m, 2H), 1.90-2.15 (m, 2H), 1.49 (d, J=6.0 Hz, 3H), 1.22 (t, J=9.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.77, 148.56, 142.77, 129.19, 127.31, 120.77, 115.68, 113.69, 99.48, 63.29, 57.53, 56.10, 51.11, 50.09, 26.20, 22.89, 2.69 ppm; HRMS (ESI): m/z [M+H]+. C16H22N5O3S calculated value 364.1438, measured value 364.1472.
Step 1: Dissolve (1R)-1-6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by slow dropwise addition of propylsulfonyl chloride (0.2 g, 1.1 mmol). Then stir under reflux by heating for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (1R)-1-(1-(1-(1-(propylsulfonyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol as a yellow oil (0.3 g, 80%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C23H28N5O5S2 calculated value 518.1526, measured value 518.1536.
Step 2: (1R)-1-(1-(1-(1-(propylsulfonyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.3 g, 0.6 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, and 5 mL of 1M sodium hydroxide was added, and then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS05: (1R)-1-(1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-ol (0.2 g, 91%). 1 HNMR (300 MHz, DMSO-d6): δ=11.93 (s, 1H), 8.77 (s, 1H), 7.58 (s, 1H), 6.67 (s, 1H), 4.21-4.68 (m, 1H), 3.51-3.73 (m, 1H), 3.21-3.28 (m, 2H), 3.18 (t, J=9.0 Hz, 2H), 2.79-2.99 (m, 2H), 1.83-2.25 (m, 2H), 1.52-1.68 (m, 2H), 1.49 (d, J=9.0 Hz, 3H), 0.90 (t, J=6.0 Hz, 3H) ppm; 13C NMR (75 MHz, DMSO-d6) δ 148.66, 148.59, 142.19, 129.02, 127.15, 120.73, 115.61, 113.69, 99.31, 63.66, 60.40, 57.59, 56.19, 50.02, 26.28, 22.84, 13.33, 12.44 ppm; HRMS (ESI): m/z [M+H]+. C17H24NSO3S calculated value 378.1594, measured value 378.1599.
Step 1: Dissolve (1R)-1-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by the slow dropwise addition of butylsulfonyl chloride (0.2 g, 1.1 mmol). Then stir under reflux by heating for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (1R)-1-(1-(1-(1-(butylsulfonyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol as a pale white oil (0.3 g, 77%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C24H30N5O5S2 calculated value 532.1683, measured value 532.1703.
Step 2: (1R)-1-(1-(1-(1-(butylsulfonyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.3 g, 0.6 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, and 5 mL of 1 M sodium hydroxide was added, and then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS06: (1R)-1-(1-(1-(butylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-ol (0.2 g, 91%). 1 HNMR (300 MHz, DMSO-d6): δ=12.03 (s, 1H), 8.47 (s, 1H), 7.68 (s, 1H), 6.87 (s, 1H), 4.41-4.62 (m, 1H), 3.50-3.79 (m, 1H), 3.11-3.22 (m, 2H), 3.10 (t, J=6.0 Hz, 2H), 2.77-2.80 (m, 2H), 1.93-2.15 (m, 2H), 1.51-1.61 (m, 2H), 1.48 (d, J=6.0 Hz, 3H), 1.29-1.31 (m, 2H), 0.93 (t, J=9.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.65, 148.51, 142.18, 129.04, 127.11, 120.79, 115.61, 99.39, 63.68, 57.93, 57.58, 56.17, 50.01, 26.28, 22.82, 21.93, 21.05, 13.88 ppm; HRMS (ESI): m/z [M+H]+. C18H26N5O3S calculated value 392.1751, measured value 392.1755.
Step 1: Dissolve (1R)-1-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by the slow dropwise addition of o-fluorobenzenesulfonyl chloride (0.2 g, 1.1 mmol). Then stir under reflux by heating for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (1R)-1-(1-(1-(1-(((2-fluorophenyl)sulfonyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol as a light yellow oil (0.3 g, 72%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C26H25FN5O5S2 calculated value 570.1276, measured value 570.1257.
Step 2: (1R)-1-(1-(1-(1-(((2-fluorophenyl)sulfonyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.5 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, and 5 mL of 1 M sodium hydroxide was added, and then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS07: (1R)-1-(1-(1-((2-fluorophenyl)sulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-ol (0.1 g, 44%). 1 HNMR (300 MHz, DMSO-d6): δ=12.16 (s, 1H), 8.87 (s, 1H), 7.80-7.84 (m, 1H), 7.63-7.75 (m, 1H), 7.60 (s, 1H), 7.48-7.52 (m, 1H), 7.21-7.43 (m, 1H), 6.77 (s, 1H), 4.51-4.70 (m, 1H), 3.63-3.81 (m, 1H), 3.09-3.33 (m, 2H), 2.63-2.83 (m, 2H), 1.89-2.19 (m, 2H), 1.93 (d, J=9.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 158.22, 148.66, 148.57, 142.18, 133.53, 129.03, 128.91, 127.11, 126.33, 124.69, 120.74, 115.84, 115.64, 99.38, 63.69, 57.58, 55.70, 49.63, 26.24, 22.89 ppm; HRMS (ESI): m/z [M+H]+. C20H21FN5O3S calculated value 430.1344, measured value 430.1350.
Step 1: Dissolve (1R)-1-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) and slowly add p-toluenesulfonyl chloride (0.2 g, 1.4 mmol) dropwise, then raise the temperature while maintaining reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (1R)-1-(6-(phenylsulfonyl)-1-(1-toluenesulfonylpyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol as a yellow oil (0.3 g, 73%). The product can be used directly in the next step without further purification. HRMS (EST): m/z [M+H]+. C27H28N5O5S2 calculated value 566.1526, measured value 566.1523.
Step 2: (1R)-1-(6-(phenylsulfonyl)-1-(1-toluenesulfonylpyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.3 g, 0.5 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol. Then, 5 mL of 1 M sodium hydroxide was added and the mixture was stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS08: (1R)-1-(1-(1-tosylpyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-ol (0.2 g, 89%). 1 HNMR (300 MHz, DMSO-d6): δ=11.95 (s, 1H), 8.84 (s, 1H), 7.74 (d, J=7.5 Hz, 2H), 7.68 (s, 1H), 7.40 (d, J=7.5 Hz, 2H), 6.87 (s, 1H), 4.53-4.68 (m, 1H), 3.55-3.81 (m, 1H), 3.10-3.25 (m, 2H), 2.79-2.89 (m, 2H), 2.34 (s, 3H), 1.91-2.25 (m, 2H), 1.43 (d, J=4.5 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.63, 148.59, 143.33, 142.17, 137.66, 129.39, 129.00, 128.33, 127.16, 120.71, 115.65, 99.39, 63.64, 57.58, 55.74, 49.63, 26.23, 22.81, 21.39 ppm. HRMS (EST): m/z [M+H]. C21H24N5O3S calculated value 426.1594, measured value 426.1596.
Step 1: Dissolve (1R)-1-6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) and slowly add p-phenylbenzenesulfonyl chloride (0.3 g, 1.1 mmol) dropwise. Raise the temperature while maintaining reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (1R)-1-(1-(1-(1-([[1,1′-biphenyl]-4-yl]sulfonyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol as a yellow oil (0.3 g, 66%), 66%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C32H30N5O5S2 calculated value 628.1683, measured value 628.1686.
Step 2: (1R)-1-(1-(1-(1-([[1,1′-Biphenyl]-4-yl sulfonyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d ]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.5 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol. After adding 5 mL of 1 M sodium hydroxide, the mixture was stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate =1:1) to yield LXS09: (1R)-1-(1-(1-([1,1′-biphenyl]-4-ylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-ol (0.2 g, 86%). 1 HNMR (300 MHz, DMSO-d6): δ=12.35 (s, 1H), 8.74 (s, 1H), 7.88-7.92 (m, 4H), 7.67 (s, 1H), 7.45-7.52 (m, 4H), 7.28-7.41 (m, 1H), 6.87 (s, 1H), 4.43-4.71 (m, 1H), 3.52-3.79 (m, 1H), 3.11-3.35 (m, 2H), 2.77-2.88 (m, 2H), 1.90-2.15 (m, 2H), 1.49 (d, J=6.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.69, 148.53, 142.19, 140.88, 138.62, 133.06, 129.21, 129.04, 127.93, 127.88, 127.11, 127.10, 120.73, 115.63, 99.38, 63.63, 57.58, 55.70, 49.62, 26.25, 22.81 ppm: HRMS (EST): m/z [M+H]+. C26H26N5O3S calculated value 488.1751, measured value 488.1757.
Step 1: Dissolve (1R)-1-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by slow dropwise addition of naphthalenesulfonyl chloride (0.3 g, 1.1 mmol). Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (1R)-1-(1-(1-(1-naphthalen-2-yl sulfonyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol as a pale yellow oil (0.3 g, 68%). The product can be used directly in the next step without further purification. HRMS (ESI); m/z [M+H]+. C30H28N5O5S2 calculated value 602.1526, measured value 602.1540.
Step 2: (1R)-1-(1-(1-(1-naphthalen-2-yl sulfonyl)pyrrolidin-3-yl)-6-(benzenesulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.3 g, 0.5 mmol) was dissolved in a mixture of 5 mL tetrahydrofuran and 5 mL methanol, and 5 mL of 1 M sodium hydroxide was added, and then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS10: (1R)-1-(1-(1-(naphthalen-2-ylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-ol (0.2 g, 87%). 1 HNMR (300 MHz, DMSO-d6): δ=12.04 (s, 1H), 8.87 (s, 1H), 8.80 (s, 1H), 8.30-8.41 (m, 1H), 8.03-8.15 (m, 1H), 8.00 (d, J=7.5 Hz, 2H), 7.59 (d, J=7.5 Hz, 2H=7.5 Hz, 2H), 7.45 (s, 1H), 6.87 (s, 1H), 4.55-4.69 (m, 1H), 3.45-3.91 (m, 1H), 3.15-3.35 (m, 2H), 2.69-2.74 (m, 2H), 1.96-2.28 (m, 2H), 1.49 (d, J=6.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.62, 148.51, 142.17, 137.03, 136.71, 134.11, 129.44, 129.09, 128.11, 127.11, 126.25, 126.04, 123.41, 120.74, 115.60, 99.38, 63.69, 57.57, 55.76, 49.60, 26.22, 22.81 ppm; HRMS (ESI): m/z [M+H]+. C24H24N5O3S calculated value 462.1594, measured value 462.1607.
Step 1: Dissolve (1R)-1-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) and then slowly add p-trifluoromethoxy-benzenesulfonyl chloride (0.3 g, 1.1 mmol) dropwise, Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (1R)-1-(6-(phenylsulfonyl)-1-(1-((4-(trifluoromethoxy)phenyl)sulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol as a yellow oil (0.3 g, 65%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C27H25F3N5O6S2 calculated value 636.1193, measured value 636.1195.
Step 2: (1R)-1-(6-(phenylsulfonyl)-1-(1-((4-(trifluoromethoxy)phenyl)sulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.5 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, and 5 mL of 1 M sodium hydroxide was added and then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS11: (1R)-1-(1-(1-((4-(trifluoromethoxy)phenyl)sulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-ol (0.2 g, 86%). 1 HNMR (300 MHz, DMSO-d6): δ=12.15 (s, 1H), 8.79 (s, 1H), 7.64 (d, J=7.5 Hz, 2H), 7.59 (s, 1H), 7.12 (d, J=7.5 Hz, 2H), 6.74 (s, 1H), 4.33-4.58 (m, 1H), 3.65-3.83 (m, 1H), 3.15-3.31 (m, 2H), 2.74-2.86 (m, 2H), 1.95-2.35 (m, 2H), 1.46 (d, J=9.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 153.88, 148.61, 148.59, 142.10, 132.00, 129.79, 129.04, 127.17, 126.11, 120.73, 115.61, 114.67, 99.38, 63.69, 57.58, 55.70, 49.66, 26.25, 22.81 ppm: HRMS (ESI): m/z [M+H]+. C21H21F3N5O4S calculated value 496.1261, measured value 496.1262.
Step 1: Dissolve (1R)-1-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by the slow dropwise addition of thiophene sulfonyl chloride (0.2 g, 1.1 mmol). Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (1R)-1-(6-(phenylsulfonyl)-1-(1-(thiophen-2-yl sulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol as a yellow oil (0.3 g, 74%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C24H24N5O5S3 calculated value 558.0934, measured value 558.0949.
Step 2: (1R)-1-(6-(phenylsulfonyl)-1-(1-(thiophen-2-yl sulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.3 g, 0.5 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, and 5 mL of 1 M sodium hydroxide was added and then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS12: (1R)-1-(1-(1-(thiophen-2-ylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-ol (0.1 g, 45%). 1 HNMR (3AX MHz, DMSO-d6): δ=12.02 (s, 1H), 8.84 (s, 1H), 7.70 (s, 1H), 7.20 (d, J=7.5 Hz, 1H), 6.96 (d, J=9.0 Hz, 2H), 6.77 (s, 1H), 4.51-4.69 (m, 1H), 3.57-3.83 (m, 1H), 3.17-3.31 (m, 2H), 2.72-2.85 (m, 2H), 1.93-2.31 (m, 2H), 1.07 (d, J=9.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.61, 148.57, 142.11, 129.09, 127.21, 127.11, 126.33, 120.73, 115.60, 99.31, 63.61, 57.59, 55.74, 49.62, 26.22, 22.81 ppm; HRMS (ESI): m/z [M+H]+. C18H20N5O3S2 calculated value 418.1002, measured value 418.1012.
Step 1: Dissolve 2-(3-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (0.3 g, 0.5 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.0 mmol) followed by slow addition of ethylsulfonyl chloride (0.1 g, 0.8 mmol) dropwise. Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-(1-(ethylsulfonyl)-3-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile as a yellow oil (0.2 g, 57%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C29H27N8O6S2 calculated value 647.1489, measured value 647.1482.
Step 2: 2-(1-(Ethylsulfonyl)-3-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (0.2 g, 0.3 mmol) was dissolved in a mixture of solvent of 5 mL tetrahydrofuran and 5 mL methanol, 5 mL of 1 M sodium hydroxide was added and then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS13: 2-(1-(ethylsulfonyl)-3-(4-(2-(5-(hydroxymethyl)furan-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (0.1 g, 64%). 1 HNMR (300 MHz, DMSO-d6): δ=11.94 (s, 1H), 8.90 (s, 1H), 8.69 (s, 1H), 8.14 (s, 1H), 7.37 (s, 1H), 6.41 (s, 2H), 5.95 (s, 1H), 5.43 (t, J=6.0 Hz, 1H), 4.62 (d, J=9.0 Hz, 2H), 4.47 (d, J=6.0 Hz, 2H), 4.34 (d, J=9.0 Hz, 2H), 3.78 (s, 2H), 3.25-3.32 (m, 2H), 1.28 (t, J=7.5 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 153.88, 151.11, 148.67, 142.16, 141.57, 130.77, 129.79, 129.04, 127.12, 120.74, 117.77, 115.65, 107.93, 104.05, 100.53, 99.38, 59.23, 57.39, 51.60, 50.81, 23.20, 2.62 ppm: HRMS (ESI): m/z [M+H]+. C23H23N5O4S calculated value 507.1557, measured value 507.1549.
Step 1: Dissolve 2-(3-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (0.3 g, 0.5 mmol) in 10 mL of tetrahydrofuran. After the addition of DIPEA (0.2 g, 1.0 mmol), propylsulfonyl chloride (0.1 g, 0.8 mmol) was added slowly dropwise. Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-(1-(propylsulfonyl)-3-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile as a yellow oil (0.3 g, 84%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C30H29N5O6S2 calculated value 661.1646, measured value 661.1649.
Step 2: 2-(1-(propylsulfonyl)-3-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (0.3 g, 0.5 mmol) was dissolved in a solvent mixture of 5 mL of tetrahydrofuran and 5 mL of methanol. Then, 5 ml of 1 M sodium hydroxide was added and the mixture was stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS14: 2-(1-(propylsulfonyl)-3-(4-(2-(5-(hydroxymethyl)furan-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (0.1 g, 42%). 1 HNMR (300 MHz, DMSO-d6): δ=11.91 (s, 1H), 8.87 (s, 1H), 8.66 (s, 1H), 8.11 (s, 1H), 7.34 (s, 1H), 6.38 (s, 2H), 5.93 (s, 1H), 5.40 (t, J=6.0 Hz, 1H), 4.58 (d, J=9.0 Hz, 2H), 4.45 (d, J=3.0 Hz, 2H), 4.31 (d, J=9.0 Hz, 2H), 3.75 (s, 2H), 3.24 (t, J=7.5 Hz, 2H), 1.69-1.77 (m, 2H), 1.18-1.30 (m, 2H), 1.00 (t, J=7.5 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 157.85, 145.73, 143.25, 143.07, 139.10, 135.78, 135.68, 134.04, 128.63, 124.75, 119.74, 117.03, 113.11, 109.22, 104.52, 96.44, 58.75, 57.15, 56.11, 50.29, 27.30, 16.89, 13.16 ppm; HRMS (ESI): m/z [M+H]+. C24H25N8O4S calculated value 521.1714, measured value 521.1695.
Step 1: Dissolve 2-(3-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (0.3 g, 0.5 mmol) in 10 mL of tetrahydrofuran. After the addition of DIPEA (0.2 g, 1.0 mmol), butylsulfonyl chloride (0.1 g, 0.8 mmol) was added slowly dropwise. Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-(1-(butylsulfonyl)-3-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b ]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile as a yellow oil (0.2 g, 55%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C31H31N8O6S2 calculated value 675.1802, measured value 675.1811.
Step 2: 2-(1-(Butylsulfonyl)-3-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b ]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (0.2 g, 0.3 mmol) was dissolved in a solvent mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, 5 ml of 1 M sodium hydroxide was added and the mixture was then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS15: 2-(1-(butylsulfonyl)-3-(4-(2-(5-(hydroxymethyl)furan-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (0.1 g, 63%). 1 HNMR (300 MHz, DMSO-d6): δ=11.95 (s, 1H), 8.90 (s, 1H), 8.69 (s, 1H), 8.14 (s, 1H), 7.37 (s, 1H), 6.41 (s, 2H), 5.96 (s, 1H), 5.44 (t, J=6.0 Hz, 1H), 4.62 (d, J=9.0 Hz, 2H), 4.48 (d, J=6.0 Hz, 2H), 4.35 (d, J=9.0 Hz, 2H), 3.78 (s, 2H), 3.29 (t, J=7.5 Hz, 2H), 2.53 (s, 2H), 1.66-1.76 (m, 2H), 1.38-1.50 (m, 2H), 0.92 (t, J=7.5 Hz, 3H) ppm: 1 C NMR (75 MHz, DMSO-d6) δ 157.86, 145.75, 143.27, 143.09, 139.10, 135.80, 135.69, 134.05, 128.65, 124.76, 119.77, 117.06, 113.13, 109.23, 104.54, 96.46, 58.78, 57.16, 56.13, 48.45, 27.33, 25.08, 21.36, 13.94 ppm: HRMS (ESI): m/z [M+H]+. C25H27N5O4S calculated value 535.1870, measured value 535.1870.
Step 1: Dissolve 5-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)furan-2-methanol (0.3 g, 0.6 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.2 mmol) followed by slow dropwise addition of propylsulfonyl chloride (0.1 g, 0.9 mmol). Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (5-(6-(phenylsulfonyl)-1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl furan-2-yl) methanol as a yellow oil (0.2 g, 54%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C26H28N5O6S2 calculated value 570.1476, measured value 570.1488.
Step 2: Dissolve (5-(6-(phenylsulfonyl)-1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl furan-2-yl) methanol (0.2 g, 0.4 mmol) in a solvent mixture of 5 mL of tetrahydrofuran and 5 mL of methanol. Then, add 5 mL of 1 M sodium hydroxide. Stir at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS16: (5-(1-(1-(thiophen-2-ylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)furan-2-yl)methanol (0.1 g, 66%). 1HNMR (300 MHz, DMSO-d6): δ=12.11 (s, 1H), 8.69 (s, 1H), 7.62 (s, 1H), 7.20 (d, J=3.0 Hz, 1H), 6.83 (s, 1H), 6.64 (d, J=3.0 Hz, 1H), 5.58-5.92 (m, 1H), 5.52 (t, J=6.64 (d, J=3.0 Hz, 1H), 5.58-5.92 (m, 1H), 5.52 (t, J=6.0 Hz, 1H), 4.59 (d, J=6.0 Hz, 2H), 4.04 (t, J=10.5 Hz, 1H), 3.79-3.87 (m, 2H), 3.53-3.62 (m, 1H), 3.24-3.29 (m, 2H), 2.69-2.83 (m, 1H), 1.76-1.88 (m, 2H), 1.07 (t, J=7.5 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 158.09, 144.98, 143.27, 143.04, 136.34, 135.19, 132.29, 125.05, 114.29, 109.37, 105.02, 98.97, 56.21, 54.67, 49.36, 48.97, 46.90, 30.36, 16.90, 13.35 ppm: HRMS (ESI): m/z. [M+H]+. C20H24N5O4S calculated value 430.1544, measured value 430.1550.
Step 1: Dissolve (5-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)furan-2-methanol (0.3 g, 0.6 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.2 mmol) followed by slow dropwise addition of cyclopropanecarbonyl chloride (0.1 g, 0.9 mmol). Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield cyclopropyl (3-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)methanone as a brown oil (0.2 g, 58%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C27H26N5O5S calculated value 532.1649, measured value 532.1661.
Step 2: Cyclopropyl (3-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl) methanone (0.2 g, 0.4 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol. Then, 5 mL of 1 M sodium hydroxide was added and the mixture was stirred for 5 h at room temperature. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS17: Cyclopropyl(3-(2-(5-(hydroxymethyl)furan-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl) methanone (0.1 g, 68%). 1 HNMR (300 MHz, DMSO-d6): δ=12.10 (s, 1H), 8.69 (d, J=3.0 Hz, 1H), 7.60 (t, J=4.5 Hz, 1H), 7.22 (dd, J1=9.0 Hz, J2=3.0 Hz, 1H), 6.61 (t, J=4.5 Hz, 1H), 6.46 (d, J=9.0 Hz, 1H), 5.84-6.03 (m, 1H), 5.50 (t, J=4.5 Hz, 1H), 4.57 (d, J=6.0 Hz, 2H), 4.20-4.42 (m, 1H), 4.02-4.12 (m, 1H), 3.81-3.95 (m, 1H), 2.65-2.86 (m, 1H), 1.72-2.01 (m, 1H), 1.25 (s, 2H), 0.84 (t, J=7.5 Hz, 2H), 0.74 (d, J=9.0 Hz, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 171.92, 158.10, 158.05, 144.98, 143.47, 142.97, 136.41, 135.27, 132.40, 125.22, 114.32, 109.43, 105.09, 105.01, 98.67, 56.20, 45.58, 12.55, 7.68 ppm; HRMS (ESI): m/z [M+H]+. C21H22N5O3 calculated value 392.1717, measured value 392.1727.
Step 1: Dissolve 2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by the slow dropwise addition of propylsulfonyl chloride (0.2 g, 1.0 mmol). Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)-1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine as a yellow oil (0.2 g, 54%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C24H26N7O4S2 calculated value 540.1482, measured value 540.1485.
