Patent Application
20150307520
This invention relates generally to the field of medicine and, more specifically, to novel heteroaryl and heterocycle compounds and pharmaceutical compositions comprising them, uses and methods thereof for inhibiting the activity of PI3K and for treating inflammatory and autoimmune diseases and cancer.
Phosphoinositide 3-kinases (PI 3-kinases or PI3Ks) are a family of enzymes involved in cellular functions such as cell growth, proliferation, differentiation, motility, survival and intracellular trafficking. After exposure of cells to various biological stimuli, PI3Ks primarily phosphorylate phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2, PIP2) at the 3′-OH position of the inositol ring to generate phosphatidylinositol-3,4,5-trisphosphate (PtdIns(3,4,5)P3, PIP3) which has an important role as second messengers by working as a docking platform for lipid-binding domains, such as the pleckstrin homology (PH) domains of various cellular proteins. These include kinases (such as 3-phosphoinositide-dependent protein kinase 1 (PDK1) and protein kinase B (PKB)/Akt) that trigger downstream kinase cascades, and guanine-nucleotide exchange factors (such as Vav and P-Rex) that control the activity of small GTPases (T Rückle, M. K. et al. Nature Reviews Drug Discovery, 2006, 5, 903-9018).
Based on sequence homology and lipid substrate specificity, the PI3K family is divided into three classes: I, II, and III. The most studied and the focus of this invention, the class I PI3Ks, are heterodimeric proteins, each containing a smaller regulatory domain and a larger 110 kDa catalytic domain which occur in four isoforms differentiated as p110α, p110β, p110γ and p110δ (T. J. Sundstrom. et al Org. Biomol. Chem., 2009, 7, 840-850). Among them, p110α, p110β and p110δ together, termed as the class IA PI3K, bind to p85 regulatory subunit and are primarily activated by protein tyrosine kinase-coupled receptors (RTK) and/or Ras proteins, whereas PI3Kγ as the sole class IB member, binds to one of two noncatalytic subunits, p101 or p87, is activated by G-protein coupled receptors (GPCRs) through direct interaction with G-protein βγ dimers and Ras proteins, which are widely implicated in various aspects of immune function and regulation.
All four class I catalytic PI3K isoforms show a characteristic expression pattern in vivo. p110α and p110β are ubiquitously expressed, while p110γ and p110δ are found predominantly in leukocytes, endothelial cells and smooth muscle cells (T. J. Sundstrom. et al Org. Biomol. Chem., 2009, 7, 840-850). Deletion of the class IA isoform p110α or β induces embryonic lethality (E9.5-E10) (Bi L, Okabe I. et al. J Biol Chem, 1999, 274: 10963-8; Bi L, Okabe I. et al. Mamm Genome. 2002, 13, 169-72) p110γ-deficient mice develop and reproduce normally, although they have suboptimal immune responses because of defects in T-cell activation as well as in neutrophil and macrophage migration. The loss of p110δ mice are also viable and fertile but exhibit significant defects in T, B cell activation (A Ghigo. et al. BioEssays 2010, 32: 185-196).
Dysregulation and overactivation of the PI3K/AKT pathway has been firmly established in cancer cells. In principle, modulating PI3K and thus controlling PIP3 levels should regulate AKT activity and ultimately suppress tumor growth. The expression of PI3Kδ is generally restricted to hematopoietic cell types. The p110δ isoform is constitutively activated in B cell tumors. Genetic and pharmacologic approaches that specifically inactivate the p110δ isoform have demonstrated its important role for the treatment of B cell malignancy (B. J. Lannutti. et al. Blood. 2011, 117, 591-594). Previous studies have shown that CAL-101, a potent and selective p110 inhibitor, has broad antitumor activity against cancer cells of hematologic origin. (Lannutti B. J. Am Soc Hematol. 2008; 112. Abstract 16; Flinn I. W. et al. J. Clin. Oncol. 2009; 27(A3543))
In addition to cancer, PI3K has also been suggested as a target for inflammatory and autoimmune disorders. The isoforms p110δ and p110γ are mainly expressed in cells of the immune system and contributes to innate and adaptive immunity. p110δ and p110γ regulate diverse immune cell function. For example, inhibition of p110δ leads to suppression of B-cell activation and function, suppression of T-lymphocyte proliferation, T-cell trafficking, and Th1-Th2 differentiation and Treg function. Inhibition of both p110δ and p110γ results in inhibition of neutrophil (leukocyte) chemotaxis, inhibition of mast cell activation, intact macrophage phagocytosis and endothelium activation. Inhibition of p110γ could activate microglial (C. Rommel. et al. Current Topics in Microbiology and Immunology, 2010, 1, 346, 279-299). So isoform-specific p110δ or p110γ inhibitors are expected to have therapeutic effects on these diseases without interfering with general PI3K signaling critical to the normal function of other cellular systems. p110δ and p110γ supporting the hypothesis that p110γ alone, p110δ alone, or dual-blockade of both, all present a unique therapeutic opportunity in that pharmacological inhibition, but the two PI3K isoforms simultaneously may yield more superior clinical results in the treatment of a variety of complex immune-mediated inflammatory diseases. In the case of RA, Phosphoinositide 3-kinases (PI3Ks), most notably PI3Kδ and PI3Kγ, have crucial and specific roles at all stages of disease progression: in antigen signalling in B and T cells, and in signalling downstream of FcRs, cytokine receptors and chemokine receptors in mast cells, macrophages, neutrophils and synoviocytes (C. Rommel. et al. Nature Reviews Immunology, 2007, 7, 191-201). Although the pathogenesis of RA is not yet completely understood, chemokines and other chemoattractants have been detected in the inflamed joint and are responsible for the recruitment of leukocytes into the joints. Amongst these, neutrophils constitute the most abundant population and are capable of inducing inflammatory response and tissue damage (T Riickle, M. K. et al. Nature Reviews Drug Discovery, 2006, 5, 903-9018). Blockade of hematopoietic PI3Kγ and/or PI3Kδ can potently suppresses neutrophil chemotaxis and, in turn, the progression of joint inflammation and cartilage erosion.
Novel compounds are disclosed which in some instances are inhibitors of PI3Ks kinase activity including p110δ, p110γ, p110α, and p110β. These compounds therefore have potential therapeutic benefit in the treatment of a variety of diseases associated with inappropriate p110δ, p110γ, p110α, and p110β activity, such as cancer, inflammatory, allergic and autoimmune diseases and leukemia etc, in particular systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA), allergic disorders, respiratory diseases like asthma and chronic obstructive pulmonary disease (COPD), multiple sclerosis, all pathologic conditions whose onset and/or progression is driven by an inflammatory insult, such as myocardial infarction and cancer.
The present invention provides a compound of formula I-1, I-2 or I-3:
and/or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or mixtures of any ratio or pharmaceutically acceptable salts thereof, wherein all substituents are as defined in the detailed description.
Also provided is a pharmaceutical composition, comprising at least one compound of formula I-1, I-2 or I-3 and/or at least one pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier.
Also provided is a method of inhibiting the activity of PI3K kinase, comprising contacting the kinase with an effective amount of at least one compound of formula I-1, I-2 or I-3 and/or at least one pharmaceutically acceptable salt thereof.
Also provided is a method of treating a disease responsive to inhibition of PI3K in a subject, comprising administering a therapeutically effective amount of at least one compound of formula I-1, I-2 or I-3 and/or at least one pharmaceutically acceptable salt thereof.
Also provided is at least one compound and/or at least one pharmaceutically acceptable salt described herein for use in the treatment of diseases responsive to inhibition of PI3K.
Also provided is a use of at least one compound and/or at least one pharmaceutically acceptable salt described herein in the manufacture of a medicament for use in the treatment of diseases responsive to inhibition of PI3K.
The subject described herein can be human.
Provided is at least one compound of formula I-1, I-2 or I-3:
and/or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or mixtures of any ratio, or pharmaceutically acceptable salts thereof, wherein
wherein each optionally substituted group above for which the substituent(s) is (are) not specifically designated, can be unsubstituted or independently substituted with one or more, such as one, two, or three, substituents independently chosen from halo, —OH, —CN, —CF3, —SO2R′, —NR′R″, alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocycle, aryl, and heteroaryl, in which alkoxy, cycloalkyl, heterocycle, aryl and heteroaryl can be further optionally substituted with one or more groups selected from halo, —OH, —CN, —CF3, —SO2R′, —NR′R″, alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocycle, aryl, and heteroaryl.
In some embodiments, the each optionally substituted group can be unsubstituted or independently substituted with one or more, such as one, two, or three, substituents independently chosen from halogen, —OH, —CN, —CF3, —SO2R′, —NR′R″, C1-C10 alkyl (preferably C1-C6 alkyl, more preferably C1-C4 alkyl), C2-C10 alkenyl (preferably C2-C6 alkenyl, more preferably C2-C4 alkenyl), C2-C10 alkynyl (preferably C2-C6 alkynyl, more preferably C2-C4 alkynyl), C1-C10 alkoxy (preferably C2-C6 alkoxy, more preferably C2-C4 alkoxy), C3-C12 cycloalkyl, 3-12 membered heterocycle, aryl and heteroaryl, in which alkoxy, cycloalkyl, heterocycle, aryl and heteroaryl can be further optionally substituted with one or more groups selected from halo, —OH, —CN, —CF3, —SO2R′, —NR′R″, alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, heterocycle, aryl, and heteroaryl.
In some embodiments, the each optionally substituted group can be unsubstituted or independently substituted with one or more, such as one, two, or three, substituents independently chosen from halogen, —OH, —CN, —CF3, —SO2CH3, —N(C1-C4 alkyl) (C1-C4 alkyl), C1-C4 alkyl, C1-C4 alkoxy, C3-C6 cycloalkyl, morpholinyl, phenyl and pyrimidinyl, in which morpholinyl, phenyl and pyrimidinyl can be further optionally substituted with one or more groups selected from halo, —OH, —CN, —CF3, and C1-C4 alkyl.
In some embodiments, optionally substituted alkyl can be unsubstituted or independently substituted with one or more substituents independently chosen from: halogen, —OH, —CN, —CF3, C1-C4 alkoxy, C3-C6 cycloalkyl, 4-6 membered heterocycle, 5-6 membered aryl, 5-6 membered heteroaryl, —N(C1-C4 alkyl) (C1-C4 alkyl), and SO2R′; wherein R′ is selected from C1-6 alkyl and C3-6 cycloalkyl.
In some embodiments, optionally substituted alkenyl can be unsubstituted or independently substituted with one or more substituents independently chosen from: C1-C4 alkoxy and C1-C4 alkyl.
In some embodiments, optionally substituted alkynyl can be unsubstituted or independently substituted with one or more substituents independently chosen from: —OH, C1-C4 alkoxy and C1-C4 alkyl.
In some embodiments, optionally substituted cycloalkyl can be unsubstituted or independently substituted with one or more substituents independently chosen from: halogen, —OH, —CN, —CF3, C1-C4 alkoxy, and C1-C4 alkyl.
In some embodiments, optionally substituted heteroaryl can be unsubstituted or independently substituted with one or more substituents independently chosen from: halogen, —CN, —CF3, —NO2, —OR′, —NR′R″, —NR′COR″, —COR′, —CONR′R″, —SO2R′, —SR′, and —C(═NOR′)—R″, C1-C4 alkyl, C3-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C1-C4 alkoxy, 4-6 membered heterocycle, and 5-6 membered heteroaryl; wherein
In some embodiments, optionally substituted aryl can be unsubstituted or independently substituted with one or more substituents independently chosen from: halogen, —CN, C1-C4 alkoxy, C1-C4 alkyl, and SO2R′; wherein R′ is selected from C1-6 alkyl and C3-6 cycloalkyl.
In some embodiments, optionally substituted heterocycl can be unsubstituted or independently substituted with one or more substituents independently chosen from: halogen, —OH, —CN, —CF3, —SO2R′, oxo, C1-C4 alkyl, and C1-C4 alkoxy; wherein C1-C4 alkoxy is optionally substituted by C1-C4 alkoxy, R′ is selected from C1-6 alkyl and C3-6 cycloalkyl.
In some embodiments, provided is at least one compound of formula I-1,
In some embodiments, provided is at least one compound of formula I-1, wherein Z═N, R3, R5 and the atoms they are attached to form an heterocyclic ring, which is optionally substituted
In some embodiments, provided is at least one compound of formula I-1, wherein Z═N, R3, R5 and the atoms they are attached to form an heterocyclic ring, which is optionally substituted
In some embodiments, provided is at least one compound of formula I-1, wherein Z═N, R3, R5 and the atoms they are attached to form an optionally substituted 5 membered saturated or partially unsaturated monocyclic heterocyclic ring, which contains one or more, preferably one or two heteroatoms selected from N, O, and S; R1, R2, R4, and W are as defined herein.
In some embodiments, the said 5 membered monocyclic saturated or partially unsaturated heterocyclic ring, which is formed by R3, R5 and the atoms they are attached to, is selected from
each of which is optionally substituted.
In some embodiments, the said 5 membered monocyclic saturated or partially unsaturated heterocyclic ring, which is formed by R3, R5 and the atoms they are attached to, is
which is optionally substituted.
In some embodiments, provided is at least one compound of formula I-1, wherein Z═N, R3, R5 and the atoms they are attached to form an optionally substituted 6 membered mono- or bicyclic saturated or partially unsaturated heterocyclic ring, which contains one or more, preferably one or two heteroatoms selected from N, O, and S; R1, R2, R4, and W are as defined herein.
In some embodiments, the said 6 membered mono- or bicyclic saturated heterocyclic ring, which is formed by R3, R5 and the atoms they are attached to, is
each of which is optionally substituted.
In some embodiments, the said 6 membered mono- or bicyclic saturated heterocyclic ring, which is formed by R3, R5 and the atoms they are attached to, is
which is optionally substituted.
In some embodiments, provided is at least one compound of formula I-1, Z═N, the said heterocyclic ring, which is formed by R3, R5 and the atoms they are attached to, can be optionally substituted with one or more groups selected from halo, —OH, —CN, oxo, —SO2Ra, —ORa and optionally substituted C1-6 alkyl; wherein Ra is C1-6 alkyl, which is optional substituted with C1-C6 alkoxy.
In some embodiments, provided is at least one compound of formula I-1, Z═N, the said heterocyclic ring, which is formed by R3, R5 and the atoms they are attached to, can be optionally substituted with one or more groups selected from oxo, —SO2Ra, and —ORa; or can be optionally substituted with one or more groups selected from methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, each of which is optionally substituted;
In some embodiments, provided is at least one compound of formula I-1, Z═N, R3 and R5 are as defined above; R2 is hydrogen.
In some embodiments, provided is at least one compound of formula I-1, Z═N, R3 and R5 are as defined above; R4 is selected from halo, C1-6 alkyl, C3-C6 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, —C(O)NR′R″, wherein C1-C6 alkyl is optionally substituted with one or more groups selected from: C1-C4 alkoxyl, —OH, and halo.
In some embodiments, provided is at least one compound of formula I-1, Z═N, R3 and R5 are as defined above; R4 is selected from halo, —CF3, and C1-4 alkyl.
In some embodiments, provided is at least one compound of formula I-1, Z═N, R3 and R5 are defined as above; R4 is F, Cl or Br.
In some embodiments, m is 1.
In some embodiments, the said formula I-1 is
In some embodiments, provided is at least one compound of formula I-1, I-2 or I-3, wherein Z═CH; R3, R5 and the atoms they are attached to form an optionally substituted 4-6 membered mono- or bi-cyclic saturated or partially unsaturated heterocyclic ring, which contains one or more, preferably one or two heteroatoms selected from N, O, and S; R1, R2, R4, and W are as defined herein.
In some embodiments, provided is at least one compound of formula I-1, I-2 or I-3, Z═CH; R3, R5 and the atoms they are attached to form an optionally substituted heterocycle selected from:
In some embodiments, provided is at least one compound of formula I-1, I-2 or I-3, Z═CH; the said heterocyclic ring, which is formed by R3, R5 and the atoms they are attached to, can be optionally substituted with one or more groups selected from halo, —OH, —CN, oxo, —SO2Ra, —ORa and optionally substituted C1-6 alkyl; wherein Ra is C1-6 alkyl, which is optional substituted with C1-C6 alkoxy.
In some embodiments, provided is at least one compound of formula I-1, I-2 or I-3, Z═CH; the said heterocyclic ring, which is formed by R3, R5 and the atoms they are attached to, can be optionally substituted with one or more groups selected from oxo, —SO2Ra and —ORa and optionally substituted C1-4 alkyl; wherein Ra is C1-4 alkyl, which is optionally substituted with C1-4 alkoxyl.
In some embodiments, provided is at least one compound of formula I-1, I-2 or I-3, Z═CH; R3 and R5 are as defined above; R2 is hydrogen.
In some embodiments, provided is at least one compound of formula I-1, I-2 or I-3, Z═CH; R2 and R3 are each independently H, methyl or ethyl.
In some embodiments, provided is at least one compound of formula I-1, I-2 or I-3, Z═CH; R5 is hydrogen.
In some embodiments, provided is at least one compound of formula I-1, I-2 or I-3, Z═CH; R1, R2, R3, R5, and W are as defined above; R4 is selected from hydrogen, halo, optionally substituted C1-C6 alkyl, and optionally substituted 5-6 membered monocyclic heteroaryl.
In some embodiments, provided is at least one compound of formula I-1, I-2 or I-3, Z═CH; R1, R2, R3, R5, and W are as defined above; R4 is selected from hydrogen, halo, C1-C4 alkyl and 5-6 membered monocyclic heteroaryl, wherein 5-6 membered monocyclic heteroaryl is optionally substituted with C1-4 alkyl.
In some embodiments, m is 0, 1 or 2.
In some embodiments, m is 1.
In some embodiments, the said formula I-1, I-2 and I-3 are II-1, II-2 and II-3 respectively.
wherein R1, R2, R3, R4, R5, and W are as defined herein.
In some embodiments, R1 is selected from, C1-C6 alkyl, C3-C6 cycloalkyl, —(CR′R″)n-morpholinyl, —(CR′R″)n-phenyl, —(CR′R″)n-pyridinyl, or —(CR′R″)n-pyrimidinyl, in which each of alkyl, morpholinyl, phenyl, pyridinyl and pyrimidinyl independently are optionally substituted with one or more groups selected from halo, C1-C4 alkyl, C1-C4 alkoxyl, —CN, —CF3, and —SO2R′. n, R′ and R″ are as defined herein.
In some embodiments, R1 is (CR′R″)n-aryl, n is 0 and said aryl can be optionally substituted with one or more groups selected from halo, —CN, C1-C4 alkoxyl and —SO2R′. n. R′ and R″ are as defined herein. In some embodiments, R1 is C1-4 alkyl, which is optionally substituted with one or more groups selected from halo, —OH, —NR′R″, —CN, —CF3, —SO2R′, C3-C6 cycloalkyl, 5-6 membered heteroaryl and 5-6 membered heterocycle.
In some embodiments, R1 is selected from C3-C6 cycloalkyl, phenyl, pyridyl, and pyrimidinyl, each of which is optionally substituted with one or more groups selected from halo, C1-4 alkyl, —CN, —CF3 and —SO2R′; R′ and R″ are each independently hydrogen or C1-C4 alkyl.
In some embodiments, R1 is (CR′R″)n-phenyl, n is 0 and said phenyl can be optionally substituted with one or more groups selected from halo, —CN, C1-C4 alkoxyl, and —SO2R′.
In some embodiments, R1 is phenyl optionally substituted with one or more halo.
In some embodiments, R′ and R″ are each independently selected from hydrogen, C1-6 alkyl, C3-6 cycloalkyl and 4-6 membered heterocycle. In some embodiments, R′ and R″ are each independently selected from hydrogen, halo, —CN, —OH, and —CF3.
In some embodiments, n is 0, 1 or 2.
In some embodiments, W is selected from IV-1 to IV-22,
In some embodiments, W is selected from IV-1 to IV-22, which is optionally substituted with one or more groups selected from halo, —CN, —CF3, —NO2, —OR′, —NR′R—C(O)NR′R″, —NR′COR″, —C(O)R′, —C(═N—OR′)—R″, —S(O)pR′, —SR′, C1-6 alkyl, C2-6 alkenyl, C2-6alkynyl, C1-6alkoxy, 5-6 membered monocyclic heterocycle and 5-6 membered monocyclic heteroaryl; wherein alkyl, alkenyl, alkynyl, heterocycle and heteroaryl is optionally substituted with one or more groups selected from —OH, —CN, C1-4alkoxy, C1-4 alkyl, and —NR′R″;
R′ and R″ are each independently hydrogen, C1-4 alkyl, C3-6 cycloalkyl or 4-6 membered heterocycle; wherein alkyl is optionally substituted with one or more groups selected from —OH, halo and C1-4alkoxy.
In some embodiments, W is IV-2, which is substituted with one or more groups selected from —CN, —NH2, C1-C6 alkyl and —C(O)R′; R′ is C1-C6 alkyl optionally substituted with one or more halo, or R′ is C3-6 cyclcoalkyl optionally substituted with one or more halo.
In some embodiments, W is IV-2, which is substituted with —C(O)R′; R′ is C1-C4 alkyl optionally substituted with one or more halo.
In some embodiments, W is IV-2, which is substituted with —C(O)CF3.
In some embodiments, W is IV-2, which is substituted with —C(O)R′; R′ is C1-C4 alkyl.
In some embodiments, W is IV-4, which is substituted with one or more groups selected from —CN, halo and —C(O)R′.
In some embodiments, W is IV-4, which is substituted with —CN.
In some embodiments, W is selected from IV-1 to IV-22, which is optionally substituted with halo, —CN, —CF3, —NH2, —S(O)CH3, —C(O)CH3, —C(O)NH2, —C(O)NHCH3, —C(O)N(CH3)2, —NHCOCH3, ethenyl, —CH≡CCH2OH, morpholinyl, 1H-pyrazolyl, pyridyl, pyrimidyl, wherein pyridyl and pyrimidyl can be optionally substituted with methyl, halo, —NH2 or methoxyl.
In some embodiments, m is 0, 1, or 2.
In some embodiments, Z═N.
In some embodiments, Z═CH.
In some embodiments, provided is at least one compound of formula I-1, I-2 or I-3, Z═CH. R2 and R3 are each independently H, methyl and ethyl; and R5 is hydrogen.
In some embodiments, provided is at least one compound of formula I-1, wherein Z═N; R1 is selected from 5-6 membered monocyclic aryl and heteroaryl, which are optionally substituted with one or more groups selected from halo and C1-6 alkyl; R2, R3, R4, R5, and W are as defined herein.
In some embodiments, provided is at least one compound of formula I-1, wherein Z═N; R1 is phenyl or pyridyl, which are optionally substituted with one or more groups selected from halo and C1-6 alkyl; R2, R3, R4, R5, and W are as defined herein.
In some embodiments, provided is at least one compound of formula I-1, wherein Z═N; R3, R5 and the atoms they are attached to form an heterocyclic ring, which is optionally substituted
R1, R2, R4, and W are as defined above.
In some embodiments, provided is at least one compound of formula I-1, wherein Z═N; R3, R5 and the atoms they are attached to form
which is optionally substituted with one or more groups selected from C1-6 alkyl and C1-C4 alkoxy; R1, R2, R4, and W are as defined above.
In some embodiments, provided is at least one compound of formula I-1, wherein Z═N; R3, R5 and the atoms they are attached to form
which is optionally substituted with one or more groups selected from methyl and ethyl; R1, R2, R4, and W are as defined above.
In some embodiments, provided is at least one compound of formula I-1, wherein Z═N; R4 is selected from halo, —CN, C1-6 alkyl, C1-C6 haloalkyl, and C2-C6 alkynyl; R1, R2, R3, R5, and W are as defined herein. In some embodiments, said C1-C6 haloalkyl is —CF3.
In some embodiments, provided is at least one compound of formula I-1, wherein Z═N; R1, R2, R3, R4, and R5 are as defined herein; W is selected from the formula of IV-2, IV-3, IV-4, IV-6, and IV-16, each of which is optionally substituted with one or more groups selected from halo, —CN, —NR′R″, C1-6 alkyl, and —C(O)R′, wherein R′ and R″ are each independently selected from hydrogen, C1-6 alkyl, and C1-C6 haloalkyl.
In some embodiments, provided is at least one compound of formula I-1, wherein Z═N; R1, R2, R3, R4, and R5 are as defined herein; W is selected from the formula of IV-2, IV-3, IV-4, IV-6, and IV-16, each of which is optionally substituted with one or more groups selected from halo, —CN, —NH2, —CH3, —C(O)CH3, and —C(O)CHF2.
Also provided is at least one compound selected from compounds 1 to 521 and/or at least one its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or mixtures of any ratio, or pharmaceutically acceptable salt thereof.
Also provided is a composition comprising at least one compound of formula I-1, I-2 or I-3, and/or at least one pharmaceutically acceptable salt described herein, and at least one pharmaceutically acceptable carrier.
Also provided is a method of inhibiting the activity of PI3K kinase comprising contacting the kinase with an effective amount of at least one compound of formula I-1, I-2 or I-3 and/or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or mixtures of any ratio, or pharmaceutically acceptable salts thereof described herein to the subject in need thereof.
Also provided is a method of treating a disease responsive to inhibition of PI3K comprising administering to a subject in need thereof a therapeutically effective amount of at least one compound of formula I-1, I-2 or I-3 and/or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or mixtures of any ratio, or pharmaceutically acceptable salts thereof described herein.
Also provided is at least one compound of formula I-1, I-2 or I-3 and/or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or mixtures of any ratio, or pharmaceutically acceptable salts thereof described herein for use in the treatment of diseases responsive to inhibition of PI3K.
Also provided is a use of at least one compound of formula I-1, I-2 or I-3 and/or its solvates, racemic mixture, enantiomers, diasteromers, tautomers, or mixtures of any ratio, or pharmaceutically acceptable salts thereof described herein in the manufacture of a medicament for treating diseases responsive to inhibition of PI3K.
In some embodiments, the disease responsive to inhibition of PI3K described above is immune-based disease or cancer.
In some embodiments, the said immune-based disease is rheumatoid arthritis, COPD, multiple sclerosis, asthma, glomerulonephritis, lupus, or inflammation related to any of the aforementioned; the said cancer is lymphoma or acute myeloid leukemia, multiple myeloma and chronic lymphocytic leukemia.
In some embodiments, the said compound described herein can be administered in combination with another kinase inhibitor that inhibits a kinase activity other than a PI3K kinase.
As used in the present specification, the following words, phrases and symbols are generally intended to have the meanings as set forth below, except to the extent that the context in which they are used indicates otherwise. The following abbreviations and terms have the indicated meanings throughout:
A dash (“-”) that is not between two letters or symbols is used to indicate a point of attachment for a substituent. For example, —CONH2 is attached through the carbon atom.
The term “alkyl” herein refers to a C1-10 straight or branched hydrocarbon. Preferably “alkyl” refers to a straight or branched hydrocarbon, containing 1-6 carbon atoms. More preferably “alkyl” refers to a straight or branched hydrocarbon, containing 1-4 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, and t-butyl. “Hydroxylalkyl” refers to the alkyl which is substituted with OH. “Haloalkyl” refers to the alkyl which is substituted with halogen. “Alkoxylalkyl” refers to the alkyl which is substituted with alkoxy. “Aminoalkyl” refers to the alkyl which is substituted with NRaRb, Ra and Rb can be hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclo, aryl, heteroaryl.
By “alkoxy” is meant a straight or branched alkyl group of the indicated number of carbon atoms attached through an oxygen bridge. Alkoxy groups will usually have from 1 to 10 carbon atoms attached through the oxygen bridge. Preferably “alkoxy” refers to a straight or branched alkoxy, wherein the alkyl portion contains 1-6 carbon atoms. More preferably “alkoxy” refers to a straight or branched alkoxy, wherein the alkyl portion contains 1-4 carbon atoms. Examples of alkyl groups include, but not limited to, methoxy, ethoxy, propoxy, i-propoxy, n-butoxy, s-butoxy, t-butoxy, pentoxy, 2-pentyloxy, i-pentoxy, neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy, 3-methylpentoxy, and the like.
The term “alkenyl” herein refers to a C2-10 straight or branched hydrocarbon, containing one or more C═C double bonds. Preferably “alkenyl” refers to a C2-6 straight or branched hydrocarbon, containing one or more C═C double bonds. More preferably “alkenyl” refers to a C2-4 straight or branched hydrocarbon, containing one or more C═C double bonds. Examples of alkenyl groups include, but are not limited to, vinyl, 1-propenyl, and 1-butenyl.
The term “alkynyl” herein refers to a C2-10 straight or branched hydrocarbon, containing one or more C≡C triple bonds. Preferably “alkynyl” refers to a C2-6 straight or branched hydrocarbon, containing one or more CC triple bonds. More preferably “alkynyl” refers to a C2-4 straight or branched hydrocarbon, containing one or more C≡C triple bonds. Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, and 1-butynyl.
The term “cycloalkyl” refers to a saturated and partially unsaturated monocyclic or bicyclic hydrocarbon group having 3 to 12 carbons. The ring may be saturated or have one or more double bonds (i.e. partially unsaturated), but not fully conjugated. Examples of bicycle cycloalkyl groups include, but are not limited to octahydropentalene, decahydronaphthalene, bicyclo[3.2.0]heptane, octahydro-1H-indene. Examples of single cycle cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.
Cycloalkyl also includes 3- to 12-membered monocyclic or bicyclic carbocyclic ring fused with a 5- or 6-membered aromatic ring, and the point of the attachment is on the cycloalkyl ring.
“Aryl” encompasses: 5- and 6-membered C5-6 carbocyclic aromatic rings, for example, benzene; 8- to 12-membered bicyclic ring systems wherein at least one ring is carbocyclic and aromatic, for example, naphthalene; and 11- to 14-membered tricyclic ring systems wherein at least one ring is carbocyclic and aromatic, for example, fluorene.
For bi- or tricyclic rings, wherein one or two carbocyclic aromatic rings are fused with other rings (such as carbocyclic, heterocyclic or heterocyclic aromatic ring), the resulting ring system is aryl, provided that the point of attachment is at the carbocyclic aromatic ring.
For example, aryl includes 5- and 6-membered C5-6 carbocyclic aromatic rings fused to a 5- to 7-membered non-aromatic carbocyclic or heterocyclic ring containing one or more heteroatoms selected from N, O, and S, or a 3- to 12-membered cycloalkyl, provided that the point of the attachment is on the carbocyclic aromatic rings.
Bivalent radicals formed from substituted benzene derivatives and having the free valences at ring atoms are named as substituted phenylene radicals. Bivalent radicals derived from univalent polycyclic hydrocarbon radicals whose names end in “-yl” by removal of one hydrogen atom from the carbon atom with the free valence are named by adding “-idene” to the name of the corresponding univalent radical, e.g., a naphthyl group with two points of attachment is termed naphthylidene. Aryl, however, does not encompass or overlap in any way with heteroaryl, separately defined below.
The term “halo” includes fluoro, chloro, bromo, and iodo, and the term “halogen” includes fluorine, chlorine, bromine, and iodine.
The term “heteroaryl” refers to
For bi- or tricyclic rings, wherein one or two heterocyclic aromatic rings are fused with other rings (such as carbocyclic, heterocyclic or carbocyclic aromatic ring), the resulting ring system is heteroaryl, provided that the point of attachment is at the heteroaromatic ring.
For example, heteroaryl includes 5- to 6-membered heterocyclic aromatic ring fused to a 5- to 7-membered heterocyclic ring containing one or more heteroatoms selected from N, O, and S, or a 5- to 7-membered cycloalkyl ring, provided that the point of the attachment is on the heterocyclic aromatic ring.
When the total number of S and O atoms in the heteroaryl group exceeds 1, those heteroatoms are not adjacent to one another. In some embodiments, the total number of S and O atoms in the heteroaryl group is not more than 2. In some embodiments, the total number of S and O atoms in the aromatic heterocycle is not more than 1.
Examples of heteroaryl groups include, but are not limited to, pyridyl, pyrazinyl, pyridazinyl, pyrimidinyl, triazinyl, pyrrolyl, pyrazolyl, imidazolinyl, isoxazolyl, oxazolyl, thiazolyl, thiadiazolyl, tetrazolyl, thienyl, benzothienyl, furyl, benzofuryl, benzoimidazolinyl, indazolyl, indolyl, triazolyl, quinolinyl, quinoxalinyl, pyrido[3,2-d]pyrimidinyl, quinazolinyl, naphthyridinyl, benzothiazolyl, benzoxazolyl, purinyl, pyrrolopyridinyl, pyrrolopyrimidinyl, imidazolopyridinyl, imidazolopyrimidinyl, imidazolotriazinyl, triazolopyridinyl, triazolopyrimidinyl and triazolotriazinyl.
Bivalent radicals derived from univalent heteroaryl radicals whose names end in “-yl” by removal of one hydrogen atom from the atom with the free valence are named by adding “-idene” to the name of the corresponding univalent radical, e.g., a pyridyl group with two points of attachment is a pyridylidene. Heteroaryl does not encompass or overlap with aryl as defined above.
Substituted heteroaryl also includes ring systems substituted with one or more oxide substituents, such as pyridinyl N-oxides.
The terms “heterocycle” refers to 3- to 12-membered monocyclic, bicyclic and tricyclic rings containing one or more, for example, from 1 to 5, or, in some embodiments, from 1 to 4, heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon; The rings may be saturated or partially unsaturated (i.e. have one or more double bonds), but not fully conjugated. In some embodiments “heterocycle” refers to 4-6 membered monocyclic rings containing one or more heteroatoms selected from N, O, and S, with the remaining ring atoms being carbon.
Heterocycle also includes 5- to 7-membered heterocyclic ring containing one or more heteroatoms selected from N, O, and S fused with a 5- or 6-membered carbocyclic aromatic ring or a 5- or 6-membered heterocyclic aromatic ring, and the point of the attachment is on the cycloalkyl ring. The point of the attachment may be on a carbon or heteroatom in the heterocyclic ring. The heterocycle can be substituted by oxo.
Heterocycle also refers to an aliphatic spirocyclic ring containing one or more heteroatoms selected from N, O, and S, provided that the point of attachment is at the heterocyclic ring.
Suitable heterocycles include, but not limited to, azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl, morpholinyl, piperazinyl, tetrahydropyranyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, oxazolidinyl, thiazolidinyl and thiomorpholinyl.
By “optional” or “optionally” is meant that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances in which it does not. For example, “optionally substituted alkyl” encompasses both “unsubstituted alkyl” and “substituted alkyl” as defined below. It will be understood by those skilled in the art, with respect to any group containing one or more substituents, that such groups are not intended to introduce any substitution or substitution patterns that are sterically impractical, synthetically non-feasible and/or inherently unstable.
The term “substituted”, as used herein, means that any one or more hydrogens on the designated atom or group is replaced with a selection from the indicated group, provided that the designated atom's normal valence is not exceeded. When a substituent is oxo (i.e., ═O) then 2 hydrogens on the atom are replaced. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds or useful synthetic intermediates. A stable compound or stable structure is meant to imply a compound that is sufficiently robust to survive isolation from a reaction mixture, and subsequent formulation as an agent having at least practical utility. Unless otherwise specified, substituents are named into the core structure. For example, it is to be understood that when (cycloalkyl)alkyl is listed as a possible substituent, the point of attachment of this substituent to the core structure is in the alkyl portion.
Compounds described herein include, but are not limited to, their optical isomers, racemates, and other mixtures thereof. In those situations, the single enantiomers or diastereomers, i.e., optically active forms, can be obtained by asymmetric synthesis or by resolution of the racemates or mixtures of diastereomers. Resolution of the racemates or mixtures of diastereomers can be accomplished, for example, by conventional methods such as crystallization in the presence of a resolving agent, or chromatography, using, for example a chiral high-pressure liquid chromatography (HPLC) column. In addition, such compounds include R- and S-forms of compounds with chiral centers. Such compounds also include crystal forms including polymorphs and clathrates. Similarly, the term “salt” is intended to include all isomers, racemates, other mixtures, R- and S-forms, tautomeric forms and crystal forms of the salt of the compound.
The invention includes also pharmaceutically acceptable salts of the compounds represented by Formula I-1, I-2 or I-3, preferably of those described below and of the specific compounds exemplified herein, and methods using such salts.
A “pharmaceutically acceptable salt” is intended to mean a salt of a free acid or base of a compound represented by Formula I-1, I-2 or I-3 that is non-toxic, biologically tolerable, or otherwise biologically suitable for administration to the subject. See, generally, S. M. Berge, et al., “Pharmaceutical Salts”, J. Pharm. Sci., 1977, 66:1-19, and Handbook of Pharmaceutical Salts, Properties, Selection, and Use, Stahl and Wermuth, Eds., Wiley-VCH and VHCA, Zurich, 2002.
Preferred pharmaceutically acceptable salts are those that are pharmacologically effective and suitable for contact with the tissues of patients without undue toxicity, irritation, or allergic response. A compound of Formula I-1, I-2 or I-3 may possess a sufficiently acidic group, a sufficiently basic group, or both types of functional groups, and accordingly react with a number of inorganic or organic bases, and inorganic and organic acids, to form a pharmaceutically acceptable salt. Examples of pharmaceutically acceptable salts include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogen-phosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caproates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyne-1,4-dioates, hexyne-1,6-dioates, benzoates, chlorobenzoates, methyl benzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates, sulfonates, xylenesulfonates, phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, γ-hydroxybutyrates, glycolates, tartrates, methane-sulfonates, propanesulfonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates, and mandelates.
If the compound of Formula I-1, I-2 or I-3 contains a basic nitrogen, the desired pharmaceutically acceptable salt may be prepared by any suitable method available in the art, for example, treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, nitric acid, boric acid, phosphoric acid, and the like, or with an organic acid, such as acetic acid, phenylacetic acid, propionic acid, stearic acid, lactic acid, ascorbic acid, maleic acid, hydroxymaleic acid, isethionic acid, succinic acid, valeric acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, oleic acid, palmitic acid, lauric acid, a pyranosidyl acid, such as glucuronic acid or galacturonic acid, an alpha-hydroxy acid, such as mandelic acid, citric acid, or tartaric acid, an amino acid, such as aspartic acid or glutamic acid, an aromatic acid, such as benzoic acid, 2-acetoxybenzoic acid, naphthoic acid, or cinnamic acid, a sulfonic acid, such as laurylsulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, any compatible mixture of acids such as those given as examples herein, and any other acid and mixture thereof that are regarded as equivalents or acceptable substitutes in light of the ordinary level of skill in this technology.
If the compound of Formula I-1, I-2 or I-3 is an acid, such as a carboxylic acid or sulfonic acid, the desired pharmaceutically acceptable salt may be prepared by any suitable method, for example, treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary or tertiary), an alkali metal hydroxide, alkaline earth metal hydroxide, any compatible mixture of bases such as those given as examples herein, and any other base and mixture thereof that are regarded as equivalents or acceptable substitutes in light of the ordinary level of skill in this technology. Illustrative examples of suitable salts include organic salts derived from amino acids, such as glycine and arginine, ammonia, carbonates, bicarbonates, primary, secondary, and tertiary amines, and cyclic amines, such as benzylamines, pyrrolidines, piperidine, morpholine, and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum, and lithium.
A “solvate,” such as a “hydrate,” is formed by the interaction of a solvent and a compound. The term “compound” is intended to include solvates, including hydrates, of compounds. Similarly, “salts” includes solvates, such as hydrates, of salts. Suitable solvates are pharmaceutically acceptable solvates, such as hydrates, including monohydrates and hemi-hydrates.
As used herein the terms “group”, “radical” or “fragment” are synonymous and are intended to indicate functional groups or fragments of molecules attachable to a bond or other fragments of molecules.
The term “active agent” is used to indicate a chemical substance which has biological activity. In some embodiments, an “active agent” is a chemical substance having pharmaceutical utility.
The terms “treating” or “treatment” or “alleviation” refers to administering at least on compounds/or at least one pharmaceutically acceptable salt described herein to a subject to slow down (lessen) an undesired physiological change or disorder, such as the development or spread of inflammation or cancer. For purposes of this invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of disease state, and remission (whether partial or total), whether detectable or undetectable. “Treatment” can also mean prolonging survival as compared to expected survival if not receiving treatment. Those in need of treatment include those with the condition or disorder.)
The term “effective amount” means an amount or dose of a PI3K-inhibiting agent sufficient to generally bring about a therapeutic benefit in patients in need of treatment for a disease, disorder, or condition mediated by PI3K activity. Effective amounts or doses of the active agents of the present invention may be ascertained by routine methods such as modeling, dose escalation studies or clinical trials, and by taking into consideration routine factors, e.g., the mode or route of administration or drug delivery, the pharmacokinetics of the agent, the severity and course of the disease, disorder, or condition, the subject's previous or ongoing therapy, the subject's health status and response to drugs, and the judgment of the treating physician. An exemplary dose is in the range of from about 0.0001 to about 200 mg of active agent per kg of subject's body weight per day, preferably about 0.001 to 100 mg/kg/day, or about 0.01 to 35 mg/kg/day, or about 0.1 to 10 mg/kg daily in single or divided dosage units (e.g., BID, TID, QID). For a 70-kg human, an illustrative range for a suitable dosage amount is from about 0.05 to about 7 g/day, or about 0.2 to about 5 g/day. Once improvement of the patient's disease, disorder, or condition has occurred, the dose may be adjusted for maintenance treatment. For example, the dosage or the frequency of administration, or both, may be reduced as a function of the symptoms, to a level at which the desired therapeutic effect is maintained. Of course, if symptoms have been alleviated to an appropriate level, treatment may cease. Patients may, however, require intermittent treatment on a long-term basis upon any recurrence of symptoms.
The term “inhibition” indicates a decrease in the baseline activity of a biological activity or process. “Inhibition of PI3K activity” refers to a decrease in the activity of PI3K as a direct or indirect response to the presence of at least one at least one compound and/or at least one pharmaceutically acceptable salt described herein, relative to the activity of PI3K in the absence of the at least one compound and/or the at least one pharmaceutically acceptable salt thereof. The decrease in activity may be due to the direct interaction of the at least one compound and/or at least one pharmaceutically acceptable salt described herein with PI3K, or due to the interaction of the at least one compound and/or at least one pharmaceutically acceptable salt described herein, with one or more other factors that in turn affect PI3K activity. For example, the presence of at least one compound and/or at least one pharmaceutically acceptable salt described herein, may decrease PI3K activity by directly binding to the PI3K, by causing (directly or indirectly) another factor to decrease PI3K activity, or by (directly or indirectly) decreasing the amount of PI3K present in the cell or organism.
In addition, the active agents of the invention may be used in combination with additional active ingredients in the treatment of the above conditions. The additional active ingredients may be coadministered separately with an active agent of Formula I-1, I-2 or I-3 or included with such an agent in a pharmaceutical composition according to the invention. In an exemplary embodiment, additional active ingredients are those that are known or discovered to be effective in the treatment of conditions, disorders, or diseases mediated by PI3K activity, such as another PI3K modulator or a compound active against another target associated with the particular condition, disorder, or disease. The combination may serve to increase efficacy (e.g., by including in the combination a compound potentiating the potency or effectiveness of an active agent according to the invention), decrease one or more side effects, or decrease the required dose of the active agent according to the invention.
The active agents of the invention are used, alone or in combination with one or more additional active ingredients, to formulate pharmaceutical compositions of the invention. A pharmaceutical composition of the invention comprises: (a) an effective amount of at least one active agent in accordance with the invention; and (b) a pharmaceutically acceptable excipient.
A “pharmaceutically acceptable excipient” refers to a substance that is non-toxic, biologically tolerable, and otherwise biologically suitable for administration to a subject, such as an inert substance, added to a pharmacological composition or otherwise used as a vehicle, carrier, or diluent to facilitate administration of a agent and that is compatible therewith. Examples of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.
Delivery forms of the pharmaceutical compositions containing one or more dosage units of the active agents may be prepared using suitable pharmaceutical excipients and compounding techniques known or that become available to those skilled in the art. The compositions may be administered in the inventive methods by a suitable route of delivery, e.g., oral, parenteral, rectal, topical, or ocular routes, or by inhalation.
The preparation may be in the form of tablets, capsules, sachets, dragees, powders, granules, lozenges, powders for reconstitution, liquid preparations, or suppositories. Preferably, the compositions are formulated for intravenous infusion, topical administration, or oral administration.
For oral administration, the active agents of the invention can be provided in the form of tablets or capsules, or as a solution, emulsion, or suspension. To prepare the oral compositions, the active agents may be formulated to yield a dosage of, e.g., from about 5 mg to 5 g daily, or from about 50 mg to 5 g daily, in single or divided doses. For example, a total daily dosage of about 5 mg to 5 g daily may be accomplished by dosing once, twice, three, or four times per day.
Oral tablets may include the active ingredient(s) mixed with compatible pharmaceutically acceptable excipients such as diluents, disintegrating agents, binding agents, lubricating agents, sweetening agents, flavoring agents, coloring agents and preservative agents. Suitable inert fillers include sodium and calcium carbonate, sodium and calcium phosphate, lactose, starch, sugar, glucose, methyl cellulose, magnesium stearate, mannitol, sorbitol, and the like. Exemplary liquid oral excipients include ethanol, glycerol, water, and the like. Starch, polyvinyl-pyrrolidone (PVP), sodium starch glycolate, microcrystalline cellulose, and alginic acid are exemplary disintegrating agents. Binding agents may include starch and gelatin. The lubricating agent, if present, may be magnesium stearate, stearic acid or talc. If desired, the tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate to delay absorption in the gastrointestinal tract, or may be coated with an enteric coating.
Capsules for oral administration include hard and soft gelatin capsules. To prepare hard gelatin capsules, active ingredient(s) may be mixed with a solid, semi-solid, or liquid diluent. Soft gelatin capsules may be prepared by mixing the active ingredient with water, an oil such as peanut oil or olive oil, liquid paraffin, a mixture of mono and di-glycerides of short chain fatty acids, polyethylene glycol 400, or propylene glycol.
Liquids for oral administration may be in the form of suspensions, solutions, emulsions or syrups or may be lyophilized or presented as a dry product for reconstitution with water or other suitable vehicle before use. Such liquid compositions may optionally contain: pharmaceutically-acceptable excipients such as suspending agents (for example, sorbitol, methyl cellulose, sodium alginate, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel and the like); non-aqueous vehicles, e.g., oil (for example, almond oil or fractionated coconut oil), propylene glycol, ethyl alcohol, or water; preservatives (for example, methyl or propyl p-hydroxybenzoate or sorbic acid); wetting agents such as lecithin; and, if desired, flavoring or coloring agents.
The active agents of this invention may also be administered by non-oral routes. For example, compositions may be formulated for rectal administration as a suppository. For parenteral use, including intravenous, intramuscular, intraperitoneal, or subcutaneous routes, the agents of the invention may be provided in sterile aqueous solutions or suspensions, buffered to an appropriate pH and isotonicity or in parenterally acceptable oil. Suitable aqueous vehicles include Ringer's solution and isotonic sodium chloride. Such forms may be presented in unit-dose form such as ampules or disposable injection devices, in multi-dose forms such as vials from which the appropriate dose may be withdrawn, or in a solid form or pre-concentrate that can be used to prepare an injectable formulation. Illustrative infusion doses range from about 1 to 1000 μg/kg/minute of agent admixed with a pharmaceutical carrier over a period ranging from several minutes to several days.
For topical administration, the agents may be mixed with a pharmaceutical carrier at a concentration of about 0.1% to about 10% of drug to vehicle. Another mode of administering the agents of the invention may utilize a patch formulation to affect transdermal delivery.
Active agents may alternatively be administered in methods of this invention by inhalation, via the nasal or oral routes, e.g., in a spray formulation also containing a suitable carrier.
The compounds described herein, and/or the pharmaceutically acceptable salts thereof, can be synthesized from commercially available starting materials by methods well known in the art. The following schemes illustrate methods for most of compound preparation. In each of the schemes, R1, R2, R3, R4, R5 and W are as defined herein.
The compounds thus obtained can be further modified at their peripheral positions to provide the desired compounds. Synthetic chemistry transformations are described, for example, in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley and Sons (1999); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995) and subsequent editions thereof.
The examples below are intended to be purely exemplary and should not be considered to be limiting in any way. Efforts have been made to ensure accuracy with respect to numbers used (for example, amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric. All MS data were checked by agilent 6120 or agilent 1100. All NMR data were generated using a Varian 400-MR machine. All reagents, except intermediates, used in this invention are commercially available. All compound names except the reagents were generated by Chemdraw 10.0.
In the following examples, the abbreviations below are used:
At 55-60° C. with vigorous stirring to a mixture of NCS (107 g, 800 mmol) in THF (250 mL) in a 2 L flask was added 5-methyl-3,4-dihydro-2H-pyrrole (83 g, 1000 mmol) in one-portion. After addition, the reaction spontaneously heated to reflux for about 5 min, then reacted at 60-70° C. for another 1.5 hours. After cooled to r.t., hexane (300 mL) and water (300 mL) were added to the mixture. The organic layer was separated, collected and concentrated. The residue was used in the next step without further purification. To a mixture of the crude 4,4-dichloro-5-(trichloromethyl)-3,4-dihydro-2H-pyrrole (240 g, 941 mmol) in MeOH (2 L) in an ice-bath was added a solution of NaOMe (324 g, 6 mol) in MeOH (1.5 L) drop-wise over an hour. After addition, the mixture was stirred at r.t. for another one hour. Then 2N HCl aq. was added to adjust its pH to 2 and the resulting was stirred at room temperature for 15 minutes. The mixture was concentrated and diluted with EtOAc (2.5 L) and water (2 L). The organic layer was separated, concentrated and purified by column chromatography eluting with EtOAc/PE and then crystallize upon standing. Methyl 3-chloro-1H-pyrrole-2-carboxylate was obtained as an orange solid (91.3 g, yield: 61%). MS (m/z): 160.1 (M+H)+. 1H NMR (400 MHz, DMSO-d6) δ 12.05 (s, 1H), 6.98 (m, 1H), 6.21 (t, J=2.6 Hz, 1H), 3.75 (s, 3H).
To a solution of ethyl 3-amino-1H-pyrrole-2-carboxylate hydrochloride (953 mg, 5.0 mmol) in 48% HBr aq. (3 mL, 26.0 mmol) and water (20 mL) was added NaNO2 (966 mg, 14.0 mmol) in water (3 mL) at −5° C. The resulting mixture was then stirred at −5° C. for another 30 minutes. CuBr (2.01 g, 14.0 mmol, fine powder) was added portion-wise at this temperature, and the mixture was stirred at r.t. for 30 minutes and refluxed for 2 hours. The reaction mixture was then extracted with EtOAc. The organic layer was separated, concentrated and purified by flash column chromatography, eluting with EtOAc/PE to afford ethyl 3-bromo-1H-pyrrole-2-carboxylate as a yellow solid (562 mg, yield: 52%). MS (m/z): 218.0, 220.0 (M+H)+. 1H NMR (400 MHz, DMSO-d6) δ 9.22 (s, 1H), 6.86 (t, J=2.8 Hz, 1H), 6.34 (t, J=2.8 Hz, 1H), 4.36 (q, J=7.1 Hz, 2H), 1.39 (t, J=7.1 Hz, 3H).
To a mixture of 60% NaH (12 g, 0.3 mol) in DMF (100 mL) at 0° C. was added methyl 3-chloro-1H-pyrrole-2-carboxylate (32 g, 0.2 mol) in DMF (100 mL) dropwise over one hour. After stirred at 0° C. for another 2.5 hours, to the light brown mixture was added a solution of O-(2,4-dinitrophenyl)hydroxylamine (48 g, 0.24 mol) in DMF (100 mL) slowly over 30 minutes. The reaction was stirred at 0° C. for 2.5 hours and warmed to room temperature overnight. The mixture was quenched by Na2S2O3 aq. and extracted with EtOAc and washed with 10% LiCl aq. The organic layer was separated, concentrated and purified by flash column chromatography eluting with MeOH/water to give methyl 1-amino-3-chloro-1H-pyrrole-2-carboxylate as a yellow solid (30 g, yield: 86%). MS (m/z): 174.9 (M+H)+.
A mixture of methyl 1-amino-3-chloro-1H-pyrrole-2-carboxylate (30 g, 0.172 mol) in 7N NH3/MeOH (300 mL) was allowed to heat to 130° C. in a sealed tube overnight. After concentrated, the residue was purified by flash column chromatography over silica gel eluting with EtOAc/PE to give 1-amino-3-chloro-1H-pyrrole-2-carboxamide as a white solid (16 g, yield: 58%). MS (m/z): 160.1 (M+H)+.
To a solution of 60% NaH (2.88 g, 72 mmol) in dry DMF (90 mL) was drop-wise added a solution of ethyl 3-bromo-1H-pyrrole-2-carboxylate (13.08 g, 60 mmol) in dry DMF (30 mL) at 0-5° C. over 30 min, then the reaction was stirred at 0-5° C. for 30 min. Subsequently, O-(2,4-dinitrophenyl)hydroxylamine (14.34 g, 72 mmol) in dry DMF (30 mL) was added drop-wise and the reaction was stirred at r.t. for another 16 hours. The mixture was poured into water and extracted with EtOAc. The combined layers were washed with brine, concentrated and purified by flash column chromatography eluting with PE/EA to afford ethyl 1-amino-3-bromo-1H-pyrrole-2-carboxylate as a yellow oil (12.5 g, yield: 89%). MS (m/z): 233.0, 235.0 (M+H)+.
A mixture of ethyl 1-amino-3-bromo-1H-pyrrole-2-carboxylate (12.5 g, 53.6 mol) in 7N NH3/MeOH (80 mL) was heat at 130° C. overnight in a sealed tube. After concentration, the residue was purified by flash column chromatography eluting with MeOH/H2O, and further purified by flash column chromatography over silica gel eluting with EtOAc/PE to give 1-amino-3-bromo-1H-pyrrole-2-carboxamide as a yellow solid (6.0 g, yield: 55%). MS (m/z): 203.9, 205.9 (M+H)+. 1H NMR (400 MHz, DMSO-d6) δ 7.71 (s, 1H), 7.47 (s, 1H), 6.89 (d, J=2.9 Hz, 1H), 6.47 (s, 2H), 6.09 (d, J=2.9 Hz, 1H).
To a solution of CuBr (7.25 g, 50 mmol) and Cs2CO3 (16.25 g, 50 mmol) in DMF (150 mL) was added cyclopropylacetylene (3.3 g, 50 mmol) at r.t. under N2. The reaction was stirred at 120° C. for 15 min, then ethyl isocyanoacetate (11.4 g, 100 mmol) in DMF (20 mL) was added drop-wise and the reaction was stirred at 120° C. for 2 h. The mixture was concentrated and purified by flash column chromatography to give ethyl 3-cyclopropyl-1H-pyrrole-2-carboxylate as a white solid (4.0 g, yield: 49.9%). MS (m/z): 180.1 (M+H)+.
To a mixture of NaH (210 mg, 60%, 5.25 mmol) in DMF (10 mL) was added ethyl 3-cyclopropyl-1H-pyrrole-2-carboxylate (626 mg, 3.5 mol) in DMF (8 mL) dropwise at 0° C., the reaction was stirred at 0° C. for 1 h, then O-(2,4-dinitrophenyl)hydroxylamine (836 mg, 4.2 mmol) in DMF (5 mL) was added dropwise, the reaction was continued at 0° C. for 2 h. The mixture was poured into water and extracted with EtOAc. The organic layers were washed with brine, dried over Na2SO4, concentrated and purified by flash column chromatography to give ethyl 1-amino-3-cyclopropyl-1H-pyrrole-2-carboxylate as a yellow solid (679 mg). MS (m/z): 195.1 (M+H)+.
Ethyl 1-amino-3-cyclopropyl-1H-pyrrole-2-carboxylate (679 mg, 3.5 mmol) was dissolved in MeOH (5 mL), 5 mL of aq. LiOH solution (1 N) was added. The reaction was stirred at reflux for 1 h. The mixture was concentrated, the resulting aqueous mixture was adjusted to pH˜7.0 using 1 N HCl, then extracted with EtOAc, the organic layer was dried over Na2SO4, concentrated to give the crude product 1-amino-3-cyclopropyl-1H-pyrrole-2-carboxylic acid (581 mg) which was used in the next step without further purification.
The mixture of 1-amino-3-cyclopropyl-1H-pyrrole-2-carboxylic acid (581 mg, about 3.5 mmol), NH4Cl (1855 mg, 35 mmol), HATU (1330 mg, 3.5 mmol) and DIPEA (2 mL, 11.5 mmol) in DMF (4 mL) was stirred at r.t. overnight. The reaction mixture was poured into water, extracted with EtOAc, dried over Na2SO4, concentrated and purified by flash column chromatography to give the title product (166 mg, yield: 28%) as a white solid. MS (m/z): 166.1 (M+H)+.
These intermediates were prepared according to the procedure of Intermediate 5 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art.
The mixture of 5-amino-3-(methylthio)-1H-pyrazole-4-carboxamide (516 mg, 3 mmol) and formamide (1 mL) was stirred at 180° C. for 1 h. The reaction was cooled to r.t., and added water. The precipitate was collected and recrystallized from MeOH to give 3-(methylthio)-1H-pyrazolo[3,4-d]pyrimidin-4-ol as a white solid. Yield: 99%. MS (m/z): 182.9 (M+1)+.
The mixture of 3-(methylthio)-1H-pyrazolo[3,4-d]pyrimidin-4-ol (540 mg, 3 mmol) and POCl3 (3 mL) was stirred at reflux for 4 h. The reaction was concentrated, and added ice-cold water, the resulting precipitate was filtered and washed with water to give the desired product as a yellow solid, which was used for the next step without further purification. MS (m/z): 200.8 (M+1)+.
To a solution of 4,6-dichloropyrimidin-2-amine (5.4 g, 33 mmol) and tert-butyl 3-aminopropanoate hydrochloride (6.0 g, 33 mmol) in DMF (3 mL) was added Et3N (5 mL). The reaction was stirred at 60° C. overnight. The mixture was poured into water, extracted with EtOAc, the organic layers were washed with brine, dried over Na2SO4, and concentrated to give tert-butyl 3-((2-amino-6-chloropyrimidin-4-yl)amino) propanoate as a white solid, which was used for the next step without further purification. MS (m/z): 273.0 (M+1)+.
The mixture of tert-butyl 3-((2-amino-6-chloropyrimidin-4-yl)amino)propanoate (6.0 g, 22 mmol) and TFA (20 mL) was stirred at r.t. for 1 h, then concentrated, and adjusted to pH=3-4 with 1N NaOH solution. The precipitate was filtered and washed with water to give 3-((2-amino-6-chloropyrimidin-4-yl)amino)propanoic acid as a white solid, which was used for the next step without further purification. Yield: 61%. MS (m/z): 217.0 (M+1)+.
The mixture of 3-((2-amino-6-chloropyrimidin-4-yl)amino)propanoic acid (2.9 g, 13.4 mmol) and Eaton's reagent (30 mL) was stirred at 75° C. for 3 h. The reaction mixture was poured into iced NH4OH, extracted with EtOAc, the organic layers were washed with brine, dried over Na2SO4, concentrated to give the desired title compound as a yellow solid, which was used for the next step without further purification. MS (m/z): 199.0 (M+1)+.
To a solution of (S)-methyl 2-amino-3-methylbutanoate (6.0 g, 35.9 mmol) in DCM (150 mL) were added HOBT (5.34 g, 39.5 mmol), EDCI.HCl (7.55 g, 39.5 mmol) and picolinic acid (4.86 g, 39.5 mmol) followed with DIEA (14 g, 108 mmol). The reaction was stirred at r.t. overnight. The mixture was concentrated and purified by flash chromatography to afford (S)-methyl 3-methyl-2-(picolinamido)butanoate as a colorless oil. Yield: 52.3%. MS (m/z): 237.0 (M+1)+.
To a solution of (S)-methyl 3-methyl-2-(picolinamido)butanoate (1.5 g, 6.36 mmol) in toluene (15 mL) were added Pd(OAc)2 (36 mg, 0.16 mmol), PhI(OAc)2 (5.12 g, 15.9 mmol) and AcOH (71163 mg, 12.72 mmol) under N2, the mixture was bubbled with N2 for 5 min. The reaction was stirred at 110° C. for 24 h in a sealed tube. After cooling to the r.t., the reaction was concentrated and purified by flash chromatography to afford (2S)-methyl 3-methyl-1-picolinoylazetidine-2-carboxylate as a yellow oil. Yield: 57%. MS (m/z): 234.9 (M+1)+.
To a solution of (2S)-methyl 3-methyl-1-picolinoylazetidine-2-carboxylate (1.3 g, 5.56 mmol) in THF (7 mL) was added a solution of NaOH (267 mg, 6.67 mmol) in H2O (7 mL) at r.t. The reaction was stirred at r.t for 2 h, then adjusted to pH=6 with aq. HCl solution (1N). The mixture was concentrated and purified by flash chromatography to afford the title compound as a white solid. MS (m/z): 221.1 (M+1)+.
To a solution of ethyl 4-chloro-2-(methylthio)pyrimidine-5-carboxylate (2.32 g, 10 mmol) in THF (60 mL) was added DIBAL-H (1N in hexane, 30 mL) dropwise at 0° C., the reaction was stirred at 0° C. for 30 min, then H2O was added followed by 2N HCl solution (45 mL). The mixture was extracted with EtOAc, the organic layers were washed with brine, dried over Na2SO4, concentrated to give (4-chloro-2-(methylthio)pyrimidin-5-yl)methanol as a yellow solid, which was used for the next step without further purification. Yield: 60%, MS (m/z): 190.9 (M+1)+.
To a solution of (4-chloro-2-(methylthio)pyrimidin-5-yl)methanol (1.14 g, 6 mmol) in DCM (200 mL) was added MnO2 (8.7 g, 100 mmol), the reaction was stirred at r.t. overnight, then filtered, the filtrate was concentrated to give 4-chloro-2-(methylthio)pyrimidine-5-carbaldehyde as a yellow solid, which was used for the next step without purification. Yield: 72.7%, MS (m/z): 188.9 (M+1)+.
To a solution of 4-chloro-2-(methylthio)pyrimidine-5-carbaldehyde (376 mg, 2 mmol) in THF (5 mL) was added EtMgBr (3.0 M in hexane, 0.7 mL) dropwise at −78° C. The reaction was stirred at −78° C. for 30 min, then 1N HCl (2 mL) was added. The mixture was extracted with EtOAc, the organic layers were washed with brine, dried over Na2SO4, and concentrated to give 1-(4-chloro-2-(methylthio)pyrimidin-5-yl)propan-1-ol as a colorless oil, which was used for the next step without purification. MS (m/z): 219.0 (M+1)+.
To a solution of 1-(4-chloro-2-(methylthio)pyrimidin-5-yl)propan-1-ol (436 mg, 2 mmol) in DCM (10 mL) was added PCC (537 mg, 2.5 mmol), the mixture was stirred at r.t. under N2 for 2 h, then filtered, the filtrate was concentrated to give 1-(4-chloro-2-(methylthio)pyrimidin-5-yl)propan-1-one as a yellow oil, which was used for next step without purification. MS (m/z): 217.0 (M+1)+.
Intermediate 12 and Intermediate 13 were prepared according to the procedures described in Intermediate 11 using the corresponding reagents and intermediates.
Intermediate 12: MS (m/z): 256.8 (M+1)+.
Intermediate 13: MS (m/z): 229.0 (M+1)+.
To a solution of 1a (3.0 g, 24.0 mmol) and (S)-1-(tert-butoxycarbonyl) pyrrolidine-2-carboxylic acid (7.1 g, 28.8 mmol) in THF (150 mL) was added EDC (5.52 g, 28.8 mmol). The reaction mixture was stirred at room temperature for 3.5 hours, then the mixture was diluted in water and extracted with EtOAc three times. The combined organic layers were separated, dried over anhydrous Na2SO4, filtered and concentrated to afford 1b as a white solid (4.6 g, yield: 60%). MS (m/z): 322.7 (M+H)+. It was used in the next step without further purification
Ethanol (50 ml) was added to 1b (3.1 g, 9.6 mmol), then to the mixture was added a solution of KOH (2.88 g, 49.6 mmol) in water (50 mL). The reaction mixture was heated to 100° C. for 3 days. After cooling to room temperature, the reaction mixture was diluted in water and adjusted to pH=3-4 with 1N HCl aq. A precipitate was filtered off and dried to afford 1c as a white solid (1.7 g, yield: 58%). MS (m/z): 304.7 (M+H)+
A mixture of 1c (604 mg, 2.0 mmol), phenylboronic acid (0.49 g, 4.0 mmol), 4AMS (2 g), Cu(OAc)2 (0.73 g, 4.0 mmol) and Pyridine (0.8 mL, 10.0 mmol) in dry DCM (30 mL) was stirred for 18 hours at room temperature under dry air atmosphere. The mixture was concentrated in vacuo and purified by flash column chromatography eluting with MeOH/water to get 1d as a white solid (150 mg, yield: 20%). MS (m/z): 380.7 (M+H)+
A solution of 1d (150 mg, 0.395 mmol) in 6N HCl/MeOH (20 mL) was stirred for 2.5 hours at room temperature, then concentrated under reduced pressure to afford 1e as a yellow oil which was used directly in next step without further purification.
A mixture of 1e (30 mg, 0.095 mmol), 4-chloro-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (22 mg, 0.128 mmol) and TEA (0.05 ml, 0.360 mmol) in n-BuOH (3 mL) was stirred at reflux for 1.5 h. The reaction mixture was concentrated and purified by flash column chromatography eluting with MeOH/DCM to afford Compound 1 as a white solid (29 mg, yield: 64%). MS (m/z): 422.6 (M+H)+. 1H NMR (400 MHz, DMSO-d6) δ: 12.81 (s, 1H), 8.27-8.26 (m, 2H), 7.72-7.68 (m, 1H), 7.64-7.41 (m, 5H), 6.88 (dd, J=4.3, 1.7 Hz, 1H), 6.47 (dd, J=4.3, 2.7 Hz, 1H), 4.72-4.65 (m, 1H), 4.12-4.06 (m, 1H), 3.96-3.89 (m, 1H), 2.35-2.15 (m, 2H) 2.06-1.83 (m, 2H).
The following Compounds were prepared according to the procedure of Compound 1 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ: 8.24 (m, 2H), 7.58 (m, 3H), 7.50 (m, 2H), 7.43 (m, 1H), 6.93 (m, 1H), 6.53 (m, 1H), 5.10 (m, 1H), 4.35 (m, 1H), 4.14 (m, 1H), 2.63 (m, 1H), 2.06 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 8.18 (s, 1H), 8.07 (m, 1H), 7.57-7.52 (m, 5H), 7.42 (m, 1H), 6.92 (m, 1H), 6.51 (m, 1H), 5.13 (m, 1H), 4.10 (m, 2H), 2.63 (m, 1H), 2.21 (m, 1H).
1H NMR (400 MHz, CDCl3) δ: 8.12 (s, 1H), 7.55 (m, 4H), 7.39 (s, 1H), 7.13 (m, 2H), 6.56 (m, 1H), 5.43 (s, 2H), 5.15 (m, 1H), 4.46 (m, 1H), 4.19 (m, 1H), 2.39 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 8.29 (s, 1H), 8.26 (s, 1H), 7.78-7.72 (m, 1H), 7.63-7.47 (m, 5H), 6.55 (d, J = 3.0 Hz, 1H), 4.68-4.60 (m, 1H), 4.12-4.04 (m, 1H), 3.96-3.88 (m, 1H), 2.36-2.16 (m, 2H), 2.03-1.86 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 12.95 (s, 1H), 8.65-8.05 (m, 3H), 7.72-7.40 (m, 5H), 6.57-6.50 (m, 1H), 5.34-5.26 (m, 0.5H), 4.67-4.59 (m, 0.5H), 4.33-4.25 (m, 0.5H), 4.11-4.03 (m, 0.5H), 3.89-3.83 (m, 0.5H), 3.62-3.58 (m, 0.5H), 2.35-2.15 (m, 2H), 1.98-1.81 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 7.86 (s, 1H), 7.66-7.42 (m, 6H), 6.64-6.54 (m, 3H), 4.53-4.43 (m, 1H), 4.08-3.98 (m, 1H), 3.88-3.80 (m, 1H), 2.11-1.99 (m, 2H), 1.84-1.74 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 8.24-8.23 (m, 2H), 7.70-7.41 (m, 6H), 6.61 (s, 1H), 5.13-5.05 (m, 1H), 4.38-4.28 (m, 1H), 4.15-4.09 (m, 1H), 2.66-2.58 (m, 1H), 2.10-1.98 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 12.93 (s, 1H), 8.18 (s, 1H), 8.09 (s, 1H), 7.72-7.41 (m, 6H), 6.59 (s, 1H), 5.18-5.04 (m, 1H), 4.19-4.03 (m, 2H), 2.68-2.60 (m, 1H), 2.24-2.16 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 7.81 (s, 1H), 7.70-7.62 (m, 1H), 7.7-7.30 (m, 5H), 6.75-6.51 (m, 3H), 4.91-4.81 (m, 1H), 4.20-4.10 (m, 1H), 4.00-3.90 (m, 1H), 2.46-2.38 (m, 1H), 2.01-1.89 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 8.91-6.98 (m, 9H), 6.80-6.48 (m, 1H), 5.03-4.80 (m, 1H), 4.08-3.90 (m, 2H), 2.47-2.37 (m, 1H), 2.10-1.90 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 8.03 (s, 1H), 7.63-7.47 (m, 6H), 7.21 (s, 2H), 6.61-6.55 (m, 1H), 4.61-4.53 (m, 1H), 4.02-3.94 (m 1H), 3.82-3.74 (m 1H), 2.24-2.03 (m, 2H), 1.99-1.71 (m, 2H).
1H NMR (400 MHz, CDCl3) δ 8.36 (s, 1H), 7.95-7.68 (br, 1H), 7.51-6.60 (m, 5H), 6.45-6.20 (m, 1H), 5.50-5.20 (m, 1H), 4.61-4.16 (m, 2H), 2.75-2.25 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 12.78 (s, 1H), 8.30-8.18 (m, 2H), 7.65-7.57 (m, 1H), 7.52-7.38 (m, 1H), 7.26-6.93 (m, 3H), 6.61 (s, 1H), 5.18-5.02 (m, 1H), 4.48-4.18 (m, 1H), 4.14-4.08 (m, 1H), 3.78 (s, 1.5H), 3.74 (s, 1.5H), 2.72-2.56 (m, 1H), 2.15-2.07 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 7.98 (s, 1H), 7.71 (d, J = 2.7 Hz, 1H), 7.59-7.21 (m, 3H), 7.18-6.90 (m, 3H), 6.64 (d, J = 2.6 Hz, 1H), 5.15-4.95 (br, 1H), 4.13-3.93 (m, 2H), 3.76 (s, 1.5H), 3.73 (s, 1.5H), 2.65-2.50 (m, 1H), 2.15-2.03 (m, 1H).
1H NMR (400 MHz, CD3OD) δ: 8.24 (s, 1H), 8.10-7.91 (m, 1H), 7.55-7.38 (m, 1H), 7.41-7.15 (m, 2H), 7.14-6.96 (m, 2H), 6.50-6.35 (m, 1H), 5.68-5.60 (m, 0.5H), 5.38-5.20 (m, 0.5H), 4.41-4.33 (m, 0.5H), 4.20-4.12 (m, 0.5H), 4.03-3.95 (m, 0.5H), 3.91-3.80 (m, 3H), 3.82-3.74 (m, 0.5H), 2.48-1.98 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ: 8.24-8.23 (m, 2H), 7.69-7.59 (m, 2H), 7.58-7.29 (m, 3H), 6.66-6.56 (m, 1H), 5.24-5.00 (m, 1H), 4.36-4.26 (m, 1H), 4.16-4.08 (m, 1H), 2.67-2.57 (m, 1H), 2.15-2.03 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 12.96 (s, 1H), 8.24-8.20 (m, 1H), 8.10 (s, 1H), 7.66-7.39 (m, 5H), 6.60-6.52 (m, 1H), 5.36-5.30 (m, 0.5H), 4.68-4.62 (m, 0.5H), 4.35-4.29 (m, 0.5H), 4.12-4.06 (m, 0.5H), 3.92-3.86 (m, 0.5H), 3.73-3.67 (m, 0.5H), 2.28-2.22 (m, 1H), 2.05-1.86 (m, 3H).
1H NMR (400 MHz, CDCl3) δ: 8.22 (s, 1H), 7.77 (s, 1H), 7.67 (s, 1H), 7.48 (d, J = 7.5 Hz, 1H), 7.37 (t, J = 7.6 Hz, 1H), 7.32-7.26 (m, 2H), 6.75 (d, J = 2.0 Hz, 1H), 5.46-5.38 (m, 1H), 4.07-3.99 (m, 1H), 3.90-3.80 (m, 1H), 2.40-2.18 (m, 2H), 2.11-2.03 (m, 2H).