Step 2: 2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)-1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine (0.2 g, 0.4 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol. 5 mL of 1 M sodium hydroxide was then added and the mixture was stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhdrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS18: 2-(1H-imidazol-2-yl)-1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine (0.1 g, 68%). 1 HNMR (300 MHz, DMSO-d6): δ=13.34 (s, 1H), 12.09 (d, J=9.0 Hz, 1H), 8.72 (d, J=6.0 Hz, 1H), 7.58 (t, J=12.0 Hz, 2H), 7.41 (s, 1H), 7.25 (d, J=6.0 Hz, 2H), 6.83 (s, 1H), 4.08 (s, 2H), 3.83-3.97 (m, 3H), 3.20-3.32 (m, 2H), 2.79 (t, J=9.0 Hz, 1H), 2.11 (s, 2H), 1.76-1.86 (m, 2H), 1.25 (s, 1H), 1.08 (t, J=6.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 157.04, 148.61, 142.46, 138.11, 129.09, 127.92, 122.74, 120.71, 115.15, 99.49, 60.42, 56.83, 56.19, 50.01, 36.27, 13.20, 12.62 ppm; HRMS (ESI): m/z [M+H]+. C18H22N7O2S calculated value 400.1550, measured value 400.1561.
Step 1: Dissolve 2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by the slow dropwise addition of trifluoromethylsulfonyl chloride (0.2 g, 1.0 mmol). Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)-1-(1-(((trifluoromethyl)sulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine as a yellow oil (0.3 g, 77%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C22H19F3N7O4S2 calculated value 566.0887, measured value 566.0894.
Step 2: 2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)-1-(1-(((trifluoromethyl)sulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine (0.3 g, 0.5 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol. Then, 5 ml of 1M sodium hydroxide was added and then the mixture was stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS19: 2-(1H-imidazol-2-yl)-1-(1-((trifluoromethyl)sulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine (0.1 g, 44%). 1 HNMR (300 MHz, DMSO-d6): δ=13.36 (s, 1H), 12.16 (s, 1H), 8.72 (s, 1H), 7.66 (s, 1H), 7.41 (s, 1H), 7.26 (s, 2H), 6.65 (s, 1H), 4.17 (d, J=9.0 Hz, 2H), 3.90 (d, J=6.0 Hz, 1H), 2.57-2.72 (m, 1H), 1.25 (s, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 144.80, 142.52, 138.37, 136.04, 134.54, 132.85, 125.37, 119.82, 104.82, 98.39, 48.52 ppm: HRMS (ESI): m/z [M+H]+. C16H15F3N7O2S calculated value 426.0955, measured value 426.0959.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((R-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add carbonyl diimidazole (0.2 g, 1.4 mmol) and slowly add trifluoromethyl ethylamine (0.2 g, 1.4 mmol) dropwise. Then, stir at room temperature for 3 h and monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (R)-3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-N-(2,2,2-trifluoroethyl)pyrrolidine-1-carboxamide as a pale yellow oil (0.2 g, 51%).). The product can be used directly in the next step without further purification. HRMS (ESI); m/z [M+H]. C23H24F3N6O4S calculated value 537.1526, measured value 537.1511.
Step 2: Dissolve (R)-3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-N-(2,2,2-trifluoroethyl)pyrrolidine-1-carboxamide (0.2 g, 0.4 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, add 5 mL of 1 M sodium hydroxide and stir the mixture at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS20: (R)-3-(2-((R)-1-Hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-N-(2,2,2-trifluoroethyl)pyrrolidine-1-carboxamide (0.1 g, 44%). 1 HNMR (300 MHz, DMSO-d6): δ=11.96 (s, 1H), 8.62 (s, 1H), 7.51 (s, 1H), 7.09 (s, 1H), 6.37 (s, 1H), 5.80 (d, J=6.0 Hz, 2H), 5.22 (t, J=6.0 Hz, 1H), 3.87-3.96 (m, 5H), 3.49 (d, J=6.0 Hz, 1H), 2.64-2.76 (m, 11H), 2.43 (s, 1H), 1.68 (d, J=3.0 Hz, 3H), 1.53 (s, 1H), 1.26 (s, 3H), 0.89 (d, J=6.0 Hz, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 155.73, 148.66, 148.57, 142.76, 129.09, 127.12, 124.28, 120.70, 115.65, 99.38, 63.64, 57.99, 54.17, 48.74, 41.88, 26.60, 22.82 ppm: HRMS (ESI): m/z [M+H]+. C17H20F3N6O2 calculated value 397.1594, measured value 397.1603.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((R-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) and then slowly add propyl sulfonyl chloride (0.2 g, 1.4 mmol) dropwise. Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (R)-1-(6-(phenylsulfonyl)-1-((R)-1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol as a yellow oil (0.3 g, 80%). The product can be used directly in the next step without further purification. HRMS (EST): m/z [M+H]+. C23H28N5O5S2 calculated value 518.1526, measured value 518.1534.
Step 2: (R)-1-(6-(Phenylsulfonyl)-1-((R)-1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.3 g, 0.6 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol. Then, 5 mL of 1 M sodium hydroxide was added and the mixture was then stirred for 5 h at room temperature. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS21: (R)-1-(1-((R)-1-(Propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.2 g, 91%). 1 HNMR (300 MHz, DMSO-d6): δ=11.97 (s, 1H), 8.62 (s, 1H), 7.56 (s, 1H), 6.72 (s, 1H), 5.81 (d, J=6.0 Hz, 2H), 5.22 (t, J=6.0 Hz, 11H), 3.82-3.94 (m, 2H), 3.74 (t, J=9.0 Hz, 1H), 3.51-3.59 (m, 1H), 3.28 (t, J=7.5 Hz, 2H), 2.65-2.75 (m, 1H), 1.78-1.85 (m, 2H), 1.68 (d, J=6.0 Hz, 3H), 1.53 (s, 1H), 1.26 (s, 1H), 1.08 (t, J=7.5 Hz, 3H) ppm; —C NMR (75 MHz, DMSO-d6) δ 155.12, 144.83, 136.22, 134.22, 132.43, 124.69, 104.91, 98.76, 62.52, 53.69, 49.72, 49.02, 46.86, 31.15, 22.08, 16.98, 13.32 ppm; HRMS (ESI): m/z [M+H]+. C17H24N5O3S calculated value 378.1594, measured value 378.1609.
Step 1: Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by the addition of 5-hydroxymethylfurfural (0.2 g, 1.1 mmol) dropwise. Then, raise the temperature to 90° C. and stir for 12 hours. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was then purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield trans-4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile as a yellow solid (0.2 g, 53%). HRMS (ESI): m/z [M+H]+. C26H24N5O4S calculated value 502.1544, measured value 502.1556.
Step 2: Dissolve trans-4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.2 g, 0.4 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, add 5 mL of 1 M sodium hydroxide and stir at room temperature for 5 hours. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS22: trans-4-(2-(5-(hydroxymethyl)furan-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 69%), 69%). 1 HNMR (300 MHz, DMSO-d6): δ=11.99 (s, 1H), 8.60 (s, 1H), 7.51 (s, 1H), 7.03 (s, 1H), 6.87 (s, 1H), 6.57 (s, 1H), 5.47 (s, 1H), 4.88 (s, 1H), 4.55 (s, 1H), 3.11 (s, 1H), 2.35 (d, J=9.0 Hz, 2H), 2.24 (d, J=12.0 Hz, 2H), 1.98 (s, 3H), 1.88 (d, J=12.0 Hz, 2H), 1.19 (t, J=9.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 153.81, 150.11, 146.67, 144.93, 142.06, 129.05, 127.92, 122.75, 120.71, 115.65, 105.93, 104.75, 99.68, 64.56, 57.69, 30.22, 26.91, 24.60 ppm: HRMS (ESI): m/z [M+H]+. C20H20N5O2 calculated value 362.1612, measured value 362.1625.
Step 1: Dissolve (R)-(5-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) furan-2-yl methanol (0.3 g, 0.6 mmol) in 10 mL of tetrahydrofuran, add carbonyl diimidazole (0.2 g, 1.2 mmol) and then slowly add trifluoromethyl ethylamine (0.2 g, 1.2 mmol) dropwise. Then, stir the mixture at room temperature for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (R)-3-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-N-(2,2,2-trifluoroethyl)pyrrolidine-1-carboxamide as a pale yellow oil (0.2 g (52%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C26H24F3N6O5S calculated value 589.1475, measured value 589.1482.
Step 2: (R)-3-(2-(5-(Hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-N-(2,2,2-trifluoroethyl)pyrrolidine-1-carboxamide (0.2 g, 0.3 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol. After adding 5 mL of 1M sodium hydroxide, the mixture was stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate =1:1) to yield LXS23: (R)-3-(2-(5-(hydroxymethyl)furan-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-N-(2,2,2-trifluoroethyl)pyrrolidine-1-carboxamide (0.1 g, 66%). 1 HNMR (300 MHz, DMSO-d6): δ=12.08 (s, 1H), 8.66 (s, 1H), 7.57 (s, 1H), 7.19 (s, 1H), 7.09 (s, 1H), 6.61 (s, 1H), 6.47 (s, 1H), 5.94 (d, J=9.0 Hz, 1H), 5.49 (s, 1H), 4.57 (d, J=6.0 Hz, 2H), 3.95 (d, J=9.0 Hz, 6H), 2.71 (t, J=12.0 Hz, 1H), 1.26 (s, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 155.75, 153.18, 151.71, 148.07, 144.94, 142.96, 129.54, 127.72, 124.74, 120.77, 115.35, 107.53, 104.35, 99.68, 57.79, 54.88, 53.59, 48.73, 41.81, 26.20 ppm, HRMS (ESI): m/z [M+H]+. C20H20F3N6O3 calculated value 449.1543, measured value 449.1550.
Step 1: Dissolve (R)-2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add carbonyl diimidazole (0.2 g, 1.4 mmol) and then slowly add trifluoromethyl ethylamine (0.2 g, 1.4 mmol) dropwise. Then, stir the mixture at room temperature for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (R)-3-(2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-N(2,2,2-trifluoroethyl)pyrrolidine-1-carboxamide as a pale yellow oil (0.2 g, 52%).). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C24H22F3N5O3S calculated value 559.1482, measured value 559.1479.
Step 2: Dissolve (R)-3-(2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-N-(2,2,2-trifluoroethyl)pyrrolidine-1-carboxamide (0.2 g, 0.4 mmol) in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol. Then, add 5 ml of 1 M of sodium hydroxide and stir the mixture at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate =1:1) to yield LXS24: (R)-3-(2-(1H-imidazol-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-N-(2,2,2-trifluoroethyl) pyrrolidine-1-carboxamide (0.1 g, 67%). 1 HNMR (300 MHz, DMSO-d6): δ=13.34 (s, 1H), 12.10 (s, 1H), 8.72 (s, 1H), 7.58 (s, 1H), 7.41 (s, 1H), 7.25 (s, 2H), 7.10 (s, 1H), 6.47 (s, 1H), 3.89-4.00 (m, 6H), 2.77 (t, J=12.0 Hz, 1H), 2.12 (s, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 157.08, 155.78, 148.66, 142.49, 138.11, 129.00, 127.19, 124.27, 122.77, 120.75, 115.68, 99.33, 57.26, 54.17, 48.70, 41.80, 26.63 ppm: HRMS (ESI): m/z [M+H]+. C18H18F3N8O calculated value 419.1550, measured value 419.1551.
Step 1 Dissolve (R)-2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by the slow dropwise addition of acryloyl chloride (0.1 g, 1.4 mmol). Then, stir the mixture at room temperature for 3 h and monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (R)-1-(3-(2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyrrolidin-1(6H)-yl)pyrrolidin-1-yl)prop-2-en-1-one as a pale yellow oil (0.2 g, 59%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C24H22N7O3S calculated value 488.1499, measured value 488.1511.
Step 2: (R)-1-(3-(2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)prop-2-en-1-one (0.2 g, 0.4 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol. Then, 5 ml of 1 M sodium hydroxide was added and the mixture was stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS25: (R)-1-(3-(2-(1H-imidazol-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)prop-2-en-1-one (0.1 g, 70%). 1 HNMR (300 MHz, DMSO-d6): δ=13.34 (s, 1H), 12.10 (s, 1H), 8.72 (s, 1H), 7.58 (d, J=9.0 Hz, 1H), 7.41 (s, 1H), 7.25 (s, 1H), 6.46 (s, 1H), 4.02-4.22 (m, 2H), 3.95 (t, J=9.0 Hz, 1H), 3.67 (d, J=6.0 Hz, 2H), 3.61 (d, J=6.0 Hz, 1H), 3.34 (s, 2H), 3.23 (s, 1H), 2.74-2.88 (m, 1H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 166.30, 157.05, 148.66, 142.96, 138.17, 131.16, 129.05, 127.17, 126.88, 122.75, 120.74, 115.65, 99.38, 57.73, 52.07, 46.68, 27.17 ppm; HRMS (ESI): m/z [M+H]+. C18H18N7O calculated value 348.1567, measured value 348.1557.
Step 1 Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add NaS2O5 (0.8 g, 4 mmol) followed by the dropwise addition of 5-formyl-2-methylthiazole (0.2 g, 1.1 mmol). Stir the mixture for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice with dichloromethanel. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(2-(2-methylthiazol-5-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile as a yellow oil (0.2 g, 52%). HRMS (ESI): m/z [M+H]+. C25H23N6O2S2 calculated value 503.1318, measured value 503.1322.
Step 2: Trans-4-(2-(2-methylthiazol-5-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.2 g, 0.4 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, and then stirred at room temperature for 5 h after the addition of 5 mL of 1M sodium hydroxide. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS26: trans-4-(2-(2-Methylthiazol-5-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 69%).). 1 HNMR (300 MHz, DMSO-d6): δ=12.05 (s, 1H), 8.65 (s, 1H), 8.10 (s, 1H), 7.56 (s, 1H), 6.89 (s, 1H), 4.76 (s, 1H), 3.20 (t, J=12.0 Hz, 1H), 2.80 (s, 3H), 2.43 (t, J=12.0 Hz, 2H)=12.0 Hz, 2H), 2.27 (d, J=12.0 Hz, 2H), 2.10 (d, J=15.0 Hz, 2H), 1.86-1.98 (m, 2H) ppm; —C NMR (75 MHz, DMSO-d6) δ 168.60, 144.99, 143.26, 143.05, 136.18, 135.29, 132.94, 127.33, 124.63, 123.32, 104.45, 100.98, 54.58, 31.18, 29.48, 29.16, 28.49, 26.19, 19.36 ppm; HRMS (ESI): m/z [M+H]+. C19H19N6S calculated value 363.1386, measured value 363.1395.
Step 1: Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add NaS2O5 (0.8 g, 4 mmol) followed by the addition of 2-aminothiazole-5-carboxaldehyde (0.2 g, 1.1 mmol) dropwise. Then, stir the mixture for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice with dichloromethanel. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(2-(2-aminothiazol-5-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile as a yellow oil (0.2 g, 52%). HRMS (ESI): m/z [M+H]+. C24H22N7O2S2 calculated value 504.1271, measured value 504.1277.
Step 2: Trans-4-(2-(2-aminothiazol-5-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.2 g, 0.4 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, and then stirred at room temperature for 5 h after the addition of 5 mL of 1 M sodium hydroxide. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS27: trans-4-(2-(2-aminothiazol-5-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 69%).). 1 HNMR (300 MHz, DMSO-d6): δ=11.95 (s, 1H), 8.57 (s, 1H), 7.51 (s, 2H), 7.45 (s, 1H), 6.84 (s, 1H), 4.78 (s, 1H), 3.22 (d, J=12.0 Hz, 1H), 2.44 (t, J=13.5 Hz, 2H), 2.28 (d, J=12.0 Hz, 2H), 1.01 (d, J=12.0 Hz, 2H), 1.91 (t, J=13.5 Hz, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 168.94, 148.64, 144.94, 142.16, 137.74, 129.05, 127.15, 122.75, 122.29, 120.75, 115.67, 99.39, 65.97, 30.88, 26.95, 24.66 ppm; HRMS (ESI): m/z [M+H]+. C18H18N7S calculated value 364.1339, measured value 364.1365.
Step 1: Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by the addition of 5-methyl-2-thiophenecarboxaldehyde (0.2 g, 1.1 mmol) dropwise, and stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice with dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(2-(5-methylthiophen-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile as a yellow oil (0.2 g, 53%). HRMS (ESI): m/z [M+H]+. C26H24N5O2S2 calculated value 502.1366, measured value 502.1377.
Step 2: Dissolve trans-4-(2-(5-methylthiophen-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile (0.2 g, 0.4 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, add 5 mL of 1 M sodium hydroxide and then stir for 5 h at room temperature. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS28: trans-4-(2-(2-methylthiophen-5-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 69%).). 1 HNMR (300 MHz, DMSO-d6): δ=12.00 (s, 1H), 8.63 (s, 1H), 7.54 (s, 1H), 7.38 (d, J=3.0 Hz, 1H), 7.02 (s, 1H), 6.87 (s, 1H), 4.84 (d, J=12.0 Hz, 1H), 3.21 (t, J=12.0 Hz, 1H), 2.58 (s, 3H), 2.45 (t, J=12.0 Hz, 2H), 2.29 (d, J=12.0 Hz, 2H), 2.03 (d, J=6.0 Hz, 2H), 1.81-1.93 (m, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 145.90, 144.93, 143.26, 136.04, 135.11, 132.96, 129.68, 129.49, 127.19, 124.51, 123.28, 104.53, 100.88, 54.70, 31.17, 29.17, 28.65, 26.21, 15.41 ppm; HRMS (ESI): m/z [M+H]+. C20H20N5S calculated value 362.1434, measured value 362.1432.
Step 1: Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by the addition of 5-methyl-2-furanformaldehyde (0.2 g, 1.1 mmol) dropwise, and stir the mixture for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice with dichloromethanel and the organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(2-(5-methylfuran-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile as a yellow oil (0.2 g, 54%). HRMS (ESI): m/z [M+H]+. C26 (H24N5O3S calculated value 486.1594, measured value 486.1601.
Step 2: Dissolve trans-4-(2-(5-methylfuran-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile (0.2 g, 0.4 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, add 5 mL of 1M sodium hydroxide and stir for 5 h at room temperature. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane and the organic phase were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS29: trans-4-(2-(2-Methylfuran-5-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 70%).). 1 HNMR (300 MHz, DMSO-d6): δ=12.00 (s, 1H), 8.62 (s, 1H), 7.53 (s, 1H), 7.01 (s, 1H), 6.89 (s, 1H), 6.41 (s, 1H), 4.89 (t, J=10.5 Hz, 1H), 3.15 (t, J=12.0 Hz, 1H), 2.53 (s, 3H), 2.40 (d, J=12.0 Hz, 2H), 2.29 (d, J=12.0 Hz, 2H), 2.00 (d, J=10.0 Hz, 2H), 1.91 (d, J=12.0 Hz, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 154.49, 145.07, 143.04, 142.81, 136.20, 135.14, 132.90, 124.49, 123.34, 114.47, 108.68, 104.57, 100.44, 54.96, 31.17, 29.36, 28.78, 26.41, 22.57, 13.91 ppm; HRMS (ESI): m/z [M+H]+. C20H20N5O calculated value 346.1662, measured value 346.1664.
Step 1: Dissolve tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (0.3 g, 0.7 mmol) in 10 mL of DMF, add Na2S2O5 (0.7 g, 3.5 mmol) followed by addition of o-hydroxybenzaldehyde (0.2 g, 1.4 mmol). Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane and the organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield tert-butyl 3-(2-(2-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate as a yellow solid (0.2 g, 55%). HRMS (ESI): m/z [M+H]+. C29H30N5O5S calculated value 560.1962, measured value 560.1965.
Step 2: Dissolve tert-butyl 3-(2-(2-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate (0.2 g, 0.4 mmol) in 10 mL dichloromethane, slowly add trifluoroacetic acid (0.5 g, 4.0 mmol) and stir at room temperature for 12 h. The product was concentrated under vacuum to yield 2-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)phenol as a light yellow oil (0.1 g, 61%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C24H22N5O3S calculated value 460.1438, measured value 460.1441.
Step 3: Dissolve 2-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)phenol (0.1 g, 0.2 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.1 g, 0.4 mmol) followed by the slow dropwise addition of propylsulfonyl chloride (0.1 g, 0.3 mmol) and stir for 3 h at room temperature. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane and the organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-(6-(phenylsulfonyl)-1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)phenol as a pale yellow oil (0.1 g, 81%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C27H28N5O5S2 calculated value 566.1526, measured value 566.1533.
Step 4: Dissolve 2-(6-(phenylsulfonyl)-1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)phenol (0.1 g, 0.2 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol. Then, add 5 mL of 1 M sodium hydroxide and then stir the mixture at room temperature for 5 hours. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane and the organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS30: 2-(1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)phenol (65.0 mg, 86%). 1 HNMR (300 MHz, DMSO-d6): δ=12.06 (s, 1H), 8.67 (s, 1H), 7.59 (t, J=3.0 Hz, 1H), 7.49-7.52 (m, 1H), 7.41-7.47 (m, 1H), 7.19 (d, J=9.0 Hz, 1H), 7.01 (t, J=7.5 Hz, 1H), 6.80 (d, J=3.0 Hz, 1H), 5.12-5.24 (m, 1H), 3.98-4.10 (m, 1H), 3.74-3.87 (m, 2H), 3.19-3.25 (m, 2H), 2.69-2.83 (m, 1H), 2.33-2.42 (m, 1H), 1.77-1.84 (m, 2H), 1.05 (t, J=9.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 170.82, 156.82, 150.94, 144.75, 136.04, 135.50, 132.54, 131.85, 131.74, 124.82, 119.58, 117.92, 116.59, 104.91, 98.80, 60.22, 55.10, 49.24, 46.75, 30.10, 21.24, 16.91, 14.55, 13.32 ppm; HRMS (ESI): m/z [M+H]+. C21H24N5O3S calculated value 426.1594, Measured value 426.1462.
Step 1: Dissolve tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (0.3 g, 0.7 mmol) in 10 mL of DMF, add Na2S2O5 (0.7 g, 3.5 mmol) followed by addition of 2,4-dihydroxybenzaldehyde (0.2 g, 1.4 mmol). Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane and the organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield tert-butyl 3-(2-(2,4-dihydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate as a yellow solid (0.3 g, 79%). HRMS (ESI): m/z [M+H]+. C29H30N5O6S calculated value 576.1911, measured value 576.1919.
Step 2: Dissolve tert-butyl 3-(2-(2,4-dihydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate (0.3 g, 0.5 mmol) in 10 mL dichloromethane, slowly add trifluoroacetic acid (0.6 g, 5.0 mmol) and stir at room temperature for 12 hours. The product is then concentrated under vacuum to yield 4-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)benzene-1,3-diol as a light yellow oil (0.2 g, 81%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]. C24H22N5O4S calculated value 476.1387, measured value 476.1391.
Step 3: Dissolve 4-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)benzene-1,3-diol (0.2 g, 0.4 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.1 g, 0.8 mmol) followed by the slow dropwise addition of propylsulfonyl chloride (0.1 g, 0.6 mmol). Stir at room temperature for 3 h. The reaction was monitored by TLC for completion. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane and the organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 4-(6-III (phenylsulfonyl)-1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) benzene-1,3-diol as a pale yellow oil (0.2 g, 82%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C27H28N5O6S2 calculated value 582.1476, measured value 582.1501.