1H NMR (400 MHz, CDCl3) δ: 8.59-8.09 (m, 1H), 7.98 (s, 1H), 7.86-7.55 (m, 2H), 7.52-7.32 (m, 3H), 6.74 (s, 1H), 5.41-5.29 (m, 1H), 4.35-3.76 (m, 2H), 2.49-2.25 (m, 2H), 2.08-1.98 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 13.14-12.79 (m, 1H), 8.24-8.08 (m, 2H), 7.91-7.29 (m, 5H), 6.63-6.45 (m, 1H), 5.34-5.22 (m, 0.5H), 4.66-4.58 (m, 0.5H), 4.41-4.25 (m, 0.5H), 4.15-4.01 (m, 0.5H), 3.91-3.83 (m, 0.5H), 3.70-3.62 (m, 0.5H), 2.30-2.16 (m, 1H), 2.06-1.78 (m, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 8.31-8.21 (m, 2H), 7.75-7.69 (m, 1H), 7.62-7.48 (m, 4H), 7.33 (d, J = 2.5 Hz, 1H), 6.28 (s, 1H), 4.69-4.61 (m, 1H), 4.11-4.03 (m, 1H), 3.96-3.88 (m, 1H), 2.34 (s, 3H), 2.32-2.24 (m, 1H), 2.20-2.12 (m, 1H), 2.00-1.93 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 8.29-8.08 (m, 2H), 7.73-7.47 (m, 5H), 7.31-7.23 (m, 1H), 6.31-6.20 (m, 1H), 5.38-5.28 (m, 0.5H), 4.68-4.58 (m, 0.5H), 4.34-4.24 (m, 0.5H), 4.13-4.03 (m, 0.5H), 3.89-3.83 (m, 0.5H), 3.69-3.63 (m, 0.5H), 2.37-2.29 (m, 3H), 2.19-1.83 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ: 8.19 (s, 1H), 7.73-7.49 (m, 4H), 7.34-7.28 (m, 1H), 6.31-6.23 (m, 1H), 5.72-5.56 (m, 2H), 5.33-5.23 (m, 0.5H), 4.69-4.59 (m, 0.5H), 4.27-4.17 (m, 0.5H), 4.02-3.94 (m, 0.5H), 3.79-3.73 (m, 0.5H), 3.64-3.58 (m, 0.5H), 2.35 (s, 1.5H), 2.32 (s, 1.5H), 2.26-1.67 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ: 7.85 (s, 1H), 7.63-7.42 (m, 5H), 7.40-7.30 (m, 1H), 6.76-6.52 (br, 2H), 6.35-6.25 (m, 1H), 4.56-4.44 (m, 1H), 4.08-3.98 (m, 1H), 3.87-3.77 (m, 1H), 2.33 (s, 3H), 2.13-1.95 (m, 2H), 1.78-1.70 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 8.22 (s, 1H), 7.93 (s, 1H), 7.76-7.68 (m, 1H), 7.64-7.51 (m, 3H), 7.40-7.29 (m, 1H), 7.30-7.17 (m, 1H), 6.28 (d, J = 3.2 Hz, 1H), 5.34-5.24 (m, 1H), 4.63-4.57 (m, 1H), 4.29-4.19 (m, 1H), 2.63-2.53 (m, 1H), 2.25-2.15 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.22-8.10 (m, 2H), 7.70-7.35 (m, 6H), 6.53-6.47 (m, 1H), 5.54-4.85 (m, 2H), 4.52-4.44 (m, 1H), 4.03-3.66 (m, 1H), 2.27-1.93 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.39 (s, 1H), 8.33 (s, 1H), 7.80 (d, J = 7.8 Hz, 1H), 7.64-7.54 (m, 5H), 6.60 (d, J = 3.0 Hz, 1H), 5.67-5.53 (m, 1H), 4.77-4.73 (m, 1H), 4.39-4.27 (m, 1H), 4.19-4.10 (m, 1H), 2.59-2.29 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.22-8.18 (m, 1H), 8.06-7.71 (m, 1H), 7.68-7.61 (m, 5H), 6.91-6.81 (m, 2H), 6.67 (s, 1H), 4.59-4.51 (m, 1H), 3.81-3.73 (m, 1H), 3.59-3.51 (m, 1H), 2.22-2.07 (m, 2H), 1.93-1.81 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.17 (s, 1H), 7.92-7.84 (m, 1H), 7.75-7.48 (m, 5H), 6.83-6.59 (m, 3H), 4.70-4.62 (m, 1H), 3.72-3.62 (m, 1H), 3.58-3.48 (m, 1H), 2.17-1.97 (m, 2H), 1.89-1.79 (m, 1H), 1.73-1.63 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 8.25 (s, 1H), 8.03 (s, 1H), 7.68-7.53 (m, 5H), 7.21 (s, 1H), 6.10 (d, J = 2.4 Hz, 1H), 4.38-4.04 (m, 2H), 3.43 (br, 1H), 2.64-2.59 (m, 1H), 2.36-2.25 (m, 2H), 2.11-2.03 (m, 2H), 1.02-1.00 (m, 2H), 0.71-0.70 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.55 (s, 1H), 8.51 (s, 1H), 8.01 (d, J = 7.6 Hz, 1H), 7.88-7.77 (m, 5H), 7.57 (d, J = 2.4 Hz, 1H), 6.31 (d, J = 2.8 Hz, 1H), 4.95-4.92 (m, 1H), 4.37-4.32 (m, 1H), 4.22-4.16 (m, 1H), 2.57-2.54 (m, 1H), 2.47-2.41 (m, 1H), 2.29-2.19 (m, 3H), 1.15-1.13 (m, 2H), 0.84 (m, 2H)
1H NMR (400 MHz, CD3OD) δ 7.76-7.63 (m, 5H), 7.53-7.52 (m, 1H), 7.26 (d, J = 2.4 Hz, 1H), 6.11 (d, J = 2.8 Hz, 1H), 4.37 (br, 1H), 4.11 (br, 1H), 3.44 (br, 1H), 2.67-2.62 (m, 1H), 2.33 (br, 1H), 2.20-2.17 (m, 1H), 2.08-2.06 (m, 1H), 2.00-1.90 (m, 1H), 1.02-1.00 (m, 2H), 0.71 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 7.75-7.65 (m, 2H), 7.62-7.52 (m, 2H), 7.48-7.35 (m, 2H), 7.15 (d, J = 2.7 Hz, 1H), 6.72 (d, J = 8.5 Hz, 1H), 6.43 (d, J = 2.7 Hz, 1H), 6.23 (d, J = 7.9 Hz, 1H), 5.73-5.67 (m, 1H), 3.85-3.77 (m, 1H), 3.59-3.51 (m, 1H), 2.20-2.08 (m, 2H), 1.98-1.90 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.42 (s, 2H), 7.85-7.60 (m, 6H), 6.86 (d, J = 2.9 Hz, 1H), 5.32-5.20 (br, 1H), 4.55-4.45 (m, 1H), 4.36-4.26 (m, 1H), 2.84-2.78 (m, 1H), 2.27-2.17 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.21 (s, 1H), 8.13 (s, 1H), 7.68-7.40 (m, 6H), 6.70 (s, 1H), 5.22-5.08 (m, 1H), 4.25-4.08 (m, 2H), 2.73-2.63 (m, 1H), 2.28-2.18 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.35-8.29 (m, 1H), 8.23-8.10 (m, 1H), 7.86-7.42 (m, 6H), 6.62-6.52 (m, 1H), 5.66-5.56 (m, 0.5H), 4.94-4.82 (br, 1H), 4.62-4.52 (br, 0.5H), 4.41-4.31 (br, 0.5H), 4.21-4.11 (br, 0.5H), 3.03-2.91 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.28-8.24 (m, 1H), 8.12-8.05 (m, 1H), 7.83-7.75 (m, 1H), 7.68-7.54 (m, 5H), 6.66-6.60 (m, 1H), 4.96-4.81 (m, 1H), 4.64-4.36 (m, 2H), 3.03-2.83 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 8.14 (s, 1H), 7.64-7.47 (m, 8H), 6.64 (d, J = 3.0 Hz, 1H), 4.92-4.84 (m, 1H), 4.45-4.29 (m, 2H), 2.93-2.81 (m, 1H), 2.47-2.41 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 11.79 (s, 1H), 8.20 (s, 1H), 7.76-7.56 (m, 6H), 7.24 (s, 1H), 6.74 (s, 1H), 6.41 (s, 1H), 5.05-4.99 (br, 1H), 4.22-4.10 (m, 2H), 2.78-2.72 (m, 1H), 2.26-2.16 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 12.86 (s, 1H), 8.32 (s, 1H), 8.00 (s, 1H), 7.75-7.31 (m, 6H), 6.60 (d, J = 3.0 Hz, 1H), 5.33-5.10 (m, 1H), 4.69-4.59 (m, 1H), 4.02-3.81 (m, 1H), 3.41 (s, 3H), 2.58-2.48 ( m, 1H), 1.89-1.79 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 8.26 (s, 1H), 7.99 (s, 1H), 7.72-7.53 (m, 6H), 7.39 (d, J = 2.9 Hz, 1H), 6.58 (d, J = 2.9 Hz, 1H), 4.83 (t, J = 7.0 Hz, 1H), 4.31-4.21 (m, 1H), 3.65-3.61 (m, 1H), 3.44 (s, 3H), 2.13-2.03 (m, 2H), 1.93-1.89 (m, 1H), 1.72-1.68 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 8.14 (s, 1H), 7.75 (d, J = 8.1 Hz, 1H), 7.65-7.51 (m, 6H), 7.15 (d, J = 3.4 Hz, 1H), 6.61 (d, J = 3.6 Hz, 1H), 6.59 (d, J = 2.9 Hz, 1H), 4.66 (d, J = 7.3 Hz, 1H), 4.13-4.05 (m, 1H), 3.87-3.79 (m, 1H), 2.30-2.19 (m, 2H), 2.05-2.01 (m, 1H), 1.90-1.84 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 8.22-8.12 (m, 2H), 7.64-7.49 (m, 6H), 6.61-6.55 (m, 1H), 5.34-4.60 (m, 1H), 4.33-4.10 (m, 1H), 3.84-3.65 (m, 1H), 2.29-2.23 (m, 1H), 2.01-1.89 (m, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 8.17 (s, 1H), 7.76 (d, J = 8.0 Hz, 1H), 7.61-7.53 (m, 6H), 7.17 (s, 1H), 6.58 (d, J = 2.9 Hz, 1H), 4.70-4.64 (m, 1H), 3.92-3.86 (m, 1H), 3.74-3.68 (m, 1H), 2.29-2.19 (m, 2H), 2.02-1.98 (m, 1H), 1.95-1.89 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 11.62 (s, 1H), 8.19 (s, 1H), 7.73-7.50 (m, 6H), 7.23-7.13 (m, 1H), 6.75-6.65 (m, 1H), 5.06-4.98 (m, 1H), 4.23-4.15 (m, 1H), 4.12-4.04 (m, 1H), 2.75-2.67 (m, 1H), 2.25-2.16 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 8.28 (d, J = 7.1 Hz, 0.5H), 8.20 (s, 1H), 8.01(s, 0.5H), 7.98(s, 0.5H), 7.24(s, 0.5H), 7.16(s, 0.5H), 7.77-7.41 (m, 5H), 6.49(s, 0.5H), 6.45(s, 0.5H), 5.58(d, J = 2.4 Hz, 0.5H), 4.99-4.96 (m, 0.5H), 4.59 (s, 2H), 4.44-4.33 (m, 0.5H), 4.21-4.10 (m, 0.5H), 4.04-3.94 (m, 0.5H), 3.80-3.72 (m, 0.5H), 3.31 (s, 3H), 2.35-1.93 (m, 4H).
1H NMR (400 MHz, CD3OD) δ 8.24 (s, 1H), 7.97 (s, 1H), 7.77 (d, J = 7.9 Hz, 1H), 7.65-7.53 (m, 4H), 7.43 (d, J = 7.3 Hz, 1H), 7.29 (d, J = 2.7 Hz, 1H), 6.52 (d, J = 2.6 Hz, 1H), 4.93-4.92 (m, 1H), 4.51 (s, 2H), 4.29-4.25 (m, 1H), 4.09-4.05 (m, 1H), 3.35 (s, 3H), 2.47-2.40 (m, 1H), 2.19-2.18 (m, 1H), 2.11-2.04 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 8.15 (s, 1H), 7.76 (d, J = 7.5 Hz, 1H), 7.66-7.56 (m, 3H), 7.43 (d, J = 7.1 Hz, 1H), 7.31 (d, J = 2.3 Hz, 1H), 6.56 (d, J = 2.0 Hz, 1H), 4.95-4.93 (m, 1H), 4.53 (s, 2H), 3.86-3.82 (m, 1H), 3.72-3.67 (m, 1H), 3.37 (s, 3H), 2.26-2.17(m, 1H), 2.07-2.02 (m, 1H), 1.93-1.84 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 8.14 (s, 1H), 8.00(s, 1H), 7.59 (d, J = 8.0 Hz, 1H), 7.52(d, J = 3.2 Hz, 1H), 7.48-7.41 (m, 2H), 7.29-7.26(m, 1H), 7.16-7.12(m, 1H), 6.59(d, J = 2.8 Hz, 1H), 5.07-5.05 (m, 1H), 4.29-4.24 (m, 2H), 4.01-3.97 (m, 2H), 3.84-3.79 (m, 1H), 3.72-3.68(m, 1H).
1H NMR (400 MHz, CD3OD) δ 8.11 (s, 1H), 7.87(s, 1H), 7.72-7.70 (m, 1H), 7.51-7.47(m, 1H), 7.44-7.37(m, 4H), 6.54(d, J = 2.8 Hz, 1H), 5.01-4.92 (m, 1H), 4.30-4.19 (m, 2H), 4.07-4.03 (m, 1H), 3.69-3.63 (m, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 12.39 (s, 1H), 8.23 (s, 1H), 8.22(s, 1H), 7.88-7.30 (m, 7H), 6.56 (d, J = 3.0 Hz, 1H), 6.55 (d, J = 3.0 Hz, 1H), 4.76-4.60 (m, 1H), 4.15-3.63 (m, 2H), 2.90 (s, 1.5H), 2.85 (s, 1.5H), 2.31-2.15 (m, 1H), 2.01-1.69 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 8.24(s, 1H), 7.76 (d, J = 8.0 Hz, 1H), 7.67-7.54 (m, 5H), 7.39 (d, J = 2.8 Hz, 1H), 6.60 (d, J = 2.8 Hz, 1H), 4.78 (t, J = 7.1 Hz, 1H), 3.87-3.79(m, 2H), 2.93(s, 3H), 2.15-2.07 (m, 2H), 2.00-1.94 (m, 1H), 1.85-1.73(m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 8.23(s, 1H), 7.85(s, 1H), 7.77 (d, J = 8.0 Hz, 1H), 7.64-7.53 (m, 4H), 7.49 (d, J = 2.8 Hz, 1H), 6.58 (d, J = 2.8 Hz, 1H), 4.68-4.65 (m, 1H), 4.25-4.18(m, 1H), 3.69-3.63(m, 1H), 2.88(s, 3H), 2.29-2.18 (m, 2H), 1.97-1.88 (m, 2H).
1compound was purified by flash column chromatography
2and 3compounds were purified by preparative TLC
1H NMR (400 MHz, CD3OD) 8.24 (s, 1H), 8.03 (s, 1H), 7.84-7.41 (m, 5H), 7.15-7.09 (m, 1H), 6.30-6.15 (m, 1H), 5.65-5.50 (m, 0.5H), 4.91-4.85 (m, 0.5H), 4.42-4.37 (m, 0.5H), 4.23-4.13 (m, 0.5H), 4.05-3.95 (m, 0.5H), 3.85-3.78 (m, 0.5H), 2.37-1.97 (m, 4H).
1H NMR (400 MHz, CD3OD) δ 8.18 (s, 1H), 7.97 (s, 1H), 7.65-7.45 (m, 2H), 7.34-7.20 (m, 2H), 7.10-7.03 (m, 1H), 6.23-6.10 (m, 1H), 5.58-5.48(m, 0.5H), 4.87-4.78 (m, 0.5H), 4.35-4.28 (m, 0.5H), 4.17-4.07 (m, 0.5H), 3.99-3.89 (m, 0.5H), 3.80-3.70 (m, 0.5H), 2.30-1.94 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ: 12.87 (s, 1H), 8.46-8.23 (m, 3H), 8.16-8.11 (m, 1H), 7.98-7.88 (m, 2H), 7.60-7.57 (m, 1H), 6.65-6.59 (m, 1H), 4.72-4.51 (m, 1H), 4.23-4.07 (m, 1H), 3.97-3.91(m, 1H), 3.32-3.28 (m, 3H), 2.43-2.21 (m, 2H), 2.13-1.96 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 12.95 (s, 1H), 9.03-7.75 (m, 6H), 7.54-7.45 (m, 1H), 6.57-6.54 (m, 1H), 5.35-5.13 (m, 0.5H), 4.53-4.31 (m, 0.5H), 4.05-3.65 (m, 2H), 3.25-3.20(m, 3H), 2.38-1.84 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ: 8.08 (s, 1H), 7.82 (d, J = 2.9 Hz, 1H), 7.74-7.60 (m, 3H), 7.53 (d, J = 7.2 Hz, 2H), 7.34 (br, 2H), 6.81 (d, J = 2.9 Hz, 1H), 5.09 (s, 1H), 4.16 (s, 2H), 2.72-2.60 (m, 1H), 2.25-2.08 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.02 (s, 1H), 7.61-7.49 (m, 6H), 7.22 (brs, 2H), 6.50 (d, J = 2.8 Hz, 1H), 4.60 (s, 3H), 4.00-3.94 (m, 1H), 3.81-3.75 (m, 1H), 3.340(brs, 1H), 3.22 (s, 2H), 2.19-2.07 (m, 2H), 1.97-1.90 (m, 1H), 1.83-1.73 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 12.51 (br, 1H), 8.23 (d, J = 1.9 Hz, 1H), 8.04-7.35 (m, 7H), 6.69-6.53 (m, 1H), 5.15-4.98 (m, 1H), 4.50-4.28 (m, 1H), 3.97-3.90 (m, 1H), 2.89 (d, J = 4.2 Hz, 3H), 2.62-2.55 (m, 1H), 2.04-1.84 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 12.39 (s, 1H), 8.23 (d, J = 4.1 Hz, 1H), 7.85-7.31 (m, 7H), 6.56-6.53 (m, 1H), 4.75-4.67 (m, 1H), 4.13-3.64 (m, 2H), 2.90 (s, 1.5H), 2.85 (s, 1.5H), 2.23-1.71 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ: 12.41 (br, 1H), 8.25-8.22 (m, 1H), 7.91-7.32 (m, 6H), 6.60-6.55 (m, 1H), 4.87-4.52 (m, 1H), 4.23-3.61 (m, 2H), 2.90 (s, 1.5H), 2.85 (s, 1.5H), 2.24-1.78 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ: 12.48 (br, 1H), 8.21 (d, J = 2.0 Hz, 1H), 8.05-7.30 (m, 6H), 6.74-6.51 (m, 1H), 5.10-5.03 (m, 1H), 4.52-4.25 (m, 1H), 3.96-3.93 (m, 1H), 2.88 (d, J = 6.3 Hz, 3H), 2.68-2.54 (m, 1H), 2.14-1.93 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 8.08-7.78 (m, 1H), 7.67-7.37 (m, 5H), 7.35 (s, 1H), 6.51-6.48 (m, 1H), 4.68-4.58 (m, 1H), 3.81-3.73 (m, 1H), 3.60-3.53 (m, 1H), 2.85 (s, 3H), 2.20-2.10 (m, 2H), 2.00-1.87 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 7.91 (s, 0.5H), 7.84-7.81 (m, 0.5H), 7.61-7.46 (m, 5H), 6.73 (s, 1H), 6.59 (d, J = 3.2 Hz, 0.5H), 6.58 (d, J = 2.8 Hz, 0.5H), 4.58-4.57(m, 0.5H), 4.51-4.49(m, 0.5H), 3.77-3.60 (m, 1H), 3.45-3.38 (m, 1H), 2.14 1.92 (m, 5H), 1.83-1.70 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 7.74-7.52 (m, 4H), 7.42-7.07 (m, 2H), 6.31 (d, J = 2.5 Hz, 1H), 5.44-5.22 (m, 1H), 4.48-4.26 (m, 1H), 3.55-3.35 (m, 2H), 3.25-3.04 (m, 1H), 2.64-2.42 (m, 2H), 2.37-2.18 (m, 1H), 0.85-0.44 (m, 3H).
1H NMR (400 MHz, CD3OD) δ 8.03 (s, 1H), 7.92 (s, 1H), 7.39 (d, J = 7.2 Hz, 1H), 7.36 (d, J = 3.2 Hz, 2H), 7.26 (s, 2H), 7.11-7.04 (m, 2H), 6.45 (d, J = 2.8 Hz, 1H), 5.25 (br, 1H), 4.48 (br, 1H), 3.60 (br, 1H), 2.12-2.03 (m, 2H), 1.74-1.40 (m, 4H)
1H NMR (400 MHz, CD3OD) δ 8.24 (d, J = 2.6 Hz, 1H), 7.93 (s, 1H), 7.78 (s, 1H), 7.66-7.62 (m, 1H), 7.60-7.52 (m, 2H), 7.39-7.33 (m, 1H), 7.21 (dd, J = 7.5, 4.3 Hz, 1H), 6.30 (dd, J = 3.2, 2.0 Hz, 1H), 5.28-5.22 (m, 1H), 4.79-4.68 (m, 1H), 4.11-4.04 (m, 1H), 3.01 (s, 3H), 2.61-2.51 (m, 1H), 2.20-2.07 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 7.72 (d, J = 2.9 Hz, 1H), 7.60-7.36 (m, 5H), 6.79 (br, 2H), 6.65 (d, J = 3.0 Hz, 1H), 4.61 (s, 1H), 3.83-3.74 (m, 2H), 2.45-2.40 (m, 1H), 2.10-1.97 (m, 1H), 2.05 (s, 3H).
1H NMR (400 MHz, CDCl3) δ 8.37 (s, 1H), 7.67 (s, 1H), 7.31 (t, J = 8.4 Hz, 1H), 7.21 (dd, J = 8.2, 2.3 Hz, 1H), 7.18-7.12 (m, 1H), 7.10 (s, 1H), 6.44 (d, J = 2.0 Hz, 1H), 5.47 (s, 2H), 5.09 (br, 1H), 4.50-4.24 (m, 4H), 3.58-3.34 (m, 1H), 2.40 (br, 1H), 2.22 (s, 3H), 0.80 (d, J = 6.7 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 8.18 (s, 1H), 7.85 (d, J = 7.7 Hz, 1H), 7.60-7.46 (m, 5H), 6.97 (br, 2H), 6.59 (d, J = 3.0 Hz, 1H), 4.57-4.56 (m, 1H), 3.93 (br, 1H), 3.77-3.73 (m, 1H), 2.07-2.04 (m, 2H), 1.89 (br, 1H), 1.70-1.60 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 8.16 (s, 1H), 8.03 (s, 1H), 7.79-7.75 (m, 1H), 7.67-7.61 (m, 1H), 7.45-7.36 (m, 3H), 6.57 (d, J = 3.0 Hz, 1H), 4.50-4.44 (m, 1H), 3.86-3.82 (m, 1H), 3.68-3.64 (m, 1H), 2.48 (s, 3H), 2.30-1.94 (m, 3H), 1.67-1.59 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 12.43 (s, 1H), 8.25 (s, 1H), 7.94-7.57 (m, 3H), 7.53-7.31 (m, 3H), 6.58-4.55 (m, 1H), 4.75-4.62 (m, 1H), 4.19-4.12 (m, 0.5H), 3.85-3.80 (m, 0.5H), 3.70-3.51 (m, 1H), 290 (s, 1.5H), 2.86 (s, 1.5H), 2.33-2.04 (m, 2H), 2.01-1.73 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 12.28 (br, 1H), 8.17 (s, 1H), 8.04 (s, 1H), 7.66-7.31 (m, 5H), 6.58 (s, 1H), 4.59-4.38 (m, 1H), 3.94-3.62 (m, 2H), 2.48 (s, 3H), 2.15-1.89 (m, 3H), 1.67-1.64 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 12.29 (br, 1H), 8.16 (s, 1H), 8.03 (s, 1H), 7.80-7.35 (m, 6H), 6.57 (s, 1H), 4.47-4.44 (m, 1H), 3.81-3.64 (m, 2H), 2.48 (s, 3H), 2.11-1.93 (m, 3H), 1.65-1.56 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 7.72 (d, J = 6.0 Hz, 1H), 7.63-7.49 (m, 6H), 6.62 (d, J = 3.0 Hz, 1H), 5.96 (s, 2H), 4.56-4.55 (m, 1H), 3.86-3.81 (m, 1H), 3.63-3.56 (m, 1H), 2.16-1.99 (m, 2H), 1.87-1.71 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.25 (s, 1H), 8.17 (d, J = 5.4 Hz, 1H), 7.67-7.35 (m, 5H), 6.63 (d, J = 3.0 Hz, 1H), 5.32-4.06 (m, 1H), 4.82-4.70 (m, 1H), 4.22-4.06 (m, 1H), 4.03-3.87 (m, 1H), 2.55-2.51 (m, 3H), 2.43-2.26 (m, 2H).
1H NMR (400 MHz, CDCl3) δ 11.60 (s, 1H), 8.23 (s, 1H), 7.76 (d, J = 7.9 Hz, 1H), 7.64 (s, 1H), 7.62-7.46 (m, 3H), 7.30-7.26 (m, 1H), 7.08-7.00 (m, 1H), 6.16 (d, J = 3.1 Hz, 1H), 4.81 (t, J = 6.8 Hz, 1H), 4.04-3.92 (m, 1H), 3.88-3.72 (m, 1H), 2.59 (s, 3H), 2.18-2.09 (m, 1H), 2.08-1.93 (m, 3H).
1H NMR (400 MHz, DMSO-d6) δ 12.33 (brs, 1H), 8.26 (s, 0.5H), 8.25(s, 0.5H), 7.87(s, 0.5H), 7.77-7.72 (m, 1H), 7.66-7.52 (m, 4.5H), 7.38-7.36(m, 0.5H), 7.27-7.25(m, 0.5H), 6.42 (d, J = 3.2 Hz, 0.5H), 6.40 (d, J = 3.2 Hz, 0.5H), 4.81-4.77(m, 0.5H), 4.68-4.65 (m, 0.5H), 4.19-4.14 (m, 0.5H), 3.84-3.80 (m, 1H), 3.71-3.65 (m, 0.5H), 2.93 (s, 1.5H), 2.88(s, 1.5H), 2.33-1.764 (m, 4H).
1H NMR (400 MHz, CD3OD) δ 8.64 (d, J = 8.0 Hz, 0.5H), 8.14 (s, 1H), 7.82 (d, J = 7.2 Hz, 0.5H), 7.66-7.62 (m, 1H), 7.58-7.53 (m, 2H), 7.47-7.43 (m, 1H), 7.27 (s, 0.5H), 7.162 (s, 0.5H), 6.43 (s, 0.5H), 6.38 (s, 0.5H), 5.72(br, 0.5 Hz), 4.71(br, 0.5 Hz), 4.48-4.424 (m, 0.5H), 4.27-4.22 (m, 0.5H), 4.02-3.96 (m, 0.5H), 3.82-3.75 (m, 0.5H), 2.41-2.23 (m, 0.5H), 2.29-2.24 (m, 0.5H), 2.21-2.15 (m, 1H), 2.10-1.95 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.21 (s, 1H), 7.78 (d, J = 8.4 Hz, 1H), 7.63-7.52 (m, 5H), 6.59 (d, J = 3.2 Hz, 1H), 4.67-4.64 (m, 1H), 4.23-4.18(m, 1H), 4.03-3.97 (m, 1H), 2.59 (s, 3H), 2.33-2.15 (m, 2H), 2.03-1.89 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.18 (s, 1H), 7.63-7.48 (m, 6H), 6.64 (d, J = 2.8 Hz, 1H), 5.08 (br, 1H), 4.49 (br, 1H), 4.15-4.09 (m, 1H), 2.68-2.61 (m, 1H), 2.55 (s, 3H), 2.14-2.07 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 7.71 (d, J = 2.8 Hz, 1H), 7.70 (s, 1H), 7.62-7.49 (m, 4H), 7.39-7.37 (m, 1H), 6.64 (d, J = 3.0 Hz, 1H), 6.28 (s, 2H), 4.81-4.77 (m, 1H), 4.18-4.12 (m, 1H), 4.02-3.96 (m, 1H), 2.46-2.39 (m, 1H), 2.01-1.95 (m, 1H).
1H NMR (400 MHz, CDCl3) δ 8.00 (s, 1H), 7.60-7.47 (m, 3H), 7.40 (d, J = 7.5 Hz, 1H), 7.30 (d, J = 2.9 Hz, 1H), 7.15-7.10 (m, 1H), 6.49 (d, J = 3.0 Hz, 1H), 5.13-5.03 (m, 1H), 4.85 (s, 2H), 4.39-4.34 (m, 1H), 4.16-4.07 (m, 1H), 3.14 (s, 1H), 2.38-2.18 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.33 (s, 1H), 8.31 (s, 1H), 7.69 (d, J = 8.7 Hz, 1H), 7.64-7.46 (m, 4H), 7.41-7.39 (m, 1H), 6.37 (d, J = 3.2 Hz, 1H), 5.55(br, 0.5H), 5.42(br, 0.5H), 4.87 (dd, J = 9.6, 3.0 Hz, 1H), 4.42-4.22 (m, 2H), 2.60-2.50 (m, 1H), 2.32-2.12 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.28 (t, J = 5.5 Hz, 1H), 8.15 (s, 1H), 7.73 (d, J = 8.0 Hz, 1H), 7.64-7.49 (m, 4H), 7.46 (d, J = 2.9 Hz, 1H), 7.31 (s, 1H), 6.56 (d, J = 2.8 Hz, 1H), 4.53 (dd, J = 7.6, 4.1 Hz, 1H), 3.97-3.86 (m, 1H), 3.77-3.68 (m, 1H), 3.44-3.38 (m, 4H), 3.23 (s, 3H), 2.17-2.04 (m, 2H), 1.93-1.82 (m, 1H), 1.79-1.68 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.23 (s, 1H), 7.84 (d, J = 7.8 Hz, 1H), 7.64-7.41 (m, 5H), 7.15 (s, 2H), 6.42 (d, J = 3.1 Hz, 1H), 5.38 (br, 0.5H), 5.26(br, 0.5H), 4.83 (br, 1H), 4.34-3.97 (m, 2H), 2.40-2.28 (m, 1H), 2.08-1.90 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.17-8.11 (m, 2H), 7.62-7.55 (m, 3H), 7.54-7.48 (m, 2H), 7.47-7.41 (m, 2H), 6.62 (d, J = 3.0 Hz, 1H), 5.05 (t, J = 7.8 Hz, 1H), 4.22-4.21 (m, 1H), 3.75-3.74 (m, 1H), 3.43-3.36 (m, 4H), 3.19 (s, 3H), 2.07-1.67 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.23 (s, 1H), 7.84 (d, J = 7.8 Hz, 1H), 7.66-7.45 (m, 5H), 7.10 (s, 2H), 6.59 (d, J = 3.0 Hz, 1H), 5.39-5.26(m, 1H), 4.83 (br, 1H), 4.22-3.99 (m, 2H), 2.42-2.29 (m, 1H), 2.10-1.93 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.18 (s, 1H), 7.85 (d, J = 7.7 Hz, 1H), 7.62-7.40 (m, 5H), 6.96 (s, 2H), 6.41 (d, J = 3.2 Hz, 1H), 4.60 (br, 1H), 3.94 (br, 1H), 3.76-3.74 (m, 1H), 2.15-1.99 (m, 2H), 1.96-1.82 (m, 1H), 1.70-1.60 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.93-7.89 (m, 1H), 7.61-7.58 (m, 2H), 7.40-7.35 (m, 2H), 7.05 (brs, 2H), 6.59 (d, J = 3.0 Hz, 1H), 4.57 (d, J = 7.4 Hz, 1H), 3.95 (brs, 1H), 3.78-3.72 (m, 1H), 2.07-2.00(m, 2H), 1.98-1.92 (m, 1H), 1.71-1.68 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.21 (d, J = 2.0 Hz, 1H), 7.92 (d, J = 9.6 Hz, 0.5H), 7.78 (d, J = 7.6 Hz, 0.5H), 7.67-7.6274 (m, 1H), 7.60-7.55 (m, 2H), 7.46-7.38 (m, 2H), 7.04 (s, 1H), 6.64-6.63 (m, 1H), 4.64-4.53 (m, 1H), 4.01-3.92 (m, 1H), 3.80-3.74 (m, 1H), 2.16-2.06 (m, 2H), 1.99-1.90 (m, 1H), 1.78-1.68 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.21 (s, 1H), 8.16 (s, 1H), 7.76-7.74 (m, 1H), 7.63- 7.51 (m, 5H), 6.56 (d, J = 3.2 Hz, 1H), 4.37 (d, J = 0.8 Hz, 1H), 4.12-4.08 (m, 1H), 4.02- 3.96 (m, 1H), 2.64-2.59 (m, 1H), 2.54-2.50 (m, 1H), 1.81-1.75 (m, 1H), 0.62 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 8.13 (s, 1H), 7.98 (s, 1H), 7.62-7.49 (m, 6H), 6.55 (s, 1H), 5.07 (br, 0.5H), 4.36 (br, 0.5H), 3.92 (br, 1H), 3.69 (br, 1H), 2.62-2.58 (m, 1H), 2.17 (br, 1H), 1.60 (br, 1H), 0.38 (br, 3H).
1H NMR (400 MHz, CD3OD) δ 8.04 (s, 1H), 7.86 (d, J = 6.9 Hz, 1H), 7.83 (s, 1H), 7.65-7.59 (m, 1H), 7.59-7.54 (m, 2H), 7.44-7.37 (m, 1H), 7.34 (d, J = 3.0 Hz, 1H), 6.49 (d, J = 3.1 Hz, 1H), 4.91-4.89 (m, 1H), 4.71-4.65 (m, 1H), 3.75 (dd, J = 8.2, 4.4 Hz, 1H), 2.78-2.60 (m, 1H), 0.54 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 8.22 (s, 1H), 7.91 (d, J = 2.4 Hz, 1H), 7.63 (d, J = 2.8 Hz, 1H), 7.60-7.59 (m, 3H), 7.56-7.52 (m, 1H), 7.11 (br, 2H), 6.63 (d, J = 2.8 Hz, 1H), 4.328 (br, 1H), 4.024 (br, 1H), 3.875 (br, 1H), 2.277 (br, 1H), 2.026-1.988 (m, 0.5H), 1.683 (br, 1H), 1.453-1.386 (m, 0.5H), 0.420 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 8.18 (s, 1H), 8.10 (s, 1H), 7.75-7.47 (m, 6H), 6.59 (d, J = 2.4 Hz, 1H), 4.90-4.60 (m, 1H), 4.48-4.24 (m, 1H), 3.70-3.60 (m, 1H), 2.96-2.84 (m, 1H), 0.71 (d, J = 6.4 Hz, 3H).
1H NMR (400 MHz, CD3OD) δ 7.90 (s, 1H), 7.76 (d, J = 7.8 Hz, 1H), 7.65-7.51 (m, 3H), 7.44-7.39 (m, 1H), 7.38 (d, J = 3.0 Hz, 1H), 6.49 (d, J = 3.0 Hz, 1H), 4.64-4.52 (m, 1H), 4.35-4.20 (m, 1H), 4.14-4.06 (m, 1H), 3.56 (s, 1H), 2.22-2.10 (m, 1H), 2.05-2.01 (m, 1H), 1.96-1.86 (m, 1H), 1.84-1.72 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 8.30-8.12 (m, 0.5H), 7.96-7.93 (m, 1H), 7.77-7.75 (m, 0.5H), 7.58-7.45 (m, 4H), 7.17-7.09 (m, 1H), 6.25-6.20 (m, 1H), 5.53(br, 0.3H), 4.90 (br, 0.8H), 4.37 (br, 0.6H), 4.12 (br, 0.6H), 4.00 (br, 0.3H), 3.82 (br, 0.3H), 3.50 (s, 3H), 2.35 (br, 0.5H), 2.19 (br, 1H), 2.06 (br, 1H), 1.95 (br, 1.5H).
1H NMR (400 MHz, DMSO-d6) δ 8.59 (s, 1H), 7.87 (s, 1H), 7.54-7.49 (m, 5H), 7.25 (br, 2H), 6.59 (d, J = 2.8 Hz, 1H), 4.56 (br, 1H), 3.13-2.96 (m, 1H), 2.06-1.92 (m, 3H), 1.87-1.76 (m, 1H), 1.71-1.63 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 7.62-7.30 (m, 7H), 6.52 (d, J = 3.0, 1H), 5.59 (br, 0.5H), 5.02 (br, 1H), 4.63 (br, 0.5H), 4.28 (br, 0.5H), 3.90-3.84 (m, 0.5H), 3.61-3.51 (m, 2H), 2.48-1.98 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ 7.88 (br, 1H), 7.57-7.50 (m, 5H), 7.37 (br, 1H), 6.60 (d, J = 3.0, 1H), 6.26 (br, 2H), 4.49 (br, 1H), 3.62 (br, 1H), 3.25-3.24 (m, 3H), 3.17- 3.16 (m, 1H), 2.37-2.25 (m, 2H), 2.03-1.94 (m, 2H), 1.84-1.77 (m, 1H), 1.63 (br, 1H).
1H NMR (400 MHz, CD3OD) δ 8.24 (s, 1H), 7.94 (s, 1H), 7.81 (d, J = 7.4 Hz, 1H), 7.66-7.56 (m, 3H), 7.47-7.38 (m, 1H), 7.17 (br, 1H), 6.29 (d, J = 3.2 Hz, 1H), 4.98 (br, 1H), 4.68 (br, 1H), 3.84-3.81 (m, 1H), 2.79 (br, 1H), 0.64 (d, J = 6.7 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 7.82 (d, J = 8.0, 1H), 7.59-7.47 (m, 5H), 6.64 (d, J = 2.9, 1H), 5.65 (s, 2H), 4.52-4.49 (m, 1H), 3.90- 3.85 (m, 1H), 3.68-3.61 (m, 1H), 3.03-2.90 (m, 2H), 2.53-2.50 (m, 2H), 2.09-1.97 (m, 2H), 1.87-1.79 (m, 3H), 1.67-1.56 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 7.82 (d, J = 8.0, 1H), 7.59-7.47 (m, 5H), 6.58 (d, J = 3.0, 1H), 5.65 (s, 2H), 4.52-4.49 (m, 1H), 3.90- 3.85 (m, 1H), 3.02-2.90 (m, 1H), 3.04-2.90 (m, 2H), 2.54-2.50 (m, 2H), 2.09-1.99 (m, 2H), 1.87-1.79 (m, 3H), 1.64-1.60 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 7.73 (d, J = 2.7, 1H), 7.62-7.51 (m, 4H), 7.39 (br, 1H), 6.65 (d, J = 2.8, 1H), 5.82 (s, 2H), 4.69-4.66 (m, 1H), 3.94-3.83 (m, 2H), 2.71-2.45 (m, 4H), 2.07-1.70 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ 7.75-7.60 (m, 5H), 7.43 (br, 1H), 6.52 (s, 1H), 4.82 (br, 1H), 4.49 (br, 1H), 3.75-3.70 (m, 1H), 3.37-3.34 (m, 2H), 2.41-2.38 (m, 3H), 1.87 (br, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.49 (br, 0.4H), 7.83 (br, 0.6H), 7.53-7.47(m, 4H), 7.36-7.32 (m, 2H), 6.39 (d, J = 3.2, 1H), 6.20 (s, 2H), 4.49 (br, 1H), 3.62 (br, 1H), 3.23 (br, 3H), 2.30-2.22 (m, 2H), 1.97 (br, 2H), 1.77-1.76 (m, 1H), 1.61 (br, 1H).
1H NMR (400 MHz, DMSO-d6) δ 7.56-7.48 (m, 5H), 7.41 (br, 1H), 7.38-7.36 (m, 1H), 6.41 (d, J = 3.2, 1H), 6.34 (s, 2H), 5.24 (br, 0.5H), 5.10 (br, 0.5H), 4.81 (br, 1H), 4.08-4.02 (m, 2H), 2.40-2.24 (m, 4H), 2.12-1.97 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 8.29 (s, 1H), 7.99 (s, 1H), 7.71-7.54 (m, 2H), 7.40-7.25 (m, 3H), 6.54-6.40 (m, 1H), 4.95-3.88 (m, 1H), 4.48-4.41(m, 1H), 4.39-4.32 (m, 1H), 4.13-4.07 (m, 1H), 2.42-2.32 (m, 1H), 2.24-2.11 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 8.24-8.07 (m, 2H), 7.76-7.35 (m, 6H), 6.60-6.44 (m, 1H), 5.45-5.35 (br, 0.5H), 4.81-4.71 (br, 0.5H), 4.58-4.47 (br, 0.5H), 4.28-4.09 (br, 2H), 3.76-3.72 (br, 0.5H), 3.52-3.48 (br, 1H), 2.20-1.98 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.15 (s, 1H), 7.58-7.19 (m, 6H), 7.09 (s, 2H), 6.48 (d, J = 3.0, 1H), 4.71 (br, 1H), 4.21 (br, 1H), 3.52 (br, 1H), 2.75 (br, 1H), 0.61 (br, 3H).
1H NMR (400 MHz, CD3OD) δ 8.09 (s, 1H), 7.61-7.57 (m, 4H), 7.40 (br, 1H), 7.38-7.35 (m, 1H), 6.55-6.53(m, 1H), 4.82 (s, 1H), 4.38 (br, 1H), 3.64 (br, 1H), 2.68 (br, 1H), 0.71 (d, J = 6.6, 3H).
1H NMR (400 MHz, DMSO-d6) δ 8.14 (s, 1H), 7.69-7.40 (m, 5H), 7.08 (s, 2H), 6.64 (d, J = 3.0, 1H), 4.74 (br, 1H), 4.29 (br, 1H), 3.58 (br, 1H), 2.79 (br, 1H), 0.71 (br, 3H).
1H NMR (400 MHz, CD3OD) δ 8.25 (br, 1H), 7.97 (s, 1H), 7.70 (d, J = 7.6 Hz, 1H), 7.63-7.53 (m, 3H), 7.36-7.30 (m, 1H), 6.91-6.90 (m, 1H), 5.41-5.26 (m, 1H), 4.56-4.44 (m, 1H), 4.31-4.17 (m, 1H), 2.61-2.52 (m, 1H), 2.47-2.37 (m, 1H), 2.06 (s, 3H).