Step 4: Dissolve 4-(6-(phenylsulfonyl)-1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) benzene-1,3-diol (0.2 g, 0.3 mmol) in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, after adding 5 mL of 1 M sodium hydroxide, stir the mixture for 5 h at room temperature and monitor the reaction for completion by TLC. Saturated sodium bicarbonate was then added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was then extracted twice using dichloromethane and the organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS31: 4-(1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)benzene-1,3-diol (0.1 g, 66%). 1 HNMR (300 MHz, DMSO-d6): δ=11.97 (s, 1H), 10.15 (s, 1H), 9.84 (s, 1H), 8.63 (s, 1H), 7.57 (s, 1H), 7.29 (d, J=9.0 Hz, 1H), 6.76 (d, J=3.0 Hz, 1H), 6.53 (d, J=3.0 Hz, 1H), 6.42-6.46 (m, 1H), 5.12-5.34 (m, 1H), 3.95 (t, J=9.0 Hz, 1H), 3.73-3.81 (m, 2H), 3.17-3.23 (m, 2H), 2.66-2.80 (m, 1H), 2.30-2.38 (m, 1H), 1.72-1.85 (m, 2H), 1.28-1.31 (m, 2H), 1.08-1.19 (m, 2H), 1.02 (t, J=9.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 160.74, 157.62, 151.30, 144.70, 135.81, 135.33, 133.33, 131.70, 124.76, 108.69, 107.72, 104.91, 102.94, 98.78, 54.98, 53.91, 49.43, 46.70, 18.47, 16.95, 13.34 ppm: HRMS (ESI): m/z [M+H]+. C21H24N5O4S calculated value 442.1544, measured value 442.1405.
Step 1: Dissolve tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (0.3 g, 0.7 mmol) in 10 mL of DMF, add Na2S2O5 (0.7 g, 3.5 mmol) followed by adding 3-hydroxybenzaldehyde (0.2 g, 1.4 mmol). Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield tert-butyl 3-(2-(3-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate as a yellow solid (0.3 g, 82%). HRMS (ESI): m/z [M+H]+. C29H30N5O5S calculated value 560.1962, measured value 560.1969.
Step 2: Dissolve tert-butyl 3-(2-(3-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate (0.3 g, 0.5 mmol) in 10 mL of dichloromethane, slowly add trifluoroacetic acid (0.6 g, 5.0 mmol) and stir at room temperature for 12 h. The product was concentrated under vacuum to yield 3-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) phenol as a light yellow oil (0.2 g, 81%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C24H22N5O3S calculated value 460.1438, measured value 460.1441.
Step 3: Dissolve 3-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)phenol (0.2 g, 0.4 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.1 g, 0.8 mmol) followed by slow dropwise addition of propylsulfonyl chloride (0.1 g, 0.6 mmol) and then stir for 3 h at room temperature. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 3-(6-(phenylsulfonyl)-1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) phenol as a pale yellow oil (0.2 g, 81%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C27H28N5O5S2 calculated value 566.1526, measured value 566.1529.
Step 4: Dissolve 3-(6-(phenylsulfonyl)-1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) phenol (0.2 g, 0.4 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, add 5 mL of 1M sodium hydroxide and then stir at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS32: 3-(1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)phenol (0.1 g, 66%). 1 HNMR (300 MHz, DMSO-d6): δ=12.06 (s, 1H), 10.04 (s, 1H), 8.68 (s, 1H), 7.60 (t, J=3.0 Hz, 1H), 7.43 (t, J=7.5 Hz, 1H), 7.20 (t, J=6.0 Hz, 1H), 7.00-7.03 (m, 1H), 6.80 (d, J=3.0 Hz, 1H), 5.41-5.53 (m, 1H), 3.99-4.09 (m, 1H), 3.76-3.89 (m, 2H), 3.46-3.55 (m, 1H), 3.16-3.32 (m, 2H), 2.71-2.85 (m, 1H), 2.40-2.49 (m, 1H), 1.72-1.84 (m, 2H), 1.05 (t, J=9.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 158.03, 152.38, 144.82, 136.27, 135.14, 132.22, 131.63, 130.44, 120.62, 117.30, 116.85, 111.64, 105.14, 55.07, 49.05, 30.03, 16.88, 13.35 ppm; HRMS (ESI): m/z [M+H]+. C21H24N5O3S calculated value 426.1594, measured value 426.1464.
Step 1: Dissolve tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (0.3 g, 0.7 mmol) in 10 mL of DMF, add Na2S2O5 (0.7 g, 3.5 mmol) followed by adding 2,4-dihydroxybenzaldehyde (0.2 g, 1.4 mmol). Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield tert-butyl 3-(2-(2,4-dihydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate as a yellow solid (0.3 g, 79%). HRMS (ESI): m/z [M+H]+. C29H30N5O6S calculated value 576.1911, measured value 576.1918.
Step 2: Dissolve tert-butyl 3-(2-(2,4-dihydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate (0.3 g, 0.5 mmol) in 10 mL dichloromethane, slowly add trifluoroacetic acid (0.6 g, 5.0 mmol) and stir at room temperature for 12 hours. The product was concentrated under vacuum to yield 4-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)benzene-1,3-diol as a light yellow oil (0.2 g, 81%). The product can be used directly in the next step without further purification. HRMS (EST): m/z [M+H]+. C24H22N5O4S calculated value 476.1387, measured value 476.1375.
Step 3: Dissolve 4-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)benzene-1,3-diol (0.2 g, 0.4 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.1 g, 0.8 mmol) followed by slow dropwise addition of acryloyl chloride (0.1 g, 0.6 mmol). Stir at room temperature for 3 h. The reaction was monitored by TLC for completion. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 1-(3-(2-(2,4-dihydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)prop-2-en-1-one as a pale yellow oil (0.2 g, 90%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C27H24N5O5S calculated value 530.1493, measured value 530.1488.
Step 4: Dissolve 1-(3-(2-(2,4-dihydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)prop-2-en-1-one (0.2 g, 0.4 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, add 5 mL of 1 M sodium hydroxide. Then, stir the mixture for 5 h at room temperature, the reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS33: 1-(3-(2-(2,4-Dihydroxyphenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)prop-2-en-1-one (0.1 g, 68%). 1 HNMR (300 MHz, DMSO-d6): δ=11.96 (s, 1H), 8.62 (s, 1H), 7.52 (d, J=6.0 Hz, 1H), 7.26 (d, J=6.0 Hz, 1H), 6.55 (s, 1H), 6.42 (d, J=9.0 Hz, 1H), 4.03-4.10 (m, 4H), 3.55-3.66 (m, 3H), 2.68-2.79 (m, 1H), 2.32 (s, 1H), 2.02 (s, 4H), 1.21 (t, J=6.0 Hz, 4H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 166.39, 159.94, 156.62, 153.78, 148.64, 142.18, 131.16, 130.11, 129.05, 127.14, 126.89, 120.74, 115.69, 110.95, 109.05, 105.64, 99.38, 57.77, 52.05, 46.69, 27.17 ppm; HRMS (ESI): m/z [M+H]+. C21H20N5O3 calculated value 390.1561, measured value 390.1692.
Step 1: Dissolve tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (0.3 g, 0.7 mmol) in 10 mL of DMF, add Na2S2O5 (0.7 g, 3.5 mmol) followed by adding 3-hydroxybenzaldehyde (0.2 g, 1.4 mmol). Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield tert-butyl 3-(2-(3-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate as a yellow solid (0.3 g, 82%). HRMS (ESI): m/z [M+H]+. C29H30N5O5S calculated value 560.1962, measured value 560.1977.
Step 2: Dissolve tert-butyl 3-(2-(3-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate (0.3 g, 0.5 mmol) in 10 mL of dichloromethane, slowly add trifluoroacetic acid (0.6 g, 5.0 mmol) and then stir at room temperature for 12 h. The product was concentrated under vacuum to yield 3-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) phenol as a light yellow oil (0.2 g, 81%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C24H22N5O3S calculated value 460.1438, measured value 460.1441.
Step 3: Dissolve (R)-2-(1H-imidazol-2-yl)-6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine (0.2 g, 0.4 mmol) in 10 mL of tetrahydrofuran, add carbonyl diimidazole (0.2 g, 1.0 mmol) and then slowly add trifluoromethyl ethylamine (0.2 g, 1.0 mmol) dropwise. Then, stir the mixture at room temperature for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 3-(2-(3-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-N-(2,2,2-trifluoroethyl)pyrrolidine-1-carboxamide as a pale yellow oil (0.2 g, 79%). The product can be used directly in the next step without further purification. HRMS (EST): m/z [M+H]+. C27H24F3N6O4S calculated value 585.1526, measured value 585.1531.
Step 4: Dissolve 3-(2-(3-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-N-(2,2,2-trifluoroethyl)pyrrolidine-1-carboxamide (0.2 g, 0.3 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, followed by adding 5 mL of 1M sodium hydroxide. Stir for 5 h at room temperature. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS34: 3-(2-(3-hydroxyphenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-N-(2,2,2-trifluoroethyl)pyrrolidine-1-carboxamide (0.1 g, 66%). 1 HNMR (300 MHz, DMSO-d6): δ=12.04 (s, 1H), 8.69 (s, 1H), 7.56 (t, J=3.0 Hz, 1H), 7.43 (t, J=7.5 Hz, 1H), 7.15 (t, J=4.5 Hz, 2H), 6.99-7.08 (m, 2H), 6.43 (d, J=3.0 Hz, 1H), 5.46 (t, J=9.0 Hz, 1H), 3.86-3.98 (m, 6H), 2.71-2.78 (m, 1H), 2.30-2.41 (m, 1H), 1.26 (s, 1H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 158.03, 156.37, 152.81, 152.40, 144.80, 136.37, 135.20, 132.31, 131.66, 130.50, 125.03, 120.61, 117.57, 117.33, 116.78, 111.67, 105.18, 98.76, 91.18, 76.29, 73.64, 73.34, 67.45, 30.10 ppm; HRMS (ESI): m/z [M+H]+. C21H20F3N6O2 calculated value 445.1594, measured value 445.1459.
Step 1: Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by adding 3-hydroxybenzaldehyde (0.2 g, 1.6 mmol) dropwise, then raise the temperature to 90° C. and stir for 12 hours. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice with dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(2-(3-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile as a yellow oil (0.3 g, 79%). HRMS (ESI): m/z [M+H]+. C27H24N5O3S calculated value 498.1594, measured value 498.1588.
Step 2: Dissolve trans-4-(2-(3-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.3 g, 0.6 mmol) in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol. Then add 5 mL of 1 M sodium hydroxide and stir at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS35: trans-4-(2-(3-Hydroxyphenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 46%). 1HNMR (300 MHz, DMSO-d6): δ=11.98 (s, 1H), 9.86 (s, 1H), 8.64 (s, 1H), 7.54 (t, J=3.0 Hz, 1H), 7.41 (t, J=7.5 Hz, 2H), 7.10 (t, J=4.5 Hz, 2H), 6.98-7.01 (m, 1H), 6.87 (s, 1H), 4.48-4.56 (m, 1H), 3.21 (s, 1H), 3.19 (s, 1H), 2.41 (t, J-=12.0 Hz, 2H), 2.26 (d, J=12.0 Hz, 2H), 2.02 (t, J=4.5 Hz, 2H), 1.67-1.79 (m, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 157.90, 152.05, 144.92, 136.19, 135.11, 132.69, 132.29, 130.35, 124.39, 123.15, 120.52, 117.15, 116.76, 104.60, 100.64, 55.37, 54.78, 49.05, 29.34, 28.75, 26.27 ppm; HRMS (ESI): m/z [M+H]+. C21H20N5O calculated value 358.1662, measured value 358.1655.
Step 1: Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by adding 2,4-dihydroxybenzaldehyde (0.2 g, 1.6 mmol) dropwise. Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(2-(2,4-dihydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile as a yellow oil (0.3 g, 77%). HRMS (ESI): m/z [M+H]+. C27H24N5O4S calculated value 514.1544, measured value 514.1560.
Step 2: Trans-4-(2-(2,4-dihydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.3 g, 0.6 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, and then stirred at room temperature for 5 h after the addition of 5 mL of 1M sodium hydroxide. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS36: trans-4-(2-(2,4-Dihydroxyphenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 46%). 1 HNMR (300 MHz, DMSO-d6): δ=11.92 (s, 1H), 8.58 (s, 1H), 7.49 (s, 1H), 7.19 (d, J=9.0 Hz, 1H), 6.84 (s, 1H), 6.58 (s, 1H), 6.41-6.44 (m, 1H), 4.23 (d, J=9.0 Hz, 1H), 3.14 (t, J=12.0 Hz, 1H), 2.79 (d, J=18.0 Hz, 1H), 2.36 (d, J=9.0 Hz, 2H), 2.26 (d, J=15.0 Hz, 2H), 2.02 (s, 1H), 1.62-1.74 (m, 2H), 1.18-1.26 (m, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 160.50, 157.73, 150.96, 144.77, 135.36, 133.04, 132.21, 124.09, 123.21, 109.32, 107.49, 104.42, 85.32, 73.67, 60.22, 54.89, 29.20, 28.95, 26.39, 14.55 ppm; HRMS (ESI): m/z [M+H]+. C21H20N5O2 calculated value 374.1612, measured value 374.1600.
Step 1. Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by adding 4-(trifluoromethyl)benzaldehyde (0.3 g, 1.6 mmol) dropwise. Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice with dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(6-(phenylsulfonyl)-2-(4-(trifluoromethyl)phenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile as a yellow oil (0.3 g, 72%). HRMS (ESI): m/z [M+H]+. C28H23F3N5O2S calculated value 550.1519, measured value 550.1521.
Step 2: Trans-4-(6-(phenylsulfonyl)-2-(4-(trifluoromethyl)phenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile (0.3 g, 0.5 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol. 5 mL of 1 M sodium hydroxide was added and then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS37: trans-4-(2-(4-(trifluoromethyl)phenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 45%).). 1 HNMR (300 MHz, DMSO-d6): δ=12.04 (s, 1H), 8.68 (s, 1H), 7.97 (d, J=3.0 Hz, 4H), 7.56 (t, J=3.0 Hz, 1H), 6.89 (s, 1H), 4.45-4.53 (m, 1H), 3.17 (t, J=13.5 Hz, 1H), 2.40 (t, J=10.5 Hz, 2H), 2.23 (d, J=12.0 Hz, 2H), 2.03 (t, J=10.5 Hz, 2H), 1.74-1.86 (m, 2H), 1.25 (s, 1H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 150.58, 145.02, 136.44, 135.27, 132.89, 131.00, 130.63, 129.90, 126.38, 126.10, 124.55, 123.21, 104.58, 100.85, 72.19, 54.92, 29.32, 28.50, 26.22 ppm; HRMS (ESI): m/z [M+H]+. C22H19F3N5 calculated value 410.1587, measured value 410.1590.
Step 1. Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by adding 2-hydroxybenzaldehyde (0.2 g, 1.6 mmol) dropwise, raise the temperature to 90° C. and stir for 12 hours. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice with dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(2-(2-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile as a yellow oil (0.3 g, 79%). HRMS (ESI): m/z [M+H]+. C27H24N5O3S calculated value 498.1594, measured value 498.1600.
Step 2: Dissolve trans-4-(2-(2-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.3 g, 0.6 mmol) in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, add 5 mL of 1 M sodium hydroxide and stir at room temperature for 5 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS38: trans-4-(2-(2-Hydroxyphenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 46%). 1HNMR (300 MHz, DMSO-d6): δ=11.94 (s, 1H), 10.17 (s, 1H), 8.62 (s, 1H), 7.52 (t, J=3.0 Hz, 1H), 7.40-7.46 (m, 2H), 6.98-7.08 (m, 2H), 6.87 (d, J=3.0 Hz, 1H), 4.18-4.26 (m, 1H), 3.14 (t, J=12.0 Hz, 1H), 2.38 (t, J=12.0 Hz, 2H), 2.26 (d, J=10.5 Hz, 2H), 1.60-1.72 (m, 2H), 1.18-1.26 (m, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 156.49, 150.28, 144.85, 135.95, 135.45, 132.47, 132.24, 131.75, 124.19, 123.16, 119.68, 118.54, 116.23, 104.41, 100.53, 72.30, 55.03, 29.22, 28.89, 26.37 ppm: HRMS (ESI): m/z [M+H]+. C21H20N5O calculated value 358.1662, measured value 358.1658.
Step 1: Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by the addition of 2-fluoro-4-hydroxybenzaldehyde (0.2 g, 1.6 mmol) dropwise and then stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice with dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(2-(2-fluoro-4-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile as a pale yellow oil (0.3 g, 77%). HRMS (ESI): m/z [M+H]+. C27H23FN5O3S calculated value 516.1500, measured value 516.1511.
Step 2: Dissolve trans-4-(2-(2-fluoro-4-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile (0.3 g, 0.6 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, add 5 mL of 1 M sodium hydroxide and then stir for 5 h at room temperature. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS39: trans-4-(2-(2-Fluoro-4-hydroxyphenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 46%).). 1 HNMR (300 MHz, DMSO-d6): δ=11.99 (s, 1H), 8.63 (s, 1H), 7.54 (t, J=3.0 Hz, 1H), 7.43-7.49 (m, 1H), 6.88 (d, J=3.0 Hz, 1H), 6.82-6.85 (m, 1H), 6.80 (d, J=3.0 Hz, 1H), 4.24 (d, J=12.0 Hz, 1H), 3.13 (t, J=13.5 Hz, 1H), 2.34 (d, J 12.0 Hz, 2H), 2.23 (d, J=12.0 Hz, 2H), 2.02 (s, 1H), 1.94 (d, J=9.0 Hz, 2H), 1.70-1.82 (m, 2H), 1.18-1.26 (m, 1H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 160.11, 159.75, 153.77, 148.62, 142.15, 130.54, 129.04, 127.16, 122.75, 120.77, 116.16, 115.64, 112.04, 104.54, 99.38, 65.93, 30.88, 26.91, 23.69 ppm: HRMS (ESI): m/z [M+H]+. C21H19FN5O calculated value 376.1568, measured value 376.1562.
Step 1: Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by the addition of 2-fluorobenzaWdehyde (0.2 g, 1.6 mmol) dropwise, raise the temperature to 90° C. and stir for 12 hours. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(2-(2-fluorophenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile as a pale yellow oil (0.3 g, 79%). HRMS (EST): m/z [M+H]+. C27H21FN5O2S calculated value 500.1551, measured value 500.1563.
Step 2: Dissolve trans-4-(2-(2-fluorophenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile (0.3 g, 0.6 mmol) in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, add 5 mL of 1 M sodium hydroxide and then stir at room temperature for 5 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS40: trans-4-(2-(2-Fluorophenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 46%). 1HNMR (300 MHz, DMSO-d6): δ=12.06 (s, 1H), 8.70 (s, 1H), 7.70 (t, J=6.0 Hz, 2H), 7.57 (s, 1H), 7.49 (t, J=9.0 Hz, 2H), 6.91 (s, 1H), 4.27 (s, 1H), 3.13 (s, 1H), 2.35 (d, J=12.0 Hz, 2H), 2.24 (d, J=9.0 Hz, 2H), 2.01 (d, J=6.0 Hz, 2H), 1.75 (d, J=12.0 Hz, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 162.07, 158.80, 146.49, 145.00, 136.27, 135.41, 133.08, 132.92, 132.42, 125.44, 124.54, 123.11, 119.40, 119.19, 116.62, 116.34, 104.40, 100.51, 60.22, 55.18, 29.42, 28.59, 26.29, 14.51 ppm; HRMS (EST): m/z [M+H]+. C21H19FN5 calculated value 360.1619, measured value 360.1615.
Step 1: Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by adding 2-fluoro 4-methoxybenzaldehyde (0.2 g, 1.6 mmol) dropwise, and then stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice with dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(2-(2-hydroxy-4-methoxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile as a yellow oil (0.3 g, 75%). HRMS (ESI): m/z [M+H]+. C28H26N5O4S calculated value 528.1700, measured value 528.1711.
Step 2: Trans-4-(2-(2-hydroxy-4-methoxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile (0.3 g, 0.6 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, and then stirred at room temperature for 5 h after the addition of 5 mL of 1 M sodium hydroxide. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS41: trans-4-(2-(2-hydroxy-4-methoxyphenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 46%), 46%). 1 HNMR (300 MHz, DMSO-d6): δ=11.91 (s, 1H), 10.26 (s, 1H), 8.60 (s, 1H), 7.51 (t, J=3.0 Hz, 1H), 7.34 (d, J=9.0 Hz, 1H), 6.86 (d, J=3.0 Hz, 1H), 6.58-6.61 (m, 2H), 4.24 (t, J=13.5 Hz, 1H), 3.83 (s, 3H), 3.15 (t, J=12.0 Hz, 1H), 2.39 (t, J=12.0 Hz, 2H), 2.25 (d, J=12.0 Hz, 2H), 2.01 (d, J=12.0 Hz, 2H), 1.62-1.74 (m, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 162.03, 157.72, 150.38, 144.83, 135.83, 135.37, 133.21, 132.24, 124.14, 123.19, 111.08, 105.80, 104.42, 101.63, 55.60, 54.94, 29.20, 28.92, 26.38 ppm; HRMS (EST): m/z [M+H]+. C22H22N5O2 calculated value 388.1768, measured value 366.1782.
Step 1: Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by adding 4-(methylthio)benzaldehyde (0.2 g, 1.6 mmol) dropwise. Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice with dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(2-(4-(methylthio)phenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile as a yellow oil (0.3 g, 75%). HRMS (ESI): m/z [M+H]+. C28H26N5O2S2 calculated value 528.1522, measured value 528.1533.
Step 2: Trans-4-(2-(4-(methylthio)phenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile (0.3 g, 0.6 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, 5 mL of 1 M sodium hydroxidewas added and then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS42: trans-4-(2-(4-(methylthio)phenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 46%). 1 HNMR (300 MHz, DMSO-d6): δ=11.98 (s, 1H), 8.65 (s, 1H), 7.66 (d, J 9.0 Hz, 2H), 7.54 (t, J=3.0 Hz, 1H), 7.48 (d, J=9.0 Hz, 2H), 6.88 (d, J=3.0 Hz, 1H), 4.49 (t, J=13.5 Hz, 1H), 3.17 (t, J=12.0 Hz, 1H), 2.61 (s, 3H), 2.45 (t, J=12.0 Hz, 2H), 2.25 (d, J=12.0 Hz, 2H), 2.03 (d, J=6.0 Hz, 2H), 1.71-1.83 (m, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 153.77, 148.43, 142.89, 139.43, 129.04, 127.77, 127.32, 127.17, 127.01, 122.74, 120.73, 115.64, 99.49, 65.93, 31.96, 26.94, 24.68, 14.88 ppm; HRMS (ESI): m/z [M+H]+. C22H22N5O2 calculated value 388.1590, measured value 388.1599.
Step 1. Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by the addition of 4-fluorobenzaldehyde (0.2 g, 1.6 mmol) dropwise, raise the temperature to 90° C. and stir for 12 hours. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(2-(4-fluorophenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile as a yellow oil (0.3 g, 79%). HRMS (ESI): m/z [M+H]+. C27H23FN5O2S calculated value 500.1551, measured value 500.1559.
Step 2: Dissolve trans-4-(2-(4-fluorophenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile (0.3 g, 0.6 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, followed by the addition of 5 mL of 1M sodium hydroxide. Then, stir the mixture at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS43: trans-4-(2-(4-Fluorophenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 46%). 1HNMR (300 MHz, DMSO-d6): δ=11.99 (s, 1H), 8.65 (s, 1H), 7.74-7.79 (m, 2H), 7.54 (t, J=3.0 Hz, 1H), 7.46 (t, J=9.0 Hz, 2H), 6.88 (d, J=3.0 Hz, 1H), 4.44 (t, J=12.0 Hz, 1H), 3.15 (t, J=12.0 Hz, 1H), 2.34-2.47 (m, 2H), 2.24 (d, J=12.0 Hz, 2H), 2.04 (d, J=9.0 Hz, 2H), 1.70-1.81 (m, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 162.99, 153.74, 148.65, 142.11, 129.19, 129.03, 127.43, 126.22, 122.74, 120.75, 116.43, 115.64, 99.40, 65.93, 30.82, 26.94, 21.54 ppm, HRMS (ESI): m/z [M+H]+. C21H19FN5 calculated value 360.1619, measured value 360.1683.
Step 1: Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by the addition of 4-methylsulfonylbenzaldehyde (0.3 g, 1.6 mmol) dropwise. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated, the aqueous phase was extracted twice with dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(2-(4-(methylsulfonyl)phenyl)-6-(phenylsulfonyl)-2,3-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile as a yellow oil (0.3 g, 70%). HRMS (ESI): m/z [M+H]+. C28H28N5O4S2 calculated value 562.1577, measured value 562.1588.
Step 2: Trans-4-(2-(4-(methylsulfonyl)phenyl)-6-(phenylsulfonyl)-2,3-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl) cyclohexanecarbonitrile (0.3 g, 0.5 mmol) was dissolved in a solvent mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, followed by the addition of 5 mL of 1 M sodium hydroxide. The mixture is then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhvdrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS44: trans-4-(2-(4-(Methylsulfonyl)phenyl)-2,3-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.2 g, 89%), 1 HNMR (300 MHz, DMSO-d6): δ=11.40 (s, 1H), 8.93 (s, 1H), 8.22 (d, J=9.0 Hz, 2H), 8.14 (s, 1H), 8.06 (d, J=6.0 Hz, 2H), 7.22 (t, J=3.0 Hz, 1H), 6.63 (d, J=3.0 Hz, 1H), 6.07 (d, J=9.0 Hz, 1H), 4.02-4.10 (m, 1H), 3.30 (s, 3H), 2.73-2.81 (m, 1H), 2.12 (d, J=9.0 Hz, 2H), 2.03 (s, 2H), 1.76-1.87 (m, 2H), 1.44-1.55 (m, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 144.28, 139.38, 138.74, 134.46, 131.18, 128.94, 128.11, 127.19, 123.38, 122.74, 108.17, 99.36, 83.56, 61.28, 47.74, 28.48, 26.94, 24.93 ppm; HRMS (ESI): m/z [M+H]+. C22H24N5O2S calculated value 422.1645, measured value 422.1679.
Step 1: Dissolve 5-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)furan-2-yl) methanol (0.3 g, 0.6 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.2 mmol) and then slowly add cyclopropylsulfonyl chloride (0.1 g, 0.9 mmol) dropwise. Then raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (5-(6-(phenylsulfonyl)-1-(1-(cyclopropylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) furan-2-yl) methanol as a yellow oil (0.3 g, 82%). The product can be used directly in the next step without further purification. HRMS (EST): m/z [M+H]+. C26H26N5O6S2 calculated value 568.1319, measured value 568.1321.
Step 2: Dissolve (5-(6-(phenylsulfonyl)-1-(1-(cyclopropylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) furan-2-yl) methanol (0.3 g, 0.5 mmol) in a solvent mixture of 5 mL oftetrahydrofuran and 5 mL of methanol, followed by the addition of 5 mL 1 M sodium hydroxide. Stir the mixture for 5 h at room temperature and the reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS45: (5-(1-(1-(cyclopropylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)furan-2-yl)methanol (0.1 g, 44%). 1 HNMR (300 MHz, DMSO-d6): δ=12.09 (s, 1H), 8.67 (s, 1H), 7.61 (t, J=3.0 Hz, 1H), 7.19 (d, J=3.0 Hz, 1H), 6.85 (s, 1H), 6.63 (d, J=3.0 Hz, 1H), 5.80-5.92 (m, 1H), 5.51 (t, J=6.0 Hz, 1H), 4.58 (d, J=6.0 Hz, 2H), 4.04-4.10 (m, 1H), 3.81-3.90 (m, 2H), 3.58-3.67 (m, 1H), 2.89-2.97 (m, 1H), 2.71-2.80 (m, 2H), 1.26 (s, 1H), 1.06 (t, J=6.0 Hz, 4H) ppm: —C NMR (75 MHz, DMSO-d6) δ 153.88, 151.18, 148.60, 144.53, 142.17, 129.04, 127.16, 120.88, 115.63, 107.95, 104.05, 99.38, 57.39, 56.43, 54.43, 49.75, 37.55, 24.66, 4.07 ppm; HRMS (ESI): m/z [M+H]+. C20H22N O4S calculated value 428.1387, measured value 428.1384.
Step 1: Dissolve 3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazo-1-yl)propanenitrile (0.3 g, 0.7 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by adding 5-hydroxymethylfurfural (0.2 g, 1.4 mmol) and then stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 3-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile as a yellow oil (0.3 g, 79%). HRMS (ESI): m/z [M+H]+. C25H20N7O4S calculated value 514.1292, measured value 514.1299.
Step 2: Dissolve 3-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.3 g, 0.6 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, then add 5 mL of 1 M sodium hydroxide. After stirring for 5 h at room temperature, the reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS46: 3-(4-(2-(5-(Hydroxymethyl)furan-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl) propanenitrile (0.1 g, 46%). 1 HNMR (300 MHz, DMSO-d6): δ=11.96 (s, 1H), 8.79 (s, 1H), 8.06 (s, 1H), 7.94 (s, 1H), 7.59 (s, 1H), 7.02 (d, J=6.0 Hz, 1H), 6.78 (s, 1H), 6.59 (d, J=6.0 Hz, 1H), 5.04 (t, J=7.5 Hz, 2H), 4.39 (s, 2H), 3.28 (t, J=7.5 Hz, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 153.89, 151.18, 148.69, 142.17, 141.89, 130.74, 129.74, 129.04, 127.17, 120.73, 117.77, 115.63, 107.94, 104.05, 100.05, 99.49, 57.40, 49.27, 15.93 ppm; HRMS (ESI): m/z [M+H]+. C19H16N7O2 calculated value 374.1360, measured value 374.1377.
Step 1: Dissolve tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (0.3 g, 0.7 mmol) in 10 mL of DMF, add Na2S2O5 (0.7 g, 3.5 mmol) followed by adding 2-amino-3-pyridinecarboxaldehyde (0.2 g, 1.4 mmol). Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield tert-butyl 3-(2-(2-aminopyridin-3-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate as a yellow solid (0.3 g, 82%). HRMS (ESI): m/z [M+H]+. C28H30N7O4S calculated value 560.2074, measured value 560.2077.
Step 2: Dissolve tert-butyl 3-(2-(2-aminopyridin-3-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate (0.3 g, 0.5 mmol) in 10 mL dichloromethane, slowly add trifluoroacetic acid (0.6 g, 5.0 mmol) and stir at room temperature for 12 hours. The product was concentrated under vacuum and 3-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)pyridin-2-amine was obtained as a light yellow oil (0.2 g, 81%). The product can be used directly in the next step without further purification. HRMS (EST): m/z [M+H]+. C23H22N7O2S calculated value 460.1550, measured value 460.1566.
Step 3: Dissolve 3-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)pyridin-2-amine (0.2 g, 0.4 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.1 g, 0.8 mmol) followed by the slow dropwise addition of propylsulfonyl chloride (0.1 g, 0.6 mmol) and then stir for 3 h at room temperature. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 3-(6-(phenylsulfonyl)-1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) pyridin-2-amine as a pale yellow oil (0.2 g, 81%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C26H28N7O4S2 calculated value 566.1639, measured value 566.1645.
Step 4: 3-(6-(Phenylsulfonyl)-1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) pyridin-2-amine (0.2 g, 0.4 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, followed by adding 5 mL of 1 M sodium hydroxide. Stir the mixture at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS47: 3-(1-(1-(propylsulfonyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-]pyridin-2-yl)pyridin-2-amine (0.1 g, 67%). 1 HNMR (300 MHz, DMSO-d6): δ=12.06 (s, 1H), 8.83 (s, 1H), 8.03 (t, J=7.5 Hz, 1H), 7.74 (s, 2H), 7.66 (d, J=6.0 Hz, 1H), 7.56 (s, 1H), 6.81 (s, 1H), 6.68 (t, J=7.5 Hz, 1H), 3.65-3.79 (m, 1H), 3.12-3.36 (m, 2H), 3.10 (t, J=9.0 Hz, 2H), 2.70-2.81 (m, 2H), 1.85-2.14 (m, 2H), 1.69 (m, 2H), 0.93 (t, J=7.5 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 156.68, 153.77, 148.64, 146.69, 142.19, 134.84, 129.04, 127.16, 120.74, 118.74, 115.63, 113.59, 99.49, 60.47, 56.83, 56.18, 50.04, 26.25, 13.38, 12.48 ppm; HRMS (ESI): m/z [M+H]+. C20H24N7O2S calculated value 426.1707, measured value 426.1705.
Step 1: Dissolve (1R)-1-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by the slow dropwise addition of 2-bromoacetonitrile (0.2 g, 1.1 mmol). Raise the temperature to maintain reflux and stir for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-(3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)acetonitrile as a pale yellow oil (0.2 g, 61%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C22H23N6O3S calculated value 451.1547, measured value 451.1550.
Step 2: Dissolve 2-(3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)acetonitrile (0.2 g, 0.4 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, and stir at room temperature for 5 hours after adding 5 mL of 1 M sodium hydroxide. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS48: 2-(3-(2-((R)-1-Hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)acetonitrile (0.1 g, 73%). 1 HNMR (300 MHz, DMSO-d6): δ=11.86 (s, 1H), 8.87 (s, 1H), 7.59 (s, 1H), 6.89 (s, 1H), 4.53-4.68 (m, 1H), 3.68-3.78 (m, 1H), 3.48 (s, 2H), 2.56-2.81 (m, 2H), 2.20-2.30 (m, 2H), 1.90-2.15 (m, 2H), 1.93 (t, J=6.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.59, 142.17, 129.04, 127.16, 120.75, 115.63, 114.78, 99.28, 63.69, 58.18, 57.39, 54.83, 50.04, 26.97, 22.86 ppm; HRMS (ESI): m/z [M+H]+. C16H19N6O calculated value 311.1615, measured value 311.1618.
Step 1 Dissolve (1R)-1-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by the slow dropwise addition of 3-bromopropionitrile (0.2 g, 1.1 mmol). Raise the temperature to maintain reflux and stir for 3 h. The reaction was monitored by TLC for completion. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-(3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-2-yl)propanenitrile as a pale yellow oil (0.2 g, 59%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C23H25N6O3S calculated value 465.1703, measured value 465.1709.
Step 2: Dissolve 2-(3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)propanenitrile (0.2 g, 0.4 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, and stir at room temperature for 5 hours after adding 5 mL of 1 M sodium hydroxide. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS49: 3-(3-(2-((R)-1-Hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)propanenitrile (0.1 g, 72%). 1 HNMR (300 MHz, DMSO-d6): δ=12.14 (s, 1H), 8.97 (s, 1H), 7.53 (s, 1H), 6.91 (s, 1H), 4.58-4.72 (m, 1H), 3.69-3.81 (m, 1H), 3.01 (t, J=9.0 Hz, 2H), 2.72-2.85 (m, 2H), 2.70 (t, J=9.0 Hz, 2H), 2.22-2.36 (m, 2H), 1.93-2.19 (m, 2H), 1.58 (t, J=7.5 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.66, 142.98, 129.03, 127.17, 120.77, 119.04, 115.69, 99.38, 63.69, 58.94, 57.69, 55.16, 27.64, 22.85, 17.17 ppm; HRMS (ESI): m/z [M+H]+. C17H21N6O calculated value 325.1771, measured value 325.1777.
Step 1: Dissolve 3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)propanenitrile (0.3 g, 0.7 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by adding 5-methylfurfural (0.2 g, 1.4 mmol). Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 3-(4-(2-(5-methylfuran-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile as a yellow oil (0.3 g, 82%). HRMS (ESI): m/z [M+H]+. C25H20N7O3S calculated value 498.1343, measured value 498.1350.
Step 2: Dissolve 3-(4-(2-(5-methylfuran-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.3 g, 0.6 mmol) in a solvent mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, followed by adding 5 mL of 1 M sodium hydroxide. The mixture was then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate =1:1) to yield LXS50: 3-(4-(2-(5-methylfuran-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.1 g, 46%). 1 HNMR (300 MHz, DMSO-d6): δ=11.87 (s, 1H), 8.65 (s, 11H), 8.48 (s, 1H), 7.95 (s, 1H), 7.34 (s, 1H), 6.32 (d, J=3.0 Hz, 1H), 6.21 (s, 1H), 5.95 (s, 1H), 4.61 (t, J=4.5 Hz, 2H), 3.26 (t, J=6.0 Hz, 2H), 2.36 (s, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 154.31, 145.68, 143.29, 142.72, 138.24, 135.85, 135.54, 134.05, 129.75, 124.57, 118.82, 118.75, 113.43, 108.51, 104.57, 96.45, 48.05, 19.20, 13.73 ppm; HRMS (ESI): m/z [M+H]+. C19H16N7O calculated value 358.1411, measured value 358.1414.
Step 1: Dissolve (5-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)furan-2-methanol (0.3 g, 0.6 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.2 mmol) followed by the slow dropwise addition of 3-bromopropionitrile (0.2 g, 0.9 mmol). Raise the temperature to maintain reflux and stir for 3 h. The reaction was monitored by TLC for completion. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 3-(3-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)propanenitrile as a pale yellow oil (0.2 g, 60%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C26H25N6O4S calculated value 517.1653, measured value 517.1659.
Step 2: Dissolve 3-(3-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)propanenitrile (0.2 g, 0.4 mmol) in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, followed by adding 5 mL of 1 M sodium hydroxide. Stir for 5 h at room temperature and monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS51: 3-(3-(2-(5-(Hydroxymethyl)furan-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)propanenitrile (0.1 g, 69%). 1 HNMR (300 MHz, DMSO-d6); δ=12.03 (s, 1H), 8.88 (s, 1H), 7.59 (s, 1H), 7.02 (d, J=9.0 Hz, 1H), 6.84 (s, 1H), 6.59 (d, J=9.0 Hz, 1H), 4.39 (s, 2H), 3.64-3.78 (m, 1H), 3.03 (t, J=9.0 Hz, 2H), 2.74-2.86 (m, 2H), 2.73 (t, J=9.0 Hz, 2H), 2.20-2.35 (m, 2H), 1.90-2.15 (m, 2H) ppm; 13 CNMR (75 MHz, DMSO-d6) δ 153.88, 151.18, 148.64, 144.94, 142.17, 129.04, 127.18, 120.79, 119.04, 115.63, 107.94, 104.06, 99.38, 57.39, 57.01, 55.84, 55.19, 27.05, 17.16 ppm, HRMS (ESI): m/z [M+H]+. C20H21N6O2 calculated value 377.1721, measured value 377.1728.
Step 1: Dissolve (5-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)furan-2-methanol (0.3 g, 0.6 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.2 mmol) followed by the slow dropwise addition of 2-bromoacetonitrile (0.2 g, 0.9 mmol). Raise the temperature to maintain reflux and stir for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 3-(3-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl) acetonitrile as a pale yellow oil (0.2 g, 61%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C25H23N6O4S calculated value 503.1496, measured value 503.1500.
Step 2: Dissolve 3-(3-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)acetonitrile (0.2 g, 0.4 mmol) in a solvent mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, followed by adding 5 mL of 1 M sodium hydroxide. Stir for 5 h at room temperature and monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXSS2: 2-(3-(2-(5-(hydroxymethyl)furan-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)acetonitrile (0.1 g, 69%). 1 HNMR (300 MHz, DMSO-d6): δ=11.99 (s, 1H), 8.67 (s, 1H), 7.58 (s, 1H), 7.06 (d, J=7.5 Hz, 1H), 6.88 (s, 1H), 6.54 (d, J=7.5 Hz, 1H), 4.33 (s, 2H), 3.69-3.83 (m, 1H), 3.48 (s, 2H), 2.56-2.81 (m, 2H), 2.20-2.30 (m, 2H), 1.93-2.18 (m, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 153.97, 151.19, 148.63, 144.95, 142.18, 129.03, 127.17, 120.75, 115.69, 114.84, 107.94, 104.05, 99.39, 57.36, 56.72, 55.09, 54.81, 50.05, 26.29 ppm; HRMS (EST): m/z [M+H]+. C19H19N6O2 calculated value 363.1564, Measured value 363.1569.
Step 1: Dissolve 3-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)phenol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by slow dropwise addition of 2-bromoacetonitrile (0.2 g, 1.1 mmol). Raise the temperature to maintain reflux and stir for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-(3-(2-(3-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl) acetonitrile as a yellow oil (0.2 g, 61%). The product can be used directly in the next step without further purification. HRMS (EST): m/z [M+H]+. C26H23N6O3S calculated value 499.1547, measured value 499.1550.
Step 2: Dissolve (3-(2-(3-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)acetonitrile (0.2 g, 0.4 mmol) in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, add 1 M sodium hydroxide 5 mL and stir at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether; ethyl acetate=1:1) to yield LXS53: 2-(3-(2-(3-Hydroxyphenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)acetonitrile (0.1 g, 70%). 1HNMR (300 MHz, DMSO-d6): δ=12.09 (s, 1H), 8.83 (s, 1H), 7.84 (d, J=6.0 Hz, 1H), 7.66 (s, 1H), 7.34 (t, J=6.0 Hz, 1H), 7.04 (s, 1H), 6.91 (d, J=6.0 Hz, 1H), 6.83 (s, 1H), 3.71-3.82 (m, 1H), 3.49 (s, 2H), 2.58-2.83 (m, 2H), 2.17-2.36 (m, 2H), 1.90-2.16 (m, 2H) ppm; —CNMR (75 MHz, DMSO-d6) δ 157.59, 153.75, 148.63, 142.17, 132.05, 130.65, 129.05, 127.16, 120.75, 120.11, 115.94, 115.65, 114.85, 112.94, 99.38, 57.3, 54.84, 50.04, 26.84 ppm; HRMS (EST): m/z [M+H]+. C20H19N6O calculated value 359.1615, measured value 359.1619.
Step 1: Triethyloxonium tetrafluoroborate (0.5 g, 2.4 mmol) and (R)-lactamide (0.2 g, 2.4 mmol) were dissolved in 10 mL of tetrahydrofuran, stirred at room temperature for 3 h and concentrated under vacuum to obtain the mixture as an oil, followed by the addition of 10 mL of ethanol to dissolve and the addition of 4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)benzonitrile (0.3 g, 0.8 mmol). Then, raise the temperature while maintaining reflux and stir for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (R)-4-(2-(1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)benzonitrile as a light green oil (0.3 g, 88%). HRMS (ESI): m/z [M+H]+. C23H18N5O3S calculated value 444.1125, measured value 444.1129.
Step 2: Dissolve (R)-4-(2-(1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)benzonitrile (0.3 g, 0.7 mmol) in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, add 1 M sodium hydroxide 5 mL and stir at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS54: (R)-4-(2-(1-Hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)benzonitrile (0.1 g, 49%). 1HNMR (300 MHz, DMSO-d6): δ=12.01 (s, 1H), 8.91 (s, 1H), 7.80 (d, J=9.0 Hz, 2H), 7.64 (d, J=9.0 Hz, 2H), 7.56 (s, 1H), 6.88 (s, 1H), 3.98-4.68 (m, 1H), 1.49 (d, J=7.5 Hz, 3H) ppm; 13 CNMR (75 MHz, DMSO-d6) δ 151.75, 148.69, 142.16, 140.48, 134.27, 129.04, 127.18, 122.84, 120.73, 118.64, 115.67, 112.17, 99.38, 62.95, 22.84 ppm; HRMS(ESI): m/z [M+H]+. C17H14N5O calculated value 304.1193, measured value 304.1203.
Step 1: Dissolve 4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)benzonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by the addition of 5-hydroxymethylfurfural (0.2 g, 1.6 mmol) dropwise, then raise the temperature to 90° C. and stir for 12 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice with dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)benzonitrile as a yellow oil (0.3 g, 79%). HRMS(ESI): m/z [M+H]+. C26H15N5O4S calculated value 496.1074, measured value 496.1080.
Step 2: Dissolve 4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)benzonitrile (0.3 g, 0.6 mmol) in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, add 5 mL of 1 M sodium hydroxide and then stir at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXSS5: 4-(2-(5-(hydroxymethyl)furan-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)benzonitrile (0.1 g, 47%). 1HNMR (300 MHz, DMSO-d6): δ 11.91 (s, 1H), 8.73 (s, 1H), 7.83 (d, J=6.0 Hz, 2H), 7.68 (d, J=6.0 Hz, 2H), 7.56 (s, 1H), 7.02 (d, J=9.0 Hz, 1H), 6.83 (s, 1H), 6.59 (d, J=9.0 Hz, 1H), 4.39 (s, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ153.85, 151.17, 148.64, 142.59, 142.31, 134.27, 129.05, 127.17, 122.85, 120.73, 118.65, 115.69, 112.15, 107.94, 104.06, 99.38, 57.39 ppm; HRMS (ESI): m/z [M+H]+. C20H14N5O2 calculated value 356.1142, measured value 356.1149.
Step 1: Dissolve 4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)benzonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by the addition of 3-hydroxybenzaldehyde (0.2 g, 1.6 mmol) dropwise, raise the temperature to 90° C. and stir for 12 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice with dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 4-(2-(3-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)benzonitrile as a yellow oil (0.3 g, 79%). HRMS(ESI): m/z [M+H]+. C27H18N5O3S calculated value 492.1125, measured value 492.1134.
Step 2: 4-(2-(3-Hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)benzonitrile (0.3 g, 0.6 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, and the mixture was stirred at room temperature for 5 h after the addition of 5 mL 1 M sodium hydroxide. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXSS6: 4-(2-(3-hydroxyphenyl)imidazo[4,5-d]pvrrolo[2,3-b]pyridin-1(6H)-yl)benzonitrile (0.1 g, 47%). 1HNMR (300 MHz, DMSO-d6): δ=12.06 (s, 1H), 8.83 (s, 1H), 7.84-7.93 (m, 1H), 7.81 (d, J=7.5 Hz, 2H), 7.64 (d, J=7.5 Hz, 2H), 7.58 (s, 1H), 7.34 (t, J=9.0 Hz, 1H), 7.08 (s, 1H), 6.91 (d, J=9.0 Hz, 1H), 6.88 (s, 1H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 157.89, 148.99, 144.47, 142.59, 142.16, 134.27, 132.04, 130.64, 129.05, 127.49, 122.84, 120.75, 120.16, 118.69, 115.96, 115.61, 112.94, 112.15, 99.54 ppm; HRMS (ESI): m/z [M+H]+. C21H14N5O calculated value 352.1193, measured value 352.1199.
Step 1: Dissolve 4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)benzonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by adding 4-methylsulfonyl benzaldehyde (0.3 g, 1.6 mmol) dropwise, then raise the temperature to 90° C. and stir for 12 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 4-(2-(4-(methylsulfonyl)phenyl)-6-(phenylsulfonyl)-2,3-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)benzonitrile as a yellow oil (0.3 g, 70%). HRMS (ESI): m/z [M+H]+. C28H22N5O4S2 calculated value 556.1108, measured value 556.1113.