1H NMR (400 MHz, CDCl3) δ 7.60-7.48 (m, 1H), 7.39 (s, 1H), 7.30-7.26 (m, 1H), 7.25-7.21 (m, 1H), 6.97 (dd, J = 24.5, 8.1, 1H), 6.49 (d, J = 2.9, 1H), 5.06 (s, 2H), 4.89-4.68 (m, 1H), 4.64-4.29 (m, 1H), 3.89- 3.55 (m, 1H), 2.72-2.46 (m, 1H), 2.37 (s, 3H), 0.87 (d, J = 6.6, 3H).
1H NMR (400 MHz, CD3OD) δ 7.68-7.55 (m, 1H), 7.48 (d, J = 7.9, 1H), 7.42-7.29 (m, 2H), 7.27-7.19 (m, 1H), 6.34 (d, J = 3.2, 1H), 4.88-4.79 (m, 1H), 4.50-4.35 (m, 1H), 3.88-3.50 (m, 1H), 2.90-2.57 (m, 1H), 2.30 (s, 3H), 0.81 (dd, J = 6.8, 2.6, 3H).
1H NMR (400 MHz, CD3OD) δ 7.72-7.61 (m, 1H), 7.43-7.38 (m, 1H), 7.37-7.28 (m, 3H), 6.33 (d, J = 3.2, 1H), 4.88-4.79 (m, 1H), 4.49-4.35 (m, 1H), 3.78-3.57 (m, 1H), 2.84-2.57 (m, 1H), 2.30 (s, 3H), 0.80 (d, J = 6.8, 3H).
1H NMR (400 MHz, DMSO-d6) δ 7.68 (s, 1H), 7.61- 7.51 (m, 5H), 7.45-7.43 (m, 1H), 6.45 (d, J = 3.2, 1H), 6.25 (s, 2H), 4.49 (br, 1H), 4.36-4.31(m, 1H), 3.58 (br, 1H), 2.75-2.66 (m, 1H), 0.68 (d, J = 6.8, 3H).
1H NMR (400 MHz, CD3OD) δ 7.57-7.43 (m, 5H), 7.29 (d, J = 3.0, 1H), 7.28-7.29 (m, 1H), 6.45-6.43 (m, 1H), 4.64-4.62 (m, 1H), 4.45-4.41 (m, 1H), 3.61- 3.57 (m, 1H), 2.62-2.53 (m, 1H), 0.71 (d, J = 6.9, 3H).
4664
1H NMR (400 MHz, CD3OD) δ 7.63-7.45 (m, 5H), 7.37 (d, J = 2.4, 1H), 7.27 (d, J = 6.4, 1H), 6.46 (d, J = 3.0, 1H), 4.81 (br, 1H), 4.00 (br, 1H), 3.75 (br, 1H), 1.15 (s, 3H), 0.72 (s, 3H).
1H NMR (400 MHz, CD3OD) δ 8.33 (s, 1H), 7.61- 7.49 (m, 1H), 7.47-7.44 (m, 1H), 7.29-7.22 (m, 2H), 7.16-7.10 (m, 1H), 6.43 (d, J = 3.0, 1H), 4.84 (br, 1H), 4.31 (br, 1H), 3.30 (br, 1H), 2.50 (br, 1H), 2.17 (s, 3H), 0.69 (d, J = 6.8, 3H).
1H NMR (400 MHz, CD3OD) δ 8.15 (s, 1H), 7.83 (s, 1H), 7.78-7.73 (m, 1H), 7.39-7.33 (m, 1H), 7.30- 7.17 (m, 3H), 6.39 (d, J = 3.0, 1H), 4.89-4.87 (m, 1H), 4.65-4.57 (m, 1H), 3.78-3.74 (m, 1H), 2.78- 2.71 (m, 1H), 0.67 (d, J = 6.8, 3H).
1H NMR (400 MHz, CD3OD) δ 7.54-7.46 (m, 4H), 7.32 (d, J = 2.8, 1H), 7.33-7.26 (m, 1H), 6.46-6.44 (m, 1H), 4.42 (br, 1H), 4.17-4.12 (m, 1H), 3.42-3.38 (m, 1H), 2.58 (br, 1H), 2.12 (s, 3H), 0.63 (d, J = 6.4, 3H).
1H NMR (400 MHz, CD3OD) δ 8.34 (s, 1H), 7.58- 7.54 (m, 2H), 7.51-7.45 (m, 2H), 7.28 (d, J = 2.7 Hz, 1H), 7.24-7.21 (m, 1H), 6.45 (d, J = 3.0 Hz, 1H), 5.15 (brs, 1H), 4.25-4.19 (m, 1H), 3.69 (brs, 1H), 2.32- 2.24 (m, 1H), 2.19 (s, 3H), 2.08-1.98 (m, 1H).
4714
1H NMR (400 MHz, CD3OD) δ 8.10 (s, 1H), 7.75 (s, 1H), 7.63-7.54 (m, 3H), 7.47 (s, 1H), 7.38 (d, J = 6.4, 1H), 6.56 (dd, J = 3.0, 1.7, 1H), 5.34-4.84 (m, 1H), 4.25-3.60 (m, 2H), 1.23 (s, 3H), 0.76 (s, 3H).
1H NMR (400 MHz, CD3OD) δ 8.61-8.58 (m, 1H), 8.02-7.98 (m, 1H), 7.64 (d, J = 7.6, 1H), 7.53-7.51 (m, 1H), 7.32 (br, 1H), 6.45 (d, J = 2.8, 1H), 4.73 (br, 2H), 4.50 (br, 1H), 3.25 (br, 2H), 2.59 (br, 1H), 2.39 (br, 1H), 2.25 (br, 1H), 0.75 (br, 3H).
1H NMR (400 MHz, CD3OD) δ 7.61-7.59 (m, 2H), 7.55-7.52 (m, 2H), 7.45-7.43 (m, 1H), 7.33-7.30 (m, 1H), 6.55-6.53 (m, 1H), 5.18-5.13 (m, 1H), 4.21- 4.15 (m, 1H), 3.62-3.50 (m, 1H), 2.49 (s, 3H), 2.46- 2.39 (m, 1H), 2.05-1.95 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 7.74-7.26 (m, 7H), 6.66-6.65 (m, 1H), 6.33 (s, 2H), 6.30 (s, 1H), 4.65 (d, J = 5.2, 0.5H), 4.58 (d, J = 5.2, 0.5H), 4.22-4.16 (m, 1H), 3.06-3.00 (m, 1H), 2.74 (br, 1H), 2.35 (s, 1.5H), 2.34 (s, 1.5H), 0.58 (d, J = 6.8, 1.5H), 0.54 (d, J = 6.4, 1.5H).
1H NMR (400 MHz, CD3OD) δ 7.90 (s, 1H), 7.54- 7.50 (m, 4H), 7.33 (br, 1H), 7.30 (br, 1H), 6.46 (d, J = 3.0, 1H), 4.75 (br, 1H), 4.39 (br, 1H), 3.61 (br, 1H), 2.62 (br, 1H), 0.63 (d, J = 6.8, 3H).
1H NMR (400 MHz, CD3OD) δ 8.43 (s, 1H), 7.41-7.36 (m, 2H), 7.26-7.19 (m, 1H), 7.09- 7.06 (m, 1H), 6.54-6.52 (m, 1H), 5.25 (br, 1H), 4.35-4.28 (m, 1H), 3.78 (br, 1H), 2.43- 2.18 (m, 5H).
1H NMR (400 MHz, CD3OD) δ 8.25 (bs, 1H), 7.97 (s, 1H), 7.70 (d, J = 7.6 Hz, 1H), 7.63-7.53 (m, 3H), 7.36-7.30 (m, 1H), 6.91-6.90 (m, 1H), 5.41-5.26 (m, 1H), 4.56-4.44 (m, 1H), 4.31-4.17 (m, 1H), 2.61-2.52 (m, 1H), 2.47-2.37 (m, 1H), 2.06 (s, 3H).
4prepared from (S)-methyl 3,3-dimethylazetidine-2-carboxylate
To a mixture of 2a (740 mg, 2.28 mmol) (2a was prepared according to the procedure of Example 1 using 1-amino-3-chloro-1H-pyrrole-2-carboxamide and (S)-azetidine-2-carboxylic acid instead of 1a and (S)-1-(tert-butoxycarbonyl) pyrrolidine-2-carboxylic acid) and Cs2CO3 (1.6 g, 4.92 mmol) in DMF (7 mL) was added 2-bromo-1,1-difluoroethane (0.4 mL, 5.02 mmol). The reaction was heated to 50° C. for one hour and 120° C. for another 1.5 hours. Then the mixture was diluted with water and extracted with EtOAc three times. The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated to give the crude product which was further purified by flash column chromatography eluting with EtOAc/PE. 230 mg of 2b was obtained (yield: 26%) and 110 mg of 2a were recovered. MS (m/z): 289.0 (M-Boc+H)+.
To a mixture of 2b (230 mg, 0.59 mmol) in MeOH (2 mL) was added conc. HCl aq. (2 mL), then the reaction was stirred at room temperature for about 3 hours. After concentration, 2c was obtained as a pale yellow solid which was used in the next step without further purification. MS (m/z): 289.0 (M+H)+.
A mixture of 2c (0.59 mmol), 4-chloro-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (105 mg, 0.59 mmol) and TEA (0.41 mL, 2.95 mmol) in n-BuOH (9 mL) was heated at 130° C. for 2 hours. After concentration, the residue was washed with water and dried, then purified by preparative TLC and Compound 59 as a pale yellow solid was obtained (160 mg, yield: 63%). MS (m/z): 431.1 (M+H)+. 1H NMR (400 MHz, DMSO-d6) δ: 12.94 (s, 1H), 8.32 (m, 2H), 7.67 (s, 1H), 6.67 (s, 1H), 6.45 (t, J=55.2 Hz, 1H), 5.92-5.82 (m, 1H), 4.80-4.54 (m, 2H), 4.52-4.26 (m, 2H), 3.06-2.96 (m, 1H), 2.78-2.66 (m, 1H)
The following Compounds were prepared according to the procedure of Compound 59 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ: 8.51-8.41 (m, 1H), 8.25-8.14 (m, 2H), 7.83-7.73 (m, 1H), 7.62-7.62 (m, 1H), 7.50-7.40 (m, 1H), 7.31-7.20 (m, 1H), 6.84 (s, 1H), 6.64-6.56 (m, 1H), 5.76-5.64 (m, 1H), 5.45-5.31 (m, 1H), 5.31-5.25 (m, 1H), 4.52-4.46 (m, 1H), 4.30-4.24 (m, 1H), 2.58-2.52 (m, 1H), 2.03-1.88 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 11.67 (s, 1H), 8.67-8.57 (m, 1H), 8.52-8.40 (m, 1H), 8.25-8.11 (m, 2H), 7.83-7.71 (m, 1H), 7.64-7.54 (m, 1H), 7.44-7.32 (m, 1H), 6.66-6.54 (m, 1H), 5.74-5.62 (m, 1H), 5.43-5.33 (m, 1H), 5.10-5.00 (m, 1H), 4.56-4.46 (m, 1H), 4.34-4.26 (m, 1H), 2.74-2.62 (m, 1H), 1.99-1.87 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 12.84 (s, 1H), 8.26 (s, 1H), 8.01 (s, 1H), 7.52-7.33 (m, 5H), 7.26 (dd, J = 17.6, 10.1 Hz, 1H), 6.58-6.45 (m, 1H), 5.47 (d, J = 16.6 Hz, 1H), 5.40-5.34 (m, 1H), 5.28 (d, J = 16.6 Hz, 1H), 4.19-4.07 (m, 1H), 4.00 (q, J =7.3 Hz, 1H), 2.33-2.20 (m, 1H), 2.20-2.03 (m, 2H), 2.03-1.91 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 12.85 (s, 1H), 8.28 (s, 1H), 8.11 (s, 1H), 7.39 (d, J = 2.9 Hz, 1H), 6.48 (d, J = 2.9 Hz, 1H), 5.55 (dd, J = 7.8, 3.0 Hz, 1H), 4.18-4.02 (m, 2H), 3.58 (s, 3H), 2.44-2.36 (m, 1H), 2.33-2.11 (m, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 8.28 (s, 1H), 8.03 (s, 1H), 7.38 (d, J = 2.9 Hz, 1H), 6.48 (d, J = 2.9 Hz, 1H), 5.44 (d, J = 5.6 Hz, 1H), 4.22 (dd, J = 14.2, 8.4 Hz, 1H), 4.17-4.10 (m, 1H), 4.04 (d, J = 9.0 Hz, 1H), 3.65 (d, J = 6.9 Hz, 1H), 2.44-2.36 (m, 1H), 2.35-2.23 (m, 1H), 2.22-2.03 (m, 2H), 2.01-1.81 (m, 1H), 1.02 (d, J = 6.7 Hz, 3H), 0.94 (d, J = 6.6 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 8.51 (s, 1H), 8.26 (s, 1H), 7.98-7.82 (m, 2H), 7.53-7.48 (m, 2H), 7.36-7.24 (br, 1H), 6.55 (s, 1H), 5.72-5.30 (m, 3H), 4.15-3.95 (m, 2H), 2.28-2.07 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ: 12.85 (s, 1H), 8.78-8.22 (m, 1H), 8.48 (s, 1H), 8.28 (s, 1H), 8.07-7.85 (br, 2H), 7.57-7.35 (br, 2H), 6.54 (s, 1H), 5.60-5.15 (m, 3H), 4.18-4.00 (m, 2H), 2.28-2.08 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ: 8.16 (s, 1H), 8.03 (s, 1H), 7.53 (s, 1H), 6.58 (s, 1H), 5.39 (s, 2H), 5.22-5.12 (m, 1H), 4.15-4.05 (m, 2H), 2.42-2.32 (m, 2H), 2.19-2.09 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 8.55-8.50 (m, 1H), 8.15-7.90 (m, 2H), 7.88-7.84 (m, 1H), 7.59-7.57 (m, 1H), 7.47-7.45 (m, 1H), 7.35-7.32 (m, 1H), 6.56-6.55 (m, 1H), 5.71-5.67 (m, 1H), 5.62-5.52 (m, 1H), 4.16-4.04 (m, 2H), 2.10-1.97 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ: 8.49 (s, 1H), 7.85-7.70 (m, 2H), 7.58-7.50 (br, 1H), 7.51-7.41 (m, 1H), 7.31-7.08 (m, 3H), 6.62-6.54 (br, 1H), 5.54-5.28 (m, 3H), 3.97- 3.86 (m, 2H), 2.11-1.98 (m, 4H).
1H NMR (400 MHz, CD3OD) δ: 8.82 (d, J = 4.9 Hz, 2H), 8.06 (s, 1H), 8.01 (s, 1H), 7.44 (t, J = 4.9 Hz, 1H), 7.33 (d, J = 2.9 Hz, 1H), 6.50 (d, J = 2.9 Hz, 1H), 5.95 (d, J = 17.7 Hz, 1H), 5.77 (d, J = 17.7 Hz, 1H), 5.50 (t, J = 5.6 Hz, 1H), 4.40-4.28 (m, 1H), 4.21-4.07 (m, 1H), 2.55-2.46 (m, 1H), 2.26-2.18 (m, 3H).
1H NMR (400 MHz, CD3OD) δ: 8.77 (s, 1H), 8.76 (s, 1H), 8.20-7.77 (m, 2H), 7.39 (t, J = 4.9 Hz, 1H), 7.19 (s, 1H), 6.40 (s, 1H), 5.88-5.76 (m, 2H), 5.70-5.64 (m, 0.5H), 5.42-5.32 (m, 0.5H), 4.40-4.34 (m, 0.5H), 4.23-4.17 (m, 0.5H), 4.07-4.01 (m, 0.5H), 3.85-3.75 (m, 0.5H), 2.35-2.11 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ: 8.58 (s, 1H), 8.40-8.10 (m, 2H), 7.92 (t, J = 7.3 Hz, 1H), 7.62 (d, J = 7.9 Hz, 1H), 7.55-7.45 (m, 1H), 7.42-7.33 (m, 1H), 6.58 (s, 1H), 5.76-5.23 (m, 4H), 4.75-4.50 (br, 1H), 4.35-4.15 (br, 1H), 2.74-2.64 (m, 1H), 2.47-2.37 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 8.60-8.56 (m, 1H), 8.36 (s, 1H), 8.06 (s, 1H), 7.91 (t, J = 7.4 Hz, 1H), 7.63-7.47 (m, 2H), 7.39-7.33 (m, 1H), 6.62-6.56 (m, 1H), 5.72-5.62 (m, 2H), 5.60-5.50 (m, 1H), 5.31-5.23 (m, 1H), 4.55-4.34 (m, 2H), 2.74-2.66 (m, 1H), 2.59-2.55 (m, 1H).
1H NMR (400 MHz, CD3OD) δ: 8.54 (d, J = 4.8 Hz, 1H), 8.40-8.02 (m, 2H), 7.89 (td, J = 7.8, 1.6 Hz, 1H), 7.58 (d, J = 7.9 Hz, 1H), 7.40-7.33 (m, 1H), 7.23 (s, 1H), 6.45 (s, 1H), 5.78-5.58 (m, 3H), 4.65-4.29 (m, 2H), 2.82-2.62 (m, 2H).
1H NMR (400 MHz, CD3OD) δ: 8.51 (d, J = 4.7 Hz, 1H), 8.13 (s, 1H), 8.06 (s, 1H), 7.87 (td, J = 7.7, 1.6 Hz, 1H), 7.53 (d, J = 7.9 Hz, 1H), 7.36-7.33 (m, 2H), 6.48 (d, J = 3.0 Hz, 1H), 5.82-5.61 (m, 3H), 4.70-4.54 (m, 2H), 2.86-2.74 (m, 1H), 2.69-2.62 (m, 1H).
1H NMR (400 MHz, CD3OD) δ: 8.75 (d, J = 4.9 Hz, 2H), 8.01 (s, 1H), 7.82 (s, 1H), 7.38 (t, J = 5.0 Hz, 1H), 7.28 (d, J = 3.0 Hz, 1H), 6.46 (d, J = 3.0 Hz, 1H), 5.93-5.72 (m, 3H), 4.56-4.49 (m, 1H), 2.84-2.77 (m, 2H).
1H NMR (400 MHz, CD3OD) δ: 8.11 (s, 1H), 7.99 (s, 1H), 7.12 (d, J = 2.9 Hz, 1H), 6.36 (d, J = 2.9 Hz, 1H), 5.79-5.71 (m, 1H), 5.06-5.00 (m, 1H), 4.31-4.25 (m, 1H), 4.21-4.15 (m, 1H), 3.28-3.14 (m, 2H), 2.58-2.48 (m, 2H), 2.45-2.35 (m, 2H), 2.28-2.22 (m, 1H), 2.18-2.12 (m, 1H), 2.06-2.00 (m, 1H), 1.94-1.86 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 7.97 (s, 1H), 7.78 (s, 1H), 7.16 (d, J = 3.2 Hz, 1H), 6.37 (d, J = 2.8 Hz, 1H), 4.75-4.65 (m, 2H), 4.38-4.30 (m, 0.5H), 4.19-4.1 (m, 0.5H), 3.99-3.92 (m, 1H), 3.33-3.32 (m, 1H), 2.82-2.75 (m, 1H), 2.52-2.44 (m, 1H), 2.40 (s, 6H), 2.33-2.24 (m, 2H), 2.19-2.12 (m, 2H).
Synthesis of Compound 70 was carried out according to the procedure of Example 1 and the following Step 3-3 using 1-amino-3-chloro-1H-pyrrole-2-carboxamide as the starting material. Compound 70 was got as a pale yellow solid. MS (m/z): 472.6 (M+H)+; 1H NMR (400 MHz, CD3OD) δ: 8.29 (s, 1H), 7.99 (s, 1H), 7.80 (d, J=7.1 Hz, 1H), 7.67-7.61 (m, 1H), 7.58 (d, J=3.1 Hz, 2H), 7.41 (d, J=6.7 Hz, 1H), 7.35-7.25 (m, 1H), 5.01-4.97 (m, 1H), 4.69-4.65 (m, 1H), 4.34 (dd, J=10.7, 4.1 Hz, 1H), 4.01 (d, J=10.8 Hz, 1H), 2.38-2.28 (m, 1H), 2.20-2.11 (m, 1H).
To a solution of 3b (610 mg, 1.72 mmol) in DCM (30 mL) was added DHP (173 mg, 2 mmol) and TsOH.H2O (65 mg, 0.34 mmol). The reaction mixture was stirred at room temperature for 5 hours. The resulting mixture was concentrated and purified by column chromatography eluting with EtOAc/PE to afford Compound 3c as a pale yellow oil (730 mg, yield: 97%). MS (m/z): 438.7 (M+H)+
Compound 71 was prepared according to the procedure of Compound 70 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 8.39-8.19 (m, 3H), 7.76-7.70 (m, 1H), 7.62-7.46 (m, 5H), 6.60-6.52 (m, 1H), 4.55-4.51 (m, 1H), 4.22-4.18 (m, 1H), 4.17-4.13 (m, 1H), 3.79-3.75 (m, 1H), 2.24-2.20 (m, 1H), 2.07-1.95 (m, 1H).
Step 4-1 was carried out according to the procedure in Example 1.
Silver oxide (72 mg, 0.33 mmol) and methyl iodide (62 mg, 0.44 mmol) were added to a solution of 4a (56 mg, 0.11 mmol) in acetone (10 mL) at room temperature. The reaction mixture was stirred in the dark at 60° C. overnight. Then the reaction mixture was filtered and the filtrate was concentrated in vacuo to provide the crude 4b without further purification which is used in the next step reaction. MS (m/z): 547 (M+H)+
To a solution of 4b (60 mg, 0.11 mmol) in MeOH (2 mL) was added conc.HCl aq (2 mL). The resulting mixture was stirred at 50° C. for one hour. Then the reaction was concentrated and 7N NH3 in MeOH (5 mL) was added. After concentration in vacuo, the crude product was purified by preparative TLC eluting with MeOH/DCM to afford Compound 72 as a pale yellow solid (16 mg, yield: 31%). MS (m/z): 462.9 (M+H)+; 1H NMR (400 MHz, DMSO-d6) δ: 8.23-8.08 (m, 2H), 7.73-7.40 (m, 6H), 6.57-6.49 (m, 1H), 5.34-5.24 (m, 1H), 4.64-4.51 (m, 1H), 4.19-4.05 (m, 2H), 3.09 (s, 3H), 2.37-2.29 (m, 1H), 2.04-1.96 (m, 1H).
Compounds 263 and Compounds 265-266 were prepared according to the procedure of Compound 72 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ: 13.09-12.75 (br, 1H), 8.27-8.09 (m, 2H), 7.76-7.39 (m, 6H), 6.59-6.48 (m, 1H), 5.48-5.38 (m, 0.5H), 4.93-4.81 (br, 0.5H), 4.67-4.55 (m, 0.5H), 4.33-4.22 (m, 0.5H), 4.08-3.99 (m, 0.5H), 3.96-3.89 (br, 0.5H), 3.86-3.77 (m, 0.5H), 3.68-3.59 (m, 0.5H), 3.18 (s, 3H), 2.31-2.06 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.05 (s, 1H), 7.82 (s, 1H), 7.62-7.39 (m, 6H), 6.51 (s, 1H), 4.07-3.98 (m, 1H), 3.49-3.13 (m, 10H), 2.08 (br, 2H).
To a solution of 3b (400 mg, 1.13 mmol) in DCM (50 mL) was added DAST (726 mg, 4.52 mmol) at 0° C. The resulting mixture was stirred at 0° C. for one hour, then at room temperature for another one hour. LC-MS showed the starting material disappeared, then NaHCO3 aq. (10 mL) was added and extracted with DCM three times. The organic layers were combined, dried over Na2SO4 and concentrated to give Compound 5a which was used in the next step without further purification. MS (m/z): 257 (M-Boc+H)+
Steps 5-2 to 4 were carried out according to the procedure of Example 1. Compound 73 was got as a white solid. MS (m/z): 451.1 (M+H)+; 1H NMR (400 MHz, DMSO-d6) δ: 8.38-8.10 (m, 3H), 7.71-7.52 (m, 4H), 7.46 (s, 1H), 6.59-6.49 (m, 1H), 5.39-5.29 (m, 1H), 4.88-4.34 (m, 1H), 4.24-3.93 (m, 2H), 2.31-2.17 (m, 2H).
Compound 74 and Compounds 267-268 was prepared according to the procedure of Compound 73 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, CD3OD) δ: 8.23 (s, 1H), 7.98 (s, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.65-7.55 (m, 3H), 7.48 (d, J = 7.6 Hz, 1H), 7.30 (d, J = 3.0 Hz, 1H), 6.46 (d, J = 3.0 Hz, 1H), 5.39-5.31 (m, 1H), 5.22-5.16 (m, 1H), 4.56-4.41 (m, 2H), 2.51-2.41 (m, 1H), 2.22-2.16 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 8.32 (d, J = 5.8 Hz, 1H), 7.96-7.65 (m, 6H), 7.56-7.30 (m, 1H), 6.57 (dd, J = 5.9, 3.0 Hz, 1H), 5.50-5.21 (m, 1H), 4.91-4.82 (m, 1H), 4.15-3.72 (m, 2H), 2.97 (d, J = 2.5 Hz, 3H), 2.31-1.91 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 12.42 (br, 1H), 8.23 (s, 1H), 8.13 (s, 1H), 7.64-7.55 (m, 1H), 7.54-7.45 (m, 5H), 6.59 (d, J = 3.0 Hz, 1H), 5.24-5.02 (m, 1H), 4.74-4.63 (m, 1H), 4.19-3.97 (m, 1H), 3.92-3.83 (m, 1H), 2.51 (s, 3H), 2.44-2.21 (m, 2H).
To a solution of 6a (4.8 g, 30.0 mmol) in DMF (40 mL) was added 60% NaH (1.2 g, 30.0 mmol) at 0-5° C. and stirred at 0-5° C. for 30 minutes. Then 1-bromobutan-2-one (5.0 g, 33 mmol) was added and stirred at room temperature for 2 hours. After concentration in vacuo, the residue was used in the next step without further purification. MS (m/z): 230.1 (M+H)+
A mixture of the obtained 6b (30.0 mmol) in 7M NH3/MeOH (80 mL) was stirred in a sealed tube at 130° C. for 16 hours. After concentration, the residue was purified by flash column chromatography eluting with MeOH/H2O to afford 6c as a white solid (2.67 g, yield: 45%). MS (m/z): 197.1 (M+H)+
A mixture of 6c (1.97 g, 10.0 mmol), 3-fluorophenylboronic acid (2.80 g, 20.0 mmol), 4AMS (24 g), Cu(OAc)2, (3.63 g, 20.0 mmol) and pyridine (3.96 g, 50.0 mmol) in dry DCM (80 mL) was stirred under dry air at room temperature for 16 hours. The mixture was filtered through celite and washed with MeOH/DCM. The filtrate was concentrated and purified by flash column chromatography eluting with MeOH/DCM to afford 6d as a yellow solid (1.53 g, yield: 53%). MS (m/z): 291.0 (M+H)+
To a solution of 6d (1.53 g, 5.26 mmol) in dioxane (25 mL) was added SeO2 (584 mg, 5.26 mmol) and stirred under reflux for one hour. After concentration, the residue was purified by flash column chromatography eluting with EtOAc/PE to afford 6e as a yellow solid (1.60 g, yield: 99%). MS (m/z): 307.0 (M+H)+
To a solution of 6e (1.60 g, 5.2 mmol) in THF (30 mL) was added DPPA (2.86 g, 10.4 mmol) and DBU (1.58 g, 10.4 mmol), then the mixture was stirred at 50-60° C. overnight. After concentration, the residue was purified by flash column chromatography eluting with EtOAc/PE to afford 6f as a yellow oil (680 mg, yield: 39%). MS (m/z): 332.0 (M+H)+
To a mixture of 6f (680 mg, 2.05 mmol) in THF (20 mL) was added PPh3 (1.08 g, 4.10 mmol) and the reaction was stirred at room temperature for 10 minutes. Then conc. NH3.H2O aq. (5 mL) was added and the reaction was stirred at 50-60° C. for another 4 hours. The reaction mixture was concentrated in vacuo and the residue was purified by flash column chromatography eluting with MeOH/H2O to afford 6g as a white solid (320 mg, yield: 51%). MS (m/z): 306.1 (M+H)+
A mixture of 6g (61 mg, 0.20 mmol), 6-chloro-9H-purine (37 mg, 0.24 mmol) and TEA (40 mg, 0.40 mmol) in n-BuOH (1 mL) was stirred under nitrogen at reflux for 16 hours. The reaction mixture was concentrated in vacuo, and the residue was purified by flash column chromatography eluting with MeOH/H2O to afford Compound 75 as a yellow solid (44.4 mg, yield: 50%). MS (m/z): 424.1 (M+H)+. 1H NMR (400 MHz, DMSO-d6) δ: 8.03-7.94 (m, 2H), 7.79 (s, 1H), 7.47 (s, 2H), 7.35-7.12 (m, 3H), 7.00 (s, 2H), 6.60 (s, 1H), 4.81 (m, 1H), 1.35 (br, 3H).
The following Compounds were prepared according to the procedure of Compound 75 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ: 8.09 (d, J = 4.4 Hz, 1H), 8.09-7.99 (m, 1H), 7.64 (d, J = 9.5 Hz, 1H), 7.50 (dd, J = 6.4, 2.9 Hz, 1H), 7.42-7.36 (m, 1H), 7.23-7.13 (m, 1H), 7.09-6.93 (m, 2H), 6.67-6.61 (m, 1H), 6.57-6.47 (m, 1H), 4.95-4.85 (m, 1H), 1.40 (d, J = 6.0 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 7.74 (d, J = 19.3 Hz, 1H), 7.57 (d, J = 2.7 Hz, 1H), 7.53 (d, J = 11.0 Hz, 1H), 7.45-7.35 (m, 2H), 7.29-7.03 (m, 5H), 6.67-6.66 (m, 1H), 4.87-4.79 (m, 1H), 1.32 (d, J = 6.6 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 7.91 (s, 1H), 7.67 (s, 2H), 7.54-7.46 (m, 1H), 7.44 (d, J = 2.8 Hz, 1H), 7.40 (s, 1H), 7.30 (td, J = 8.7, 2.9 Hz, 1H), 7.15 (td, J = 8.7, 2.9 Hz, 1H), 6.93 (d, J = 6.8 Hz, 1H), 6.58 (d, J = 2.8 Hz, 1H), 4.90-4.78 (m, 1H), 1.28 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 7.78 (s, 1H), 7.56 (d, J = 2.9 Hz, 1H), 7.51 (s, 1H), 7.47 (d, J = 7.3 Hz, 1H), 7.41-7.39 (m, 1H), 7.26-7.19 (m, 4H), 7.05 (td, J = 8.7, 3.0 Hz, 1H), 6.66 (d, J = 2.8 Hz, 1H), 4.82-4.75 (m, 1H), 1.31 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, CD3OD) δ: 8.03 (s, 2H), 7.57 (s, 1H), 7.38 (d, J = 2.9 Hz, 1H), 7.07 (d, J = 8.8 Hz, 1H), 6.84 (d, J = 9.3 Hz, 1H), 6.72 (t, J = 8.8 Hz, 1H), 6.59 (d, J = 2.8 Hz, 1H), 5.25-5.13 (m, 1H), 1.54 (d, J = 6.7 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 7.74 (s, 1H), 7.55 (d, J = 2.7 Hz, 1H), 7.53 (s, 1H), 7.39 (d, J = 7.6 Hz, 1H), 7.26 (d, J = 9.2 Hz, 1H), 7.14 (s, 2H), 7.10 (d, J = 9.5 Hz, 1H), 6.91 (d, J = 9.4 Hz, 1H), 6.65 (d, J = 2.7 Hz, 1H),4.89 (m, 1H), 1.32 (d, J = 6.5 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 8.07 (s, 1H), 8.05 (s, 1H), 7.68 (s, 1H), 7.47 (dd, J = 2.5, 1.5 Hz, 1H), 7.42-7.38 (m, 1H), 7.36-7.34 (m, 2H), 7.19 (t, J = 7.4 Hz, 1H), 7.08 (t, J = 8.1 Hz, 1H), 6.89 (d, J = 4.0 Hz, 1H), 6.56 (dd, J = 3.9, 2.6 Hz, 1H), 6.41 (d, J = 7.1 Hz, 1H), 4.85-4.79 (m, 1H), 1.39 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 12.90 (s, 1H), 8.08-7.93 (m, 3H), 7.50-7.47 (m, 2H), 7.41-7.34 (m, 3H), 7.23 (s, 1H), 7.10 (s, 1H), 6.87 (s, 1H), 6.54 (s, 1H), 4.85-4.75 (m, 1H), 1.34 (d, J = 6.2 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 7.95-7.89 (m, 1H), 7.42-7.17 (m, 4H), 7.09-6.99 (m, 2H), 6.67-6.46 (m, 4H), 5.03-4.93 (m, 1H), 1.33-1.30 (m, 3H).
1H NMR (400 MHz, CD3OD) δ 9.16 (d, J = 7.6 Hz, 1H), 8.02 (s, 1H), 7.85 (s, 1H), 7.44 (s, 1H), 7.37-7.33 (m, 1H), 7.28 (d, J = 2.8 Hz, 1H), 7.26-7.24 (m, 1H), 7.15-7.10 (m, 2H), 6.28-6.87 (m, 1H), 6.49 (d, J = 2.9 Hz, 1H), 2.44 (s, 3H), 1.39 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, CD3OD) δ 8.47 (s, 1H), 7.50-7.47 (m, 2H), 7.36 (d, J = 2.8 Hz, 1H), 7.33-7.15 (m, 4H), 6.58 (d, J = 2.8 Hz, 1H), 5.00-4.95 (m, 1H), 2.42 (s, 3H), 1.38 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, CD3OD) δ 7.70 (s, 1H), 7.49-7.54 (m, 1H), 7.44 (s, 1H), 7.35-7.31 (m, 3H), 7.28-7.24 (m, 1H), 7.20-7.18 (m, 1H), 6.58 (dd, J = 2.8, 0.6, 1H), 4.95 (q, J = 8.0 Hz, 1H), 2.56 (s., 3H), 1.40 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, CD3OD) δ 9.22 (d, J = 7.2 Hz, 0H), 7.50-7.45 (m, 1H), 7.42 (d, J = 0.8 Hz, 1H), 7.35 (d, J = 3.2. Hz 1H), 7.33-7.28 (m, 3H), 7.23-7.20 (m, 1H), 6.57 (d, J = 2.8 Hz, 1H), 4.91-4.78 (m, 1H), 3.51-3.41 (m, 2H), 2.64-2.45 (m, 2H), 1.37 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 12.44 (br, 1H), 9.02-8.98 (m, 1H), 8.26 (s, 0.5H), 8.25 (s, 0.5H), 8.01 (s, 0.5H), 7.96 (s, 0.5H), 7.59 (s, 0.5H), 7.55 (s, 0.5H), 7.53-7.50 (m, 1H), 7.46-7.37 (m, 1H), 7.24 (d, J = 8.0 Hz, 0.5H), 7.09-7.00 (m, 2H), 6.94 (d, J = 9.7 Hz, 0.5H), 6.65-6.60 (m, 1H), 4.79-4.74 (m, 1H), 1.40-1.37 (m, 3H).
1H NMR (400 MHz, DMSO-d6) δ 8.09-7.97 (m, 2H), 7.63 (d, J = 10.0 Hz, 1H), 7.50 (br, 1H), 7.46-7.35 (m, 1H), 7.24-7.02 (m, 3H), 6.62 (br, 1H), 6.37-6.31 (m, 1H), 4.87 (br, 1H), 1.39 (d, J = 6.1 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 7.68 (br, 1H), 7.58-7.36 (m, 5H), 7.32 (d, J = 2.6, 1H), 7.02 (s, 1H), 6.55 (d, J = 2.8, 1H), 6.32 (s, 2H), 4.92 (t, J = 7.5, 1H), 3.88 (br, 1H), 3.34 (br, 2H), 2.95 (br, IH), 2.44-2.37 (m, 1H), 2.28-2.23 (m, 1H), 1.97-1.45 (m, 4H).
To a solution of 6a (5.85 g, 36.7 mmol) in DMF (70 mL) was added 60% NaH (1.61 g, 40.3 mmol) at 0-5° C. and stirred at 0-5° C. for 30 minutes. Then a solution of 1-bromopropan-2-one (7.54 g, 55 mmol) in DMF (10 mL) was added dropwise at 0-5° C., and the reaction was stirred at room temperature for 30 minutes. After concentration in vacuo, the residue 7b was used in the next step without further purification.
A mixture of obtained 7b (36.7 mmol) in 7M NH3 in MeOH (80 mL) was stirred in a sealed tube at 130° C. for 16 hours. After concentration in vacuo, the residue was purified by flash column chromatography eluting with MeOH/DCM to afford 7c as a yellow solid (3.59 g, yield: 54%). MS (m/z): 183.1 (M+H)+
A mixture of 7c (910 mg, 5.0 mmol), 3-fluorophenylboronic acid (1.40 g, 10.0 mmol), 4AMS (25g), Cu(OAc)2, (1.82 g, 10.0 mmol) and pyridine (1.98 g, 25.0 mmol) in dry DCM (80 mL) was stirred under dry air at room temperature for 16 hours. The mixture was filtered through celite and washed with MeOH/DCM. The filtrate was concentrated and the residue was purified by flash column chromatography eluting with MeOH/H2O to afford 7d as a yellow solid (1.38 g, yield: 83%). MS (m/z): 277.1 (M+H)+
To a solution of 7d (1.38 g, 5.0 mmol) in dioxane (30 mL) was added SeO2 (1.11 g, 10 mmol) and the reaction was stirred at reflux for 2 hours. The mixture was diluted with EtOAc, and filtered through celite. The filtrate was collected, concentrated and purified by flash column chromatography eluting with EtOAc/PE to afford 7e as a yellow solid (1.45 g, yield: 100%). MS (m/z): 291.0 (M+H)+
To a solution of 7e (1.01 g, 3.5 mmol) in dry THF (50 mL) was added 3M EtMgBr in THF (7 mL, 21 mmol) at 0-5° C. and the reaction was stirred at room temperature for 30 minutes. The mixture was poured into sat. NH4Cl aq, and extracted with EtOAc. The organic layer was collected, concentrated and purified by flash column chromatography eluting with EtOAc/PE to afford 7f as a yellow solid (1.06 g, yield: 94%). MS (m/z): 321.0 (M+H)+
To a solution of 7f (1.06 g, 3.3 mmol) in THF (50 mL) was added DPPA (1.82 g, 6.6 mmol) and DBU (1.0 g, 6.6 mmol), then the reaction was stirred at 50-60° C. overnight. After concentration in vacuo, the residue was purified by flash column chromatography eluting with EtOAc/PE to afford 7g as a yellow oil (853 mg, yield: 75%). MS (m/z): 346.1 (M+H)+
To a mixture of 7g (853 mg, 2.46 mmol) in THF (10 mL) was added PPh3 (1.293 g, 4.92 mmol) and conc. NH3.H2O aq. (4.2 mL), then the reaction was stirred at 50-60° C. for 16 hours. After concentration in vacuo, the residue was purified by flash column chromatography eluting with MeOH/H2O to afford 7h as a yellow solid (600 mg, yield: 76%). MS (m/z): 320.1 (M+H)+
A mixture of 7h (143 mg, 0.45 mmol), 6-chloro-9H-purine (77 mg, 0.50 mmol) and TEA (136 mg, 1.35 mmol) in n-BuOH (2 mL) was stirred under nitrogen at reflux for 16 hours. The reaction mixture was concentrated in vacuo. The residue was purified by flash column chromatography eluting with MeOH/H2O and further purified by preparative TLC eluting with MeOH/DCM to afford Compound 85 as a yellow solid (16.1 mg, yield: 8.2%). MS (m/z): 438.1 (M+H)+. 1H NMR (400 MHz, DMSO-d6) δ: 8.00-7.97 (m, 2H), 7.41-7.40 (m, 2H), 7.25-7.23 (m, 1H), 7.13-7.07 (m, 2H), 7.03-6.94 (m, 2H), 6.48-6.47 (m, 1H), 1.93-1.84 (m, 1H), 1.75-1.68 (m, 1H), 0.85-0.82 (m, 3H).