Step 2: Dissolve 4-(2-(4-(methylsulfonyl)phenyl)-6-(phenylsulfonyl)-2,3-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)benzonitrile (0.3 g, 0.5 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, add 5 mL of 1 M sodium hydroxide and then stir at room temperature for 5 hours. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS57: 4-(2-(4-(methylsulfonyl)phenyl)-2,3-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)benzonitrile (0.1 g, 45%). 1HNMR (300 MHz, DMSO-d6): δ=11.86 (s, 1H), 7.89 (s, 1H), 7.74 (d, J=6.0 Hz, 2H), 7.63 (s, 1H), 7.54 (d, J=6.0 Hz, 2H), 7.42 (d, J=9.0 Hz, 2H), 6.88 (s, 1H), 6.78 (d, J=9.0 Hz, 2H), 5.08 (s, 1H), 3.32 (s, 1H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 153.77, 149.47, 139.04, 138.79, 134.41, 133.05, 131.18, 128.15, 127.93, 127.16, 123.38, 118.66, 114.28, 108.15, 101.04, 99.37, 87.84, 47.72 ppm; HRMS (ESI): m/z [M+H]+. C22H18N5O2S calculated value 416.1176, measured value 416.1180.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((R-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) and then slowly add 3-bromopropanenitrile (0.2 g, 1.1 mmol) dropwise. Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were then combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 3-((R)-3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl propanenitrile as a pale yellow oil (0.2 g, 59%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C23H25N6O3S calculated value 465.1703, measured value 465.1709.
Step 2: 3-((R)-3-(2-((R)-1-Hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl) propanenitrile (0.2 g, 0.4 mmol) was dissolved in a mixture of 5 mL tetrahydrofuran and 5 mL methanol, followed by adding 5 mL of 1 M sodium hydroxide. Stir the mixture at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS58: 3-((R)-3-(2-((R)—I-Hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)propanenitrile (0.1 g, 72%). 1HNMR (300 MHz, DMSO-d6): δ=11.98 (s, 1H), 8.73 (s, 1H), 7.56 (s, 1H), 6.78 (s, 1H), 4.61-4.75 (m, 1H), 3.65-3.83 (m, 1H), 3.04 (t, J=6.0 Hz, 2H), 2.70-2.83 (m, 2H), 2.73 (t, J=6.0 Hz, 2H), 2.20-2.31 (m, 2H), 1.90-2.15 (m, 2H), 1.49 (t, J=7.5 Hz, 3H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 148.64, 148.59, 142.19, 129.03, 127.16, 120.74, 119.04, 115.69, 99.39, 63.62, 58.91, 57.63, 55.16, 27.63, 22.84, 17.19 ppm; HRMS (ESI): m/z [M+H]+. C17H21N6O calculated value 325.1771, measured value 325.1777.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((S-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) and then slowly add 3-bromopropanenitrile (0.2 g, 1.1 mmol) dropwise. Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 3-((S)-3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl propanenitrile as a pale yellow oil (0.2 g, 59%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C23H25N6O3S calculated value 465.1703, measured value 465.1711.
Step 2: 3-((S)-3-(2-((R)-1-Hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl propanenitrile (0.2 g, 0.4 mmol) was dissolved in a mixture of 5 mL tetrahydrofuran and 5 mL methanol, followed by adding 5 mL of 1 M sodium hydroxide, stir at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS59: 3-((S)-3-(2-((R)-1-hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)propanenitrile (0.1 g, 72%). 1HNMR (300 MHz, DMSO-d6): δ=11.99 (s, 1H), 8.83 (s, 1H), 7.59 (s, 1H), 6.88 (s, 1H), 4.51-4.69 (m, 1H), 3.65-3.85 (m, 1H), 3.05 (t, J=9.0 Hz, 2H), 2.76-2.89 (m, 2H), 2.73 (t, J=9.0 Hz, 2H), 2.20-2.41 (m, 2H), 1.95-2.22 (m, 2H), 1.43 (t, J=7.5 Hz, 3H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 148.61, 148.57, 142.11, 128.64, 126.11, 120.73, 118.57, 115.62, 99.38, 63.62, 58.93, 57.62, 55.17, 27.64, 22.89, 16.14 ppm; HRMS (ESI): m/z [M+H]+. C17H21N6O calculated value 325.1771, measured value 325.1780.
Step 1: Dissolve tert-butyl 3-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (0.3 g, 0.7 mmol) in 10 mL of DMF, add Na2S2O5 (0.7 g, 3.5 mmol) followed by adding 4-methylsulfonylbenzaldehyde (0.2 g, 1.4 mmol). Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield 3-(2-(4-(methylsulfonyl)phenyl)-6-(phenylsulfonyl)-2,3-dihydroimidazo[4,5-d]pvrrolo[2,3-b]pyridin-1(6H)-pyrrolidine-1-carboxylate as a yellow solid (0.2 g, 49%). HRMS(ESI): m/z [M+H]+. CaoH34N5O6S2 calculated value 624.1945, measured value 624.1955.
Step 2: Dissolve 3-(2-(4-(methylsulfonyl)phenyl)-6-(phenylsulfonyl)-2,3-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-pyrrolidine-1-carboxylate (0.2 g, 0.3 mmol) in 10 mL of dichloromethane, slowly add trifluoroacetic acid (0.4 g, 3.0 mmol) and then stir for 12 h at room temperature. The product was then concentrated under vacuum to yield 2-(4-(methylsulfonyl)phenyl)-6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,2,3,6-tetrahydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine as a light yellow oil (0.1 g, 60%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C25H26N5O4S2 calculated value 524.1421, measured value 524.1427.
Step 3: Dissolve 2-(4-(methylsulfonyl)phenyl)-6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,2,3,6-tetrahydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine (0.1 g, 0.2 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.1 g, 0.4 mmol) followed by slow dropwise addition of 3-bromopropyl cyanide (0.1 g, 0.3 mmol). Stir at room temperature for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 3-(3-(2-(4-(methylsulfonyl)phenyl)-6-(phenylsulfonyl)-2,3-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-vl propanenitrile as a pale yellow oil (0.1 g, 91%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C28H29N6O4S2 calculated value 577.1686, measured value 577.1688.
Step 4: Dissolve 3-(3-(2-(4-(methylsulfonyl)phenyl)-6-(phenylsulfonyl)-2,3-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl propanenitrile (0.1 g, 0.2 mmol) in a mixture of 5 mL tetrahydrofuran and 5 mL methanol, add 5 mL of 1 M sodium hydroxide and then stir at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS60: 3-(3-(2-(4-(Methylsulfonyl)phenyl)-2,3-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)propanenitrile (60.0 mg, 79%). 1HNMR (300 MHz, DMSO-d6): δ=11.46 (s, 1H), 7.89 (s, 1H), 7.74 (d, J=9.0 Hz, 2H), 7.56 (s, 1H), 7.51 (d, J=9.0 Hz, 2H), 6.88 (s, 1H), 5.04 (s, 1H), 3.32 (s, 3H), 3.03 (t, J=9.0 Hz, 2H), 2.75 (t, J=9.0 Hz, 2H), 2.66-2.71 (m, 1H), 2.31-2.56 (m, 2H), 2.19-2.33 (m, 2H), 1.66-2.02 (m, 2H) ppm; 13 CNMR (75 MHz, DMSO-d6) δ 144.28, 139.28, 138.73, 134.49, 131.18, 128.95, 128.15, 127.15, 123.38, 119.04, 108.16, 99.38, 83.28, 62.18, 59.28, 55.41, 55.14, 47.72, 33.58, 17.10 ppm; HRMS (ESI): m/z [M+H]+. C22H25N6O2S calculated value 437.1754, measured value 437.1759.
Step 1: Dissolve (1R)-1-(6-(phenylsulfonyl)-1-(pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by slow dropwise addition of 3-bromopropacyanine (0.2 g, 1.1 mmol). Stir at room temperature for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 1-(3-(2-(R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-carbonyl)cyclopropanecarbonitrile as a yellow oil (0.3 g, 82%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C25H25N6O4S calculated value 505.1653, measured value 505.1660.
Step 2: 1-(3-(2-((R)-1-Hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-carbonyl)cyclopropanecarbonitrile (0.3 g, 0.6 mmol) was dissolved in a mixture of 5 mL tetrahydrofuran and 5 mL methanol, followed by the addition of 5 mL of 1 M sodium hydroxide. Stir at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS61: 1-(3-(2-((R)-1-Hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-carbonyl)cyclopropanecarbonitrile (0.1 g 46%). 1HNMR (300 MHz, DMSO-d6): δ=12.06 (s, 1H), 7.55 (s, 1H), 6.87 (s, 1H), 4.33-4.68 (m, 1H), 3.894.17 (m, 1H), 3.77-4.03 (m, 2H), 3.41-3.51 (m, 2H), 2.29-2.54 (m, 2H), 1.48 (d, J=9.0 Hz, 3H), 0.67-0.92 (m, 4H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 180.77, 148.66, 148.53, 142.17, 129.00, 127.17, 120.73, 115.63, 114.47, 99.39, 63.69, 58.42, 51.88, 46.48, 27.15, 22.84, 13.63, 10.06 ppm; HRMS (ESI): m/z [M+H]+. C19H21N6O2 calculated value 365.1721, measured value 365.1711.
Step 1: Tert-butyl 3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)-3-(cyanomethyl)azetidine-1-carboxylate (0.3 g, 0.6 mmol) was dissolved in 10 mL of DMF, Na2S2O5 (0.6 g, 3.0 mmol) was added, followed by the addition of 5-methyl-2-thiophenecarboxaldehyde (0.2 g, 0.9 mmol) dropwise and stirring for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield tert-butyl 3-(cyanomethyl)-3-(4-(2-(5-methylthien-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl) azetidine-1-carboxylate as a yellow solid (0.2 g, 56%). HRMS (ESI): m/z [M+H]+. C32H31N5O4S2 calculated value 655.1904, measured value 655.1909.
Step 2: Tert-butyl 3-(cyanomethyl)-3-(4-(2-(5-methylthien-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidine-1-carboxylate (0.2 g, 0.3 mmol) was dissolved in 10 mL of dichloromethane and then trifluoroacetic acid (0.4 g, 3.0 mmol) was slowly added and stirred at room temperature for 12 h. The product was concentrated under vacuum to yield 2-(3-(4-(2-(5-methylthien-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile as a light yellow oil (0.1 g, 59%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C27H23N5O2S2 calculated value 555.1380, measured value 555.1390.
Step 3: Dissolve 2-(3(4-(2-(5-methylthien-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (0.1 g, 0.2 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.1 g, 0.4 mmol), followed by the slow dropwise addition of ethylsulfonyl chloride (0.1 g, 0.3 mmol), stir the mixture slowly for 3 h at room temperature. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-(1-(ethylsulfonyl)-3-(4-(2-(5-methylthien-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile as a light yellow oil (0.1 g, 86%). The product can be used directly in the next step without further purification. HRMS(ESI): m/z [M+H]+. C29H27N5O4S3 calculated value 647.1312, measured value 647.1318.
Step 4: 2-(1-(Ethylsulfonyl)-3-(4-(2-(5-methylthien-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (0.1 g, 0.2 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol. Add 5 mL of 1 M sodium hydroxide and then stir at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate =1:1) to yield LXS62: 2-(1-(Ethylsulfonyl)-3-(4-(2-(5-methylthien-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (65.0 mg, 83%). 1 HNMR (300 MHz, DMSO-d6): δ=12.03 (s, 1H), 8.89 (s, 1H), 8.08 (s, 2H), 7.65 (s, 1H), 7.49 (d, J=9.0 Hz, 1H), 6.89 (s, 1H), 6.84 (d, J=9.0 Hz, 1H), 3.89-4.06 (m, 4H), 3.45 (m, 2H), 2.81 (s, 2H), 2.36 (s, 3H), 1.22 (t, J=7.5 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.69, 142.17, 141.66, 141.61, 134.33, 130.77, 129.73, 129.01, 127.49, 127.11, 120.73, 117.74, 115.63, 100.54, 99.31, 59.29, 51.66, 50.83, 23.28, 15.22, 2.69 ppm. HRMS (ESI): m/[M+H]+. C23H23N5O2S2 calculated value 507.1380, measured value 507.1390.
Step 1: Dissolve tert-butyl 3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)-3-(cyanomethyl)azetidine-1-carboxylate (0.3 g, 0.6 mmol) in 10 mL of DMF, add Na2S2O5 (0.6 g, 3.0 mmol). After adding 2-thiophenecarboxaldehyde (0.2 g, 0.9 mmol) dropwise, raise the temperature to 90° C. and stir for 12 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield tert-butyl 3-(cyanomethyl)-3-(4-(6-(phenylsulfonyl)-2-(thiophen-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidine-1-carboxylate as a yellow solid (0.2 g, 57%). HRMS (ESI): m/z [M+H]+. C31H29N5O4S2 calculated value 641.1748, measured value 641.1751.
Step 2: Tert-butyl 3-(cyanomethyl)-3-(4-(6-(phenylsulfonyl)-2-(thiophen-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidine-1-carboxylate (0.2 g, 0.3 mmol) was dissolved in 10 mL ofdichloromethane, and then trifluoroacetic acid (0.4 g, 3.0 mmol) was slowly added and stirred at room temperature for 12 hours. The product was concentrated under vacuum to yield 2-(3-(4-(6-(phenylsulfonyl)-2-(thiophen-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile as a light yellow oil (0.1 g, 59%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C26H21N5O2S2 calculated value 541.1223, measured value 541.1230.
Step 3: Dissolve 2-(3-(4-(6-(phenylsulfonyl)-2-(thiophen-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (0.1 g, 0.2 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.1 g, 0.4 mmol), followed by the dropwise addition of ethylsulfonyl chloride (0.1 g, 0.3 mmol), and then stir slowly for 3 h at room temperature. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-(3-(4-(6-(phenylsulfonyl)-2-(thiophen-2-yl)imidazo[4,5-d]pyrrolo[2,3-]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile as a pale yellow oil (0.1 g, 85%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C28H25N5O4S3 calculated value 633.1155, measured value 633.1160.
Step 4: 2-(3-(4-(6-(Phenylsulfonyl)-2-(thiophen-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (0.1 g, 0.2 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, and 5 mL of 1M sodium hydroxide was added and then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS63: 2-(1-(Ethylsulfonyl)-3-(4-(2-(thiophen-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)azetidin-3-yl)acetonitrile (60.0 mg, 77%). 1HNMR (300 MHz, DMSO-d6): δ=11.96 (s, 1H), 8.74 (s, 1H), 8.10 (s, 2H), 7.85 (d, J=7.5 Hz, 1H), 7.69 (d, J=7.5 Hz, 1H), 7.67 (s, 1H), 7.13-7.21 (m, 1H), 6.79 (s, 1H), 3.88-4.16 (m, 4H), 3.55 (m, 2H), 2.86 (s, 2H), 1.22 (t, J=7.5 Hz, 3H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 148.67, 143.88, 142.19, 141.53, 130.77, 129.73, 129.04, 128.63, 128.04, 127.17, 120.73, 117.72, 115.69, 100.53, 99.38, 59.28, 51.64, 50.88, 23.37, 5.77 ppm: HRMS (ESI): m/z [M+H]+. C22H21N5O2S2 calculated value 493.1223, measured value 493.1230.
Step 1: Triethyloxonium tetrafluoroborate (0.4 g, 2.1 mmol) and 2,2,2-trifluoroacetamide (0.2 g, 2.1 mmol) were dissolved in 10 mL of tetrahydrofuran, stirred at room temperature for 3 h and concentrated under vacuum to obtain the mixture in the form of oil, followed by the addition of 10 mL of ethanol to dissolve and the addition of 3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino-1H-pyrazol-1-yl)propanenitrile (0.3 g, 0.7 mmol). Raise the temperature to maintain reflux and stir for 3 h. The reaction was monitored by TLC for completion. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 3-(4-(6-(phenylsulfonyl)-2-(trifluoromethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile as a milk white oil (0.3 g, 84%). HRMS (ESI): m/z [M+H]+. C21H15F3N7O2S calculated value 486.0955, measured value 486.0961.
Step 2: Dissolve 3-(4-(6-(phenylsulfonyl)-2-(trifluoromethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl) propanenitrile (0.3 g, 0.6 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, and stir for 5 h at room temperature after adding 5 mL of 1 M sodium hydroxide. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS64: 3-(4-(2-(Trifluoromethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.1 g, 47%). 1HNMR (300 MHz, DMSO-d6): δ=11.96 (s, 1H), 8.89 (s, 1H), 8.06 (s, 1H), 7.94 (s, 1H), 7.56 (s, 1H), 6.86 (s, 1H), 5.04 (t, J=9.0 Hz, 2H), 3.24 (t, J=9.0 Hz, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 148.69, 144.57, 142.18, 130.77, 129.84, 129.04, 127.11, 120.73, 117.77, 116.73, 115.69, 100.52, 99.67, 49.28, 16.88 ppm. HRMS (ESI): m/z [M+H]+. C15H11F3N7 calculated value 346.1023, measured value 346.1029.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((S-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) and then slowly add 4-bromobutyronitrile (0.2 g, 1.1 mmol) dropwise. Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 4-((S)-3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)butyronitrile as a pale yellow oil (0.2 g, 57%). The product can be used directly in the next step without further purification. HRMS(ESI): m/z [M+H]+. C24H27N6O3S calculated value 479.1860, measured value 479.1871.
Step 2: 4-((S)-3-(2-((R)-1-Hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)butyronitrile (0.2 g, 0.4 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, followed by the addition of 5 mL of IM sodium hydroxide. Stir at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS65: 4-((S)-3-(2-((R)-1-Hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)butanenitrile (0.1 g, 71%). 1HNMR (300 MHz, DMSO-d6): δ=12.16 (s, 1H), 8.89 (s, 1H), 7.69 (s, 1H), 6.88 (s, 1H), 4.53-4.69 (m, 11H), 3.69-3.80 (m, 1H), 2.56-2.81 (m, 2H), 2.43 (t, J=7.5 Hz, 2H), 2.21-2.31 (m, 2H), 1.92-2.16 (m, 2H), 1.87 (t, J=7.5 Hz, 2H), 1.66-1.78 (m, 2H), 1.44 (d, J=9.0 Hz, 3H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 148.65, 148.51, 142.17, 129.04, 127.19, 120.77, 119.37, 115.62, 99.74, 63.67, 58.92, 58.49, 55.91, 55.53, 27.63, 22.81, 15.92, 15.19 ppm; HRMS (ESI): m/z [M+H]+. C18H23N6O calculated value 339.1928, measured value 339.1931.
Step 1: Dissolve 4-nitropyrazole (0.5 g, 4.4 mmol) in 10 mL of tetrahydrofuran, add DIPEA (1.2 g, 8.8 mmol) followed by slow dropwise addition of 3-bromo-3-cyclopentylpropanenitrile (1.3 g, 6.6 mmol). Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 3-cyclopentyl-3-(4-nitro-1H-pyrazol-1-yl)propanenitrile as a pale yellow oil (0.8 g, 77%). The product can be used directly in the next step without further purification. HRMS(ESI): m/z [M+H]+. C11H15N4O2 calculated value 235.1190, measured value 235.1194.
Step 2: 3-Cyclopentyl-3-(4-nitro-1H-pyrazol-1-yl)propanenitrile (0.8 g, 3.4 mmol) was dissolved in 10 mL of methanol, after adding palladium carbon (0.1 g, 10%), use hydrogen to replace the air in the reaction flask more than three times and the reaction was kept in hydrogen atmosphere. Stir at room temperature for 12 h. The reaction was monitored for completion by TLC. After filtration, the filtrate was collected and concentrated under vacuum to obtain 3-cyclopentyl-3-(4-amino-1H-pyrazol-1-yl)propanenitrile as a pink foamy solid (0.7 g, 100%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C11H17N4 calculated value 205.1448, measured value 205.1552.
Step 3: Dissolve 3-cyclopentyl-3-(4-amino-1H-pyrazol-1-yl)propanenitrile (0.7 g, 3.4 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.9 g, 6.8 mmol) followed by adding 4-chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (0.8 g, 2.3 mmol). Raise the temperature to maintain reflux and stir for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice with dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. Triturate and cure with an appropriate amount of methanol to yield 3-cyclopentyl-3-(4-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)propanenitrile as a pale yellow oil (0.6 g, 50%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C24H24N7O4S calculated value 506.1605, measured value 506.1613.
Step 4: 3-Cyclopentyl-3-(4-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)propanenitrile (0.6 g, 1.2 mmol) was dissolved in 10 mL of methanol, palladium carbon (0.1 g, 10%) was added and the air in the reaction flask was replaced more than three times using hydrogen. The reaction was carried out in hydrogen atmosphere and stirred at room temperature for 12 h. The reaction was monitored for completion by TLC. After filtration, the filtrate was collected and concentrated under vacuum to yield 3-cyclopentyl-3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)propanenitrile as a pink foamy solid (0.5 g, 88%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C24H26N7O2S calculated value 476.1863, measured value 476.1870.
Step 5: Triethyloxonium tetrafluoroborate (0.6 g, 3.3 mmol) and 2,2,2-trifluoroacetamide (0.1 g, 3.3 mmol) were dissolved in 10 mL of tetrahydrofuran, stirred at room temperature for 3 h and concentrated under vacuum to obtain the mixture as an oil, followed by the addition of 10 mL of ethanol to dissolve and the addition of 3-cyclopentyl-3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)propanenitrile (0.5 g, 1.1 mmol). Raise the temperature to maintain reflux and stir for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=2:1) to yield 3-cyclopentyl-3-(4-(6-(phenylsulfonyl)-2-(trifluoromethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl) propanenitrile as a light green oil (0.2 g, 34%). HRMS(ESI): m/z [M+H]+. C26H23F3N7O2S calculated value 554.1581, measured value 554.1593.
Step 6: 3-Cyclopentyl-3-(4-(6-(phenylsulfonyl)-2-(trifluoromethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.2 g, 0.4 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, followed by the addition of 5 mL of 1 M sodium hydroxide. Stir for 5 h at room temperature and monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS66: 3-Cyclopentyl-3-(4-(2-(trifluoromethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.1 g, 67%). 1HNMR (300 MHz, DMSO-d6): δ=11.96 (s, 1H), 8.79 (s, 1H), 8.06 (s, 1H), 7.94 (s, 1H), 7.58 (s, 1H), 6.77 (s, 1H), 3.53-3.70 (m, 1H), 2.80 (d, J=9.0 Hz, 2H), 1.35-1.60 (m, 9H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 148.66, 144.67, 142.19, 130.78, 129.77, 129.03, 127.16, 120.74, 117.74, 116.74, 115.64, 100.58, 99.39, 63.95, 34.28, 30.95, 25.16, 18.88 ppm; HRMS (ESI): m/z [M+H]+. C20H19F3N7 calculated value 414.1649, measured value 414.1654.
Step 1: Dissolve 3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)propanenitrile (0.3 g, 0.7 mmol) in 10 mL of DMF, add Na2S2O5 (0.7 g, 3.5 mmol) followed by adding 4-methylsulfonylbenzaldehyde (0.2 g, 0.9 mmol) and stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline and dried over anhydrous sodium sulfate to yield 3-(4-(2-(4-(methylsulfonyl)phenyl)-6-(phenylsulfonyl)-2,3-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile as a yellow solid (0.2 g, 47%). HRMS (ESI): m/z [M+H]+. C27H24N7O4S2S calculated value 574.1326, measured value 574.1331.