The following Compounds were prepared according to the procedure of Compound 85 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ: 7.79-7.73 (m, 1H), 7.53-7.52 (m, 1H), 7.48-7.44 (m, 2H), 7.36-7.32 (m, 2H), 7.20-7.15 (m, 3H), 7.12-7.11 (m, 1H), 6.64-6.62 (m, 1H), 4.60-4.52 (m, 1H), 1.76-1.70 (m, 2H), 0.75-0.70 (m, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 8.05-8.03 (m, 2H), 7.83 (s, 1H), 7.48-7.10 (m, 8H), 6.60 (s, 1H), 4.82-4.72 (m, 1H), 1.33 (d, J = 5.9 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 8.05 (s, 2H), 7.62 (s, 1H), 7.48 (d, J = 2.8 Hz, 1H), 7.43-7.37 (m, 3H), 7.22 (t, J = 7.3 Hz, 1H), 7.10 (t, J = 7.3 Hz, 1H), 6.61 (d, J = 2.8 Hz, 1H), 6.37 (d, J = 6.8 Hz, 1H), 4.78-4.75 (m, 1H), 1.37 (d, 7 = 6.6 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 7.72 (s, 1H), 7.52 (d, J = 2.7 Hz, 1H), 7.47-7.34 (m, 3H), 7.32-7.28 (m, 1H), 7.26-7.22 (m, 3H), 7.17-7.07 (s, 2H), 6.63 (d, J = 2.7 Hz, 1H), 4.77-4.69 (m, 1H), 1.27 (d, J = 6.8 Hz, 3H).
A mixture of ethyl 2-nitroacetate (26.6 g, 200 mmol) and triethoxymethane (44.5 g, 300 mmol) in acetic anhydride (51.5 g, 500 mmol) was stirred at 100° C. for 16 hours. After concentration, the residue was further distilled under reduced pressure to afford 8a as a yellow oil (30.3 g, yield: 82%). MS (m/z): 190 (M+H)+.
To a solution of methyl 3-methyl-1H-pyrrole-2-carboxylate (13.33 g, 96 mmol) in THF (160 mL) was added 60% NaH (5.76 g, 192 mmol) at 0-5° C. under nitrogen. The mixture was stirred at 0-5° C. for half an hour. Then 8a (27.27 g, 144 mmol) was added and the reaction was stirred at room temperature for one hour. Then the mixture was diluted with EtOAc and brine. The organic layer was collected, concentrated and purified by flash column chromatography eluting with EtOAc/PE to afford 8b as a yellow oil (24.6 g, purity: 60%).
To a solution of 8b (21.3 g, 65 mmol) in MeOH (400 mL) was added CoCl2.6H2O (30.9 g, 130 mmol) followed by NaBH4 (12.3 g, 32.4 mmol) in small portions. H2 was evolved and the reaction was stirred at room temperature for one hour. 10% HCl aq. was added to dissolve the black precipitate and MeOH was removed by evaporation. Concentrated NH3.H2O aq. was added and the mixture was extracted with EtOAc. The organic layer was dried and concentrated in vacuo to afford an orange oil which was purified by flash column chromatography eluting with EtOAc/PE to give 8c as a yellow oil (9.56 g). MS (m/z): 299 (M+H)+.
A solution of the obtained 8c (9.56 g) in toluene (180 mL) was heated at reflux under nitrogen for 40 hours. The mixture was concentrated and the residue was purified by flash column chromatography eluting with EtOAc/PE to give 8d as a brown oil (1.85 g, yield: 10%). MS (m/z): 267 (M+H)+.
To a solution of 8d (1.85 g, 6.9 mmol) in dry THF (40 mL) cooled in an ice-bath was added 60% NaH (210 mg, 7.0 mmol) and stirred at 0-5° C. for 30 minutes. MeOH was added and followed by water. The mixture was extracted with EtOAc three times. The organic layers were combined and concentrated, the residue was purified by flash column chromatography eluting with PE/EA to give 8e as a white solid (1.60 g, yield: 100%). MS (m/z): 221 (M+H)+.
To a solution of 8e (110 mg, 0.50 mmol) in THF (5 mL) was added 1M BH3/THF (5 mL, 5 mmol) at 0-5° C. and stirred at room temperature for one hour. Water was added to quench the reaction. The mixture was diluted with EtOAc and brine. The organic layer was collected and concentrated. The residue as a white solid (65 mg, yield: 74%) was used in the next step without further purification. MS (m/z): 179 (M+H)+.
To a solution of 8f (1.78 g, 10 mmol) in dry THF (60 mL) was added 60% NaH (600 mg, 20 mmol) and the reaction was stirred at room temperature for 20 minutes. Then to the mixture was added tert-butylchlorodimethylsilane (3 g, 20 mmol) and the mixture was stirred at room temperature for another 40 minutes. The reaction was quenched by MeOH, and diluted with EtOAc and brine. The organic layer was collected, concentrated and purified by flash column chromatography eluting with EtOAc/PA to give 8g as a white solid (1.12 g, yield: 38%). MS (m/z): 293 (M+H)+.
A mixture of 8g (1.03 g, 3.52 mmol), phenylboronic acid (860 mg, 7.04 mmol), diacetoxycopper (1.28 g, 7.04 mmol), pyridine (1.39 g, 17.61 mmol) and 4AMS (15 g) in DCM (60 mL) was stirred at room temperature under dry air for 16 hours. Then the reaction mixture was diluted with DCM and MeOH and filtered through celite. The filtrate was collected, concentrated and purified by flash column chromatography eluting with MeOH/H2O to give 8h as a white solid (950 mg, yield: 73%). MS (m/z): 369 (M+H)+.
To a solution of 8h (950 mg, 2.58 mmol) in THF (10 mL) was added TBAF.3H2O (814 mg, 2.58 mmol) and stirred at room temperature for 15 minutes. The mixture was diluted with EtOAc and washed with brine. The organic layer was collected, dried and concentrated to give 8i as a yellow oil (585 mg, yield: 89%). MS (m/z): 255 (M+H)+.
To a solution of 8i (585 mg, 2.30 mmol) in DCM (30 mL) was added MnO2 (3.0 g, 34.4 mmol) and the reaction was stirred at room temperature overnight. The mixture was filtered through celite. The filtrate was concentrated and purified by flash column chromatography eluting with EtOAc/PE to give 8j as a white solid (366 mg, yield: 63%). MS (m/z): 252.7 (M+H)+.
To a solution of 8j (366 mg, 1.45 mmol) in THF (30 mL) was added 2M CH3MgI in Et2O (1.45 mL, 2.9 mmol) at −78° C. and stirred for 30 minutes. The mixture was quenched by adding 10 mL of saturated NH4Cl aq. and extracted with EtOAc. The organic layer was collected and concentrated to afford 8k as a yellow solid (349 mg, yield: 89.7%), which was used in the next step without further purification. MS (m/z): 269 (M+H)+.
To a solution of 8k (349 mg, 1.3 mmol) in THF (20 mL) was added DPPA (716 mg, 2.6 mmol) at 0-5° C., followed by DBU (396 mg, 2.6 mmol) at 0-5° C. The mixture was stirred at room temperature under nitrogen for 16 hours. The mixture was concentrated and purified by flash column chromatography eluting with EtOAc/PE to give 81 as a white solid (160 mg, yield: 42%). MS (m/z): 294 (M+H)+.
To a solution of 8l (160 mg, 0.54 mmol) in THF (5 mL) was added triphenylphosphine (286 mg, 1.09 mmol) and conc. NH3.H2O aq. (1 mL), then the reaction was stirred at 50° C. for 2 hours. The mixture was concentrated and purified by flash column chromatography eluting with MeOH/water to give 8m as a yellow solid (120 mg, yield: 82.6%). MS (m/z): 268 (M+H)+.
A mixture of 8m (40 mg, 0.15 mmol), 4-amino-6-chloropyrimidine-5-carbonitrile (28 mg, 0.18 mmol) and triethylamine (30 mg, 0.3 mmol) in n-BuOH (1 mL) was reacted under N2 at reflux for 16 hours. The precipitate was collected by filtration, washed with cold n-BuOH and dried to afford Compound 90 as a white solid (38.2 mg, yield: 55%). MS (m/z): 386 (M+H)+. 1H NMR (400 MHz, DMSO-d6) δ: 7.72 (s, 1H), 7.43 (d, J=7.2 Hz, 1H), 7.41-7.31 (m, 3H), 7.29-7.19 (m, 4H), 7.10 (s, 2H), 6.37 (s, 1H), 4.77-4.69 (m, 1H), 2.38 (s, 3H), 1.26 (d, J=6.7 Hz, 3H).
The following Compounds 91 and 92 were prepared according to the procedure of Compound 90 using the corresponding reagents under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, CDCl3) δ: 8.23 (s, 1H), 7.60 (s, 1H), 7.48-7.38 (m, 2H), 7.36-7.30 (m, 1H), 7.27-7.21 (m, 2H), 7.17-7.11 (m, 1H), 7.05 (s, 1H), 6.99 (d, J = 2.5 Hz, 1H), 6.37 (d, J = 2.5 Hz, 1H), 5.47 (d, J = 7.0 Hz, 1H), 5.17-5.07 (m, 1H), 2.54 (s, 3H), 1.47 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ: 8.06-8.02 (m, 2H), 7.81 (s, 1H), 7.36-7.08 (m, 8H), 6.34 (s, 1H), 4.78 (s, 1H), 2.37 (s, 3H), 1.31 (d, J = 6.7 Hz, 3H).
To a solution of 9a (900 mg, 4.4 mmol) in anhydrous DMF (30 mL) was added 60% NaH (246 mg, 6.2 mmol.) at 0° C. The resulting mixture was stirred at 0° C. for 30 min, then 1-bromobutan-2-one (3.3g, 22 mmol.) was added and the reaction was stirred at room temperature overnight. Then the solvent was removed in vacuo and the residue was dissolved in 7M NH3 in MeOH (50 mL). The resulting mixture was stirred at 130° C. in a sealed tube for 24 hours. The reaction was cooled to room temperature and the solvent was removed in vacuo. The obtained residue was purified by flash column chromatography eluting with EtOAc/PE to give compound 9b as a yellow solid (700 mg, yield: 66%). MS (m/z): 241 (M+H)+
A mixture of 9b (700 mg, 2.92 mmol), phenylboronic acid (711 mg, 5.84 mmol), 4AMS (3 g), Cu(OAc)2 (1.06 g, 5.84 mmol) and Pyridine (1.15 g, 14.6 mmol) in dry DCM (30 mL) was stirred overnight at room temperature under dry air. The mixture was filtered through celite and the filtrate was concentrated and purified by flash column chromatography eluting with MeOH/water to afford 9c as a yellow solid (520 mg, yield: 56%). MS (m/z): 317 (M+H)+
To a mixture of 9c (500 mg, 1.58 mmol) in 1,4-dioxane (30 mL) and water (3 mL) was added 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (362 mg, 1.74 mmol), Pd(PPh3)4 (91 mg, 0.079 mmol) and K2CO3 (545 mg, 3.95 mmol). The resulting mixture was heated at reflux under N2 for 1.5 hours. Then the solvent was removed in vacuo and water was added. The mixture was extracted with DCM three times. The organic layers were combined and concentrated to give the crude product which was purified by flash column chromatography eluting with EtOAc/PE to give 9d as a yellow solid (300 mg, yield: 60%). (m/z): 319 (M+H)+
Steps 9-4 to 7 were carried out according to the procedure of Example 6 using 9d instead of 6d. Compound 93 was obtained as a white solid. MS (m/z): 451.9 (M+H)+; 1H NMR (400 MHz, CD3OD) δ: 8.18 (s, 1H), 8.04 (s, 1H), 7.99 (s, 1H), 7.90 (s, 1H), 7.51 (s, 1H), 7.47-7.39 (m, 1H), 7.36 (d, J=2.2 Hz, 1H), 7.35-7.31 (m, 1H), 7.27-7.21 (m, 1H), 7.20-7.16 (m, 1H), 6.97-6.87 (m, 1H), 6.85-6.79 (m, 1H), 5.07-4.97 (m, 1H), 3.82 (s, 3H), 1.50 (d, J=6.8 Hz, 3H).
Step 10-1 was carried out according to the procedure of Example 1 using 4-chloro-7H-pyrrolo[2,3-d]pyrimidine-5-carboxylic acid instead of 4-chloro-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile.
10a (123 mg, 0.28 mmol) was dissolved in DMF (10 mL) and to the solution was added HATU (117 mg, 0.31 mmol) and NH4Cl (300 mg, 5.6 mmol). The resulting mixture was stirred at room temperature overnight. The reaction was quenched by water and extracted with DCM three times. The organic layers were combined and concentrated to give the crude product which was purified by preparative TLC eluting with DCM/MeOH to give compound 94 as a white solid (49 mg, yield: 40%). MS (m/z): 440.7 (M+H)+; 1H NMR (400 MHz, DMSO-d6) δ: 12.08 (s, 1H), 8.22 (s, 1H), 7.90-7.70 (m, 2H), 7.65-7.43 (m, 6H), 7.28 (s, 1H), 6.90 (s, 1H), 6.50 (s, 1H), 4.69-4.57 (m, 1H), 4.09-3.99 (m, 1H), 3.90-3.80 (m, 1H), 2.19-2.05 (m, 2H), 1.98-1.88 (m, 1H), 1.81-1.71 (m, 1H).
The following Compounds were prepared according to the procedure of Compound 94 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ: 12.01 (s, 1H), 8.22 (s, 1H), 7.76 (d, J = 7.3 Hz, 2H), 7.67-7.48 (m, 5H), 7.45 (d, J = 2.3 Hz, 1H), 7.26 (s, 1H), 6.60-6.59 (m, 1H), 4.65-4.55 (m, 1H), 4.14-3.97 (m, 1H), 3.90-3.80 (m, 1H), 2.24-2.06 (m, 2H), 2.01-1.85 (m, 1H), 1.85-1.71 (m, 1H).
1H NMR (400 MHz, CD3OD) δ: 8.17 (s, 1H), 7.78-7.72 (m, 1H), 7.69-7.49 (m, 3H), 7.42 (d, J = 5.3 Hz, 2H), 7.28 (d, J = 2.7 Hz, 1H), 6.45 (d, J = 2.8 Hz, 1H), 4.74-4.68 (m, 1H), 4.01-3.91 (m, 1H), 3.83-3.70 (m, 1H), 2.90 (s, 3H), 2.19-1.96 (m, 3H), 1.82-1.72 (m, 1H).
1H NMR (400 MHz, CD3OD) δ: 8.18 (s, 1H), 7.81-7.76 (m, 1H), 7.65-7.60 (m, 1H), 7.60-7.52 (m, 2H), 7.42 (dt, J = 4.3, 1.9 Hz, 1H), 7.26 (s, 1H), 7.22 (d, J = 3.0 Hz, 1H), 6.44 (d, J = 3.0 Hz, 1H), 4.81-4.77 (m, 1H), 3.80-3.70 (m, 2H), 3.09 (s, 6H), 2.23-2.15 (m, 1H), 2.11-2.01 (m, 2H), 1.84-1.74 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 8.10-7.80 (m, 3H), 7.79-7.38 (m, 6H), 7.08 (s, 1H), 6.68 (d, J = 3.1 Hz, 1H), 5.50-5.30 (m, 1H), 4.25-3.98 (br, 2H), 2.19-1.99 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.77 (d, J = 7.9 Hz, 1H), 7.63-7.53 (m, 5H), 7.41 (s, 1H), 7.33 (s, 1H), 6.61 (d, J = 3.0 Hz, 1H), 4.62-4.54 (m, 1H), 3.81-3.62 (m, 10H), 2.15-2.11 (m, 2H), 1.97-1.89 (m, 1H), 1.84-1.76 (m, 1H).
Step 11-1 was carried out according to the procedure of Example 1 using 4-chloro-5-iodo-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine instead of 4-chloro-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile.
To a solution of 11a (70 mg, 0.11 mmol) in DMF/EtOH/H2O (4 mL/1 mL/1 mL) were added 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (51 mg, 0.33 mmol), Pd(OAc)2 (1.2 mg, 0.006 mmol), PPh3 (2.8 mg, 0.011 mmol) and Na2CO3 (70 mg, 0.66 mmol). Under N2, the reaction mixture was heated at 100° C. overnight. Then the solvent was removed in reduced pressure and the residue was purified by flash column chromatography eluting with MeOH/water to give 11b as a yellow solid (20 mg, yield: 33%).
11b (20 mg, 0.036 mmol) was dissolved in TFA (3 mL) cooled in the ice bath. The resulting mixture was stirred at room temperature for 2 hours. Then the solvent was removed in vacuo. The residue was dissolved in MeOH (1 mL) and 7N NH3 in MeOH (1 mL) was added. The mixture was stirred at room temperature for 2 hours. The solvent was removed in vacuo and the residue was purified by flash column chromatography eluting with MeOH/water to give compound 98 as a white solid (7 mg, yield: 46%). MS (m/z): 423.7 (M+H)+; 1H NMR (400 MHz, CDCl3) δ: 7.82-7.76 (m, 1H), 7.60-7.52 (m, 3H), 7.28 (s, 1H), 7.26-7.20 (m, 2H), 7.08-7.02 (m, 2H), 6.95-6.88 (m, 1H), 6.51-6.40 (m, 1H), 5.53-5.43 (m, 1H), 5.22-5.12 (m, 1H), 4.99-4.93 (m, 1H), 4.05-3.94 (m, 1H), 3.81-3.71 (m, 1H), 2.31-2.21 (m, 1H), 2.12-1.95 (m, 2H), 1.91-1.82 (m, 1H).
The following Compounds were prepared according to the procedure of Example 98 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 12.16 (s, 1H), 8.73 (s, 2H), 8.25 (s, 1H), 7.67-7.41 (m, 7H), 6.71-6.61 (br, 1H), 5.08-4.98 (m, 1H), 3.95 (s, 3H), 3.30-3.25 (m, 1H), 3.10-3.00 (m, 1H), 2.44-2.36 (m, 1H), 1.75-1.67 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 11.84 (s, 1H), 8.18 (s, 1H), 8.05-8.01 (m, 1H), 7.66-7.62 (m, 1H), 7.60-7.56 (m, 1H), 7.54-7.46 (m, 4H), 7.43-7.39 (m, 1H), 7.19-7.15 (m, 1H), 6.67-6.63 (m, 1H), 6.52-6.46 (m, 1H), 5.89 (s, 2H), 5.00-4.92 (m, 1H), 3.29-3.25 (m, 1H), 3.18-3.10 (m, 1H), 2.39-2.23 (m, 1H), 1.76-1.66 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 12.01 (s, 1H), 8.37-8.17 (m, 2H), 7.86-7.78 (m, 1H), 7.69-7.65 (m, 1H), 7.63-7.45 (m, 4H), 7.43-7.39 (m, 1H), 7.33-7.29 (m, 1H), 6.89-6.85 (m, 1H), 6.67-6.63 (m, 1H), 5.02-4.94 (m, 1H), 3.88 (s, 3H), 3.28-3.24 (m, 1H), 3.07-2.98 (m, 1H), 2.40-2.29 (m, 1H), 1.74-1.64 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 12.54 (s, 1H), 9.27 (s, 2H), 8.45-8.18 (m, 1H), 7.95-7.85 (m, 1H), 7.73-7.18 (m, 6H), 6.80-6.72 (m, 1H), 5.15-4.96 (m, 1H), 3.20-3.14 (m, 2H), 2.42-2.24 (m, 1H), 1.72-1.62 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 12.04 (s, 1H), 8.37 (s, 2H), 8.22 (s, 1H), 7.68-7.64 (m, 1H), 7.62-7.58 (m, 1H), 7.58-7.48 (m, 3H), 7.46-7.42 (m, 1H), 7.35-7.31 (m, 1H), 6.76-6.59 (m, 3H), 5.06-4.98 (m, 1H), 3.24-3.14 (m, 2H), 2.44-2.38 (m, 1H), 1.78-1.68 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 11.84 (s, 1H), 8.17 (s, 1H), 7.75-7.71 (m, 1H), 7.63-7.53 (m, 5H), 7.49-7.45 (m, 1H), 6.68-6.63 (m, 1H), 5.33-5.23 (m, 1H), 5.00-4.94 (m, 1H), 4.65-4.55 (m, 1H), 4.33-4.27 (m, 2H), 4.15-4.07 (m, 1H), 2.66-2.59 (m, 1H), 2.11-2.03 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 8.14 (s, 1H), 7.70 (d, J = 3.0 Hz, 1H), 7.67-7.49 (m, 5H), 7.47-7.40 (m, 1H), 6.62 (d, J = 2.9 Hz, 1H), 4.98-4.91 (m, 1H), 4.55-4.45 (m, 1H), 4.28 (s, 2H), 4.12-4.05 (m, 1H), 3.26 (s, 3H), 2.62-2.56 (m, 1H), 2.07-2.00 (m, 1H).
To a mixture of Compound 55 (84 mg, 0.173 mmol), Pd(PPh3)2Cl2 (8 mg, 0.0116 mmol) and CuI (2.2 mg, 0.0116 mmol) in DMF (4 mL) was added Et3N (0.36 mL, 2.6 mmol) and ethynyltrimethylsilane (44 mg, 0.448 mmol). The reaction was heated under N2 at 90° C. for 4 hours, then the mixture was cooled to room temperature, filtered and concentrated. The residue was further purified by flash column chromatography eluting with MeOH/water to get 12a (60 mg, yield: 69%). MS (m/z): 505 (M+H)+.
To a solution of 12a (60 mg, 0.12 mmol) in DMF (2 mL) was added 1.0 M TBAF in THF (0.15 mL, 0.15 mmol). After 20 minutes, the reaction mixture was diluted in water and extracted with EtOAc three times. The combined organic layers were dried, filtered and concentrated to give the crude product which was purified by flash column chromatography eluting with MeOH/water to afford Compound 105 as a white solid (2.0 mg, yield: 4%). MS (m/z): 433.2 (M+H)+. 1H NMR (400 MHz, CD3OD) δ: 8.22 (s, 1H), 7.94 (s, 1H), 7.74 (d, J=7.6 Hz, 1H), 7.66-7.59 (m, 1H), 7.58-7.51 (m, 2H), 7.40-7.30 (m, 2H), 6.64 (d, J=2.8 Hz, 1H), 5.33 (dd, J=9.5, 5.2 Hz, 1H), 4.64-4.60 (m, 1H), 4.32-4.20 (m, 1H), 3.52 (s, 1H), 2.67-2.51 (m, 1H), 2.07-1.97 (m, 1H).
To a mixture of 13a (200 mg, 0.62 mmol) and Cs2CO3 (403 mg, 1.24 mmol) in DMF (5 mL) was added 1-bromo-2-methylpropane (170 mg, 1.24 mmol), then the reaction was heated to 80° C. for 2 hours. The mixture was diluted with water and extracted with EtOAc three times. The combined organic layers were washed with brine, dried over MgSO4, filtered, concentrated and purified by flash column chromatography eluting with MeOH/water to give 14a (50 mg, yield: 21%). MS (m/z): 278.8 (M-Boc+H)+.
To a mixture of 14a (50 mg, 0.132 mmol) in MeOH (5 mL) was added conc. HCl aq (5 mL), then the reaction was stirred at room temperature for 2 hours. After concentration under reduced pressure, 14b was obtained as a yellow oil which was used directly in the next step without further purification. MS (m/z): 278.8 (M+H)+
A mixture of 14b (0.132 mmol), 4-chloro-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (24 mg, 0.132 mmol) and TEA (0.09 mL, 0.66 mmol) in n-BuOH (10 mL) was heated at reflux for 2 hours. The reaction mixture was concentrated purified by flash column chromatography eluting with MeOH/water to afford compound 107 as a slight yellow solid (17 mg, yield: 31%). MS (m/z): 420.7 (M+H)+. 1H-NMR (400 MHz, DMSO-d6) δ: 8.29 (s, 1H), 8.03 (s, 1H), 6.77 (t, J=5.1 Hz, 1H), 6.16 (t, J=4.0 Hz, 1H), 5.55-5.45 (m, 1H), 4.30-4.22 (m, 1H), 4.18-4.05 (m, 2H), 3.71-3.67 (m, 1H), 2.37-2.01 (m, 5H), 1.00 (d, T=6.6 Hz, 3H), 0.93 (d, T=6.5 Hz, 3H).
A mixture of 15a (50 mg, 0.106 mmol) (15a was prepared according to the procedure of Example 1), 2-methoxy-5 tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (28 mg, 0.116 mmol), Pd(dppf)2Cl2 (9 mg, 0.0106 mmol) and Na2CO3 (23 mg, 0.212 mmol) in dioxane (20 mL) and water (2 mL) was heated at 130° C. under N2 atmosphere for 3 hours. Then the mixture was filtered, concentrated and purified by flash column chromatography eluting with MeOH/water to give Compound 108 as a white solid (30 mg, yield: 56%). MS (m/z): 500.6 (M+H)+. 1H NMR (400 MHz, DMSO-d6) δ: 8.18-7.39 (m, 8H), 7.29 (d, J=6.4 Hz, 2H), 6.73-6.57 (m, 1H), 5.82 (s, 2H), 4.55-4.45 (m, 1H), 3.81 (s, 3H), 3.22-3.08 (m, 2H), 2.29-2.19 (m, 1H), 1.80-1.70 (m, 1H).
The following Compounds were prepared according to the procedure of Compound 108 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, CD3OD) δ 8.72 (s, 1H), 8.61 (s, 1H), 8.30-8.26 (m, 1H), 8.12-8.08 (m, 1H), 8.03-7.98 (m, 1H), 7.94-7.84 (m, 2H), 7.78-7.68 (m, 2H), 7.28-7.24 (m, 1H), 6.83-6.81 (m, 1H), 5.65-5.63 (m, 1H), 4.29 (s, 3H), 3.83-3.73 (m, 1H), 3.49-3.46 (m, 1H), 2.37-2.22 (m, 4H).
1H NMR (400 MHz, CDCl3) δ: 12.06 (s, 1H), 8.70 (s, 2H), 8.39 (s, 1H), 7.79-7.73 (m, 1H), 7.68-7.60 (m, 1H), 7.57-7.49 (m, 2H), 7.30-7.22 (m, 2H), 6.45-6.41 (m, 1H), 5.32 (s, 2H), 5.10-5.02 (m, 1H), 3.43-3.35 (m, 1H), 3.28-3.20 (m, 1H), 2.06-1.94 (m, 4H).
1H NMR (400 MHz, CDCl3) δ: 12.31 (s, 1H), 8.45-8.35 (m, 2H), 7.84-7.74 (m, 2H), 7.65-7.50 (m, 3H), 7.29-7.25 (m, 1H), 7.22-7.20 (m, 1H), 6.67-6.59 (m, 1H), 6.45-6.39 (m, 1H), 5.03-4.97 (m, 1H), 4.71 (s, 2H), 3.41-3.33 (m, 1H), 3.23-3.15 (m, 1H), 2.00-1.90 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ: 8.86 (s, 2H), 8.33 (s, 1H), 7.78-7.50 (m, 7H), 6.67-6.59 (m, 1H), 4.78-4.72 (m, 1H), 4.00 (s, 3H), 3.10-3.04 (m, 2H), 2.09-2.01 (m, 1H), 1.95-1.87 (m, 1H), 1.85-1.77 (m, 1H), 1.60-1.52 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 13.61 (s, 1H), 8.26 (s, 1H), 8.09 (d, J = 2.0 Hz, 1H), 7.75-7.31 (m, 7H), 6.64 (d, J = 2.9 Hz, 1H), 6.51 (d, J = 8.5 Hz, 1H), 6.19 (s, 2H), 5.02-4.88 (m, 1H), 3.45-3.39 (m, 2H), 2.43-2.37 (m, 1H), 1.87-1.81 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 13.81 (s, 1H), 8.44-8.21 (m, 2H), 7.87 (dd, J = 8.5, 2.3 Hz, 1H), 7.71-7.33 (m, 6H), 6.93 (d, J = 8.5 Hz, 1H), 6.65 (d, J = 3.0 Hz, 1H), 4.98-4.90 (m, 1H), 3.89 (s, 3H), 3.40-3.36 (m, 2H), 2.45-2.35 (m, 1H), 1.88-1.69 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 13.76 (s, 1H), 8.40 (s, 2H), 8.28 (s, 1H), 7.65-7.54 (m, 2H), 7.51-7.36 (m, 4H), 6.93 (s, 2H), 6.64 (d, J = 3.0 Hz, 1H), 5.04-4.90 (m, 1H), 3.59-3.40 (m, 2H), 2.04-1.78 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 8.78 (s, 2H), 8.32 (s, 1H), 7.63 (d, J = 2.9 Hz, 1H), 7.60-7.40 (m, 4H), 7.39 (dd, J = 4.8, 2.2 Hz, 1H), 6.65 (d, J = 3.0 Hz, 1H), 5.02-4.90 (m, 1H), 4.12-4.02 (m, 1H), 3.96 (s, 3H), 3.52-3.41 (m, 1H), 2.03-1.76 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 8.98 (d, J = 1.3 Hz, 1H), 8.35 (s, 1H), 8.26 (dd, J = 8.0, 2.1 Hz, 1H), 8.18 (d, J = 8.1 Hz, 1H), 7.68-7.35 (m, 6H), 6.66 (d, J = 3.0 Hz, 1H), 5.02-4.90 (m, 1H), 3.48-3.36 (m, 2H), 2.04-1.73 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 8.13 (s, 1H), 8.05 (d, J = 2.0 Hz, 1H), 7.70-7.66 (m, 2H), 7.64 (dd, J = 8.6, 2.4 Hz, 1H), 7.59-7.43 (m, 4H), 7.12 (s, 1H), 6.70 (d, J = 8.5 Hz, 1H), 6.65 (s, 1H), 4.84-4.79 (m, 1H), 1.32 (d, J = 6.8 Hz, 4H).
1H NMR (400 MHz, CDCl3) δ 9.69 (s, 1H), 8.69 (s, 2H), 8.34 (s, 1H), 7.79-7.74 (m, 1H), 7.58-7.49 (m, 3H), 7.39-7.34 (m, 1H), 7.17 (s, 1H), 7.11 (d, J = 2.4 Hz, 1H), 6.50 (s, 1H), 4.85-4.78 (m, 1H), 4.74 (d, J = 5.7 Hz, 1H), 4.10 (s, 3H), 1.27 (d, J = 6.7 Hz, 3H).
1H NMR (400 MHz, CDCl3) δ 9.80 (s, 1H), 8.49 (s, 1H), 8.45 ((d, J = 2.0 Hz, 1H), 7.92 (d, J = 8.0 Hz, 1H), 7.83-7.81 (dd, J = 8.4, 2.4 Hz, 1H), 7.62-7.53 (m, 3H), 7.30-7.27 (m, 1H), 7.21 (d, J = 2.0 Hz, 1H), 7.12 (d, J = 3.2 Hz, 1H), 6.64 (d, J = 8.4 Hz, 1H), 6.39 (d, J = 2.8 Hz, 1H), 4.55 (s, 2H), 4.48-4.45 (m, 1H), 3.92-3.80 (m, 2H), 3.53-3.48 (m, 1H), 3.39-3.33 (m, 2H), 2.89-2.83 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 7.63 (d, J = 3.0 Hz, 1H), 7.58-7.38 (m, 6H), 7.34-7.29 (m, 1H), 7.23 (s, 1H), 6.65 (d, J = 3.0 Hz, 1H), 6.07 (s, 2H), 4.57 (t, J = 7.5 Hz, 1H), 3.76 (s, 3H), 3.68-3.60 (m, 1H), 2.36-2.28 (m, 1H), 1.86-1.80 (m, 1H).
A mixture of 16a (120 mg, 0.23 mmol), Zn(CN)2 (560 mg, 4.77 mmol), dppf (120 mg, 0.22 mmol), Pd2(dba)3 (120 mg, 0.13 mmol) and Zinc powder (120 mg, 1.83 mmol) in DMA (4 mL) was stirred at 150° C. for 30 min under microwave condition. The reaction mixture was diluted with 200 mL of DCM and washed with water. The organic layer was separated, concentrated and purified by preparative TLC and chromatography to give Compound 111 as a white solid (8 mg, yield: 7%). MS (m/z): 457.7 (M+H)+. 1H NMR (400 MHz, DMSO-d6) δ 8.24 (s, 1H), 7.74 (d, J=7.5 Hz, 1H), 7.64-7.47 (m, 6H), 6.56 (d, J=2.9 Hz, 1H), 4.70-4.62 (m, 1H), 4.15-4.07 (m, 1H), 3.99-3.93 (m, 1H), 2.33-2.27 (m, 1H), 2.25-2.17 (m, 1H), 2.08-2.04 (m, 1H), 1.96-1.93 (m, 1H).
Under N2 atmosphere, to a solution of 17a (300 mg, 0.63 mmol) (17a was prepared according to the procedure of Example 1) in DMF (20 mL) was added Zn(CN)2 (945 mg, 3.15 mmol), followed by Pd(PPh3)4 (655 mg, 0.567 mmol), the reaction was stirred at 140° C. overnight under N2. After concentration, the residue was purified by column chromatography to give Compound 497 as a white solid (150 mg, yield: 56%). MS (m/z): 424.4 (M+H)+. 1H NMR (400 MHz, CD3OD) δ 7.54-7.45 (m, 5H), 7.27-7.23 (m, 1H), 6.90 (d, J=3.2 Hz, 1H), 5.15-5.02 (m, 1H), 4.27-4.16 (m, 1H), 4.08-4.01 (m, 1H), 2.46-2.38 (m, 1H), 2.21 (s, 3H), 2.19-2.12 (m, 1H).
The following compounds were prepared according to the procedure of Compound 497 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 8.43 (s, 1H), 7.83-7.77 (m, 1H), 7.72- 7.66 (m, 1H), 7.61-7.59 (m, 1H), 7.55-7.52 (m, 2H), 7.43-7.40 (m, 1H), 7.11 (d, J = 3.2 Hz, 1H), 6.78 (s, 2H), 5.00-4.75 (m, 1H), 4.19-4.08 (m, 1H), 2.45-2.35 (m, 2H), 1.24 (s, 3H), 1.91-1.86 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 7.58- 7.46 (m, 5H), 7.33-7.30 (m, 1H), 6.89 (dd, J = 3.0, 0.7 Hz, 1H), 4.76 (brs, 1H), 4.35 (brs, 1H), 3.61 (brs, 1H), 2.64 (brs, 1H), 2.21 (s, 3H), 0.65 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, CD3OD) δ 8.37 (brs, 1H), 7.66-7.52 (m, 4H), 7.37- 7.31 (m, 2H), 6.87 (d, J = 3.0 Hz, 1H), 4.92 (brs, 1H), 4.34 (brs, 1H), 3.29 (brs, 1H), 2.52 (brs, 1H), 2.21 (brs, 3H), 0.62 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, CD3OD) δ 8.09 (s, 1H), 7.64-7.60 (m, 4H), 7.55 (d, J = 2.9 Hz, 1H), 7.41-7.39 (m, 1H), 6.98 (dd, J = 3.0, 0.5 Hz, 1H), 4.85 (brs, 1H), 4.41 (brs, 1H), 3.69 (brs, 1H), 2.74 (brs, 1H), 0.74 (d, J = 6.7 Hz, 3H).
To a mixture of 18a (185 mg, 0.462 mmol) (18a was prepared according to the procedure of Example 1) in MeOH (1 mL) was added conc. HCl (1 mL) at r.t. The mixture was stirred at r.t for 30 min. The mixture was concentrated to give 18b as a brown solid which was used in the next step without purification.
To a mixture of 18-b (0.462 mmol) in n-BuOH (5 mL) were added 2,6-dichloro-9H-purine (87 mg, 0.462 mmol) and DIEA (298 mg, 2.31 mmol) at r.t. The mixture was stirred at 80° C. for 3 h, then morpholine (1 mL) was added, the mixture was stirred at 130° C. overnight. The reaction was concentrated and purified by flash column chromatography to afford Compound 114 as a yellow solid (180 mg, 77%). Yield: MS (m/z): 503.8 (M+H)+. 1H NMR (400 MHz, DMSO-d6) δ 12.26 (s, 1H), 7.71 (s, 1H), 7.64 (s, 1H), 7.59-7.46 (m, 4H), 7.39 (d, J=6.6 Hz, 1H), 6.61 (d, J=2.6 Hz, 1H), 5.05 (s, 1H), 4.05 (s, 2H), 3.63-3.45 (m, 8H), 2.65-2.54 (m, 1H), 2.27-2.13 (m, 1H).