Step 2: 3-(4-(2-(4-(Methylsulfonyl)phenyl)-6-(phenylsulfonyl)-2,3-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.2 g, 0.3 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol followed by adding 1 M sodium hydroxide 5 mL and then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate =1:1) to yield LXS67: 3-(4-(2-(4-(methylsulfonyl)phenyl)-2,3-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.1 g, 66%). 1 HNMR (300 MHz, DMSO-d6): δ=11.06 (s, 11H), 7.89 (s, 1H), 7.86 (s, 1H), 7.74 (d, J=9.0 Hz, 2H), 7.58 (s, 1H), 7.51 (d, J=9.0 Hz, 2H), 7.18 (s, 1H), 6.87 (s, 1H), 5.04 (t, J=7.5 Hz, 3H), 3.34 (s, 3H), 3.20 (t, J=7.5 Hz, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 149.78, 139.07, 138.77, 134.44, 131.17, 130.48, 130.15, 128.19, 127.94, 127.18, 123.34, 117.79, 115.75, 108.15, 99.43, 87.59, 49.22, 47.78, 17.88 ppm; HRMS (ESI): m/z [M+H]+. C21H20N7O2S calculated value 434.1394, measured value 434.1400.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((S-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) and then slowly add 5-bromopentanonitrile (0.2 g, 1.1 mmol) dropwise. Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 5-((S)-3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl) pentanonitrile as a pale yellow oil (0.2 g, 56%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C25H29N6O3S calculated value 493.2016, measured value 493.2020.
Step 2: Dissolve 5-((S)-3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl) pentanonitrile (0.2 g, 0.4 mmol) in a mixture of 5 mL tetrahydrofuran and 5 mL methanol, followed by adding 5 mL of 1 M sodium hydroxide and stirring at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS68: 5-((S)-3-(2-((R)-1-Hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)pentanonitrile (0.1 g, 70%). 1HNMR (300 MHz, DMSO-d6): δ=12.03 (s, 1H), 8.87 (s, 1H), 7.58 (s, 1H), 6.85 (s, 1H), 4.594.68 (m, 1H), 3.65-3.79 (m, 1H), 2.58-2.83 (m, 2H), 2,44 (t, J=9.0 Hz, 2H), 2.19-2.28 (m, 2H), 1.90-2.13 (m, 2H), 1.90 (t, J=9.0 Hz, 2H), 1.63-1.77 (m, 2H), 1.48 (d, J=9.0 Hz, 3H), 1.25-1.30 (m, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 148.94, 148.51, 142.11, 129.75, 127.63, 120.71, 119.39, 115.67, 99.52, 62.68, 58.93, 58.41, 55.96, 55.81, 27.63, 27.21, 23.22, 22.84, 17.18 ppm; HRMS (ESI): m/z [M+H]+. C19H25N6O calculated value 353.2084, measured value 353.2088.
Step 1: Disslovetert-butyl (S)5-azaspiro[2,4]hept-7-carbamate (0.3 g, 1.4 mmol) in 10 mL of DMF, add potassium carbonate (0.4 g, 2.8 mmol) and 4-broinobutanenitrile (0.3 g, 2.1 mmol) and stir at room temperature for 12 h. Monitor the reaction for completion by MS. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted three times using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (S)-tert-butyl (5-(3-cyanopropyl)-5-azaspiro[2,4]hept-7-yl)carbamate as a pale yellow oil (0.3 g, 76%) The product can be used directly in the next step without further purification. HRMS (ESI): nm/z [M+H]+. C18H26N3O2 calculated value 280.2020, measured value 280.2029.
Step 2: Dissolve (S)-tert-butyl (5-(3-cyanopropyl)-5-azaspiro[2,4]hept-7-yl)carbamate (0.3 g, 1.1 mmol) in 10 mL dichloromethane, slowly add trifluoroacetic acid (1.3 g, 11.0 mmol) and then stir at room temperature for 12 h. Concentrate under vacuum to obtain (S)-4-(7-amino-5-azaspiro[2,4]hept-5-yl)butanenitrile as a light yellow oil (0.2 g, 100%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C10H18N3 calculated value 180.1495, measured value 180.1499.
Step 3: 4-Chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (0.3 g, 0.9 mmol) was dissolved in 100 mL of tetrahydrofuran followed by adding DIPEA (0.7 g, 5.5 mmol) and (S)-4-(7-amino-5-azaspiro[2,4]hept-5-yl)butanenitrile (0.2 g, 1.1 mmol). Raise the temperature to maintain reflux and stir for 4 h. The reaction was monitored for completion by TLC. Concentrate under vacuum to yield a yellow oily liquid. Triturate and cure with an appropriate amount of methanol to yield (S)-4-(7-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-5-azaspiro[2,4]hept-5-yl)butanenitrile as a yellow solid (0.3 g, 70%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C23H25N6O4S calculated value 481.1653, measured value 481.1659.
Step 4: Dissolve (S)-4-(7-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-5-azaspiro[2,4]hept-5-yl)butanenitrile (0.3 g, 0.6 mmol) in 10 mL of methanol, add palladium carbon (0.1 g, 10%) and replace the air in the reaction flask more than three times with hydrogen gas. The reaction was kept in hydrogen atmosphere and the reaction was stirred at room temperature for 12 h. The reaction was monitored for completion by TLC. The filtrate was collected and concentrated under vacuum to yield (S)-4-(7-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-5-azaspiro[2,4]hept-5-yl)butanenitrile as a light pink foamy solid (0.3 g, 100%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C23H27N6O2S calculated value 451.1911, measured value 451.1922.
Step 5: Triethyloxonium tetrafluoroborate (0.4 g, 2.0 mmol) and (R)-lactamide (0.2 g, 2.0 mmol) were dissolved in 20 mL of tetrahydrofuran, stirred at room temperature for 3 h and concentrated under vacuum to obtain the mixture as an oil, followed by the addition of 20 mL of ethanol to dissolve and the addition of (S)-4-(7-(((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-5-azaspiro[2.4]hept-5-yl)butanenitrile (0.3 g, 0.7 mmol). Raise the temperature to maintain reflux and stir for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield 4-((S)-7-(2-((R)-1-Hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-5-azaspiro[2,4] hept-5-yl)butanenitrile as a light yellow oil (0.2 g, 60%). HRMS (ESI): m/z [M+H]+. C26H29N6O3S calculated value 505.2016, measured value 505.2020.
Step 6: 4-((S)-7-(2-((R)-1-Hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-5-azaspiro[2,4]hept-5-yl)butanenitrile (0.2 g, 0.4 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, followed by adding 5 mL of 1 M sodium hydroxide and then stirring at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS69: 4-((S)-7-(2-((R)-1-Hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-5-azaspiro[2,4]hept-5-yl)butanenitrile (0.1 g, 69%). 1HNMR (300 MHz, DMSO-d6): δ=11.96 (s, 1H), 8.79 (s, 1H), 7.59 (s, 1H), 6.85 (s, 1H), 4.58-4.71 (m, 1H), 3.70-3.77 (m, 1H), 2.66-2.83 (m, 2H), 2,47 (t, J=7.5 Hz, 2H), 2.12-2.22 (m, 2H), 1.88 (t, J=9.0 Hz, 2H), 1.76-1.80 (m, 2H), 1.48 (d, J=9.0 Hz, 3H), 0.05-0.27 (m, 4H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 149.33, 148.58, 141.66, 129.05, 127.99, 120.78, 119.38, 115.64, 99.56, 71.22, 68.47, 63.68, 56.19, 53.48, 25.59, 22.83, 15.98, 15.11, 4.22 ppm; HRMS (ESI): m/z [M+H]+. C20H25N6O calculated value 365.2084, measured value 365.2089.
LXS70 was prepared in a similar preparation way to Example 69 and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS70: (R)-1-(1-((S)-5-(4,4,4-Trifluorobutyl)-5-azaspiro[2,4]hept-7-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.1 g, 66%). 1HNMR (300 MHz, DMSO-d6): δ=12.03 (s, 1H), 8.77 (s, 1H), 7.61 (s, 1H), 6.93 (s, 1H), 4.60-4.73 (m, 1H), 3.73-3.79 (m, 1H), 2.56-2.81 (m, 2H), 2.43 (t, J=9.0 Hz, 2H), 2.15-2.26 (m, 2H), 1.77-1.81 (m, 2H), 1.43 (d, J=4.5 Hz, 3H), 1.26-1.38 (m, 2H), 0.02-0.22 (m, 4H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 148.93, 148.51, 142.16, 129.04, 127.11, 126.83, 120.77, 115.61, 99.78, 71.21, 67.54, 62.59, 56.93, 53.76, 37.74, 25.78, 22.81, 10.16, 5.83 ppm; HRMS (EST): m/z [M+H]+. C20H25F3N5O calculated value 408.2006, measured value 408.2010.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((S-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by slow dropwise addition of 4,4,4-trifluoro-1-iodobutane (0.3 g, 1.1 mmol). Raise the temperature to maintain reflux and stir for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (R)-1-(6-(phenylsulfonyl)-1-((S)-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol as a pale yellow oil (0.3 g, 79%). The product can be used directly in the next step without further purification. HRMS (EST): m/z [M+H]+. C24H27F3N5O3S calculated value 522.1781, measured value 522.1788.
Step 2: (R)-1-(6-(phenylsulfonyl)-1-((S)-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d] pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.3 g, 0.6 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, and 5 mL of lM sodium hydroxide was added and then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhvdrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS71: (R)-1-(1-((S)-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.1 g, 46%). 1HNMR (300 MHz, DMSO-d6): δ=12.04 (s, 1H), 8.83 (s, 1H), 7.56 (s, 1H), 6.89 (s, 1H), 4.61-4.74 (m, 1H), 3.71-3.79 (m, 1H), 2.56-2.81 (m, 2H), 2.41 (t, J=4.5 Hz, 2H), 2.20-2.32 (m, 2H), 1.90-2.15 (m, 2H), 1.80-1.87 (m, 2H), 1.44 (d, J=6.0 Hz, 3H), 1.35-1.40 (m, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 150.11, 149.33, 144.34, 129.88, 127.19, 126.53, 120.77, 116.53, 98.57, 64.78, 58.93, 58.42, 56.32, 55.93, 37.77, 27.61, 22.84, 10.56 ppm; HRMS (ESI): m/z [M+H]+. C18H23F3N5O calculated value 382.1849, measured value 382.1852.
LXS72 was prepared in a similar preparation way to Example 69 and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS72: 5-((S)-7-(2-((R)-1-Hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-5-azaspiro[2,4]hept-5-yl)pentanonitrile (0.1 g, 45%). 1HNMR (300 MHz, DMSO-d6): δ=11.99 (s, 1H), 8.87 (s, 11H), 7.59 (s, 1H), 6.72 (s, 1H), 4.63-4.77 (m, 11H), 3.73-3.82 (m, 11H), 2.52-2.74 (m, 2H), 2.47 (t, J=6.0 Hz, 2H), 2.12-2.23 (m, 2H), 1.87 (t, J=6.0 Hz, 2H), 1.59-1.68 (m, 2H), 1.48 (d, J=9.0 Hz, 3H), 1.33-1.41 (m, 2H), 0.08-0.22 (m, 4H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 150.68, 149.47, 142.19, 129.33, 127.86, 120.73, 119.39, 115.62, 98.56, 71.29, 68.40, 65.27, 56.44, 53.49, 27.28, 25.58, 23.29, 22.81, 17.11, 5.88 ppm; HRMS (ESI): m/z [M+H]+. C21H27N6O calculated value 379.2241, measured value 379.2243.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((S-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by slow dropwise addition of 1-bromo-5-fluoropentane (0.2 g, 1.1 mmol), raise the temperature to reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (R)-1-(1-((S)-1-(5-fluoropentyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol as a pale yellow oil (0.3 g, 82%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C25H31FN5O3S calculated value 500.2126, measured value 500.2130.
Step 2: (R)-1-(1-((S)-1-(5—Fluoropentyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.3 g, 0.6 mmol) was dissolved in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, and 5 mL of 1 M sodium hydroxide was added, then stir for 5 h at room temperature, the reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS73: (R)-1-(1-((S)-1-(5-fluoropentyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.1 g, 46%). 1HNMR (300 MHz, DMSO-d6): δ=11.98 (s, 1H), 8.88 (s, 1H), 7.63 (s, 1H), 6.84 (s, 1H), 4.68-4.76 (m, 1H), 4.09-4.13 (m, 2H), 3.74-3.79 (m, 1H), 2.59-2.84 (m, 2H), 2.49 (t, J=9.0 Hz, 2H), 2.19-2.30 (m, 2H), 1.91-2.18 (m, 2H), 1.49 (d, J=9.0 Hz, 3H), 1.40-1.45 (m, 2H), 1.36-1.40 (m, 2H), 1.27-1.31 (m, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 151.08, 148.53, 142.19, 129.04, 127.88, 120.78, 115.62, 98.67, 83.69, 64.55, 58.92, 58.44, 56.61, 55.92, 30.71, 28.09, 27.66, 19.45 ppm; HRMS (ESI): m/z [M+H]. C19H27FN5O calculated value 360.2194, measured value 360.2199.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((S-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by slow dropwise addition of 2-bromo-N-(2,2,2-trifluoroethyl)acetamide (0.3 g, 1.1 mmol). Raise the temperature to reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-((S)-3-(2-((R)-1-Hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)-N-(2,2,2-trifluoroethyl)acetamide as a pale yellow oil (0.3 g, 75%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C24H26F3N6O4S calculated value 551.1683, measured value 552.1688.
Step 2: Dissolve 2-((S)-3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)-N-(2,2,2-trifluoroethyl)acetamide (0.3 g, 0.5 mmol) in 5 mL of a mixture of tetrahydrofuran and 5 mL of methanol. Add 5 mL of 1 M sodium hydroxide and then stir at room temperature for 5 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS74: 2-((S)-3-(2-((R)-1-Hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)-N-(2,2,2-trifluoroethyl)acetamide (0.1 g, 45%). 1HNMR (300 MHz, DMSO-d6): δ=12.11 (s, 1H), 8.89 (s, 1H), 8.03 (s, 1H), 7.58 (s, 1H), 6.91 (s, 1H), 4.65-4.78 (m, 1H), 3.72-3.79 (m, 3H), 3.25 (s, 2H), 2.58-2.83 (m, 2H), 2.21-2.34 (m, 2H), 1.94-2.18 (m, 2H), 1.49 (d, J=9.0 Hz, 3H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 170.55, 152.67, 148.55, 143.18, 129.04, 127.84, 124.79, 120.76, 115.69, 98.47, 64.87, 59.54, 58.28, 57.48, 54.94, 39.41, 27.88, 22.81 ppm: HRMS (ESI): m/z [M+H]+. C18H22F3N6O2 calculated value 411.1751, measured value 411.1777.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((S-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran. Add DIPEA (0.2 g, 1.4 mmol) and then slowly add 2-bromo-N-(2,2,2-trifluoroethyl)propanamide (0.3 g, 1.1 mmol) dropwise, raise the temperature to reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-((S)-3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)-N-(2,2,2-trifluoroethyl)propanamide as a pale yellow oil (0.3 g, 73%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C25H28F3N6O4S calculated value 565.1839, measured value 565.1841.
Step 2. Dissolve 2-((S)-3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)-N-(2,2,2-trifluoroethyl)propanamide (0.3 g, 0.5 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol. Add 5 mL of 1 M sodium hydroxide and stir at room temperature for 5 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS75: 2-((S)-3-(2-((R)-1-Hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)-N-(2,2,2-trifluoroethyl)propanamide (0.1 g, 44%). 1HNMR (300 MHz, DMSO-d6): δ=11.91 (s, 1H), 8.77 (s, 1H), 8.05 (s, 1H), 7.68 (s, 1H), 6.86 (s, 1H), 4.67-4.79 (m, 1H), 3.65-3.79 (m, 4H), 2.56-2.81 (m, 2H), 2.18-2.31 (m, 2H), 1.90-2.15 (m, 2H), 1.41 (d, J=7.5 Hz, 3H), 1.28 (d, J=7.5 Hz, 3H) ppm; 13 CNMR (75 MHz, DMSO-d6) δ 171.96, 148.67, 148.51, 142.18, 129.04, 127.18, 124.75, 120.75, 115.64, 97.56, 69.29, 63.64, 58.59, 54.92, 52.48, 39.47, 27.36, 22.86, 18.44 ppm, HRMS (EST): m/z [M+H]+. C19H24F3N6O2 calculated value 425.1907, measured value 425.1911.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((S-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by the slow dropwise addition of 2-bromo-N-(2,2,2-trifluoroethyl)butanamide (0.3 g, 1.1 mmol). Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-((S)-3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)-N-(2,2,2-trifluoroethyl)butanamide as a pale yellow oil (0.3 g, 71%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C26H30F3N6O4S calculated value 579.1996, measured value 579.2001.
Step 2: Dissolve 2-((S)-3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)-N-(2,2,2-trifluoroethyl)butanamide (0.3 g, 0.5 mmol) in a mixture of 5 mL tetrahydrofuran and 5 mL methanol. Add 5 mL of 1M sodium hydroxide and stir at room temperature for 5 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS76: 2-((S)-3-(2-((R)-1-Hydroxyethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)-N-(2,2,2-trifluoroethyl)butanamide (0.1 g, 44%). 1HNMR (300 MHz, DMSO-d6): δ=12.01 (s, 1H), 8.78 (s, 1H), 8.08 (s, 1H), 7.53 (s, 1H), 6.79 (s, 1H), 4.68-4.74 (m, 1H), 3.56-3.79 (m, 4H), 2.57-2.85 (m, 2H), 2.20-2.30 (m, 2H), 1.93-2.17 (m, 2H), 1.57-1.64 (m, 2H), 1.48 (d, J=7.5 Hz, 3H), 0.91 (t, J=9.0 Hz, 3H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 173.88, 151.77, 148.53, 142.11, 129.04, 127.17, 124.77, 120.71, 115.63, 98.56, 75.93, 64.65, 58.53, 55.22, 52.71, 39.48, 27.63, 23.66, 22.84, 12.89 ppm; HRMS (ESI): m/z [M+H]+. C20H26F3N6O2 calculated value 439.2064, measured value 439.2072.
LXS77 was prepared using a similar preparation way to Example 63 and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS77: 2-(1-(ethylsulfonyl)-3-(4-(2-(5-methylfuran-2-yl)imidazolyl[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl) azetidin-3-yl) acetonitrile (0.1 g, 53%). 1HNMR (300 MHz, DMSO-d6): δ=12.07 (s, 1H), 8.88 (s, 1H), 8.08 (s, 2H), 7.58 (s, 1H), 6.95 (d, J=7.5 Hz, 1H), 6.83 (s, 1H), 6.08 (d, J=7.5 Hz, 1H), 4.80-4.14 (m, 4H), 3.45 (t, J=9.0 Hz, 2H), 2.85 (s, 2H), 2.30 (s, 3H), 1.22 (t, J=7.5 Hz, 3H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 152.28, 151.49, 149.11, 143.19, 141.55, 130.75, 129.78, 129.01, 128.66, 121.67, 117.78, 115.61, 107.81, 107.62, 100.57, 99.56, 59.27, 51.63, 50.83, 23.27, 15.98, 4.77 ppm. HRMS (ESI): m/z [M+H]+. C2H23N5O3S calculated value 491.1608, measured value 491.1612.
Step 1: Triethyloxonium tetrafluoroborate (0.5 g, 2.4 mmol) and 2,2,2-trifluoroacetamide (0.2 g, 2.4 mmol) were dissolved in 10 mL of tetrahydrofuran, stirred at room temperature for 3 h and concentrated under vacuum to obtain the mixture in the form of oil, followed by the addition of 10 mL of ethanol to dissolve and the addition of 2-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)acetonitrile (0.3 g 0.8 mmol). Raise the temperature to maintain reflux and stir for 3 h. The reaction was monitored by TLC for complete reaction. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhvdrous sodium sulfate and concentrated under vacuum to yield 2-(4-(6-(phenylsulfonyl)-2-(trifluoromethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl) acetonitrile as a milk white oil (0.3 g, 83%). HRMS (ESI): m/z [M+H]+. C20H13F3N7O2S calculated value 472.0798, measured value 472.0802.
Step 2: Dissolve 2-(4-(6-(phenylsulfonyl)-2-(trifluoromethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)acetonitrile (0.3 g, 0.6 mmol) in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, and stir at room temperature for 5 h after adding 5 mL of 1 M sodium hydroxide. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS78: 2-(4-(2-(trifluoromethyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)acetonitrile (0.1 g, 47%). 1HNMR (300 MHz, DMSO-d6): δ=11.97 (s, 1H), 8.87 (s, 1H), 8.05 (s, 1H), 7.91 (s, 1H), 7.56 (s, 1H), 6.83 (s, 1H), 4.85 (s, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 148.11, 144.33, 130.74, 129.74, 129.04, 127.65, 120.73, 116.73, 116.32, 115.62, 100.53, 98.45, 45.65 ppm; HRMS (ESI): m/z [M+H]+. C1-4H9F3N7 calculated value 332.0866, measured value 332.0871.
Step 1: Dissolve 3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)propanenitrile (0.3 g, 0.7 mmol) in 10 mL of toluene, followed by the addition of triethyl orthoformate (0.2 g, 1.4 mmol), p-toluenesulfonic acid (30.0 mg, 10%), and raise the temperature to maintain reflux and stir for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice with dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 3-(4-(6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile as a pale yellow oil (0.3 g, 98%). HRMS (ESI): m/z [M+H]+. C20H16N7O2S calculated value 418.1081, measured value 418.1088.
Step 2: Dissolve 3-(4-(6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.3 g, 0.7 mmol) in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, add 5 mL of 1 M sodium hydroxide and then stir at room temperature for 5 h. Monitor the reaction by TLC for completion. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS79: 3-(4-(imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.1 g, 50%). 1HNMR (300 MHz, DMSO-d6): δ=11.99 (s, 1H), 8.89 (s, 1H), 8.15 (s, 1H), 7.95 (s, 1H), 7.56 (s, 1H), 7.15 (s, 1H), 6.89 (s, 1H), 5.04 (t, J=7.5 Hz, 2H), 3.21 (t, J=7.5 Hz, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 150.44, 142.88, 134.58, 130.74, 129.73, 129.04, 127.16, 120.74, 117.73, 115.60, 100.54, 98.62, 49.37, 17.50 ppm; HRMS (ESI): m/z [M+H]+. C14H12N7 calculated value 278.1149, measured value 278.1151.
Step 1: Dissolve 3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)propanenitrile (0.3 g, 0.7 mmol) in 10 mL of DMF, add Na2S2O5 (0.7 g, 3.5 mmol) followed by the addition of 2-chlorobenzaldehyde (0.2 g, 1.0 mmol). Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline and dried over anhydrous sodium sulfate to yield 3-(4-(2-(2-chlorophenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile as a yellow solid (0.3 g, 77%). HRMS (ESI): m/z [M+H]+. C26H19ClN7O2S calculated value 528.1004, measured value 528.1014.
Step 2: Dissolve 3-(4-(2-(2-chlorophenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.3 g, 0.6 mmol) in a mixture of 5 mL tetrahydrofuran and 5 mL methanol, add 5 mL of 1 M sodium hydroxide and then stir at room temperature for 5 hours. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS80: 3-(4-(2-(2-Chlorophenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.1 g, 45%). 1HNMR (300 MHz, DMSO-d6): δ=12.03 (s, 1H), 8.78 (s, 1H), 8.08 (s, 1H), 7.99 (s, 1H), 7.73 (d, J=7.5 Hz, 1H), 7.58 (s, 1H), 7.55 (d, J=7.5 Hz, 1H), 7.35-7.39 (m, 2H), 6.85 (s, 1H), 5.09 (t, J=9.0 Hz, 2H), 3.28 (t, J=9.0 Hz, 2H) ppm: 1′CNMR (75 MHz, DMSO-d6) δ 149.21, 143.48, 142.19, 138.56, 132.27, 130.77, 130.16, 129.75, 129.38, 129.05.127.30, 127.17, 120.77, 117.74, 115.64, 100.63, 98.56, 50.45, 17.54 ppm: HRMS (ESI): m/z [M+H]+. C20H15ClN7 calculated value 388.1072, measured value 388.1077.
Step 1: Dissolve 2-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)acetonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4.0 mmol) followed by adding 5-hydroxymethylfurfural (0.3 g, 1.6 mmol). Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl) acetonitrile as a pale yellow oil (0.3 g, 79%). HRMS (EST): m/z [M+H]+. C24H18N7O4S calculated value 500.1135, measured value 500.1140.
Step 2: Dissolve 2-(4-(2-(5-(hydroxymethyl)furan-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-(6H)-yl)-1H-pyrazol-1-yl)acetonitrile (0.3 g, 0.6 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, then add 5 mL of 1 M sodium hydroxide. Stir for 5 h at room temperature. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS81: 2-(4-(2-(5-(Hydroxymethyl)furan-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl) acetonitrile (0.1 g, 46%). 1HNMR (300 MHz, DMSO-d6); δ=11.93 (s, 1H), 8.91 (s, 1H), 8.05 (s, 1H), 7.91 (s, 1H), 7.56 (s, 1H), 7.02 (d, J=7.5 Hz, 1H), 6.89 (s, 1H), 6.59 (d, J=7.5 Hz, 1H), 4.85 (s, 2H), 4.39 (s, 2H) 3.65 (s, 1H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 153.88, 151.17, 149.67, 142.18, 141.33, 130.75, 129.75, 129.07, 127.19, 120.77, 116.36, 115.63, 107.96.104.07, 100.58, 98.67, 57.38, 46.17 ppm; HRMS (ESI): m/z [M+H]+. C18H14N7O2
90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline and dried over anhydrous sodium sulfate to yield 3-(4-(2-(3-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile as a yellow solid (0.3 g, 79%). HRMS (ESI): m/z [M+H]+. C26H20N7O3S calculated value 510.1343, measured value 510.1351.
Step 2: Dissolve 3-(4-(2-(3-hydroxyphenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.3 g, 0.6 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, add 5 mL of 1 M sodium hydroxide and then stir at room temperature for 5 hours. The reaction was monitored completion by TLC for 5 hours. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS82: 3-(4-(2-(3-Hydroxyphenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.1 g, 46%). 1HNMR (300 MHz, DMSO-d6): δ=11.79 (s, 1H), 8.83 (s, 1H), 8.06 (s, 1H), 7.94 (s, 1H), 7.84 (d, J=9.0 Hz, 1H), 7.59 (s, 1H), 7.34 (t, J=9.0 Hz, 1H), 7.04 (s, 1H), 6.91 (d, J=9.0 Hz, 1H), 6.89 (s, 1H), 5.35 (s, 1H), 5.03 (t, J=7.5 Hz, 2H), 3.29 (t, J=7.5 Hz, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 157.59, 149.56, 143.43, 142.19, 132.08, 130.75, 130.61, 129.77, 120.06, 127.18, 120.74, 120.15, 117.74, 115.97, 115.61, 112.95, 100.59, 98.65, 50.65, 18.66 ppm; HRMS (ESI): m/z [M+H]+. C20H16N7O calculated value 370.1411, measured value 370.1419.
Step 1: Dissolve 3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)propanenitrile (0.3 g, 0.7 mmol) in 10 mL of DMF, add Na2S2O5 (0.7 g, 3.5 mmol) followed by adding 2-fluorobenzaldehyde (0.2 g, 1.0 mmol). Stir for 12 h at 90 C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane, be organic phases were combined and washed with saturated saline and dried over anhydrous sodium sulfate to yield 3-(4-(2-(2-fluorophenyl)-6-(phenylsulfanyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile as a yellow solid (0.3 g, 80%). HRMS (EST): m/z [M+H]+. C26H19FN7O2S calculated value 512.1299, measured value 512.1301.
Step 2: Dissolve 3-(4-(2-(2-fluorophenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.3 g, 0.6 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol, add 5 mL of 1 M sodium hydroxide and then stir at room temperature for 5 hours. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS83: 3-(4-(2-(2-Fluorophenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.1 g, 46%), 46%). 1HNMR (30) MHz, DMSO-d6): δ=12.07 (s, 1H), 8.78 (s, 1H), 8.05 (s, 1H), 7.96 (s, 1H), 7.71-7.77 (m, 2H), 7.56 (s, 1H), 7.49 (d, J=7.5 Hz, 1H), 7.28 (t, J=7.5 Hz, 1H), 6.88 (s, 1H), 5.04 (t, J=9.0 Hz, 2H), 3.21 (t, J=9.0 Hz, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 158.39, 148.63, 143.39, 142.17, 130.77, 130.38, 129.76, 129.16, 129.06, 127.17, 124.88, 123.59, 120.74, 117.74, 115.69, 114.74, 101.76, 100.43, 51.98, 22.51 ppm; HRMS (ESI): m/z [M+H]+. C20H16FN7 372.1367, measured value 372.1377.
Step 1: Dissolve 3-(4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)-1H-pyrazol-1-yl)propanenitrile (0.3 g, 0.7 mmol) in 10 mL of DMF, add Na2S2O5 (0.7 g, 3.5 mmol) followed by adding 5-methyl-2-thiophenecarboxaldehyde (0.2 g, 1.0 mmol). Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline and dried over anhydrous sodium sulfate to yield 3-(4-(2-(5-methylthiophen-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile as a yellow solid (0.3 g, 80%). HRMS (ESI); m/z [M+H]+. C25H20N7O2S2 calculated value 514.1114, measured value 514.1119.
Step 2: Dissolve 3-(4-(2-(5-methylthiophen-2-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.3 g, 0.6 mmol) in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol. After the addition of 5 mL of 1M sodium hydroxide, the mixture was then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS84: 3-(4-(2-(5-methylthiophen-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.1 g, 46%). 1HNMR (300 MHz, DMSO-d6): δ=11.93 (s, 1H), 8.83 (s, 1H), 8.06 (s, 1H), 7.99 (s, 1H), 7.56 (s, 1H), 7.51 (d, J=7.5 Hz, 1H), 6.89 (s, 1H), 6.83 (d, J=7.5 Hz, 1H), 5.10 (t, J=6.0 Hz, 2H), 3.26 (t, J=6.0 Hz, 2H), 2.36 (s, 3H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 151.63, 142.17, 141.63, 141.30, 134.38, 130.74, 129.74.129.06, 127.59, 127.50, 127.14, 120.79, 117.78, 115.62, 101.86, 99.38, 51.96, 15.93, 15.22 ppm: HRMS (ESI): m/z [M+H]+. C19H16N7S calculated value 374.1182, measured value 374.1189.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((S)-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrazolo[3,4-b]pyridin-2-yl) ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by the slow dropwise addition of 4-bromobutyronitrile (0.2 g, 1.1 mmol). Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 4-((S)-3-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrazolo[3,4-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)butyronitrile as a pale yellow oil (0.2 g, 57%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C23H26N7O3S calculated value 480.1812, measured value 480.1825.
Step 2: 4-((S)-3-(2-((R)-1-Hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrazolo[3,4-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)butyronitrile (0.2 g, 0.4 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol. After the addition of 5 mL of 1 M sodium hydroxide, stir the mixture for 5 h at room temperature and monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS85: 4-((S)-3-(2-((R)-1-hydroxyethyl)imidazo[4,5-d]pyrazolo[3,4-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)butanenitrile (0.1 g, 71%). 1HNMR (300 MHz, DMSO-d6): δ=12.54 (s, 1H), 8.89 (s, 1H), 7.55 (s, 1H), 4.65-4.70 (m, 1H), 3.64-3.81 (m, 1H), 2.56-2.83 (m, 2H), 2.43 (t, J=9.0 Hz, 2H), 2.23-2.31 (m, 2H), 1.90-2.15 (m, 2H), 1.87 (t, J=9.0 Hz, 2H), 1.66-1.76 (m, 2H), 1.41 (d, J=6.0 Hz, 3H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 154.79.148.53, 140.85, 140.53, 132.46, 125.49, 119.39, 104.75, 63.68, 58.52, 58.41, 55.92, 55.51, 27.61, 22.85, 15.94, 15.11 ppm; HRMS (ESI): m/z [M+H]+. C17H22N7O calculated values 340.1880, measured value 340.1888.
Step 1: 4-Chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (1.0 g, 3.0 mmol) was dissolved in 10 mL of tetrahydrofuran. Then, DIPEA (0.8 g, 6.0 mmol) and (3R) tert-butyl 3-amino-4-fluoropyrrolidine-1-carboxylate (0.9 g, 4.5 mmol) were added and stirred for 4h while maintaining reflux at an elevated temperature. The reaction was monitored for completion by TLC. Concentrate under vacuum to obtain a yellow oily liquid. Triturate and cure with an appropriate amount of methanol to yield (4R)-tert-butyl 3-fluoro-4-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate as a yellow solid (1.2 g, 80%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C22H25FN5O6S calculated value 506.1504, measured value 506.1509.
Step 2: Dissolve (4R)-tert-butyl 3-fluoro-4-((5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate (1.2 g, 2.4 mmol) in 20 mL of methanol, add palladium carbon (0.1 g, 10%) and then use hydrogen to replace the air in the reaction flask more than three times. The reaction was carried out in hydrogen atmosphere and stirred at room temperature for 12 h. The reaction was monitored for completion by TLC. After filtration, the filtrate was collected and concentrated under vacuum to yield (4R)-tert-butyl 3-fluoro-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-carboxylate as a light pink foamy solid (1.1 g, 97%). The product can be used directly in the next step without further purification. HRMS (EST): m/z [M+H]+. C22H27FN5O4S calculated value 476.1762, measured value 476.1770.
Step 3: Dissolve triethyloxonium tetrafluoroborate (1.2 g, 6.3 mmol) and (R)-lactamideide (0.6 g, 6.3 mmol) in 10 mL of tetrahydrofuran, stir for 3 h at room temperature and then concentrate the mixture under vacuum to obtain an oily liquid, followed by the addition of 10 mL of ethanol to dissolve. Then, add (4R)-tert-butyl 3-fluoro-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)pyrrolidine-1-formate (1.0 g, 2.1 mmol). Raise the temperature to maintain reflux and stir for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield (4R)-tert-butyl 3-fluoro-4-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)pyrrolidine-1-carboxylate as a green oil (0.5 g, 45%). HRMS (EST): m/z [M+H]+. C25H29FN5O5S calculated value 530.1868, measured value 530.1870.
Step 4: Dissolve (4R)-tert-butyl 3-fluoro-4-(2-((R)-1-hydroxyethyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1 (6H)-yl)pyrrolidine-1-carboxylate (0.5 g, 0.9 mmol) in 10 mL of dichloromethane, slowly add trifluoroacetic acid (1.0 g, 9.0 mmol) and stir at room temperature for 12 h. Concentrate under vacuum to yield (1R)-1-(1-(((3R)-4-fluoropyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol as a light brown oil (0.4 g, 99%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C20H21FN5O3S calculated value 430.1344, measured value 430.1350.
Step 5: Dissolve (1R)-1-(1-(((3R)-4-fluoropyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) and then add 4,4,4-trifluoro-1-iodobutane (0.3 g, 1.1 mmol) dropwise. Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (1R)-1-(1-(((3R)-4-fluoro-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol as a pale yellow oil (0.2 g, 53%), 53%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C24H26F4N5O3S calculated value 540.1687, measured value 540.1689.
Step 6: (1R)-1-(1-(((3R)-4-fluoro-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.2 g, 0.4 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol. After adding 5 mL of 1 M sodium hydroxide, the reaction was stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate =1:1) to yield LXS86: (1R)-1-(1-((3R)-4-fluoro-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-ol (0.1 g, 68%). 1HNMR (300 MHz, DMSO-d6): δ=11.94 (s, 1H), 8.68 (s, 1H), 7.54 (s, 1H), 6.77 (s, 1H), 4.684.72 (m, 1H), 3.91-4.06 (m, 1H), 3.30-3.41 (m, 1H), 2.56-2.81 (m, 2H), 2.54 (t, J=4.5 Hz, 2H), 2.29-2.51 (m, 2H), 1.64-1.81 (m, 2H), 1.46 (d, J=9.0 Hz, 3H), 1.36-1.42 (m, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 151.89, 148.53, 142.19, 129.06, 127.18, 126.84, 120.77, 115.69.98.56, 91.20, 63.91, 61.77, 56.33, 54.88, 51.67, 37.71, 22.86, 11.67 ppm, HRMS (ESI): m/z [M+H]+. C18H22F4N50 calculated value 400.1755, measured value 400.1770.
(R)-1-(1-((3R,4R)-4-fluoro-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-olUsing (3R,4R)-tert-butyl 3-amino-4-fluoropyrrolidine-1-carboxylate instead of (3R)-tert-butyl 3-amino-4-fluoropyrrolidine-1-carboxylate, LXS 87 was prepared in a similar preparation way described in Example 86. The crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS87: (R)-1-(1-((3R,4R)-4-fluoro-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-olpyridin-2-yl) ethanol (0.1 g, 60%). 1HNMR (300 MHz, DMSO-d6): δ=11.99 (s, 1H), 8.78 (s, 1H), 7.58 (s, 1H), 6.75 (s, 1H), 4.64-4.71 (m, 1H), 3.90-4.05 (m, 1H), 3.31-3.55 (m, 1H), 2.53-2.87 (m, 2H), 2.50 (t, J=6.0 Hz, 2H), 2.23-2.47 (m, 2H), 1.73-1.88 (m, 2H), 1.41 (d, J=6.0 Hz, 3H), 1.33-1.44 (m, 2H) ppm; 13 CNMR (75 MHz, DMSO-d6) δ 148.97, 148.53, 142.19, 129.00, 127.65, 126.88, 120.73, 115.69, 97.63, 91.28, 63.63, 61.79, 56.39, 54.85, 51.63, 37.77, 22.81, 11.65 ppm: HRMS (ESI): m/z [M+H]+. C18H22F4N5O calculated value 400.1755, measured value 400.1768.
Using (3R,4R)-tert-butyl 3-amino-4-fluoropyrrolidine-1-carboxylate instead of (3R)-tert-butyl 3-amino-4-fluoropyrrolidine-1-carboxylate, LXS 88 was prepared in a similar preparation way described in Example 86. The crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS88: (R)-1-(1-((3R,4S)-4-fluoro-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-ol (0.1 g, 50%). 1HNMR (300 MHz, DMSO-d6): δ=12.13 (s, 1H), 8.81 (s, 1H), 7.60 (s, 1H), 6.57 (s, 1H), 4.58-4.70 (m, 1H), 3.90-4.05 (m, 1H), 3.33-3.42 (m, 1H), 2.51-2.86 (m, 2H), 2.42 (t, J=9.0 Hz, 2H), 2.30-2.41 (m, 2H), 1.66-1.82 (m, 2H), 1.49 (d, J=9.0 Hz, 3H), 1.39-1.46 (m, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 151.45, 158.55, 142.19, 129.05, 127.16, 126.86, 120.74, 115.64, 98.56, 91.55, 63.65, 61.75, 56.38, 54.85, 51.68, 37.77, 22.89, 11.56 ppm; HRMS (ESI): m/z [M+H]+. C18H22F4N50 calculated value 400.1755, measured value 400.1763.
(R)-1-(1-((3S,5S)-5-(Hydroxymethyl)-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-olUsing tert-butyl (2S,4S)-4-amino-2-(hydroxymethyl)pyrrolidine-1-carboxylate instead of tert-butyl (3R)-3-amino-4-fluoropyrrolidine-1-carboxylate, LXS 89 was prepared in a similar preparation way described in Example 86. The crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS89: (R)-1-(1-((3S,5S)-5-(hydroxymethyl)-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-ol (0.1 g, 51%). 1HNMR (300 MHz, DMSO-d6): δ=11.98 (s, 1H), 8.78 (s, 1H), 7.59 (s, 1H), 6.88 (s, 1H), 4.68-4.74 (m, 1H), 3.79-3.85 (m, 1H), 3.34 (d, J=6.0 Hz, 2H), 2.59-2.82 (m, 2H), 2.43 (t, J=4.5 Hz, 2H), 2.33-2.41 (m, 1H), 1.87-2.12 (m, 2H), 1.64-1.81 (m, 2H), 1.47 (d, J=6.0 Hz, 3H), 1.36-1.40 (m, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 148.94, 148.57, 142.19, 129.04, 127.11, 126.88, 120.75, 115.63, 98.67, 68.74, 63.67, 62.55, 56.71, 56.21, 54.18, 37.79, 28.31, 22.84, 10.49 ppm; HRMS (ESI): m/z [M+H]+. C19H25F3N5O2 calculated value 412.1955, measured value 412.1961.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((S)-pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrazolo[3,4-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by the slow dropwise addition of 4,4,4-trifluoro-1-iodobutane (0.3 g, 1.1 mmol). Raise the temperature to maintain reflux and stir for 3 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (R)-1-(6-(phenylsulfonyl)-1-((S)-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrazolo[3,4-b]pyridin-2-yl) ethanol as a pale yellow oil (0.2 g, 52%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C23H26F3N6O3S calculated value 523.1734, measured value 523.1740.
Step 2: (R)-1-(6-(Phenylsulfonyl)-1-((S)-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d] pyrazolo[3,4-b]pyridin-2-yl) ethanol (0.2 g, 0.4 mmol) was dissolved in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, and add 5 mL of IM sodium hydroxide and then stir at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS90: (R)-1-(1-((S)-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrazolo[3,4-b]pyridin-2-yl)ethan-1-ol (0.1 g, 68%). 1HNMR (300 MHz, DMSO-d6): δ=13.54 (s, 1H), 8.82 (s, 1H), 7.58 (s, 1H), 4.65-4.73 (m, 1H), 3.64-3.79 (m, 2H), 2.53-2.81 (m, 2H), 2.41 (t, J=6.0 Hz, 2H), 2.20-2.33 (m, 2H), 1.91-2.14 (m, 2H), 1.85 (m, 2H), 1.41 (d, J=9.0 Hz, 3H), 1.27-1.37 (m, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 154.75, 148.52, 140.88, 140.51, 132.48, 126.84, 125.47, 105.37, 63.68, 58.51, 58.43, 56.38, 55.91, 38.88, 27.91, 22.86, 9.56 ppm; HRMS (ESI): m/z [M+H]+. C17H22F3N60 calculated value 383.1802, measured value 383.1800.
(R)-1-(1-((3S,5R)-5-(hydroxymethyl)-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-olLXS91 was prepared by replacing tert-butyl (3R)-3-amino-4-fluoropyrrolidine-1-carboxylate with tert-butyl (2R,4S)-4-amino-2-(hydroxymethyl)pyrrolidine-1-carboxylate, using a preparation method similar to Example 86, and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS91: (R)-1-(1-((3S,5R)-5-(hydroxymethyl)-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-ol (0.1 g, 43%). 1 HNMR (300 MHz, DMSO-d6): δ=11.68 (s, 1H), 8.73 (s, 1H), 7.61 (s, 1H), 6.89 (s, 1H), 4.65-4.71 (m, 1H), 3.76-3.89 (m, 1H), 3.35 (d, J=6.0 Hz, 2H), 2.64-2.81 (m, 2H), 2.45 (t, J=7.5 Hz, 2H), 2.37-2.39 (m, 1H), 1.89-2.15 (m, 2H), 1.60-1.84 (m, 2H), 1.43 (d, J=9.0 Hz, 3H), 1.33-1.41 (m, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 151.87, 148.54, 142.19, 129.05, 127.18, 126.84, 120.77, 115.69, 98.56, 68.74, 63.69, 62.57, 56.71, 56.38, 54.18, 38.96, 28.54, 22.85, 13.74 ppm; HRMS (EST): m/z [M+H]+. C19H25F3N5O2 calculated value 412.1955, measured value 412.1963.
(R)-1-(1-((3S,5R)-5-(hydroxymethyl)-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-olLXS92 was prepared by replacing tert-butyl (3R)-3-amino-4-fluoropyrrolidine-1-carboxylate with tert-butyl (2R,4S)-4-amino-2-(hydroxymethyl)pyrrolidine-1-carboxylate, using a preparation method similar to Example 86, and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS92: (R)-1-(1-((3S,5R)-5-(hydroxymethyl)-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethan-1-ol (0.1 g, 33%). 1HNMR (300 MHz, DMSO-d6): δ=13.78 (s, 1H), 8.87 (s, 1H), 7.54 (s, 1H), 4.69-4.72 (m, 1H), 3.83-3.89 (m, 1H), 3.37 (d, J=6.0 Hz, 2H), 2.61-2.80 (m, 2H), 2.47 (t, J=9.0 Hz, 2H), 2.36-2.43 (m, 1H), 1.88-2.15 (m, 2H), 1.58-1.79 (m, 2H), 1.44 (d, J=10.5 Hz, 3H), 1.38-1.43 (m, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 155.75, 148.56, 140.83, 140.51, 132.48, 126.84, 125.42, 104.74, 68.75, 63.61, 62.51, 56.39, 56.29, 54.17, 38.67, 28.56, 22.87, 11.65 ppm; HRMS (ESI): m/z [M+H]+. C18H24F3N6O2 calculated value 413.1907, measured value 413.1914.
LXS93 was prepared in a preparation way similar to Example 86 and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS93: 4-((4R)-3-Fluoro-4-(2-((R)-1-hydroxyethyl)imidazo[4,5-d]pyrazolo[3,4-b]pyridin-1(6H)-yl)pyrrolidin-1-yl)butanenitrile (0.1 g, 31%). 1HNMR (300 MHz, DMSO-d6): δ=13.73 (s, 1H), 8.89 (s, 1H), 7.61 (s, 1H), 4.57-4.71 (m, 1H), 3.914.08 (m, 11H), 3.33-3.45 (m, 1H), 2.57-2.81 (m, 2H), 2.47 (t, J=6.0 Hz, 2H), 2.40-2.43 (m, 2H), 1.90 (t, J=9.0 Hz, 2H), 1.70-1.76 (m, 2H), 1.43 (d, J=7.5 Hz, 3H) ppm; 1; CNMR (75 MHz, DMSO-d6) δ 154.76, 148.54, 141.65, 140.51, 132.78, 125.47, 119.38, 104.78, 91.38, 64.89, 61.33, 55.59, 54.86, 51.69, 22.85, 17.56, 16.45 ppm: HRMS (ESI): m/z [M+H]+. C17H21FN7O calculated value 358.1786, measured value 358.1789.
LXS94 was prepared using a preparation method similar to Example 86 by replacing tert-butyl (3R)-3-amino-4-fluoropyrrolidine-1-carboxylate with tert-butyl (2S,4S)-4-amino-2-(hydroxymethyl)pyrrolidine-1-carboxylate, and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS94: 4-((2S,4S)-4-(2-((R)-1-Hydroxyethyl)imidazo[4,5-d]pyrazolo[3,4-b]pyridin-1(6H)-yl)-2-(hydroxymethyl)pyrrolidin-1-yl)butanenitrile (0.1 g, 27%). 1HNMR (300 MHz, DMSO-d6): δ=13.68 (s, 1H), 8.89 (s, 1H), 7.58 (s, 1H), 4.63-4.71 (m, 1H), 3.85-3.91 (m, 1H), 3.39 (d, J=9.0 Hz, 2H), 2.66-2.81 (m, 2H), 2.39 (t, J=6.0 Hz, 2H), 2.38-2.45 (m, 1H), 1.79-2.23 (m, 2H), 1.56-1.72 (m, 2H), 1.41 (d, J=10.5 Hz, 3H), 1.33-1.39 (m, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 155.78, 149.65, 140.89, 140.32, 132.89, 124.23, 119.38, 104.73, 69.65, 63.65, 62.56, 57.76, 56.21, 53.39, 29.65, 22.89, 18.56, 14.65 ppm; HRMS (ESI): m/z [M+H]+. C18H24N7O2 calculated value 370.1986, measured value 370.1990.