Compounds 281-284 was prepared according to the procedure of Compound 114 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1HNMR (400 MHz, DMSO-d6) δ 7.67 (d, J = 2.6 Hz, 1H), 7.65 (s, 1H), 7.61-7.55 (m, 2H), 7.52-7.50 (m, 2H), 7.38-7.35 (m, 1H), 6.63 (dd, J = 3.0, 0.5 Hz, 1H), 6.31 (s, 2H), 4.90-4.80 (m, 1H), 3.86-3.74 (m, 2H), 2.67 (s, 6H), 2.44-2.38 (m, 1H), 1.97-1.91 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.67 (s, 1H), 7.73 (s, 1H), 7.64-7.30 (m, 7H), 6.63 (s, 1H), 5.26-5.22 (m, 0.3H), 4.83-4.77 (m, 0.7H), 4.24-4.09 (m, 1H), 3.89-3.67 (m, 0.3H), 3.68 (s, 0.7H), 2.46-2.41 (m, 1H), 1.95-1.80 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 7.73 (d, J = 2.6 Hz, 1H), 7.61-7.49 (m, 4H), 7.42-7.38 (m, 1H), 7.04 (br, 2H), 6.65 (d, J = 3.0 Hz, 1H), 5.06-4.71 (m, 1H), 4.16-3.89 (m, 2H), 2.55-2.48 (m, 1H), 2.19 (s, 3H), 2.05-1.98 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 7.64-7.53 (m, 3H), 7.48 (d, J = 7.3 Hz, 1H), 7.41 (d, J = 3.0 Hz, 1H), 7.38 (s, 1H), 7.30-7.26 (m, 1H), 6.53 (d, J = 3.0 Hz, 1H), 4.96-4.94 (m, 1H), 4.23-4.17 (m, 1H), 3.99-3.95 (m, 1H), 3.60 (s, 3H), 2.47-2.39 (m, 1H), 2.31-2.18 (m, 1H).
The mixture of 19a (20.8 g, 200 mmol), KSCN (20.0 g, 206 mmol), Ac2O (20.0 mL) and AcOH (80 mL) was stirred at r.t. overnight. Then H2O (100 mL) was added and extracted with DCM: MeOH=9:1, the organic layer was dried and concentrated to give 19b as a yellow solid which was used in the next step without further purification (2.0 g, yield: 53%)
To a solution of 19b (20 g, 106 mmol) in DMF (15 mL) was added aniline (9.2 mL) at r.t., the reaction was stirred at reflux until 19b disappeared by TLC. The mixture was concentrated, the residue was washed with EtOH, and filtered to give 19c as a yellow solid (880 mg, yield: 40.7%). MS (m/z): 203.1 (M+1)+.
The solution of 19c (7.29 g, 36 mmol) in CH3CN (120 mL) was purged by NH3 for 5 min, then BOP (20.7 g, 46.8 mmol) and DBU (8.21 g, 54 mmol) were added, the reaction was stirred overnight. The mixture was filtered to give 19d was as a white solid (7.24 g). MS (m/z): 201.7 (M+1)+.
To a solution of 19d (7.24 g, 36 mmol) in EtOH (100 mL) was added 40% 2-chloroacetaldehyde in water (17.8 mL, 108 mmol), the reaction was stirred at 100° C. overnight. The mixture was concentrated and purified by flash column chromatography to give 19e as a white solid (6.2 g, yield: 77%). MS (m/z): 225.9 (M+1)+.
19e (2.25 g, 10 mmol) and NCS (700 mg, 5.26 mmol) were dissolved in DMF (10 mL), the reaction was stirred at r.t. for 3 h. The mixture was poured into H2O (100 mL), and extracted with EtOAc, the organic layers were washed with brine, dried over anhydrous Na2SO4 and concentrated. The resulting residue was washed with MeOH to give 19f as a white solid (600 mg, yield: 23%). MS (m/z): 260.1 (M+1)+.
19f (600 mg, 2.3 mmol) and SeO2 (257 mg, 2.3 mmol) were dissolved in dioxane (20 mL), the reaction was stirred at reflux overnight, then concentrated and purified by flash column chromatography to give 19g as a white solid (250 mg, yield: 39%). MS (m/z): 274.1 (M+1)+.
To a solution of 19g (250 mg, 0.9 mmol) in THF (10 mL) cooled to −78° C. was added MeMgBr (3M in ether, 1.2 mL) dropwise under N2, the reaction was stirred at −78° C. for 30 min. Then MeOH (3 mL) was added dropwise, the resulting mixture was concentrated and purified by flash column chromatography to give 19h as a white solid (220 mg, yield: 83%). MS (m/z): 290.1 (M+1)+.
To a solution of 19h (200 mg, 0.69 mmol) in THF (20 mL) was added DPPA (630 mg, 2.29 mmol), followed by DBU (300 mg, 1.97 mmol) at r.t., the reaction was stirred at reflux for 3 h, then concentrated and purified by flash column chromatography to give 19i as a yellow oil (130 mg, yield: 59.9%). MS (m/z): 315.1 (M+1)+.
To a solution of 19i (130 mg, 0.4 mmol) in THF (10 mL) was added NH3.H2O (25% aq., 1 mL), followed by PPh3 (200 mg, 0.76 mmol), the reaction was stirred at r.t. for 30 min, then warmed to 60° C. for another 2 hours. The mixture was concentrated and purified by flash column chromatography to give 19j as a white solid (60 mg, yield: 50%). MS (m/z): 288.9 (M+1)+.
To a solution of 19j (30 mg, 0.104 mmol) in n-BuOH (3 mL) were added DIEA (0.052 mL, 0.312 mmol) and 6-chloro-9H-purine (19.3 mg, 0.125 mmol), the reaction was stirred at 130° C. overnight. The mixture was concentrated and purified by preparative thin layer chromatography to give Compound 115 as a white solid (3.6 mg, yield: 9%). MS (m/z): 406.9 (M+1)+. 1H NMR (400 MHz, CD3OD) δ: 8.06 (s, 1H), 7.96 (s, 1H), 7.59-7.47 (m, 3H), 7.38 (t, J=7.3 Hz, 1H), 7.27-7.24 (m, 2H), 6.76 (s, 1H), 4.93-4.89 (m, 1H), 1.47 (d, T=6.7 Hz, 3H).
The following Compounds were prepared according to the procedure of Compound 115 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 8.11 (s, 1H), 7.95 (s, 1H), 7.70-7.68 (m, 1H), 7.59- 7.51 (m, 2H), 7.46-7.39 (m, 2H), 7.34 (s, 1H), 6.88 (s, 1H), 6.42 (d, J = 6.3 Hz, 1H), 4.60-4.57 (m, 1H), 1.36 (d, J = 6.7 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 7.88 (s, 1H), 7.67-7.32 (m, 7H), 7.20 (s, 2H), 6.75 (s, 1H), 4.64-4.54 (m, 1H), 1.29 (d, J = 6.6 Hz, 3H).
1H NMR (400 MHz, CD3OD) δ 8.09 (s, 1 H), 7.74 (s, 1H), 7.18-6.96 (m, 3H), 6.74 (s, 1H), 6.66-6.58 (m, 2H), 5.70 (s, 1H), 5.43-5.38 (m, 1H), 1.48 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, CDCl3) δ 8.07 (s, 1H), 7.55 (t, J = 7.4 Hz, 1H), 7.49-7.34 (m, 4H), 7.25 (s, 1H), 6.65 (s, 1H), 5.00 (d, J = 6.4 Hz, 1H), 4.93 (q, J = 6.9 Hz, 1H), 4.88 (s, 2H), 1.38 (d, J = 6.6 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 8.03 (s, 1H), 7.67-7.63 (m, 1H), 7.57-7.48 (m, 3H), 7.43-7.39 (m, 2H), 7.34 (s, 1H), 7.24 (d, J = 7.2 Hz, 1H), 7.06 (d, J = 2.8 Hz, 1H), 6.82 (s, 1H), 4.71-4.62 (m, 1H), 1.35 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 8.12 (s, 1H), 7.79 (s, 1H), 7.72-7.66 (m, 1H), 7.63-7.59 (m, 1H), 7.58-7.51 (m, 2H), 7.49-7.46 (m, 2H), 7.34 (s, 1H), 6.55 (s, 1H), 4.61-4.53 (m, 1H), 3.20 (s, 3H), 1.28 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 9.30 (d, J = 8.0 Hz, 1H), 8.31 (s, 1H), 8.13 (s, 1H), 7.64-7.55 (m, 3H), 7.49-7.41 (m, 2H), 7.39 (s, 1H), 6.59 (s, 1H), 4.68-4.62 (m, 1H), 2.52 (s, 3H), 1.37 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, CD3OD) δ 9.40 (d, J = 7.2 Hz, 1H), 7.57-7.52 (m, 2H), 7.49-7.45 (m, 3H), 7.25 (s, 1H), 6.60 (s, 1H), 4.82-4.79 (m, 1H), 3.54-3.41 (m, 2H), 2.64-2.48 (m, 2H), 1.34 (d, J = 6.8 Hz, 3H).
To a mixture of 20a (7.55 g, 50 mmol) in Et2O (80 mL) was added 2-oxoacetic acid.1H2O (5.05 g, 55 mmol), the reaction was stirred at r.t. overnight. The mixture was concentrated in vacuo to give 20b as a white solid which was used in the next step without further purification.
To a solution of 20b (about 11.25 g, 50 mmol) in MeOH (150 mL) was added concentrated sulfuric acid (2 mL) dropwise at 0° C. After the addition, the reaction mixture was stirred at r.t. for 90 h, then poured into the iced sat. NaHCO3 aq. (300 mL), the resulting mixture was extracted with EtOAc, the organic layers were dried over anhydrous Na2SO4, concentrated and purified by column chromatography to give 20c as a white solid (12 g, yield: 95%). MS (m/z): 275.7 (M+23)+.
To a solution of 20c (12 g, 47.4 mmol) in toluene (60 mL) was added PBr3 (12.8 g, 47.4 mmol) at 70° C., the reaction was stirred at 70° C. for 20 h, then triethyl phosphate (7.87 g, 47.4 mmol) was added dropwise and stirred at 70° C. for another 2 h. The mixture was concentrated, diluted with EtOAc, and washed with sat. NaHCO3 aq. The organic layers were dried over anhydrous Na2SO4, filtered and concentrated. The resulting residue was dissolved in EtOAc, petroleum ether was added with vigorous stirring, then filtrated to give 20d as a white solid (8 g, yield: 47%).
To a solution of 20d (8 g, 22.3 mmol) in DCM (80 mL) was added 1,1,3,3-tetramethylguanidine (2.44 g, 21.2 mmol) at r.t., the reaction was stirred at r.t for 15 min, then a solution of 1H-pyrrole-2-carbaldehyde (1.92 g, 20.2 mmol) in DCM (5 mL) was added dropwise at −30° C., the reaction mixture was stirred at −30° C. for 45 min, then warmed to r.t. and stirred for 48 h. The mixture was concentrated and purified by column chromatography to give 20e as a white solid (2 g, yield: 51%). MS (m/z): 192.9 (M+1)+.
To a solution of 20e (576 mg, 3 mmol) in DCM (20 mL) was added phenylboronic acid (732 mg, 6 mmol), copper(II) acetate (1.08 g, 6 mmol), pyridine (1.18 g, 15 mmol) and 4 Å molecular sieve at r.t., the reaction was stirred at r.t. for 20 h. The mixture was filtered, concentrated and purified by column chromatography to give 20f as a white solid (650 mg, yield: 81%). MS (m/z): 268.8 (M+1)+.
To a solution of 20f (1 g, 3.73 mmol) in EtOH (30 mL) and THF (30 mL) was added NaOH aq. (11.19 mL, 1N) at 0° C., the reaction was stirred at 0° C. for 30 min. The mixture was concentrated, diluted with H2O (10 mL), adjusted to pH=6 with HCl aq. (1N) and concentrated in vacuo to give 20g as a brown solid which was used in the next step without further purification. MS (m/z): 254.7 (M+1)+.
To a solution of 20g (about 950 mg, 3.73 mmol) in DMF (10 mL) were added DIEA (1.44 g, 11.19 mmol) and HBTU (1.70 g, 4.48 mmol), the mixture was stirred at r.t for 5 min, then N,O-dimethylhydroxylamine hydrochloride (438 mg, 4.48 mmol) was added, the reaction was stirred at r.t overnight. The mixture was concentrated and purified by column chromatography to give 20h as a white solid (550 mg, yield: 50%). MS (m/z): 297.7 (M+1)+.
To a solution of 20h (550 mg, 1.85 mmol) in THF (5 mL) was added a solution of Methylmagnesium bromide in Et2O (1.23 mL, 3N) at 0° C. under N2, the reaction was stirred at 0° C. for 1 h. The mixture was quenched with sat. NH4Cl aq., concentrated and purified by column chromatography to give 20i as a yellow solid (220 mg, yield: 47%). MS (m/z): 252.7 (M+1)+.
To a solution of 20i (50.4 mg, 0.2 mmol) in EtOH (6 mL) were added ammonium acetate (550 mg, 7.1 mmol) and sodium cyanoborohydride (126 mg, 2 mmol), the reaction was stirred at 130° C. for 2 h under Microwave condition, then another part of ammonium acetate (550 mg, 7.1 mmol) and sodium cyanoborohydride (126 mg, 2 mmol) was added, the reaction was stirred at 90° C. for 20 h. After cooling to r.t, aq. HCl (0.5 mL, 1 N) was added, the mixture was stirred for 30 min, followed by conc. NH3.H2O (3 mL), the mixture was stirred for 10 min, then NaBH4 (30 mg, 0.79 mmol) was added, the mixture was stirred for another 30 min. The mixture was concentrated and purified by flash column chromatography to give 20j as a yellow solid (32 mg, yield: 63%). MS (m/z): 236.7 (M-16)+.
To a solution of 20j (40 mg, 0.158 mmol) in n-BuOH (8 mL) was added 6-chloro-9H-purine (29 mg, 0.190 mmol) and DIEA (61 mg, 0.474 mmol) at r.t., the reaction was stirred at 130° C. overnight. The mixture was concentrated and purified by flash column chromatography to give Compound 119 as a yellow solid (10 mg, yield: 17%). MS (m/z): 371.6 (M+1)+. 1H NMR (400 MHz, DMSO-d6) δ 8.05 (s, 1H), 7.97 (s, 1H), 7.72 (s, 1H), 7.66 (s, 1H), 7.57-7.30 (m, 6H), 6.71 (s, 1H), 6.63 (s, 1H), 6.29 (s, 1H), 4.78 (s, 1H), 1.32 (d, J=6.5 Hz, 3H).
The following Compounds 120 and 121 were prepared according to the procedures of Compound 119 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 7.80 (s, 1H), 7.54 (d, J = 7.3 Hz, 1H), 7.48-7.39 (m, 4H), 7.34 (s, 2H), 7.16 (s, 2H), 6.70 (s, 1H), 6.65 (s, 1H), 6.38 (s, 1H), 4.71-4.62 (m, 1H), 1.29 (d, J = 6.6 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 8.03 (s, 1H), 7.91 (s, 1H), 7.57 (d, J = 8.2 Hz, 1H), 7.54-7.25 (m, 5H), 6.80 (s, 1H), 6.63 (s, 1H), 6.31 (s, 1H), 6.08 (s, 1H), 4.67 (m, 1H), 1.35 (d, J = 6.4 Hz, 3H).
To a solution of Compound 119 (60 mg, 0.16 mmol) in DMF (3 mL) was added NCS (21 mg, 0.16 mmol) at r.t., the reaction was stirred at 70° C. for 30 min, then another part of NCS (6 mg, 0.045 mmol) was added, the reaction was stirred at 70° C. for another 30 min. The mixture was concentrated and purified by flash column chromatography to give Compound 122 as a white solid (15 mg, yield: 23%) and Compound 123 as a white solid (5 mg, yield: 7.7%)). Compound 122: MS (m/z): 406.1 (M+1)+. 1H NMR (400 MHz, DMSO-d6) δ 8.03 (s, 1H), 7.89 (s, 1H), 7.69 (s, 1H), 7.56 (s, 1H), 7.56-7.34 (m, 5H), 6.64-6.55 (m, 2H), 6.25 (d, J=3.7 Hz, 1H), 4.87-4.57 (m, 1H), 1.28 (d, J=6.6 Hz, 3H). Compound 123: MS (m/z): 405.7 (M+1)+. 1H NMR (400 MHz, CD3OD) δ 7.90 (s, 1H), 7.83 (s, 1H), 7.49 (d, J=3.2 Hz, 1H), 7.46 (d, J=7.6 Hz, 1H), 7.42-7.35 (m, 2H), 7.28 (t, J=7.1 Hz, 1H), 7.03 (t, J=7.4 Hz, 1H), 6.77 (s, 1H), 6.65 (d, J=3.0 Hz, 1H), 1.49 (d, J=6.7 Hz, 3H).
The following Compounds were prepared according to the procedures of Compound 122 and 123 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 7.96 (s, 1H), 7.67 (s, 1H), 7.65 (d, J = 5.3 Hz, 1H), 7.58-7.35 (m, 5H), 6.64 (s, 1H), 6.60 (d, J = 3.8 Hz, 1H), 6.27 (d, J = 3.8 Hz, 1H), 5.47 (d, J = 6.7 Hz, 1H), 4.58-4.51 (m, 1H), 1.30 (d, J = 6.7 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 7.80 (s, 1H), 7.47 (d, J = 7.2 Hz, 1H), 7.44-7.36 (m, 3H), 7.35-7.28 (m, 2H), 7.13 (s, 2H), 6.62 (d, J = 3.8 Hz, 1H), 6.61 (s, 1H), 6.35 (d, J = 3.8 Hz, 1H), 4.74-4.43 (m, 1H), 1.26 (d, J = 6.7 Hz, 3H).
1H NMR (400 MHz, DMSO-d6) δ 7.88 (s, 1H), 7.60 (d, J = 7.3 Hz, 1H), 7.52-7.43 (m, 3H), 7.42-7.36 (m, 2H), 7.20 (s, 2H), 6.87 (s, 1H), 6.60 (s, 1H), 4.62-4.53 (m, 1H), 1.29 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, CD3OD) δ 7.95 (s, 0.5H), 7.93 (s, 0.5H), 7.834 (s, 0.5H), 7.83 (s, 0.5H), 7.52 (dd, J = 14.4, 8.0 Hz, 1H), 7.32-7.27 (m, 1H), 7.24-7.18 (m, 1H), 7.10-6.91 (m, 2H), 6.81 (s, 0.5H), 6.80 (s, 0.5H), 6.59 (d, J = 1.7 Hz, 0.5H), 6.58 (d, J = 1.7 Hz, 0.5H), 6.40 (d, J = 4.1 Hz, 0.5H), 6.38 (d, J = 4.1 Hz, 0.5H), 5.46-5.33 (m, 1H), 1.56 (d, J = 5.3 Hz, 1.5H)-1.54 (d, J = 5.3 Hz, 1.5H)
1H NMR (400 MHz, DMSO-d6) δ 8.29 (s, 1H), 8.13-8.07 (m, 1.5H), 8.04 (s, 0.5H), 7.57-7.40 (m, 1H), 7.39-7.02 (m, 3H), 6.80 (s, 0.5H), 6.78 (s, 0.5H), 6.64 (d, J = 3.6 Hz, 0.5H ), 6.63 (d, J = 3.6 Hz, 0.5H), 6.56 (s, 0.5H), 6.54 (s, 0.5H), 6.35 (d, J = 3.8 Hz, 0.5H), 6.33 (d, J = 3.8 Hz, 0.5H), 4.85-4.62 (m, 1H), 1.39 (d, J = 6.7 Hz, 3H)
1H NMR (400 MHz, DMSO-d6) δ 8.32 (s, 1H), 8.15 (s, 0.5H), 8.12 (br, 1H), 8.09 (s, 0.5H), 7.57-7.45 (m, 1H), 7.43-7.12 (m, 3H), 6.85 (d, J = 1.0 Hz, 0.5H), 6.84 (d, J = 1.1 Hz, 0.5H), 6.82-6.76 (m, 1H), 6.75 (br, 1H), 4.73-4.60 (m, 1H), 1.40 (d, J = 4.3 Hz, 3H)
Compound 132 was prepared according to the procedures of Example 1 and the following Steps 24-1 and 2. Compound 132 was got as a white solid. MS (m/z): 434.8 (M+H)+; 1H NMR (400 MHz, CD3OD) δ: 8.27 (s, 1H), 8.16-7.93 (m, 2H), 7.65-7.49 (m, 4H), 7.15-7.05 (br, 1H), 6.24-6.20 (m, 1H), 5.41 (s, 0.5H), 5.30-5.26 (m, 0.5H), 4.61-4.20 (br, 2H), 4.02-3.94 (m, 1H), 2.58-2.44 (m, 1H), 2.32-2.14 (m, 1H).
To a solution of 24a (400 mg, 2.94 mmol) and (2S,4S)-1-(tert-butoxycarbonyl)-4-fluoropyrrolidine-2-carboxylic acid (889 mg, 3.82 mmol) in THF (35 mL) was added EDC (729 mg, 3.82 mmol). The reaction mixture was stirred at r.t. for 2 hours, then the solvent was removed in vacuo and water was added. The mixture was extracted with EtOAc three times. The organic layers were combined, died over anhydrous Na2SO4 and concentrated to give 24b.
24b was dissolved in 7N NH3 in MeOH (100 mL) and the mixture was stirred in a sealed tube at 130° C. overnight. The solvent was removed in vacuo and the residue was purified by flash column chromatography eluting with EtOAc/PE to give 24c as a white solid (110 mg, yield: 11%). MS (m/z): 341 (M+H)+
A mixture of Compound 55 (308 mg, 0.632 mmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (200 mg, 1.265 mmol), Pd(dppf)2Cl2 (52 mg, 0.0632 mmol) and Na2CO3 (201 mg, 1.896 mmol) in dioxane (20 mL) and water (2 mL) was reacted at 130° C. under N2 atmosphere in a microwave oven for 30 min. Then the mixture was filtered, concentrated and purified by flash column chromatography eluting with MeOH/DCM to give 25a as a slight yellow solid (120 mg, yield: 44%). MS (m/z): 435.1 (M+H)+.
To a solution of 25a (60 mg, 0.138 mmol) in methanol (10 mL) was added Pd/C (6 mg), the mixture was stirred at r.t. under H2 atmosphere for 2.5 hours, then the mixture was filtered, concentrated and purified by flash column chromatography eluting with MeOH/water to give Compound 133 as a white solid (41 mg, yield: 68%). MS (m/z): 436.8 (M+H)+. 1H NMR (400 MHz, DMSO-d6) δ 8.31 (s, 2H), 7.78-7.42 (m, 6H), 6.47 (s, 1H), 5.18-5.08 (br, 1H), 4.49-4.15 (m, 2H), 2.88 (q, J=7.4 Hz, 2H), 2.73-2.63 (m, 1H), 2.19-2.09 (m, 1H), 1.21 (t, J=7.5 Hz, 3H).
The following Compounds 291-292 was prepared according to the procedure of Compound 133 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ: 8.36 (d, J = 9.3 Hz, 2H), 7.80-7.49 (m, 6H), 7.38-7.31 (m, 1H), 6.96 (d, J = 2.2 Hz, 1H), 5.87 (d, J = 17.8 Hz, 1H), 5.31 (d, J = 11.3 Hz, 1H), 5.32-5.21 (m, 1H), 4.51-4.46 (m, 1H), 4.34-4.23 (m, 1H), 2.86-2.74 (m, 1H), 2.29-2.21 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 8.50-8.40 (m, 2H), 7.72-7.47 (m, 5H), 7.39-7.33 (m, 1H), 7.20-7.15 (m, 1H), 6.73-6.70 (m, 1H), 5.68-5.62 (m, 1H), 5.26-5.23 (m, 1H), 5.13-5.10 (m, 1H), 4.78-4.71 (m, 1H), 4.07-4.01 (m, 1H), 2.23-2.11 (m, 2H), 2.00-1.85 (m, 2H).
To a solution of 26a (250 mg, 1.25 mmol) in 20 mL of dry DCM was added m-CPBA (473 mg, 2.75 mmol) and stirred at r.t. for 16 hours. The solution was used forward next step without further purification.
To the solution 26b was added 26c (63 mg, 0.18 mmol) (26c was prepared according to the procedure of Example 1) and DIEA (78 mg, 0.60 mmol), then the mixture was stirred at r.t. overnight. The mixture was concentrated and purified by flash column chromatography eluting with MeOH/H2O to afford 26d as a yellow solid (85 mg, yield: 49%). MS (m/z): 511.0 (M+H)+.
To a solution of 26d (82 mg, 0.16 mmol) in 5 mL of THF was added 4 mL of 7N NH3 in MeOH, then the mixture was stirred at r.t. overnight. After concentration, the residue was purified by flash column chromatography, eluting with MeOH/H2O, and further purified by preparative TLC, eluting with MeOH/DCM=1/80, to give Compound 134 as a white solid (28.8 mg, yield: 40%). MS (m/z): 448.1 (M+H)+. 1H NMR (400 MHz, DMSO-d6) δ 7.93-7.78 (m, 2H), 7.63-7.54 (m, 5H), 6.62-6.36 (m, 3H), 5.70-5.59 (m, 1H), 4.71-4.31 (m, 1H), 3.95-3.83 (m, 1H), 3.72-3.64 (m, 1H), 2.12-1.74 (m, 4H).
The following Compounds was prepared according to the procedure of Compound 134 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ: 7.90 (s, 1H), 7.80-7.62 (m, 5H), 7.61-7.46 (m, 2H), 6.82 (s, 1H), 6.60 (s, 2H), 5.78-5.66 (br, 1H), 4.40-4.11 (m, 2H), 2.75-2.69 (m, 1H), 2.50-2.12 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 7.98-7.77 (m, 2H), 7.63-7.54 (m, 5H), 6.61-6.55 (m, 3H), 5.92-5.71 (m, 1H), 4.98-4.81 (m, 1H), 4.28-4.19 (m, 1H), 3.13-2.90 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ: 7.98-7.48 (m, 7H), 6.82-6.53 (m, 3H), 5.88-5.61 (m, 1H), 5.61-4.95 (m, 1H), 4.68-4.06 (m, 2H), 2.72-2.64 (m, 1H), 2.52-2.05 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 8.40 (s, 1H), 7.66-7.61 (m, 1H), 7.58-7.52 (m, 3H), 7.28-7.26 (m, 1H), 6.90 (s, 1H), 5.39-5.29 (m, 1H), 4.34-4.27 (m, 1H), 3.89-3.78 (m, 1H), 2.32-2.24 (m, 1H), 2.22-2.2.19 (m, 1H), 2.17 (s, 3H), 2.14 (s, 3H).
1H NMR (400 MHz, CD3OD) δ 8.32 (br, 1H), 8.10 (br, 1H), 7.76 (d, J = 6.0 Hz, 1H), 7.59-7.47 (m, 3H), 7.28-7.26 (m, 2H), 6.46 (d, J = 3.0 Hz, 1H), 5.07 (br, 1H), 4.67 (br, 1H), 3.88 (br, 1H), 2.15 (br, 3H), 1.10 (s, 3H), 0.64 (s, 3H).)
2,4-dichloro-1,3,5-triazine (45 mg, 0.3 mmol) was added to 2 mL of NH3.H2O aq., the reaction was stirred at −20° C. for 10 min, then filtered, washed with water and dried to give 4-chloro-1,3,5-triazin-2-amine (18 mg, yield: 46%) as a yellow solid which was used in the next step without further purification. MS (m/z): 131.0 (M+H)+.
(S)-2-(1-(4-amino-1,3,5-triazin-2-yl)pyrrolidin-2-yl)-5-chloro-3-phenylpyrrolo[2,1-f][1,2,4]triazin-4(3H)-one was prepared with 4-chloro-1,3,5-triazin-2-amine as the material according to the procedure of Example 1 from 1e to Compound 1. MS (m/z): 409.1 (M+H)+. 1H NMR (400 MHz, CD3OD) δ: 8.02 (d, J=1.6 Hz, 1H), 7.81 (d, J=7.6 Hz, 1H), 7.64-7.54 (m, 3H), 7.42-7.39 (m, 1H), 7.37-7.35 (m, 1H), 6.50-6.49 (m, 1H), 4.67-4.64 (m, 1H), 3.81-3.73 (m, 1H), 3.59-3.53 (m, 1H), 2.20-2.08 (m, 2H), 1.97-1.85 (m, 2H).
To a mixture of Intermediate 7 (500 mg, 2.36 mmol) in THF (40 mL) were added BOC-L-Proline (557 mg 2.59 mmol) and EDC (497 mg 2.59 mmol) at r.t. The reaction was stirred at r.t overnight. The mixture was concentrated and purified by flash chromatography to afford 28a as a yellow oil (800 mg, yield: 83%). MS (m/z): 410.5 (M+1)+.
The mixture of 28a (800 mg 1.96 mmol) in a solution of NH3 in MeOH (7N, 50 mL) was stirred at 130° C. for 36 h in a sealed tube. The reaction was concentrated and purified by chromatography to afford 28b as a yellow solid (580 mg, yield: 75%). MS (m/z):
348.5 (M+1)+.
Compound 139 was prepared from 28b according to the procedure of Example 1.
MS (m/z): 442.2 (M+1)+. 1H NMR (400 MHz, DMSO-d6) δ 9.22 (s, 1H), 8.23-8.18 (m, 1.5H), 8.10 (s, 0.5H), 7.87-7.42 (m, 6H), 7.35 (s, 1H), 6.95 (s, 0.5H), 6.92 (s, 0.5H), 5.37-5.25 (m, 0.5H), 4.74-4.45 (m, 0.5H), 4.38-4.26 (m, 0.5H), 4.15-4.01 (m, 0.5H), 3.94-3.84 (m, 0.5H), 3.74-3.63 (m, 0.5H), 2.35-2.21 (m, 2H), 2.01-1.93 (m, 1H), 1.90-1.82 (m, 1H).
Compound 140 was prepared according to the procedure of Compound 139 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art.
1H NMR (400 MHz, DMSO-d6) δ 9.2 (s, 1H), 8.29 (s, 1H), 8.27 (s, 1H), 7.79 (d, J = 7.9 Hz, 1H), 7.65-7.52 (m, 5H), 7.35 (s, 1H), 6.96 (d, J = 2.9 Hz, 1H), 4.67 (dd, J = 8.0, 3.5 Hz, 1H), 4.16-4.05 (m, 1H), 3.94 (m, 1H), 2.32-2.19 (m, 2H), 2.04-1.92 (m, 2H).
To a solution of Compound 149 (30 mg, 0.068 mmol) in CH2Cl2 (1 mL) was added TFA (2 mL) at 0° C., the reaction was stirred at r.t. for 30 min, then concentrated at r.t. The residue was dissolved in MeOH (2 mL), and treated with 1N KOH (2 mL), then stirred at r.t. for another 1 h. The mixture was adjusted to pH=7.0, then concentrated and purified by chromatography to give the title compound as a white solid (12 mg, yield: 41%). MS (m/z): 429.6 (M+1)+1H NMR (400 MHz, CD3OD) δ 8.21 (s, 1H), 8.14 (s, 1H), 7.95 (s, 0.5H), 7.91 (s, 0.5H), 7.69-7.43 (m, 4H), 7.37 (br, 1H), 7.17 (s, 0.5H), 7.09 (s, 0.5H), 6.43 (s, 0.5H), 6.40 (s, 0.5H), 5.51 (br, 0.5H), 4.48 (s, 2H), 4.31 (br, 0.5H), 4.09 (br, 0.5H), 3.92 (br, 0.5H), 3.71 (br, 0.5H), 2.29-1.88 (m, 4H).
The following Compounds 178-179 were prepared according to the procedure of Compound 177 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art.
1H NMR (400 MHz, CD3OD) δ 8.27 (s, 1H), 7.85 (d, J = 7.8 Hz, 1H), 7.74-7.63 (m, 3H), 7.52 (d, J = 7.2 Hz, 1H), 7.39 (d, J = 2.5 Hz, 1H), 6.65 (d, J = 2.4 Hz, 1H), 5.04-5.01 (m, 1H), 4.95 (s, 2H), 3.97-3.87 (m, 1H), 3.83-3.73 (m, 1H), 2.34-2.28 (m, 1H), 2.14-2.13 (m, 1H), 2.02-1.91 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 8.23 (s, 1H), 7.97 (s, 1H), 7.77 (d, J = 7.8 Hz, 1H), 7.65-7.53 (m, 3H), 7.43 (d, J = 7.3 Hz, 1H), 7.28 (d, J = 2.6 Hz, 1H), 6.52 (d, J = 2.6 Hz, 1H), 4.92-4.90 (m, 1H), 4.56 (s, 2H), 4.30-4.24 (m, 1H), 4.10-4.04 (m, 1H), 2.47-2.41 (m, 1H), 2.20- 2.15 (m, 1H), 2.12-1.99 (m, 2H).
To a solution of 30a (409 mg, 1 mmol) in 1,4-dioxane/water (10 mL/1 mL) was added 2-aminopyrimidin-5-ylboronic acid (139 mg, 1 mmol), Pd(dppf)Cl2 (81.6 mg, 0.1 mmol) and K2CO3 (414 mg, 3 mmol). Under N2, the reaction mixture was heated at 100° C. for 2 h. Then the solvent was removed in reduced pressure and the residue was purified by flash column chromatography eluting with MeOH/DCM to give 30b as a yellow solid (310 mg, yield: 82.4%). MS (m/z): 377.1 (M+H)+
A mixture of 30c (64 mg 0.2 mmol) (The intermediate was synthesized according to the procedure of Example 1), 30b (68 mg, 0.18 mmol) and Et3N (80 mg, 0.8 mmol) in n-BuOH (2 mL) was stirred at 100° C. for 1 h. The reaction solution was concentrated and the residue was dissolved in TFA (3 mL). The resulting mixture was stirred at r.t. for 30 min. Then the solvent was removed in vacuo. To the residue was added a solution of NH3 in MeOH (7N, 3 mL). The mixture was stirred at r.t. for 30 min. The solvent was evaporated and the residue was purified by flash column chromatography eluting with MeOH/water to give Compound 180 as a white solid (37 mg, yield: 37.4%). MS (m/z): 495.1 (M+H)+; 1H NMR (400 MHz, DMSO-d6) δ: 12.00 (s, 1H), 8.37 (s, 2H), 8.23 (s, 1H), 7.66-7.57 (m, 1H), 7.57-7.48 (m, 4H), 7.43 (d, J=2.7 Hz, 1H), 7.32 (d, J=2.4 Hz, 1H), 6.65 (s, 2H), 6.49 (d, J=3.2 Hz, 1H), 5.06-5.00 (m, 1H), 3.20-3.16 (m, 1H), 3.13-2.99 (m, 1H), 2.42-2.38 (m, 1H), 1.78-1.68 (m, 1H).
Compounds 181-184 were prepared according to the procedure of Compound 180 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, CD3OD) δ: 8.75 (s, 2H), 8.23 (s, 1H), 7.62-7.45 (m, 4H), 7.38-7.25 (m, 3H), 6.38-6.26 (br, 1H), 5.17-5.09 (m, 1H), 4.03 (s, 3H), 3.43-3.33 (m, 1H), 3.23-3.17 (br, 1H), 2.37-2.17 (br, 1H), 1.86-1.76 (br, 1H).
1H NMR (400 MHz, CD3OD) δ 8.26 (d, J = 2.0 Hz, 1H), 8.19 (s, 1H), 7.83 (dd, J = 8.5, 2.3 Hz, 1H), 7.56-7.52 (m, 2H), 7.47 (d, J = 7.7 Hz, 2H), 7.35-7.30 (m, 1H), 7.26 (d, J = 7.5 Hz, 1H), 7.17 (s, 1H), 6.84 (d, J = 8.5 Hz, 1H), 6.31 (d, J = 3.2 Hz, 1H), 5.12-5.00 (m, 1H), 3.91 (s, 3H), 3.38-3.31 (m, 1H), 3.22-3.12 (m, 1H), 2.30-2.19 (m, 1H), 1.81-1.69 (m, 1H).
1H NMR (400 MHz, CD3OD) δ: 8.17 (s, 1H), 8.04 (s, 1H), 7.64-7.47 (m, 5H), 7.37-7.24 (m, 2H), 7.10 (s, 1H), 6.63 (d, J = 8.5 Hz, 1H), 6.31 (d, J = 3.1 Hz, 1H), 5.08-5.03 (m, 1H), 3.41-3.31 (m, 2H), 2.28-2.20 (m, 1H), 1.80-1.72 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ: 9.27 (s, 2H), 8.33 (s, 1H), 7.89 (s, 1H), 7.65-7.43 (m, 6H), 6.51 (d, J = 2.6 Hz, 1H), 5.20-5.08 (m, 1H), 3.20-3.14 (m, 2H), 2.49-2.43 (m, 1H), 1.75-1.61 (m, 1H).
A mixture of 31a (60 mg, 0.09 mmol) (The intermediate was synthesized according to the procedure of Example 1), CuI (10 mg, 0.05 mmol), Pd(PPh3)2Cl2 (50 mg, 0.05 mmol), DIEA (0.2 mL) and (trimethylsilyl)acetylene (0.5 mL) were stirred at r.t. in DMF (5 mL) under N2 for 3 h. The mixture was diluted with DCM and washed with water three times and brine once, dried over Na2SO4, filtered and concentrated. The residue was purified by flash chromatography to give 31b as a brown solid (30 mg, yield: 52%).
Cooled in ice-batch, to 31b (30 mg, 0.046 mmol) was added TFA (5 mL) and the mixture was stirred 0.5 h at 0° C., then 1.5 h at r.t. The reaction mixture was concentrated and the resulting residue was diluted with MeOH (10 mL). Then Conc. NH3.H2O aq. (5 mL) was added and the mixture was stirred for another 2 h. After concentration, the residue was purified by chromatography eluting with MeOH/water to give Compound 185 as a solid (12 mg, yield: 56%). MS (m/z): 460.2 (M+H)+; 1H NMR (400 MHz, DMSO-d6) δ: 12.41 (s, 1H), 8.21 (s, 1H), 8.14 (s, 1H), 7.67-7.52 (m, 5H), 7.49-7.43 (m, 1H), 6.66-6.62 (m, 1H), 5.05-4.95 (br, 1H), 4.33-4.23 (m, 1H), 3.78-3.72 (m, 1H), 2.49-2.44 (m, 1H), 2.40 (s, 3H), 1.89-1.79 (m, 1H).
A mixture of 33a (prepared according to the procedures described in Example 41 using the corresponding reagents and intermediates) (180 mg, 0.392 mmol), phenylboronic acid (96 mg, 0.784 mmol), Cu(OAc)2 (143 mg, 0.784 mmol) and pyridine (0.125 mL, 1.568 mmol) in DCM (20 mL) was stirred at r.t. overnight, then filtered and concentrated. The residue was further purified by flash chromatography eluting with water and methanol to give 33-b as a white solid. Yield: 4.6%. MS (m/z): 551.1 (M+1)+
A solution of 33b (10 mg, 0.0181 mmol) in HCl/MeOH (2 N, 2 mL) was stirred at r.t. for 15 min, then neutralized with aq. NaHCO3 and extracted with EtOAc three times. The combined organic layers were dried, concentrated and purified by flash chromatography to give Compound 293 as a white solid. Yield: 51%. 1H NMR (400 MHz, CD3OD) δ 8.34 (s, 1H), 8.19-7.89 (m, 2H), 7.82-7.44 (m, 4H), 7.36-7.23 (m, 1H), 6.48-6.41 (m, 1H), 4.59-4.51 (m, 3H), 3.36-3.32 (m, 2H). MS (m/z): 467.1 (M+H)+.