Step 1; Dissolve trans-4-((5-amino-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridin-4-yl)amino)cyclohexanecarbonitrile (0.3 g, 0.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.8 g, 4 mmol) followed by the addition of 3,4-difluorobenzaldehyde (0.2 g, 1.6 mmol) dropwise. Stir for 12 h at 90° C. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice with dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield trans-4-(2-(3,4-difluorophenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile as a yellow oil (0.3 g, 76%). HRMS (ESI): m/z [M+H]+. C27H22F2N5O2S calculated value 518.1457, measured value 518.1461.
Step 2: Dissolve trans-4-(2-(3,4-difluorophenyl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.3 g, 0.6 mmol) in a mixture of 5 mL of tetrahydrofuran and 5 mL of methanol. After adding 5 mL of 1 M sodium hydroxide, stir at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS95: trans-4-(2-(3,4-difluorophenyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)cyclohexanecarbonitrile (0.1 g, 46%). 1HNMR (300 MHz, DMSO-d6): δ=12.02 (s, 1H), 8.66 (s, 1H), 7.77-7.84 (m, 1H), 7.56-7.73 (m, 1H), 7.55 (d, J=3.0 Hz, 2H), 6.88 (s, 1H), 4.414.49 (m, 1H), 3.15 (t, J=12.0 Hz, 1H), 2.32-2.44 (m, 2H), 2.23 (d, J=12.0 Hz, 2H), 2.04 (d, J=12.0 Hz, 2H), 1.73-1.85 (m, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 153.78, 150.05, 149.64, 148.69, 142.19, 129.05, 127.83, 127.16, 124.74, 122.75, 120.75, 117.52, 115.63, 115.18, 97.58, 65.94, 31.89, 28.78, 22.63 ppm; HRMS (ESI): m/z [M+H]+. C21H18F2N5 calculated value 378.1525, measured value 378.1529.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((S)-pyrrolidin-3-yl)imidazo[4,5-d]pyrrolo[3,4-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) followed by the slow dropwise addition of 4,4,4-trifluoro-1-iodopropane (0.2 g, 1.4 mmol). Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (R)-1-(6-(phenylsulfonyl)-1-((S)-1-(3,3,3-trifluoropropyl)pyrrolidin-3-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol as a pale yellow oil (0.2 g, 54%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C23H25F3N5O3S calculated value 508.1625, measured value 508.1630.
Step 2: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((S)-1-(3,3,3-trifluoropropyl)pyrrolidin-3-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.2 g, 0.4 mmol) in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol. After the addition of 5 mL of 1M sodium hydroxide, stir the mixture for 5 h at room temperature and monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS96: (R)-1-(1-((S)-1-(3,3,3-trifluoropropyl)pyrrolidin-3-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.1 g, 69%). 1HNMR (300 MHz, DMSO-d6): δ=12.09 (s, 1H), 8.87 (s, 1H), 7.53 (s, 1H), 6.83 (s, 1H), 4.60-4.75 (m, 1H), 3.73-3.78 (m, 1H), 2.56-2.87 (m, 2H), 2.42 (t, J=4.5 Hz, 2H), 2.15-2.32 (m, 2H), 1.90-2.11 (m, 2H), 1.41 (d, J=6.0 Hz, 3H), 1.35-1.40 (m, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 148.67, 148.51, 142.18, 129.02, 127.11, 124.88, 120.74, 115.65, 99.35, 63.64, 58.94, 58.41, 55.95, 40.25, 38.45, 27.69, 22.81 ppm: HRMS (ESI): m/z [M+H]+. C17H21F3N5O calculated value 368.1693, measured value 368.1670.
Step 1: Dissolve (R)-1-(6-(phenylsulfonyl)-1-((S)-pyrrolidin-3-yl)imidazo[4,5-d]pyrrolo[3,4-b]pyridin-2-yl)ethanol (0.3 g, 0.7 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.2 g, 1.4 mmol) and then slowly add 1,1,1,2,2-pentafluoro-4-iodobutane (0.3 g, 1.1 mmol) dropwise. Then, raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield (R)-1-(1-((S)-1-(3,3,4,4,4-pentafluorobutyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol as a pale yellow oil (0.2 g, 49%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C24H25F5N5O3S calculated value 558.1593, measured value 558.1602.
Step 2: Dissolve (R)-1-(1-((S)-1-(3,3,4,4,4-pentafluorobutyl)pyrrolidin-3-yl)-6-(phenylsulfonyl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.2 g, 0.36 mmol) in a mixture of 5 mL tetrahydrofuran and 5 mL methanol, and 5 mL of 1M sodium hydroxide was added and the mixture was then stirred at room temperature for 5 h. The reaction was monitored for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS97: (R)-1-(1-((S)-1-(3,3,4,4,4-pentafluorobutyl)pyrrolidin-3-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.1 g, 67%). 1HNMR (300 MHz, DMSO-d6): δ=12.85 (s, 1H), 8.77 (s, 1H), 7.56 (s, 1H), 6.89 (s, 1H), 4.60-4.75 (m, 1H), 3.73-3.78 (m, 1H), 2.56-2.87 (m, 2H), 2.47 (t, J=4.5 Hz, 2H), 2.15-2.32 (m, 2H), 1.90-2.11 (m, 2H), 1.45 (d, J=6.0 Hz, 3H), 1.34-1.40 (m, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 148.67, 148.51, 142.18, 131.88, 129.02, 127.11, 120.93, 119.74, 114.65, 97.35, 68.64, 53.94, 51.41, 50.95, 41.55, 37.40, 22.68, 21.40 ppm; HRMS (ESI): m/z [M+H]+. C18H21F5N5O calculated value 418.1661, measured value 418.1666.
Step 1: Dissolve 98-1 (0.5 g, 2.7 mmol) in 10 mL of DMF, add K2CO3 (0.7 g, 5.4 mmol) followed by the slow dropwise addition of 4,4,4-trifluoro-1-iodobutane (0.7 g, 3.0 mmol), and then stir at room temperature for 12 h. Monitor the reaction for completion by MS. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 98-2 as a white oil (0.7 g, 88%). The product was used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C13H24F3N2O2 calculated value 297.1784, measured value 297.1788.
Step 2: Dissolve 98-2 (0.7 g, 2.4 mmol) in 10 mL of dichloromethane, add trifluoroacetic acid (2.7 g, 24.0 mmol) and stir at room temperature for 12 h. Monitor the reaction for completion by MS. The product was concentrated under vacuum to obtain 98-3 as a light yellow oil (0.4 g, 99%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C8H16F3N2 calculated value 197.1260, measured value 197.1266.
Step 3: Dissolve 98-3 (0.4 g, 2.0 mmol) in 10 mL of tetrahydrofuran, add DIPEA (0.5 g, 4.0 mmol) followed by the addition of 4-chloro-5-nitro-1-(phenylsulfonyl)-1H-pyrrolo[2,3-b]pyridine (0.6 g, 1.8 mmol). Raise the temperature to maintain reflux and stir for 3 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 98-4 as a yellow oil (0.6 g, 68%). The product was used directly in the next step without further purification. HRMS (EST): m/z [M+H]+. C21H23F3N5O4S calculated value 498.1417, measured value 498.1420.
Step 4: Dissolve 98-4 (0.6 g, 1.2 mmol) in 10 mL of methanol, add palladium carbon (0.1 g, 10%) and then use hydrogen to replace the air in the reaction flask more than three times. Keep the reaction in hydrogen atmosphere, stir for 12 h at room temperature, and monitor the reaction for completion by TLC. After filtration, the filtrate was collected and concentrated under vacuum to obtain 98-5 as a white foamy solid (0.5 g, 89%). The product can be used directly in the next step without further purification. HRMS (ESI): m/z [M+H]+. C21H25F3N5O2S calculated value 468.1676, measured value 468.1680.
Step 5: Dissolve 98-5 (0.5 g, 1.8 mmol) in 10 mL of DMF, add Na2S2O5 (0.7 g, 3.6 mmol) followed by the addition of 5-hydroxymethyl furfural (0.4 g, 3.2 mmol) dropwise, raise the temperature to 90° C. and stir for 12 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried over anhydrous sodium sulfate and concentrated under vacuum to yield 98-6 as a yellow oil (0.3 g, 49%). HRMS (ESI): m/z [M+H]+. C27H27F3N5O4S calculated value 574.1730, measured value 574.1739.
Step 6: Dissolve 98-6 (0.2 g, 0.3 mmol) in a solvent mixture of 5 mL tetrahydrofuran and 5 mL methanol, add 5 mL of 1M sodium hydroxide and then stir at room temperature for 5 h. Monitor the reaction for completion by TLC. Saturated sodium bicarbonate was added until the reaction solution was weakly basic, and the organic phase was separated. The aqueous phase was extracted twice using dichloromethane. The organic phases were combined and washed with saturated saline, dried with anhydrous sodium sulfate and concentrated under vacuum, and the residue was purified using a silica gel column (petroleum ether:ethyl acetate =1:1) to yield LXS98: (S)-(5-(1-(1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)furan-2-yl)methanol (0.1 g, 66%). 1HNMR (300 MHz, DMSO-d6): δ=12.98 (s, 1H), 8.87 (s, 1H), 7.51 (s, 1H), 7.02 (d, J=6.0 Hz, 1H), 6.84 (s, 1H), 6.59 (d, J=6.0 Hz, 1H), 4.39 (s, 1H), 3.79-3.88 (m, 1H), 2.53-2.81 (m, 2H), 2.43 (t, J=6.0 Hz, 2H), 2.20-2.30 (m, 2H), 1.90-2.15 (m, 2H), 1.81 (d, J=6.0 Hz, 2H), 1.36 (m, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 153.84, 151.12, 148.62, 144.95, 142.19, 129.08, 127.15, 126.84, 120.74, 115.64, 107.95, 104.08, 99.35, 57.85, 57.34, 56.38, 55.97, 55.84, 37.75, 27.08, 10.18 ppm; HRMS (ESI): m/z [M+H]+. C21H23F3N5O2 calculated value 434.1798, measured value 434.1788.
LXS99 was prepared using the raw material of 3-hydroxybenzaldehyde in a similar preparation way as described in Example 98, and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS99: (S)-3-(1-(1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)phenol (0.1 g, 54%). 1HNMR (300 MHz, DMSO-d6): δ=12.17 (s, 1H), 8.89 (s, 1H), 7.84 (t, J=3.0 Hz, 1H), 7.58 (s, 1H), 7.34 (t, J=7.5 Hz, 1H), 7.04 (s, 1H), 6.90-7.02 (m, 1H), 6.91 (d, J=3.0 Hz, 1H), 6.08 (d, J=7.5 Hz, 1H), 3.79-3.81 (m, 2H), 2.50-2.80 (m, 2H), 2.45 (t, J=6.0 Hz, 2H), 2.21-2.37 (m, 2H), 1.93-2.19 (m, 2H), 1.83 (d, J=6.0 Hz, 2H), 1.34 (m, 2H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 157.54, 153.74, 148.62, 142.18, 132.08, 130.68, 129.08, 127.10, 126.85, 120.77, 120.65, 115.95, 115.64, 112.97, 99.34, 58.46, 58.21, 56.37, 55.94, 37.77, 27.62, 10.18 ppm; HRMS (ESI): m/z [M+H]+. C22H23F3N5O calculated value 430.1849, measured value 430.1933.
LXS100 was prepared using a preparation method similar to Example 97 with the raw material of 2,2,3,3,4,4,4-heptafluoroiodobutane, and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS100: (R)-1-(1-((S)-1-(2,2,3,3,4,4,4-Heptafluorobutyl)pyrrolidin-3-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.1 g, 43%). 1HNMR (300 MHz, DMSO-d6): δ=12.35 (s, 1H), 8.87 (s, 1H), 7.54 (s, 1H), 6.63 (s, 1H), 4.60-4.70 (m, 1H), 3.73-3.79 (m, 1H), 2.56-2.81 (m, 2H), 2.68 (s, 2H), 2.20-2.30 (m, 2H), 1.90-2.15 (m, 2H), 1.49 (d, J=6.0 Hz, 3H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.79, 148.59, 129.16, 127.19, 120.88, 120.71, 118.74, 115.65, 104.35, 99.65, 63.64, 58.94, 58.41, 55.91, 51.95, 27.55, 22.40 ppm; HRMS (ESI): m/z [M+H]+. C18H19F7N5O calculated value 454.1472, measured value 454.1488.
(R)-1-(1-((S)-1-(Perfluoro-butyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanolLXS101 was prepared using a similar preparation method as in Example 97, with the raw material of perfluorobutane. The crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS101: (R)-1-(1-((S)-1-(Perfluoro-butyl)pyrrolidin-3-yl)-1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl)ethanol (0.1 g, 64%). 1HNMR (300 MHz, DMSO-d6): δ=12.45 (s, 1H), 8.91 (s, 1H), 7.39 (s, 1H), 6.68 (s, 1H), 4.60-4.74 (m, 1H), 3.73-3.82 (m, 1H), 2.54-2.88 (m, 2H), 1.90-2.30 (m, 4H), 1.51 (d, J=6.0 Hz, 3H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 156.61, 148.61, 148.51, 142.18, 129.01, 127.34, 120.78, 118.20, 115.28, 107.24, 99.33, 63.88, 58.99, 44.84, 42.38, 27.65, 22.81 ppm; HRMS (ESI): m/z [M+H]+. C18H17F9N5O calculated value 490.1284, measured value 490.1288.
(i) LXS102 was prepared using a similar preparation method as in Example 97 using the raw material of trifluoromethylpropylsulfonyl chloride, and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS102: (R)-1-(1-((S)-1-((3,3,3-trifluoropropyl)sulfonyl)pyrrolidin-3-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-2-yl) ethanol (0.2 g, 69%). 1HNMR (300 MHz, DMSO-d6): δ=12.71 (s, 1H), 8.56 (s, 1H), 7.38 (s, 1H), 6.74 (s, 1H), 4.68-4.78 (m, 1H), 3.73-3.84 (m, 1H), 3.20 (d, J=4.5 Hz, 2H), 3.11 (t, J=6.0 Hz, 2H), 2.70-2.84 (m, 2H), 2.37 (t, J=6.0 Hz, 2H), 1.91-2.15 (m, 2H), 1.43 (d, J=6.0 Hz, 3H) ppm; 13CNMR (75 MHz, DMSO-d6) δ 148.66, 148.51, 142.15, 129.05, 127.10, 125.47, 120.71, 115.64, 99.34, 63.64, 57.58, 56.14, 50.04, 40.08, 29.87, 26.27, 22.87 ppm; HRMS (ESI); m/z [M+H]+. C17H21F3N5O3S calculated value 432.1312, measured value 432.1318.
LXS103 was prepared using a preparation method similar to that used for the preparation of Intermediate 10 and Example 79 using the raw material of 4-bromopropyl cyanide, and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS103: 4-(4-(imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)butanenitrile (0.1 g, 57%). 1 HNMR (300 MHz, DMSO-d6): δ=12.13 (s, 1H), 8.88 (s, 1H), 8.06 (s, 1H), 7.94 (s, 1H), 7.55 (s, 1H), 7.15 (s, 1H), 6.68 (s, 1H), 4.46 (t, J=6.0 Hz, 2H), 2.10-2.21 (m, 2H), 1.87 (t, J=6.0 Hz, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.77, 142.11, 134.58, 130.77, 129.71, 129.08, 127.88, 120.74, 119.34, 115.67, 100.57, 99.37, 52.38, 22.74, 14.97 ppm; HRMS (ESI): m/z [M+H]+. C18H14N7 calculated value 292.1305, measured value 292.1316.
LXS104 was prepared using a similar preparation method as in Example 103 using the raw material of 5-methylfuran-2-formaldehyde, and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS104: 4-(4-(2-(5-methylfuran-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)butanenitrile (0.2 g, 67%). 1 HNMR (300 MHz, DMSO-d6): δ=12.33 (s, 1H), 8.85 (s, 1H), 8.11 (s, 1H), 7.98 (s, 1H), 7.55 (s, 1H), 6.95 (d, J=6.0 Hz, 1H), 6.68 (s, 1H), 6.08 (d, J=6.0 Hz, 1H), 4.38 (t, J=6.0 Hz, 2H), 2.30 (s, 3H), 2.14-2.25 (m, 2H), 1.91 (t, J=4.5 Hz, 2H) ppm; 13C NMR (75 MHz, DMSO-d6) δ 152.21, 151.47, 148.65, 142.18, 141.28, 130.75, 129.70, 129.08, 127.18, 120.78, 119.58, 115.68, 107.88, 107.67, 100.51, 99.34, 52.38, 22.74, 14.95, 13.77 ppm; HRMS (ESI): m/z [M+H]+. C20H18N7O calculated value 372.1567, measured value 352.1588.
LXS105 was prepared using a preparation method similar to Example 103 using the raw material of 5-methylthiophene-2-formaldehyde, and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS105: 4-(4-(2-(5-methylthiophen-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl butanenitrile (0.1 g, 45%). 1 HNMR (300 MHz, DMSO-d6): δ=12.28 (s, 1H), 8.68 (s, 1H), 8.06 (s, 1H), 7.94 (s, 1H), 7.63 (s, 1H), 7.49 (d, J=6.0 Hz, 1H), 6.83 (d, J=6.0 Hz, 1H), 6.61 (s, 1H), 4.46 (t, J=6.0 Hz, 2H), 2.36 (s, 3H), 2.11-2.23 (m, 2H), 1.87 (t, J=4.5 Hz, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.66, 142.18, 141.68, 141.24, 134.38, 130.75, 129.75, 129.04, 127.54, 127.45, 120.77, 119.35, 115.67, 100.57, 99.35, 52.38, 22.75, 15.29, 14.95 ppm; HRMS (ESI): m/z [M+H]+. C20H18N7S calculated value 388.1339, measured value 388.1345.
LXS106 was prepared using a similar preparation method as in Example 79 using the raw material of thiophene-2formaldehyde, and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS106: 3-(4-(2-(Thiophen-2-yl)imidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)propanenitrile (0.1 g, 64%). 1 HNMR (300 MHz, DMSO-d6): δ=12.34 (s, 1H), 8.84 (s, 1H), 8.34 (s, 1H), 7.98 (s, 1H), 7.85 (d, J=4.5 Hz, 1H), 7.69 (d, J=4.5 Hz, 1H), 7.63 (s, 1H), 7.19 (m, 1H), 6.56 (s, 1H), 5.08 (t, J=6.0 Hz, 2H), 3.20 (t, J=6.0 Hz, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.68, 143.98, 142.18, 141.28, 130.74, 129.72, 129.10, 129.00, 128.61, 128.07, 127.15, 120.75, 117.77, 115.65, 100.58, 99.38, 49.25, 15.92 ppm: HRMS (ES1): m/z [M+H]+. C18H14N7S calculated value 360.1026, measured value 360.1038.
LXS107 was prepared using a similar preparation method as in Example 103 using the raw material of cyclobutanecarboxaldehyde, and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS107: 4-(4-(2-cyclobutylimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)butanenitrile (0.1 g, 57%). 1 HNMR (300 MHz, DMSO-d6): δ=12.34 (s, 1H), 8.89 (s, 1H), 8.06 (s, 1H), 7.96 (s, 1H), 7.63 (s, 1H), 6.56 (s, 1H), 4.46 (t, J=6.0 Hz, 2H), 3.19-3.24 (m, 1H), 2.15-2.38 (m, 4H), 2.10-2.14 (m, 2H), 1.91-2.01 (m, 2H), 1.77-1.87 (m, 2H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.60, 144.89, 142.18, 130.77, 129.71, 129.08, 127.18, 120.75, 119.37, 115.67, 100.59, 99.38, 52.38, 32.69, 26.28, 22.78, 18.93, 14.92 ppm; HRMS (ESI): m/z [M+H]+. C19H20N7 calculated value 346.1775, measured value 346.1778.
LXS108 was prepared using a similar preparation method as in Example 103 using the raw material of cyclopropylformaldehyde, and the crude product was purified using a silica gel column (petroleum ether:ethyl acetate=1:1) to yield LXS108: 4-(4-(2-cyclopropylimidazo[4,5-d]pyrrolo[2,3-b]pyridin-1(6H)-yl)-1H-pyrazol-1-yl)butanenitrile (0.1 g, 62%). 1 HNMR (300 MHz, DMSO-d6): δ=12.11 (s, 1H), 8.69 (s, 1H), 8.11 (s, 1H), 7.94 (s, 1H), 7.58 (s, 1H), 6.41 (s, 1H), 4.51 (t, J=6.0 Hz, 2H), 2.10-2.18 (m, 2H), 1.77-1.87 (m, 2H), 1.35-1.52 (m, 1H), 0.99-1.24 (m, 4H) ppm; 13 C NMR (75 MHz, DMSO-d6) δ 148.68, 144.51, 142.98, 130.77, 129.75, 129.05, 127.15, 120.77, 119.35, 115.68, 100.54, 99.35, 52.38, 22.74, 14.95, 8.59, 3.48 ppm; HRMS (ESI): m/z [M+H]+. C18H18N7 calculated value 332.1618, measured value 332.1624.
Reagents and consumables
(1) Prepare 1 Kinase buffer.
(2) Compound concentration gradient preparation: The test compound was tested at 1000 nM, 3-fold dilution, 10 concentrations, two-well assay, and configured to a 100-fold final concentration in a 384-well plate. 250 nl of the test compound was then transferred to the 384 reaction plate using Echo550. 250 nl of 100% DMSO was added to each of the negative and positive control wells.
(3) Prepare 2.5 times the final concentration of kinase solution with 1×Kinase buffer.
(4) Add 10 μL of 2.5 times the final concentration of kinase solution to the compound wells and 10 μL of 1×Kinase buffer to the negative control wells.
(5) Centrifuge at 1000 rpm for 30 seconds, mix well with shaking, and incubate at room temperature for 10 minutes.
(6) Prepare a 25/15-fold final concentration of mixture of ATP and Kinase substrate with 1×Kinase buffer.
(7) Add 15 μL of a mixture of 25/15 times the final concentration of ATP and substrate to start the reaction.
(8) Centrifuge the 384-well plate at 1000 rpm for 30 seconds, mix well with shaking, and incubate at room temperature for an appropriate amount of time.
(9) Stop the kinase reaction by adding 30 μL of termination solution, centrifuge at 1000 rpm for 30 seconds, and mix well with shaking.
(10) Read the conversion rate with Caliper EZ Reader.
where: Conversion %_sample is the conversion rate reading of the sample-, Conversion %_min: the average value of the negative control wells, representing the conversion reading of the wells without enzyme activity: Conversion %-max: the average value of the positive control wells, representing the conversion reading of the wells without compound inhibition.
The log value of concentration was used as the X-axis and the percentage inhibition as the Y-axis, and the function of the analytical software GraphPad Prism 5; log(inhibitor) vs. response -Variable slope, was used to lit dose-effect curves to obtain the IC50) values of each compound on enzyme activity.
The formula is Y=Bottom+(Top-Bottom)/(1+10{circumflex over ( )}((LogIC50-X)*HillSlope))
The 1C50 data are specified in Table 1.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2021/140062 | 12/21/2021 | WO |