A mixture of 33a (2.5 g, 5.45 mmol), phenylboronic acid (1.33 g, 10.9 mmol), Cu(OAc)2 (1.98 g, 10.9 mmol), pyridine (2.2 mL, 27.25 mmol) and 4 Å molecular sieves in DCM (60 mL) was stirred at r.t. under O2 overnight, then filtered and concentrated. The residue was purified by flash chromatography to give 33b′ as a white solid. Yield: 0.7%. MS (m/z): 535.5 (M+1)+.
A solution of 33b′ (20 mg, 0.0374 mmol) in HCl/MeOH (2 N, 2 mL) was stirred at r.t. for 10 min, then neutralized with aq. NaHCO3 and concentrated and purified by flash chromatography to give Compound 294 as a white solid. Yield: 80%. 1H NMR (400 MHz, DMSO-d6) δ: 12.94 (br, 1H), 8.12-7.93 (m, 2H), 7.62-7.20 (m, 6H), 6.44-6.35 (m, 1H), 5.80-5.46 (m, 1H), 4.98-4.65 (m, 2H), 2.91-2.77 (m, 2H). MS (m/z): 451.4 (M+1)+.
Under N2, a mixture of 34a (prepared according to the procedures described in Example 1 using the corresponding reagents and intermediates) (50 mg, 0.07 mmol), tributyl(1-ethoxyvinyl)stannane (100 mg, 0.28 mmol) and Pd(PPh3)2Cl2 (100 mg, 0.14 mmol) in 5 mL of dioxane was stirred at reflux for 3 h. After cooling to r.t., to the reaction was added 0.5 mL of aq. 1N HCl. The mixture was stirred at r.t. for 3 h. Then the mixture was diluted with DCM, washed with water, brine, dried over Na2SO4, filtered and concentrated. The residue was purified by flash chromatography to give 34b as a brown solid. Yield: 46%. MS (m/z): 608.2 (M+1)+
The mixture of 34b (20 mg, 0.03 mmol) in TFA (5 mL) was stirred at 0° C. for 0.5 h, then concentrated, the resulting residue was diluted with MeOH (10 mL), followed by conc. NH3.H2O aq. (5 mL), the mixture was stirred for 2 h. After concentration, the residue was purified by p-TLC to give Compound 296 as a white solid (3 mg, yield: 19%). 1H NMR (400 MHz, DMSO-d6) δ: 8.09 (s, 1H), 8.03 (s, 1H), 7.74-7.09 (m, 5H), 6.67-6.57 (m, 1H), 4.98-4.84 (br, 1H), 4.31-4.18 (m, 1H), 3.71-3.61 (m, 1H), 2.31 (s, 3H), 1.96-1.90 (m, 1H), 1.80-1.75 (m, 1H). MS (m/z): 478.2 (M+1)+;
The following Compounds were prepared according to the procedure of Compound 296 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 7.97 (s, 1H), 7.77 (d, J = 7.6 Hz, 1H), 7.63-7.50 (m, 5H), 6.58 (d, J = 2.8 Hz, 1H), 4.46 (t, J = 6.4 Hz, 1H), 4.06-4.00 (m, 1H), 3.81- 3.75 (m, 1H), 2.61 (s, 3H), 2.11-1.93 (m, 3H), 1.63-1.58 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 8.54 (s, 1H), 8.07-8.00 (m, 1H), 7.94-7.89 (m, 2H), 7.85-7.81 (m, 2H), 7.65-7.63 (m, 1H), 6.77 (br, 1H), 4.88-4.81 (m, 0.5H), 4.32-4.22 (m, 0.5H), 2.75 (s, 3H), 2.40-2.31 (m, 1H), 1.95-1.87 (m, 0.5H), 1.67-1.62 (m, 0.5H)
1H NMR (400 MHz, DMSO-d6) δ 8.42 (s, 1H), 7.71-7.47 (m, 5H), 7.39-7.36 (m, 1H), 6.82 (s, 2H), 6.62 (d, J = 3.0 Hz, 1H), 4.87-4.75 (m, 1H), 4.15-4.08 (m, 1H), 3.29-3.28 (m, 1H), 2.43-2.35 (m, 1H), 2.23 (s, 3H), 2.03-1.75 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 7.93 (s, 1H), 7.63-7.43 (m, 6H), 7.20 (s, 2H), 6.66 (d, J = 2.8 Hz, 1H), 4.96-4.92 (m, 1H), 4.00-3.99 (m, 2H), 2.41 (s, 3H), 2.02-1.89 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 8.43 (s, 1H), 7.76-7.53 (m, 4H), 7.44-7.25 (m, 2H), 6.52 (d, J = 2.8 Hz, 1H), 4.99-4.93 (m, 1H), 4.50-4.28 (m, 1H), 3.41-3.34 (m, 1H), 2.69-2.40 (m, 1H), 2.26 (s, 3H), 0.67 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, CD3OD) δ 8.42 (s, 1H), 7.70-7.64 (m, 1H), 7.37-7.26 (m, 4H), 6.53 (d, J = 3.2 Hz, 1H), 5.31-5.14 (m, 1H), 4.33-4.27 (m, 1H), 3.83-3.59 (m, 1H), 2.39-2.31 (m, 1H), 2.27 (s, 3H), 2.19-2.10 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 7.66 (d, J = 3.0 Hz, 1H), 7.64-7.49 (m, 4H), 7.42-7.36 (m, 1H), 6.65 (d, J = 3.0 Hz, 1H), 6.43 (s, 2H), 4.73-4.69 (m, 1H), 3.80-3.75 (m, 1H), 2.49-2.39 (m, 1H), 2.32 (s, 3H), 2.10 (s, 3H), 1.93-1.86 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 8.10 (s, 1H), 7.75 (s, 1H), 7.63-7.54 (m, 3H), 7.47 (s, 1H), 7.38 (d, J = 6.4, 1H), 6.56 (dd, J = 3.0, 1.7, 1H), 5.34-4.84 (m, 1H), 4.25-3.60 (m, 2H), 1.23 (s, 3H), 0.76 (s, 3H).
A mixture of 35a (prepared according to the procedures described in Example 1 using the corresponding reagents and intermediates) (100 mg, 0.21 mmol), 2-aminoethanolin (13 mg, 0.21 mmol), HBTU (88 mg, 0.23 mmol) and DIEA (54 mg, 0.42 mmol) in DMF (25 mL) was stirred at r.t. for 6 h. Then the reaction was diluted with water and extracted with EtOAc. The organic layers were dried, concentrated and purified by flash chromatography to give 35b as a white solid. Yield: 50%. MS (m/z): 519.0 (M+1)+.
To a mixture of 35b (54 mg, 0.104 mmol), Et3N (0.115 mL, 0.832 mmol) and DMAP (25 mg, 0.208 mmol) in DCM/DMF (4 mL/1 mL) at 0° C. was added MsCl (0.021 mL, 0.260 mmol). The mixture was stirred at r.t. for 3 h, then quenched by water and extracted with EtOAc. The combined organic layer was concentrated and purified by flash chromatography to give Compound 303 as a white solid. Yield: 38%. 1H NMR (400 MHz, DMSO-d6) δ 12.12 (br, 1H), 8.17 (s, 1H), 7.57-7.46 (m, 7H), 6.55 (d, T=2.9 Hz, 1H), 4.55 (br, 1H), 4.31-4.26 (m, 1H), 3.91-3.82 (m, 2H), 3.80-3.71 (m, 1H), 2.11-1.78 (m, 6H). MS (m/z): 501.2 (M+1)+.
The following Compounds were prepared according to the procedure of Compound 303 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 11.94 (brs, 1H), 8.19 (s, 1H), 8.08 (t, J = 5.6 Hz, 1H), 7.65-7.61 (m, 3H), 7.55-7.52 (m, 2H), 7.49-7.46 (m, 2H), 6.65 (d, J = 3.0 Hz, 1H), 5.08-5.04 (m, 1H), 4.67 (brs, 1H), 4.30-4.18 (m, 1H), 3.78 (m, 1H), 3.47-3.44 (m, 2H), 3.28-3.18 (m, 2H), 2.56-2.52 (m, 1H), 1.88-1.85 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 12.22 (brs, 1H), 8.21 (s, 1H), 7.70 (d, J = 3.0 Hz, 1H), 7.64-7.59 (m, 2H), 7.58-7.52 (m, 3H), 7.47-7.44 (m, 1H), 6.66 (d, J = 3.0 Hz, 1H), 4.94-4.92 (m, 1H), 4.41-4.33 (m, 2H), 4.30-4.23 (m, 1H), 3.97-3.85 (m, 2H), 3.82-3.73 (m, 1H), 2.58-2.53 (m, 1H), 1.94-1.87 (m, 1H).
A mixture of Compound 211 (100 mg, 0.211 mmol), hydroxylamine hydrochloride (44 mg, 0.633 mmol), sodium acetate (42 mg, 0.506 mmol) in ethanol (7.5 mL) and water (5 mL) was stirred at reflux overnight, then concentrated. The residue was purified by flash chromatography to give Compound 306 (Yield: 55%) and Compound 307
Compound 306: 1H NMR (400 MHz, DMSO-d6) δ 11.81 (s, 1H), 10.80 (s, 1H), 8.15 (s, 1H), 7.73 (d, J=8.0 Hz, 1H), 7.58-7.43 (m, 4H), 7.40 (d, J=2.8 Hz, 1H), 7.16 (s, 1H), 6.56 (d, J=2.7 Hz, 1H), 4.66-4.62 (m, 1H), 3.67-3.64 (m, 2H), 2.15 (s, 3H), 2.10-2.04 (m, 2H), 1.96-1.61 (m, 2H); MS (m/z): 489.2 (M+1)+.
Compound 307: 1H NMR (400 MHz, DMSO-d6) δ 11.80 (s, 1H), 10.35 (s, 1H), 8.09 (s, 1H), 7.74-7.56 (m, 1H), 7.69-7.38 (m, 5H), 7.18 (s, 1H), 6.57 (d, J=2.9 Hz, 1H), 4.57-4.51 (m, 1H), 3.81-3.72 (m, 1H), 3.70-3.58 (m, 1H), 2.19 (s, 3H), 2.12-2.02 (m, 2H), 1.87-1.72 (m, 2H). MS (m/z): 489.2 (M+1)+.
The following Compound 308 were prepared according to the procedure of Compound 306 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ: 11.90 (s, 1H), 8.15 (s, 1H), 7.59-7.54 (m, 1H), 7.51-7.27 (m, 4H), 7.37 (d, J = 2.7 Hz, 1H), 7.27 (s, 1H), 6.56 (d, J = 2.7 Hz, 1H), 4.69-4.62 (m, 1H), 3.85 (s, 3H), 3.72-3.61 (m, 1H), 3.60-3.48 (m, 1H), 2.18 (s, 3H), 2.09-2.01 (m, 2H), 1.97-1.85 (m, 1H), 1.71-1.62 (m, 1H).
37a (prepared according to the procedures described in Example 1 using the corresponding reagents and intermediates) (407 mg, 1.25 mmol) was dissolved in DCM (3 mL), DIPEA (674 uL) was added, the mixture was stirred at r.t. for 2 min, Pyridine-N-oxide (95 mg, 1 mmol) was added, followed by PyBrOP (620 mg, 1.33 mmol), the reaction was stirred at r.t. overnight, then concentrated and purified by flash column chromatography to give product 37b as a white solid. Yield: 12%, Ms: 402.1 (M+1)+.
Compound 309 was prepared according to the procedures described in Example 1 from 37b. 1H NMR (400 MHz, DMSO-d6) δ 8.69-8.68 (m, 1H), 8.28 (s, 1H), 8.27 (s, 1H), 8.09-8.06 (m, 1H), 7.73 (d, J=2.8 Hz, 1H), 7.71 (d, J=8.0 Hz, 1H), 7.63-7.59 (m, 1H), 6.69 (d, J=3.2 Hz, 1H), 5.18-5.14 (m, 1H), 4.41-4.36 (m, 1H), 4.19-4.13 (m, 1H), 2.67-2.61 (m, 1H), 2.12-2.06 (m, 1H). MS (m/z): 444.1 (M+1)+.
The following Compounds were prepared according to the procedure of Compound 309 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 8.68-8.67 (m, 1H), 8.16 (s, 1H), 8.11-8.06 (m, 2H), 7.72-7.70 (m, 2H), 7.61-7.58 (m, 1H), 6.69 (d, J = 3.2 Hz, 1H), 5.01 (br, 1H), 4.33 (br, 1H), 3.68-3.67 (m, 1H), 2.46 (br, 1H), 2.41 (s, 3H), 1.73 (br, 1H).
1H NMR (400 MHz, CD3OD) δ 8.65 (d, J = 4.2 Hz, 1H), 8.10-8.06 (m, 1H), 7.67 (d, J = 8.0 Hz, 1H), 7.61-7.58 (m, 1H), 7.44 (brs, 1H), 6.54 (d, J = 2.8 Hz, 1H), 4.62-4.42 (m, 1H), 3.65 (br, 1H), 3.43-3.35 (m, 3H), 2.55-1.97 (m, 4H).
1H NMR (400 MHz, CD3OD) δ 8.70 (d, J = 4.1, 1H), 8.114-8.09 (m, 1H), 7.75 (d, J = 7.8, 1H), 7.64 (dd, J = 7.5, 4.9, 1H), 7.37 (s, 1H), 6.54 (d, J = 2.9, 1H), 4.89 (br, 1H), 4.46-4.41 (m, 1H), 3.36 (br, 1H), 2.68 (br, 1H), 2.29 (s, 3H).
1H NMR (400 MHz, CD3OD) δ 8.57 (dd, J = 4.9, 1.9, 1H), 8.33 (s, 1H), 8.00 (td, J = 7.8, 1.9, 1H), 7.60 (d, J = 7.9, 1H), 7.52 (dd, J = 7.5, 4.9, 1H), 7.32 (d, J = 2.8, 1H), 6.46 (d, J = 3.0, 1H), 4.94 (br, 1H), 4.26-4.19 (m, 1H), 3.63 (br, 1H), 2.38-2.29 (m, 1H), 2.20 (s, 3H), 2.14-2.07 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 8.62 (dd, J = 4.9, 1.9, 1H), 8.07-7.95 (m, 2H), 7.62 (d, J = 7.9, 1H), 7.55 (dd, J = 7.5, 4.9, 1H), 7.38 (d, J = 2.9, 1H), 6.48 (dd, J = 3.0, 0.5, 1H), 4.60-4.44 (m, 1H), 4.39- 4.30 (m, 1H), 3.75-3.53 (m, 1H), 2.86-2.65 (m, 1H), 0.81 (d, J = 6.9, 3H).
1H NMR (400 MHz, CD3OD) δ 8.61 (dd, J = 4.9, 1.2, 1H), 7.99 (td, J = 7.7, 1.9, 1H), 7.62 (d, J = 7.9, 1H), 7.54 (ddd, J = 7.5, 4.9, 0.9, 1H), 7.37 (d, J = 3.0, 1H), 6.48 (d, J = 3.2, 1H), 4.57-4.47 (m, 1H), 4.42-4.25 (m, 1H), 3.79-3.52 (m, 1H), 2.80-2.66 (m, 1H), 2.21 (s, 3H), 0.80 (d, J = 6.9, 3H).
38a (prepared according to the procedure of Example 1 using the corresponding reagents and intermediates) (40 mg, 0.08 mmol) and m-CPBA (37 mg, 75%, 0.16 mmol) were dissolved in DCM (3 mL), the reaction was stirred at r.t. overnight. The mixture was used for the next step without purification. MS (m/z): 531.0 (M+1)+.
To the mixture above was added NH3/THF (0.4 N, 3 mL), the reaction was stirred at r.t. for 2 h, then concentrated and purified by TLC to give Compound 314 as a white solid. Yield: 10.8%. 1H NMR (400 MHz, DMSO-d6) δ 7.88-7.14 (m, 1H), 7.57-7.52 (m, 5H), 7.39 (br, 1H), 6.83-6.59 (m, 3H), 5.34 (br, 0.5H), 4.88 (br, 0.5H), 4.45 (br, 0.5H), 4.17 (br, 0.5H), 4.03 (br, 0.5H), 2.64-2.52 (m, 2H), 2.33 (br, 0.5H). MS (m/z): 468.0 (M+1)+.
The following Compounds were prepared according to the procedure of Compound 314 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 8.00 (s, 1H), 7.76-7.42 (m, 6H), 7.05 (br, 2H), 6.61 (br, 1H), 5.34 (br, 0.5H), 4.90 (br, 0.5H), 4.44 (br, 1H), 4.15 (br, 1.H), 2.65-2.53 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 7.98 (s, 0.7H), 7.87 (s, 0.3H), 7.85-7.70 (m, 3H), 7.58-7.43 (m, 3H), 7.40 (d, J = 7.3 Hz, 1H), 7.32 (d, J = 8.9 Hz, 1H), 7.27-7.20 (m, 1H), 6.48-6.33 (m, 1H), 5.67-5.49 (m, 1H), 4.01-3.88 (m, 1H), 3.80-3.65 (m, 1H), 2.25-2.16 (m, 1H), 2.00-1.91 (m, 2H), 1.88-1.80 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 7.94 (d, J = 8.0 Hz, 0.5H), 7.85 (d, J = 8.0 Hz, 0.5H), 7.68-7.54 (m, 4H), 6.93 (s, 1H), 6.78 (s, 0.5H), 6.63-6.61 (m, 1H), 6.39 (d, J = 4.0 Hz, 0.5H), 5.64 (d, J = 4.0 Hz, 0.5H), 4.72 (d, J = 8.0 Hz, 0.5H), 4.54-4.42 (m, 0.5H), 4.35-4.18 (m, 0.5H), 3.96-3.88 (m, 0.5H), 3.75-3.67 (m, 0.5H), 2.37-2.28 (m, 1H), 2.21-2.11 (m, 1H), 2.04-1.88 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 7.69 (s, 1H), 7.15-7.10 (m, 1H), 6.94-6.91 (m, 1H), 6.84-6.57 (m, 5H), 6.40-6.37 (m, 2H), 5.71 (d, J = 2.9, 1H), 4.18 (t, J = 7.6, 1H), 2.85-2.79 (m, 1H), 2.09-2.00 (m, 1H), 1.70 (s, 3H), 1.13-1.08 (m, 1H), 1.00- 0.94 (m, 2H), 0.81-0.701 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 7.68-7.63 (m, 1H), 7.62-7.50 (m, 4H), 7.45-7.39 (m, 1H), 7.15 (br, 2H), 6.51 (d, J = 3.2 Hz, 1H), 5.09-4.72 (m, 1H), 4.25-3.91 (m, 2H), 2.22 (s, 3H), 2.12-1.95 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 7.67-7.47 (m, 5H), 7.33-7.28 (m, 1H), 7.25 (s, 1H), 6.32 (d, J = 3.1 Hz, 1H), 5.20 (br, 1H), 4.27 (sbr, 1H), 3.73 (br, 1H), 2.38-2.31 (m, 1H), 2.27 (s, 3H), 2.15-2.04 (m, 1H).
1H NMR (400 MHz, CDCl3) δ 8.32 (s, 1H), 7.68-7.59 (m, 2H), 7.57-7.46 (m, 2H), 7.20-7.15 (m, 1H), 6.99 (br, 1H), 6.20 (d, J = 3.1 Hz, 1H), 5.22-5.13 (m, 1H), 4.47-4.30 (m, 1H), 3.52-3.28 (m, 1H), 2.48-2.32 (m, 1H), 2.24 (s, 3H), 0.70 (d, J = 6.9 Hz, 3H).
1H NMR (400 MHz, CD3OD) δ 8.41 (br, 1H), 7.30-7.64 (m, 7H), 6.51 (s, 1H), 5.33 (br, 1H), 4.35-3.81 (m, 2H), 2.39 (br, 1H), 2.20-2.16 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.51 (s, 1H), 7.60-7.38 (m, 5H), 7.39-7.38 (m, 1H), 6.74 (s, 2H), 6.61 (d, J = 2.9 Hz, 1H), 4.89 (brs, 1H), 4.06-4.00 (m, 1H), 2.44-2.35 (m, 2H), 1.90 br (s, 1H), 0.85-0.78 (m, 4H).
1H NMR (400 MHz, CD3OD) δ 8.52 (s, 1H), 7.58-7.54 (m, 1H), 7.521-7.46 (m, 2H), 7.45-7.39 (m, 2H), 7.27 (s, 1H), 6.62 (s, 1H), 2.44 (s, 3H), 1.38 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, CD3OD) δ 8.36 (s, 1H), 7.68-7.54 (m, 4H), 7.34-7.29 (m, 2H), 6.53-6.52 (m, 1H), 5.26 (br, 1H), 4.28-4.22 (m, 1H), 3.80 (br, 1H), 2.76- 2.70 (m, 1H), 2.54 (br, 1H), 2.39-2.31 (m, 1H), 2.20-2.10 (m, 1H), 0.89 (br, 3H).
1H NMR (400 MHz, CD3OD) δ 8.46 (s, 1H), 7.49 (s, 1H), 7.46-7.43 (m, 1H), 7.37-7.35 (m, 1H), 7.24-7.14 (m, 1H), 7.06-6.97 (m, 1H), 6.88-6.85 (m, 1H), 6.59-6.57 (m, 1H), 5.06-5.01 (m, 1H), 2.41 (s, 3H), 1.42-1.40 (m, 3H).
1H NMR (400 MHz, CD3OD) δ 7.62- 7.56 (m, 4H), 7.37-7.34 (m, 1H), 7.297 (br, 1H), 6.33 (d, J = 3.1, 1H), 4.83-4.81 (m, 1H), 4.40 (br, 1H), 3.64 (br, 1H), 2.65 (br, 1H), 2.29 (s, 3H), 0.70 (d, J = 6.7, 3H).
1H NMR (400 MHz, CDCl3) δ 7.53-7.44 (m, 4H), 7.16 (br, 1H), 7.13-7.10 (m, 1H), 6.41 (d, J = 2.9, 1H), 5.09 (s, 2H), 4.75 (br, 1H), 4.38 (br, 1H), 3.60 (br, 1H), 2.47 (br, 1H), 2.31 (s, 3H), 0.70 (d, J = 6.3, 3H).
1H NMR (400 MHz, CD3OD) δ 8.41 (s, 1H), 7.71_(br, 1H), 7.40-7.28 (m, 3H), 7.18 (br, 1H), 6.30 (d, J = 2.1, 1H), 4.91 (br, 1H), 4.41-4.36 (m, 1H), 3.36 (br, 1H), 2.55 (br, 1H), 2.25 (s, 3H), 0.75 (d, J = 6.8, 3H).
1H NMR (400 MHz, CD3OD) δ 7.69-7.63 (m, 1H), 7.43-7.29 (m, 4H), 6.54 (d, J = 3.0, 1H), 4.87 (br, 1H), 4.40 (br, 1H), 3.67 (br, 1H), 2.69 (br, 1H), 2.29 (s, 3H), 0.80 (d, J = 6.8, 3H).
1H NMR (400 MHz, DMSO-d6) δ 8.42 (s, 1H), 7.66-7.49 (m, 3H), 7.40-7.36 (m, 1H), 7.26 (d, J = 7.6 Hz, 1H), 6.83 (d, J = 7.6 Hz, 2H), 6.62 (d, J = 2.8 Hz, 1H), 4.97-4.67 (m, 1H), 4.16-4.09 (m, 1H), 3.45-3.40 (m, 1H), 2.43-2.35 (m, 1H), 2.24 (s, 3H), 2.00-1.88 (m, 1H).
1H NMR (400 MHz, CDCl3) δ 7.58-7.45 (m, 1H), 7.41-7.31 (m, 1H), 7.29-7.21 (m, 2H), 6.95-6.88 (m, 1H), 6.51 (d, J = 3.2 Hz, 1H), 5.12 (s, 2H), 4.47-4.31 (m, 1H), 4.20-4.07 (m, 1H), 2.38 (s, 3H), 2.35-2.31 (m, 1H), 1.79-1.42 (m, 2H).
1H NMR (400 MHz, CDCl3) δ 7.59-7.52 (m, 1H), 7.28-7.09 (m, 4H), 6.50-6.49 (m, 1H), 5.14 (br, 2H), 4.48-4.32 (m, 1H), 421-4.07 (m, 1H), 2.37 (s, 3H), 2.34-2.31 (m, 1H), 1.60-1.49 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 7.60-7.52 (m, 4H), 7.33-7.29 (m, 1H), 6.94 (s, 1H), 5.24-5.17 (m, 1H), 4.35-4.26 (m, 1H), 4.09-4.01 (m, 1H), 2.45-2.38 (m, 1H), 2.35-2.30 (m, 1H), 2.28 (s, 3H), 2.24 (s, 3H).
1H NMR (400 MHz, CD3OD) δ 7.67 (br, 1H), 7.53-7.38 (m, 4H), 7.29 (d, J = 6.8, 1H), 6.48 (d, J = 3.0, 1H), 4.72 (br, 1H), 4.66 (br, 1H), 3.74 (br, 1H), 2.23 (s, 3H), 1.15 (s, 3H), 0.67 (s, 3H).
1H NMR (400 MHz, DMSO-d6) δ 7.71 (d, J = 2.8, 1H), 7.66 (br, 1H), 7.56-7.51 (m, 1H), 7.44-7.39 (m, 2H), 6.64 (d, J = 3.0, 1H), 6.49 (s, 2H), 6.36 (s, 2H), 4.50 (br, 1H), 4.21 (br, 1H), 3.53 (br, 1H), 2.74- 2.69 (m, 1H), 0.70 (d, J = 6.4, 3H).
1H NMR (400 MHz, DMSO-d6) δ 7.69 (d, J = 2.9, 1H), 7.57-7.52 (m, 4H), 7.47- 7.44 (m, 1H), 6.63 (d, J = 3.0, 1H), 6.47 (s, 2H), 6.34 (s, 2H), 4.51 (br, 1H), 4.21 (br, 1H), 3.51 (br, 1H), 2.69 (br, 1H), 0.61 (d, J = 6.4, 3H).
1H NMR (400 MHz, DMSO-d6) δ 7.73 (d, J = 3.0, 1H), 7.58-7.50 (m, 4H), 7.39- 7.36 (m, 1H), 6.64 (d, J = 3.0, 1H), 6.48 (s, 2H), 6.35 (s, 2H), 4.83 (br, 1H), 4.01 (br, 1H), 3.96-3.89 (m, 1H), 3.47-3.38 (m, 1H), 2.00 (br, 1H).
1H NMR (400 MHz, CD3OD) δ 7.60- 7.52 (m, 4H), 7.33-7.29 (m, 1H), 6.94 (s, 1H), 5.24-5.17 (m, 1H), 4.35-4.26 (m, 1H), 4.09-4.01 (m, 1H), 2.45-2.30 (m, 2H), 2.28 (s, 3H), 2.24 (s, 3H).
1H NMR (400 MHz, CD3OD) δ 8.40 (s, 1H), 7.66-7.61 (m, 1H), 7.58-7.52 (m, 3H), 7.28-7.26 (m, 1H), 6.90 (s, 1H), 5.39-5.29 (m, 1H), 4.34-4.27 (m, 1H), 3.89-3.78 (m, 1H), 2.32-.2.19 (m, 2H), 2.17 (s, 3H), 2.14 (s, 3H).
1H NMR (400 MHz, DMSO-d6) δ 7.87- 7.33 (m, 5H), 6.65 (d, J = 3.2, 1H), 6.49 (s, 2H), 6.38 (s, 1H), 6.36 (s, 1H), 4.48 (br, 1H), 4.22 (br, 1H), 3.54 (br, 1H), 2.75 (br, 1H), 0.75-0.69 (m, 3H).
To a mixture of 39a (prepared according to the procedure of Example 1 using the corresponding reagents and intermediates) (23 mg, 0.051 mmol) in dioxane (4 mL) were added diphenylmethanimine (18 mg, 0.102 mmol), Pd(OAc)2 (2.2 mg, 0.001 mmol), 2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (6.2 mg, 0.001 mmol) and Cs2CO3 (41 mg, 0.128 mmol) at r.t., the reaction was stirred at 110° C. overnight under N2.
After cooling to the r.t., 1M HCl (1 mL) was added to the mixture, the reaction was stirred at r.t. for 20 min, then concentrated, the resulting residue was dissolved in MeOH, and adjusted to PH˜7 with DIEA, the mixture was concentrated and purified by flash column chromatography to give Compound 329 as a yellow solid. Yield: 36%. 1H NMR (400 MHz, CDCl3) δ 7.63-7.56 (m, 1H), 7.55-7.44 (m, 3H), 7.30-7.27 (m, 1H), 7.28 (d, J=3.0 Hz, 1H), 7.18-7.13 (m, 1H), 6.48 (d, J=2.9 Hz, 1H), 6.44 (dd, J=4.4, 2.4 Hz, 1H), 6.37 (s, 1H), 5.11 (dd, J=8.5, 5.9 Hz, 1H), 4.55-4.36 (m, 1H), 4.34-4.24 (m, 1H), 4.19 (s, 2H), 2.59-2.45 (m, 1H), 2.44-2.30 (m, 1H). MS (m/z): 433.1 (M+1)+.
The following Compounds were prepared according to the procedure of Compound 329 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, CD3OD) δ 7.69 (s, 1H), 7.65-7.59 (m, 1H), 7.58-7.50 (m, 4H), 7.29-7.28 (m, 1H), 7.27 (d, J = 3.0 Hz, 1H), 6.46 (d, J = 3.0 Hz, 1H), 5.82 (s, 1H), 5.38 (dd, J = 8.6, 5.1 Hz, 1H), 5.10 (s, 1H), 4.41-4.27 (m, 1H), 4.14-3.98 (m, 1H), 2.58-2.37 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 7.54 (d, J = 8.2 Hz, 1H), 7.40 (t, J = 7.3 Hz, 1H), 7.37-7.27 (m, 2H), 7.20 (d, J = 7.3 Hz, 1H), 7.07 (br, 1H), 7.00 (br, 1H), 6.53 (br, 1H), 6.23 (br, 1H), 6.20 (br, 1H), 4.01-3.91 (m, 1H), 3.78-3.67 (m, 1H), 2.15-2.05 (m, 1H), 1.95-1.77 (m, 2H), 1.71-1.58 (m, 1H).
1H NMR (400 MHz, CDCl3) δ 7.66 (d, J = 7.9 Hz, 1H), 7.55 (br, 1H), 7.44-7.36 (m, 1H), 7.35-7.27 (m, 2H), 6.87-6.80 (m, 1H), 6.18 (d, J = 2.8 Hz, 1H), 5.83 (br, 1H), 5.75 (br, 1H), 5.62 (br, 2H), 4.96 (br, 1H), 4.38-4.18 (m, 1H), 3.64-3.40 (m, 1H), 3.37-3.21 (m, 1H), 2.04-1.67 (m, 4H).
1H NMR (400 MHz, CD3OD) δ 8.59-8.52 (m, 1H), 8.31 (d, J = 2.0 Hz, 1H), 7.65-7.63 (m, 2H), 7.60-7.58 (m, 2H), 7.36-7.33 (m, 1H), 7.27 (d, J = 2.9 Hz, 1H), 6.46 (d, J = 3.2 Hz, 1H), 5.57-5.54 (m, 1H), 4.40-4.38 (m, 1H), 4.23-4.20 (m, 1H), 2.60-2.54 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 8.36 (s, 1H), 7.63-7.55 (m, 5H), 7.42-7.26 (m, 3H), 6.45 (brs, 1H), 5.59 (brs, 1H), 4.38 (brs, 1H), 4.19 (brs, 1H), 2.57 (brs, 2H).
1H NMR (400 MHz, CDCl3) δ 8.44 (s, 1H), 7.86 (d, J = 7.9 Hz, 0.6H), 7.72-7.47 (m, 4.4H), 7.45-7.30 (m, 3H), 7.24-6.98 (m, 2H), 6.49-6.31 (m, 1H), 5.97 (d, J = 6.8 Hz, 0.6H), 4.98 (s, 0.4H), 4.77-4.63 (m, 0.4H), 4.55-4.40 (m, 0.4H), 4.12-3.97 (m, 0.6H), 3.95-3.80 (m, 0.6H), 2.14-1.84 (m, 4H).
To a solution of 41a (prepared according to the procedures described in Example 1 using the corresponding reagents and intermediates) (155 mg, 0.65 mmol) in CH3CN (15 mL) were added DIEA (0.32 mL, 1.95 mmol) and 4-chloro-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile (201 mg, 0.65 mmol), the reaction was stirred at 90° C. overnight. The mixture was concentrated and purified by flash column chromatography to give 41b as a yellow solid. Yield: 45%. MS (m/z): 511.2 (M+1)+.
To a solution of 41b (150 mg, 0.29 mmol) in CH2Cl2 (3 mL) was added DIEA (0.15 mL, 0.87 mmol), the reaction was stirred at r.t. for 3 min, then treated with the stock solution of 1M Pyridine-N-oxide in CH2Cl2 (0.232 mL, 0.232 mmol) followed by PyBrOP (135 mg, 0.29 mmol). The reaction was capped and stirred at r.t. overnight. The mixture was concentrated and purified by flash column chromatography to give 41c as a yellow solid. Yield: 17%. MS (m/z): 588.3 (M+1)+.
The solution of 41c dissolved in CF3CO2H (2 mL) was stirred at r.t. for 1 h, then concentrated, the resulting residue was dissolved in MeOH (3 mL), and treated with NH3.H2O (1 mL). The mixture was stirred at r.t. for another 1 h, then concentrated and purified by p-TLC to give Compound 337 as a white solid. Yield: 51%. 1H NMR (400 MHz, DMSO-d6) δ 8.68 (dd, J=4.8, 1.4 Hz, 1H), 8.24 (s, 2H), 8.21 (s, 0.4H), 8.147 (dd, J=4.6, 1.7 Hz, 0.4H), 8.09-8.06 (m, 1H), 8.04 (d, J=2.9 Hz, 0.3H), 8.00 (s, 0.3H), 7.82 (brs, 1H), 7.73-7.69 (m, 0.4H), 7.60-7.57 (m, 2H), 7.28-7.25 (dd, J=4.8, 1.6 Hz, 0.4H), 7.09 (d, J=8.2 Hz, 0.4H), 6.97 (d, J=2.9 Hz, 0.4H), 6.60 (d, J=3.0 Hz, 1H), 5.30-5.26 (m, 1H), 4.49 (s, 1H), 4.02-3.97 (m, 1.4H), 3.94-3.86 (m, 1.4H), 2.30-2.27 (m, 1H), 2.26-2.18 (m, 2H), 2.13-2.06 (m, 1.5H), 2.03-1.95 (m, 3H). MS (m/z): 458.1 (M+1)+.
The following Compounds were prepared according to the procedure of Compound 337 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 12.94 (s, 1H), 8.67 (s, 1H), 8.20-8.08 (m, 3.5H), 7.82-7.80 (m, 0.5H), 7.60-7.49 (m, 2H), 6.58-6.55 (m, 1H), 5.06-5.05 (m, 0.5H), 4.50 (br, 0.5H), 4.20 (br, 0.5H), 4.11-4.07 (m, 0.5H), 3.90-3.85 (m, 0.5H), 3.64-3.62 (m, 0.5H), 2.18-2.16 (m, 1H), 1.99-1.88 (m, 3H).
1H NMR (400 MHz, DMSO-d6) δ 8.08 (d, J = 4.8 Hz, 1H), 8.13-7.99 (m, 4H), 7.62-7.59 (m, 1H), 7.43 (br, 1H), 6.42 (d, J = 3.2 Hz, 1H), 5.07 (br, 1H), 4.07 (br, 2H), 2.33 (br, 1H), 2.12 (br, 1H), 1.99-1.94 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.64 (d, J = 4.8 Hz, 1H), 8.08-7.99 (m, 3H), 7.73 (d, J = 8.0 Hz, 1H), 7.55-7.50 (m, 2H), 6.59 (dd, J = 3.0, 1.0 Hz, 1H), 5.23-5.09 (m, 1H), 4.72 (brs, 1H), 4.21-4.13 (m, 1H), 3.92-3.82 (m, 1H), 2.41 (s, 3H), 2.38-2.28 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.50 (d, J = 4.8 Hz, 1H), 7.85-7.81 (m, 1H), 7.56 (d, J = 8.4 Hz, 1H), 7.42 (d, J = 2.4 Hz, 1H), 7.31 (dd, J = 6.4, 2.4 Hz, 1H), 6.52 (d, J = 6.8 Hz, 1H), 5.67- 5.53 (m, 2H), 4.00 (br, 2H), 1.98- 1.95 (m, 4H).
1H NMR (400 MHz, CD3OD) δ 9.32-9.32 (m, 1H), 9.04 (d, J = 5.2 Hz, 1H), 8.04 (br, 2H), 7.82 (s, 1H), 7.32 (d, J = 3.2 Hz, 1H), 6.47 (d, J = 2.8 Hz, 1H), 4.17 (br, 1H), 4.02 (br, 1H), 2.42 (br, 1H), 2.32-2.14 (m, 3H), 2.08-2.03 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 8.71 (dd, J = 5.1, 1.5 Hz, 1H), 8.28 (s, 1H), 8.12 (td, J = 7.7, 1.9 Hz, 1H), 8.03 (s, 1H), 7.81 (d, J = 8.0 Hz, 1H), 7.61 (dd, J = 7.5, 4.9 Hz, 1H), 7.33 (d, J = 2.9 Hz, 1H), 6.48 (d, J = 3.0 Hz, 1H), 5.52-5.39 (m, J1H), 5.03 (d, J = 7.5 Hz, 1H), 4.55-4.34 (m, 2H), 2.72-2.52 (m, 1H), 2.44-2.25 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.76-8.68 (m, 1H), 8.34 (s, 1H), 8.30 (s, 1H), 8.12 (td, J = 7.7, 1.9 Hz, 1H), 7.84 (d, J = 7.9 Hz, 1H), 7.66-7.59 (m, 1H), 7.51 (dd, J = 4.6, 3.3 Hz, 1H), 6.45 (d, J = 3.2 Hz, 1H), 5.53-5.52 (m, 1H), 4.87 (s, 1H), 4.35 (d, J = 3.7 Hz, 1H), 4.29 (d, J = 3.7 Hz, 1H), 2.47-2.27 (m, 2H).
To a solution of 42a (prepared according to the procedures described in Example 3 using the corresponding reagents and intermediates) (1.32 g, 2.48 mmol) in MeOH (10 mL) was added HCl (3 drops). The mixture was concentrated to give the product 42b as a yellow solid.
To a solution of 42b (1.1 g, 2.45 mmol) in pyridine (10 mL) was added TsCl (0.94 g, 4.9 mmol), the reaction was stirred at r.t overnight under N2, then concentrated and purified by flash column chromatography to give 42c as a yellow solid. Yield 72%. MS (m/z): 603.1 (M+1)+.
To a solution of 42c (1.07 g, 1.77 mmol) in DMSO (10 mL) was added NaCN (435 mg, 8.87 mmol). The reaction was stirred under N2 at 80° C. overnight, then poured into water, and extracted with EtOAc, the organic layers were washed with water, brine, dried, concentrated and purified by flash column chromatography to give 42d as a yellow solid. Yield 56%. MS (m/z): 458.1 (M+1)+.
Compound 347 was prepared according to the procedures described in Example 1 from 42d using the corresponding reagents and intermediates. 1H NMR (400 MHz, CD3OD) δ 8.23 (s, 0.5H), 8.22 (s, 0.5H), 8.00 (s, 0.5H), 7.99 (s, 0.5H), 7.84 (brs, 1H), 7.67-7.59 (m, 1H), 7.41-7.29 (m, 2H), 7.25 (d, J=3.0 Hz, 1H), 6.44 (d, J=3.0 Hz, 1H), 5.34-5.27 (m, 1H), 4.30-4.25 (m, 1H), 3.55-3.45 (m, 1H), 3.35-3.33 (m, 1H), 2.53-2.48 (m, 2H). MS (m/z): 476.1 (M+1)+.
The following Compounds were prepared according to the procedure of Compound 347 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, CD3OD) δ 8.20 (s, 0.5H), 8.20 (s, 0.5H), 7.97 (s, 1H), 7.61-7.53 (m, 2H), 7.37 (d, J = 2.8 Hz, 0.5H), 7.360 (d, J = 2.8 Hz, 0.5H), 7.33-7.26 (m, 2H), 6.47 (d, J = 3.0 Hz, 1H), 5.10-5.01 (m, 1H), 4.58-4.51 (m, 1H), 4.36-4.29 (m, 1H), 3.53-3.44 (m, 1H), 2.60-2.50 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.25 (s, 1H), 7.84 (dd, J = 7.6, 1.6 Hz, 1H), 7.58-7.51 (m, 5H), 7.19-7.11 (br, 2H), 6.60 (d, J = 3.0 Hz, 1H), 4.70 (brs, 1H), 4.34-4.32 (m, 1H), 3.94 (brs, 1H), 2.41-2.35 (m, 1H), 2.18-2.08 (m, 1H), 2.00-1.94 (m, 1H).
43a (prepared according to the procedures described in Example 1 using the corresponding reagents and intermediates) (40 mg, 0.08 mmol) and m-CPBA (19 mg, 75%, 0.08 mmol) were dissolved in DCM, the mixture was stirred at r.t. for 10 min, then concentrated and purified by TLC to give Compound 352 as a white solid. Yield: 61%. 1H NMR (400 MHz, DMSO-d6) δ 8.38 (d, J=2.8 Hz, 1H), 7.80-7.77 (m, 1H), 7.61-7.55 (m, 4.5H), 7.46 (d, J=2.8 Hz, 0.5H), 6.60 (d, J=2.8 Hz, 1H), 4.747-4.66 (m, 1H), 4.42-4.38 (m, 0.5H), 4.24-4.21 (m, 1H), 4.10-4.06 (m, 0.5H), 3.11 (s, 1.5H), 3.86 (s, 1.5H), 2.36-2.24 (m, 2H), 2.07-1.96 (m, 2H). MS (m/z): 495.1 (M+1)+.
The following Compound 353 and Compound 399 were prepared according to the procedure of Compound 352 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 8.19 (s, 0.5H), 8.18 (s, 0.5H), 7.65-7.54 (m, 6H), 6.63 (d, J = 3.2 Hz, 0.5H), 6.62 (d, J = 2.8 Hz, 0.5H), 5.14-5.09 (m, 1H), 4.58-4.47 (m, 1H), 4.26-4.15 (m, 1H), 3.05 (s, 1.5H), 3.018 (s, 1.5H), 2.68-2.60 (m, 1H), 2.20- 2.13 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 8.36 (s, 0.5H), 8.30 (s, 0.5H), 7.66-7.52 (m, 4H), 7.44 (d, J = 3.0 Hz, 0.5H), 7.40 (d, J = 3.0 Hz, 0.5H), 7.34-7.29 (m, 1H), 6.55 (d, J = 3.0 Hz, 0.5H), 6.54 (d, J = 3.0 Hz, 0.5H), 5.09-5.05 (m, 0.5H), 5.01-4.95 (m, 0.5H), 4.30-4.15 (m, 1H), 4.06-3.97 (m, 1H), 2.83 (s, 1.5H), 2.76 (s, 1.5H), 2.53-2.44 (m, 1H), 2.28-2.18 (m, 1H).
To a solution of Compound 299 (52 mg, 0.12 mmol) in MeOH (20 mL) was added NaBH4 (9 mg, 0.24 mmol), the reaction was stirred at r.t. overnight, then quenched with water, the mixture was concentrated and purified by flash column chromatography to give Compound 357 as a white solid. Yield: 32%. 1H NMR (400 MHz, DMSO-d6) δ 8.19 (brs, 1H), 7.84 (brs, 1H), 7.73 (d, J=7.7 Hz, 1H), 7.69 (d, J=2.9 Hz, 1H), 7.62-7.51 (m, 3H), 7.42-7.39 (m, 1H), 6.66 (d, J=2.9 Hz, 1H), 6.07 (s, 2H), 4.77-4.74 (m, 1H), 4.62-4.60 (m, 1H), 4.15-4.10 (m, 1H), 3.99-3.93 (m, 1H), 2.48-2.41 (m, 1H), 1.99-1.91 (m, 1H), 1.30 (d, J=6.3 Hz, 3H). MS (m/z): 438.3 (M+1)+.
To a solution of 48a (prepared according to the procedures described in Example 3 using the corresponding reagents and intermediates) (107 mg, 0.2 mmol) in dry THF (5 ml) was added NaH (12 mg, 0.3 mmol), the mixture was stirred at 0° C. for 0.5 h under N2, then TsCl (760 mg, 0.4 mmol) was added, the reaction was stirred for another 0.5 h. The mixture was concentrated and purified by chromatography to give 48b. Yield: 94%. MS (m/z): 687.3 (M+1)+.
The mixture of 48b (120 mg, 0.188 mmol) and NaCN (460 mg, 0.94 mmol) in dry DMSO (10 mL) was stirred at 55° C. overnight under N2. After reaction, the mixture was cooled to r.t. and poured into water, extracted with EtOAc, the organic layers were concentrated to give 48c, which was used for the next without further purification. MS (m/z): 542.1 (M+1)+.
To a mixture of 48c (100 mg, 0.185 mmol) in methanol (5 mL) was added HCl (1 mL) stirred at 60° C. for 1 h. After reaction, the mixture was concentrated and purified by flash column chromatography to give Compound 358 as a white solid. Yield: 66%. 1H NMR (400 MHz, DMSO-d6) δ 8.15 (s, 1H), 7.98 (s, 1H), 7.62-7.55 (m, 5H), 7.46 (s, 1H), 6.57 (d, J=2.8 Hz, 1H), 2.73-2.65 (m, 2H), 2.569-2.54 (m, 0.5H), 2.46-2.44 (m, 0.5H), 2.23-2.15 (m, 2H), 2.03-1.95 (m, 1H). MS (m/z): 458 (M+1)+
The following Compounds 359-361 were prepared according to the procedure of Compound 358 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, CD3OD) δ 8.10 (s, 1H), 7.94 (d, J = 7.2 Hz, 1H), 7.85 (s, 1H), 7..66-7.53 (m, 3H), 7.46-7.43 (m, 1H), 7.28 (d, J = 3.2 Hz, 1H), 6.45 (d, J = 3.2 Hz, 1H), 5.19-5.13 (m, 1H), 4.36-4.32 (m, 1H), 3.49-3.43 (m, 1H), 3.36-3.33 (m, 1H), 2.47-2.43 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 8.22 (s, 1H), 7.95 (s, 1H), 7.69-7.66 (m, 1H), 7.55-7.44 (m, 3H), 7.37-7.35 (m, 1H), 7.2553 (d, J = 3.2 Hz, 1H), 6.38 (d, J = 3.2 Hz, 1H), 4.95-4.91 (m, 1H), 4.49-4.44 (m, 1H), 4.29-4.24 (m, 1H), 3.45-3.37 (m, 1H), 2.52-2.38 (m, 2H).
1H NMR (400 MHz, CD3OD) δ 8.29 (s, 1H), 8.08 (s, 1H), 7.56-7.32 (m, 3H), 7.26-7.22 (m, 1H), 6.44 (br, 1H), 5.17 (br, 1H), 4.56-4.51 (m, 2H), 3.57-3.50 (br, 1H), 2.55-2.49 (br, 2H).
To a solution of 49a ((prepared according to the procedures described in Example 1 using the corresponding reagents and intermediates) (55 mg, 0.128 mmol) in DMF (1 mL) was added NaH (8 mg, 0.19 mmol) at 0° C., the reaction was stirred at 0° C. for 0.5 h, then 1-bromo-2-methoxyethane (36 mg, 0.256 mmol) was added, the mixture was stirred in a sealed tube at 130° C. overnight. After cooling to r.t., the reaction was quenched with water, then concentrated and purified by flash column chromatography to give 49b. Yield: 27%. MS (m/z): 489.1 (M+1)+.
Compound 264 was prepared according to the procedures described in Example 1 from 49b using the corresponding reagents and intermediates. 1H NMR (400 MHz, DMSO-d6) δ 8.31 (s, 1H), 8.28 (s, 1H), 7.73 (d, J=7.2 Hz, 1H), 7.61-7.49 (m, 5H), 6.56 (d, J=2.8 Hz, 1H), 4.59 (t, J=8.2 Hz, 1H), 4.31 (t, J=7.8 Hz, 1H), 4.17-4.10 (m, 1H), 3.83-3.79 (m, 1H), 3.54-3.48 (m, 2H), 3.42-3.38 (m, 2H), 3.19 (s, 3H), 2.41-2.28 (m, 2H). MS (m/z): 531.3 (M+1)+.
To a solution of NaH (500 mg, 60%, 12.5 mmol) in DMF was added 50a (1.59 g, 10 mmol in 10 mL of DMF) dropwise at 0° C., the reaction was stirred at r.t. for 30 min, then tert-butyl 2-(2-chloroacetyl)pyrrolidine-1-carboxylate (3.0 g, 12 mmol in 10 mL of DMF) was added dropwise at 0° C., the reaction was warmed to r.t. and stirred for 2 h. The mixture was poured into water, extracted with EtOAc, the organic layers were washed with brine, dried over Na2SO4, concentrated to give 50b as a dark oil, which was used for the next step without purification. MS (m/z): 271.1 (M-100+1)+.
50b (3.7 g, 10 mmol) was dissolved in NH3/MeOH (7 N, 100 mL), the reaction was stirred at 130° C. overnight. The mixture was concentrated to about 30 mL, the resulting precipitate was filtered, and poured into water, then 1N HCl (3 mL) was added, the resulting mixture was stirred at r.t. for 5 min, DCM was added until the precipitate was dissolved. The resulting solution was washed with water, dried over Na2SO4, concentrated to give 50c as a brown solid, which was used for next step without purification. Yield: 53%, MS (m/z): 337.9 (M+1)+.
Compound 363 was prepared according to the procedures described in Example 1 from 50c using the corresponding reagents and intermediates. 1H NMR (400 MHz, DMSO-d6) δ 12.94 (br, 1H), 8.27 (s, 1H), 8.21 (br, 1H), 7.57-7.49 (m, 5H), 7.37 (d, J=2.8, 1H), 7.08 (br, 1H), 6.54 (s, 1H), 5.41 (br, 0.5H), 4.79-4.47 (m, 0.5H), 4.10-3.97 (m, 1H), 3.62 (s, 1H), 1.94 (br, 3H), 1.70-1.65 (m, 1H). MS (m/z): 432.4 (M+1)1.
The following Compound 364 was prepared according to the procedure of Compound 363 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 8.31 (s, 1H), 8.30 (s, 1H), 7.66-7.46 (m, 5H), 7.33-7.32 (m, 2H), 6.57 (d, J = 2.8, 1H), 4.81 (dd, J = 7.9, 2.9, 1H), 4.39-4.27 (m, 1H), 3.81-3.78 (m, 1H), 2.24-2.10 (m, 1H), 2.01-1.95 (m, 1H), 1.92-1.86 (m, 1H), 1.80-1.70 (m, 1H).
To a mixture of 51a (prepared according to the procedures described in Example 48 using the corresponding reagents and intermediates) (50 mg, 0.08 mmol) in dry DCM (5 mL) was added m-CPBA (26 mg, 0.15 mmol), the reaction was stirred at r.t. for 24 h. The mixture was concentrated to give 51b as a solid, which was used for the next step without further purification. MS (m/z): 677.1 (M+1)+.
The mixture of 51b (52 mg, 0.079 mmol) in CF3COOH (1 mL) was stirred for 1 h, then concentrated, the resulting residue was added NH3.H2O (1 mL) in MeOH, the mixture was stirred for another 1 h, then concentrated and purified by flash column chromatography to give Compound 365 as a white solid. Yield: 47%. 1H NMR (400 MHz, CD3OD) δ 8.13 (s, 1H), 7.93 (s, 1H), 7.85 (d, J=7.6 Hz, 1H), 7.69-7.64 (m, 1H), 7.59-7.57 (m, 2H), 7.42-7.37 (m, 2H), 6.49 ((d, J=2.4 Hz, 1H), 4.53-4.49 (m, 1H), 4.41-4.36 (m, 1H), 4.09-4.00 (m, 1H), 3.66-3.61 (m, 1H), 3.38 (s, 3H), 2.66-2.54 (m, 2H). MS (m/z): 535.1 (M+1)+.
The following Compound 366 was prepared according to the procedure of Compound 365 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
To a solution of 2,4,6-trichloro-1,3,5-triazine (36.8 mg, 0.2 mmol) in THF (3 mL) were added DIEA (51.6 mg, 0.4 mmol) and a solution of 52a (prepared according to the procedures described in Example 1 using the corresponding reagents and intermediates, about 0.1 mmol) in THF (4 mL) at r.t. The reaction was stirred at r.t for 2 h. The mixture was used directly for next step without purification.
To the above mixture of 52b in THF was added a solution of NH3 in THF (7 N, 3 mL) at r.t., the reaction was stirred at r.t. overnight, then a solution of NH3 in MeOH (7 N, 5 mL) was added, the resulting mixture was stirred at 100° C. overnight in a sealed tube. The mixture was concentrated and purified by flash column chromatography to give 52c as a yellow solid. Yield: 94.6%. MS (m/z): 424.5 (M+1)+.
To a solution of 52c (40 mg, 0.09 mmol) in EtOH (2 mL) was added a solution of 2-chloroacetaldehyde in H2O (40%, 18.4 mg) at r.t., the reaction was stirred at 100° C. overnight. The reaction was concentrated and purified by flash column chromatography and p-TLC to give Compound 367 as a white solid. Yield: 52%. 1H NMR (400 MHz, CD3OD) δ 8.03 (s, 0.4H), 7.86 (s, 0.4H), 7.68-7.62 (m, 1H), 7.56 (br, 2H), 7.46-7.37 (m, 1H), 7.34 (br, 2H), 7.24 (m, 0.4H), 7.09 (br, 1H), 6.47 (br, 1H), 3.92-3.80 (m, 1.4H), 3.68-3.57 (m, 1.4H), 2.24-2.09 (m, 2.8H), 2.00-1.80 (m, 2.8H). MS (m/z): 448.2 (M+1)+.
The following Compound 368 was prepared according to the procedure of Compound 367 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 8.05 (s, 2H), 7.74 (d, J = 2.6 Hz, 1H), 7.63- 7.48 (m, 5H), 7.47-7.41 (m, 1H), 7.10 (s, 1H), 6.68 (d, J = 2.5 Hz, 1H), 4.75-4.64 (m, 1H), 3.92-3.81 (m, 2H), 2.20- 1.79 (m, 2H).
To a solution of 52a (about 0.2 mmol) in n-BuOH (10 mL) was added DIEA (103 mg, 0.8 mmol) and 4-chloro-5-ethynylpyrimidin-2-amine (34 mg, 0.22 mmol) at r.t., the reaction was stirred at 120° C. overnight. The mixture was concentrated and purified by flash column chromatography and p-TLC to afford Compound 369 as a white solid. Yield: 39%. 1H NMR (400 MHz, CD3OD) δ 8.40 (s, 1H), 7.79 (d, J=7.7 Hz, 1H), 7.65-7.50 (m, 3H), 7.45-7.39 (m, 1H), 7.32 (d, J=2.9 Hz, 1H), 6.48 (d, J=3.0 Hz, 1H), 4.81-4.76 (m, 1H), 3.45-3.36 (m, 1H), 3.25-3.14 (m, 1H), 2.48 (s, 3H), 2.17-1.99 (m, 2H), 1.96-1.85 (m, 1H), 1.81-1.67 (m, 1H). MS (m/z): 450.1 (M+1)+.
The mixture of 55a (prepared according to the procedures described in Example 41 using the corresponding reagents and intermediates) (99 mg, 0.2 mmol) and bromomethylcyclopropane (135 mg, 1 mmol) and Cs2CO3 (325 mg, 1 mmol) in DMF (5 mL) was stirred at 120° C. overnight in a sealed flask. After reaction, the reaction mixture was concentrated and purified by flash column chromatography to give 55b as a yellow solid. Yield: 68%. MS (m/z): 551.2 (M+1)+.
Compound 371 was prepared according to the procedures described in Example 41 using 55b instead of 41c. 1H NMR (400 MHz, CD3OD) δ 8.11 (s, 1H), 7.91 (s, 1H), 7.30 (d, J=3.2, 1H), 6.45 (d, J=3.2, 1H), 5.90-5.85 (m, 1H), 4.48-4.42 (m, 1H), 4.18-4.13 (m, 1H), 3.81-3.76 (m, 1H), 3.06-2.97 (m, 1H), 2.66-2.57 (m, 1H), 1.34-1.27 (m, 2H), 0.63-0.506 (m, 4H). MS (m/z): 421.0 (M+1)+.
The following Compounds were prepared according to the procedure of Compound 371 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, CD3OD) δ 8.23 (s, 1H), 7.90 (s, 1H), 7.15 (s, 1H), 6.38 (d, J = 2.8 Hz, 1H), 6.14-6.07 (m, 1H), 4.52-4.36 (m, 1H), 4.28-4.20 (m, 2H), 4.02-3.86 (m, 3H), 3.15 (s, 3H), 2.69- 2.53 (m, 1H), 2.39-2.26 (m, 1H), 2.24-2.09 (m, 2H).
1H NMR (400 MHz, CD3OD) 8.12 (s, 1H), 7.35 (d, J = 2.0 Hz, 1H), 6.49 (d, J = 3.2 Hz, 1H), 5.88 (s, 1H), 4.45- 4.32 (m, 1H), 4.08-4.00 (m, 1H), 3.70-3.63 (m, 1H), 3.03-2.94 (m, 1H), 2.51-2.42 (m, 1H), 1.32-1.14 (m, 2H), 0.60-0.43 (m, 4H).
1H NMR (400 MHz, CD3OD) δ 7.94 (s, 1H), 7.78 (s, 1H), 7.17 (d, J = 2.8, 1H), 6.38 (d, J = 3.2, 1H), 4.36-4.31 (m, 1H), 4.22-4.15 (m, 1H), 4.07-4.02 (m, 1H), 2.54-2.43 (m, 1H), 2.37-2.28 (m, 1H), 2.19-2.13 (m, 2H), 1.69-1.62 (m, 1H), 1.33-1.25 (m, 2H), 0.69-0.55 (m, 4H).
1H NMR (400 MHz, CD3OD) δ 8.04 (s, 1H), 7.87 (s, 1H), 7.32 (d, J = 2.8 Hz, 1H), 6.45 (d, J = 2.8 Hz, 1H), 4.79-4.74 (m, 2H), 4.44-4.38 (m, 1H), 4.17-4.12 (m, 1H), 3.82-3.76 (m, 1H), 3.04-2.95 (m, 1H), 2.638-2.558 (m, 2H), 0.628-0.494 (m, 4H).
1H NMR (400 MHz, DMSO-d6) δ 8.24 (s, 1H), 7.53 (d, J = 2.8 Hz, 1H), 6.58 (d, J = 2.8 Hz, 1H), 4.10-4.02 (m, 2H), 4.00-3.88 (m, 2H), 2.40-2.30 (m, 1H), 2.23-2.21(m, 2H), 2.03-1.96 (m, 2H), 0.87-0.84 (m, 1H), 0.64-0.43 (m, 4H).
To a solution of 56a (prepared according to the procedures described in Example 1 using the corresponding reagents and intermediates) (50 mg, 0.12 mmol) in DCM (5 mL) was added NCS (20 mg, 0.15 mmol), the reaction was stirred at r.t. for 5 h, then concentrated and purified by p-TLC to give Compound 377 as a yellow solid. Yield: 30%. 1H NMR (400 MHz, DMSO-d6) δ 7.71 (d, J=3.0 Hz, 1H), 7.65-7.50 (m, 4H), 7.41-7.34 (m, 1H), 6.64 (d, J=3.0 Hz, 1H), 6.17 (s, 2H), 4.78 (t, J=7.3 Hz, 1H), 4.20-4.15 (m, 1H), 4.00-3.94 (m, 1H), 2.45-2.38 (m, 1H), 2.13 (s, 3H), 1.98-1.87 (m, 1H). MS (m/z): 442.4 (M+1)+.
The mixture of 57a (prepared according to the procedures described in Example 56 using the corresponding reagents and intermediates) (23 mg, 0.046 mmol), CuCN (6 mg, 0.069 mmol) and CuI (1 mg, 0.005 mmol) in DMF (2 mL) was stirred at 120° C. under N2 overnight. The reaction mixture was concentrated and purified flash column chromatography to give Compound 378 as a yellow solid. Yield: 29%. 1H NMR (400 MHz, CD3OD) δ 7.61-7.53 (m, 4H), 7.48 (d, J=3.0 Hz, 1H), 7.33-7.29 (m, 1H), 6.56 (d, J=3.2 Hz, 1H), 5.08 (brs, 1H), 4.23 (brs, 1H), 4.08-4.06 (m, 1H), 3.89 (s, 3H), 2.79-2.41 (m, 1H), 2.25-2.16 (m, 1H). MS (m/z): 449.1 (M+1)+.
To a mixture of Compound 71(30 mg, 0.064 mmol) in dry DMF (25 mL) was added Dess-Martin reagent (54 mg, 0.128 mmol), the reaction was stirred at r.t. for 3 h, then filtered, the filtrate was purified by flash column chromatography to give Compound 380 as a yellow solid. Yield: 83%. 1H NMR (400 MHz, CDCl3) δ 8.38 (s, 1H), 7.78 (s, 1H), 7.67 (d, J=7.6 Hz, 1H), 7.56-7.46 (m, 3H), 7.18-7.16 (m, 1H), 7.02 (d, J=3.2 Hz, 1H), 6.35 (d, J=2.8 Hz, 1H), 5.51 (t, J=5.8 Hz, 1H), 4.66 (d, J=3.2 Hz, 2H), 2.69 (d, J=6.0 Hz, 2H). MS (m/z): 471.1 (M+1)+.
The following Compounds were prepared according to the procedure of Compound 380 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, CD3OD) δ 8.31 (s, 1H), 8.05 (s, 1H), 8.01 (br, 1H), 7.68-7.63 (m, 1H), 7.61-7.55 (m, 2H), 7.45-7.43 (m, 1H), 7.22 (d, J = 2.8 Hz, 1H), 6.43 (d, J = 3.2 Hz, 1H), 4.50-4.43 (m, 1H), 3.73-3.69 (m, 2H), 1.87-1.84 (m, 2H).
1H NMR (400 MHz, DMSO-d6) δ 8.34 (s, 1H), 8.24 (bs, 1H), 7.71-7.63 (m, 2H), 7.61-7.53 (m, 4H), 7.37 (d, J = 6.4 Hz, 1H), 6.40 (d, J = 2.8 Hz, 1H), 4.12-4.06 (m, 1H), 3.17 (s, 2H), 3.09 (d, J = 13.6 Hz, 1 H), 2.87-2.80 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.31 (s, 1H), 7.89 (d, J = 8.0 Hz, 1H), 7.64-7.60 (m, 1H), 7.56-7.53 (m, 3H), 7.50-7.48 (m, 1H), 6.45 (d, J = 3.2 Hz, 1H), 5.33-5.31 (m, 0.2H), 5.12 (d, J = 8.8 Hz, 0.8H), 4.53 (d, J = 17.2 Hz, 0.5H), 4.23 (d, J = 17.2 Hz, 1H), 4.13-4.11 (m, 0.5H), 3.17 (d, J = 4.8 Hz, 2H), 2.99 (d, J = 18.8 Hz, 1H), 2.68-2.58 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 12.52 (br, 1H), 8.26 (s, 1H), 8.25 (s, 1H), 7.64 (d, J = 7.4 Hz, 1H), 7.56-7.55 (m, 2H), 7.49-7.40 (m, 3H), 6.59 (dd, J = 2.9, 0.7 Hz, 1H), 5.28 (d, J = 8.9 Hz, 1H), 4.35-4.22 (m, 2H), 3.00 (d, J = 17.7 Hz, 1H), 2.82-2.75 (m, 1H), 2.50 (s, 3H).
1H NMR (400 MHz, DMSO-d6) δ 8.31 (d, J = 1.0 Hz, 1H), 7.90 (d, J = 7.6 Hz, 1H), 7.65-7.53 (m, 5H), 7.32-7.01 (br, 2H), 6.62 (dd, J = 3.0, 1.0 Hz, 1H), 5.10 (d, J = 9.8 Hz, 1H), 4.51 (d, J = 17.2 Hz, 1H), 4.23 (d, J = 17.3 Hz, 1H), 2.99 (d, J = 17.9 Hz, 1H), 2.64-2.57 (m, 1H).
1H NMR (400 MHz, DMSO-d6) δ 8.32 (d, J = 1.6 Hz, 1H), 7.95-7.40 (m, 5H), 7.16 (br, 2H), 6.64-6.63 (m, 1H), 5.14 (d, J = 9.5 Hz, 0.5H), 5.04 (d, J = 8.5 Hz, 0.5H), 4.52 (dd, J = 17.2, 11.5 Hz, 1H), 4.24 (dd, J = 17.6, 6.6 Hz, 1H), 3.05-2.91 (m, 1H), 2.76-2.60 (m, 1H).
1H NMR (400 MHz, DMSO-d6 + D2O) δ 8.20 (s, 2H), 7.57 (d, J = 7.6 Hz, 1H), 7.51 (brs, 2H), 7.45-7.41 (m, 1H), 7.38-7.35 (m, 1H), 7.32-7.30 (m, 1H), 6.39 (d, J = 3.2 Hz, 1H), 5.29 (d, J = 7.2 Hz, 1H), 4.29-4.17 (m, 2H), 2.77-2.70 (m, 1H), 2.52 (s, 3H).
1H NMR (400 MHz, DMSO-d6) δ 8.51 (s, 1H), 7.78 (d, J = 8.0 Hz, 1H), 7.54-7.49 (m, 4H), 7.34-7.32 (m, 1H), 6.40 (d, J = 3.2 Hz, 1H), 5.31-5.28 (m, 1H), 5.15 (d, J = 8.0 Hz, 1H), 3.79-3.65 (m, 1H), 2.93 (d, J = 19.2 Hz, 1H), 2.66-2.54 (m, 1H), 2.42 (s, 3H).
To a solution of 59a (prepared according to the procedures described in Example 1 using the corresponding reagents and intermediates) (49 mg, 0.11 mmol) in MeOH was added NH3/MeOH (7 N, 5 mL), the mixture was stirred at reflux for 1 h, then concentrated and purified by flash column chromatography to give Compound 189 as a yellow solid. Yield: 44%. 1H NMR (400 MHz, CDCl3) δ 8.05 (s, 1H), 7.71-7.44 (m, 5H), 7.16 (d, J=2.5 Hz, 1H), 6.29 (d, J=2.1 Hz, 1H), 5.56 (s, 2H), 4.88-4.87 (m, 1H), 4.30-4.20 (m, 1H), 3.96-3.89 (m, 1H), 2.49 (s, 3H), 2.40-2.30 (m, 1H), 2.00-1.89 (m, 3H). MS (m/z): 412.7 (M+1)+.
Compound 197 were resolved by chiral HPLC to produce the optically pure enantiomers Compound 382 and Compound 383. HPLC conditions: Gilson system, Column: CHIRALPAK Ia 20 mm I.D.×25 cm L; mobile phase: n-hexane/i-PrOH/DEA=7/3/0.01; flow rate, 10 mL/min; detector: UV 254 nm.
Compound 382 is the first eluent with at least 98% ee. 1H NMR (400 MHz, DMSO-d6) δ 8.24 (s, 1H), 7.74 (d, J=8.2 Hz, 1H), 7.68-7.54 (m, 5H), 7.39 (d, J=3.0 Hz, 1H), 6.59 (d, J=3.0 Hz, 1H), 4.80-4.76 (m, 1H), 3.87-3.79 (m, 2H), 2.93 (s, 1H), 2.15-2.07 (m, 2H), 2.00-1.94 (m, 1H), 1.85-1.73 (m, 1H). MS (m/z): 494.1 (M+1)+.
Compound 383 is the second eluent with at least 98% ee. 1H NMR (400 MHz, DMSO-d6) δ 8.23 (s, 1H), 7.85 (s, 1H), 7.77 (d, J=8.0 Hz, 1H), 7.64-7.53 (m, 4H), 7.49 (d, J=3.0 Hz, 1H), 6.58 (d, J=3.0 Hz, 1H), 4.68-4.65 (m, 1H), 4.25-4.18 (m, 1H), 3.69-3.63 (m, 1H), 2.88 (s, 3H), 2.29-2.18 (m, 2H), 1.97-1.88 (m, 2H). MS (m/z): 494.2 (M+1)+.
2-amino-4-((1-(3-chloro-5-oxo-6-phenyl-5,6-dihydroimidazo[1,2-c]pyrimidin-7-yl)ethyl)amino)pyrimidine-5-carbonitrile was resolved by chiral HPLC to produce the optically pure enantiomers Compound 384 and Compound 385. HPLC conditions: Gilson system, Column: CHIRALPAK Ia 20 mm I.D.×25 cm L; mobile phase: EtOH/DEA=100/0.1; flow rate, 8 mL/min; detector: UV 254 nm.
Compound 384 is the first eluent with at least 95% ee. MS (m/z): 407.0 (M+1)+.
Compound 385 is the second eluent with at least 90% ee. MS (m/z): 407.0 (M+1)+.
Compound 337 was resolved by p-TLC to produce the optically pure enantiomers Compound 386 and Compound 387 with at least 98% ee.
Under the HPLC analysis conditions below, the retention time of Compound 386 is 8.93 min, the retention time of Compound 387 is 8.61 min.
HPLC analysis conditions: Gilson system, Column: Daicel 4.6*250 mm IA; mobile phase: EtOH/DEA=100/0.1; flow rate, 0.5 mL/min; detector: UV 254 nm.
Compound 386: MS (m/z): 512.0 (M+1)+.
Compound 387: MS (m/z): 512.0 (M+1)+.
The mixture of 60a (prepared according to the procedures described in Example 41 using the corresponding reagents and intermediates) in TFA (2 mL) was stirred at r.t. for 1 h. The mixture was concentrated, the resulting residue was dissolved in MeOH (2 mL), and treated with NH3.H2O (25%), the reaction was stirred at r.t. for another 1 h. The mixture was concentrated and purified by flash column chromatography and p-TLC to give Compound 388 and Compound 389 as two yellow solids with at least 98% ee. Under the HPLC analysis conditions below, the retention time of Compound 388 is 8.91 min, the retention time of Compound 389 is 11.22 min.
HPLC analysis conditions: Gilson system, Column: Daicel 4.6*250 mm IA; mobile phase: Hexane: i-PrOH: Et2NH=70:30:0.1; flow rate, 1 mL/min; detector: UV 254 nm.
Compound 388: 1H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J=4.3 Hz, 1H), 8.11 (t, J=7.4 Hz, 1H), 8.06 (s, 1H), 7.83 (br, 1H), 7.71 (s, 1H), 7.64-7.59 (m, 1H), 7.51 (d, J=2.0 Hz, 1H), 6.63 (d, J=2.0 Hz, 1H), 4.73-4.54 (m, 1H), 3.90-3.85 (m, 2H), 2.87 (s, 3H), 2.15-2.10 (m, 2H), 2.04-1.97 (m, 1H), 1.82-1.75 (m, 1H). MS (m/z): 495.0 (M+1)+.
Compound 389: 1H NMR (400 MHz, DMSO-d6) δ 8.66 (s, 1H), 8.18 (s, 1H), 8.12-8.02 (m, 1H), 7.91-7.77 (m, 2H), 7.61-7.48 (m, 2H), 6.58 (d, J=2.9 Hz, 1H), 4.58-4.38 (m, 1H), 4.15-4.02 (m, 1H), 3.68-3.62 (m, 1H), 2.85 (s, 3H), 2.30-2.12 (m, 2H), 2.08-2.00 (m, 1H), 1.98-1.91 (m, 1H). MS (m/z): 495.1 (M+1)+.
37b (40 mg, 0.1 mmol) was dissolved in MeOH (2 mL) and conc.HCl (2 mL), the mixture was concentrated at 50° C., the resulting residue was dissolved in n-BuOH (2 mL) and DIPEA (0.5 mL), then was added 4-chloro-5-(methylsulfinyl)-7H-pyrrolo[2,3-d]pyrimidine (21 mg, 0.1 mmol), the reaction as stirred at reflux for 3 h, then concentrated and purified by flash column chromatography to give Compound 390 and Compound 391 with at least 98% ee.
Under the HPLC analysis conditions below, the retention time of Compound 390 is 10.53 min, the retention time of Compound 391 is 11.64 min.
HPLC analysis conditions: Gilson system, Column: Daicel 4.6*250 mm IA; mobile phase: EtOH/DEA=100/0.1; flow rate, 0.5 mL/min; detector: UV 254 nm.
Compound 390: 1H NMR (400 MHz, DMSO-d6) δ 8.71-8.70 (m, 1H), 8.17 (s, 1H), 8.11-8.07 (m, 1H), 7.78 (s, 1H), 7.72 (d, J=7.6 Hz, 1H), 7.64-7.60 (m, 2H), 6.67 (d, J=2.8 Hz, 1H), 5.21-5.18 (m, 1H), 4.34-4.29 (m, 1H), 3.94-3.88 (m, 1H), 2.88 (s, 3H), 2.56-2.55 (m, 1H), 1.90 (br, 1H). MS (m/z): 481.0 (M+1)+.
Compound 391: 1H NMR (400 MHz, DMSO-d6) δ 8.70 (s, 1H), 8.16 (s, 1H), 8.11-8.07 (m, 1H), 7.87 (s, 1H), 7.73-7.69 (m, 2H), 7.62-7.59 (m, 1H), 6.66 (br, 1H), 5.18 (br, 1H), 4.59 (br, 1H), 3.78-3.76 (m, 1H), 2.91 (s, 3H), 2.54 (br, 1H), 1.83 (br, 1H). MS (m/z): 481.0 (M+1)+.
Compound 348 and Compound 349 with at least 98% ee were prepared similar to Compound 390 and Compound 391.
Under the HPLC analysis conditions below, the retention time of Compound 348 is 7.99 min, the retention time of Compound 349 is 7.83 min.
HPLC analysis conditions: Gilson system, Column: Daicel 4.6*250 mm IA; mobile phase: EtOH/DEA=100/0.1; flow rate, 0.5 mL/min; detector: UV 254 nm.
Compound 348: 1H NMR (400 MHz, CD3OD) δ 8.26 (s, 0.5H), 8.25 (s, 0.5H), 7.82 (s, 0.5H), 7.81 (s, 0.5H), 7.60-7.47 (m, 2H), 7.34-7.25 (m, 3H), 6.50 (d, J=3.2 Hz, 0.5H), 6.49 (d, J=3.2 Hz, 0.5H), 5.28-5.21 (m, 1H), 4.28-4.12 (m, 2H), 3.34-3.32 (m, 1H), 3.06 (s, 1.5H), 3.06 (s, 1.5H), 2.59-2.46 (m, 2H). MS (m/z): 537.1 (M+1)+.
Compound 349: 1H NMR (400 MHz, CD3OD) δ 8.13 (s, 0.5H), 8.12 (s, 0.5H), 7.92 (s, 0.5H), 7.91 (s, 0.5H), 7.52-7.46 (m, 1H), 7.39-7.33 (m, 1H), 7.29 (d, J=2.8 Hz, 0.5H), 7.287 (d, J=2.8 Hz, 0.5H), 7.23-7.20 (m, 1H), 7.15-7.05 (m, 1H), 6.43 (d, J=2.8 Hz, 0.5H), 6.42 (d, J=3.2 Hz, 0.5H), 5.40-5.23 (m, 1H), 4.41-4.35 (m, 1H), 4.15-4.09 (m, 1H), 3.28-3.24 (m, 1H), 3.05 (s, 3H), 2.60-2.43 (m, 2H). MS (m/z): 537.1 (M+1)+.
Compound 392 and Compound 393 were prepared similar to Compound 390 and Compound 391.
Under the HPLC analysis conditions below, the retention time of Compound 392 is 7.23 min, the retention time of Compound 393 is 9.20 min.
HPLC analysis conditions: Gilson system, Column: Daicel 4.6*250 mm IA; mobile phase: Hexane: i-PrOH: Et2NH=70:30:0.1; flow rate, 1 mL/min; detector: UV 254 nm.
Compound 392: 1H NMR (400 MHz, CD3OD) δ 8.20 (d, J=0.8 Hz, 1H), 7.89 (s, 1H), 7.62-7.51 (m, 2H), 7.36-7.27 (m, 2H), 7.24 (dd, J=4.2, 3.0 Hz, 1H), 6.46 (dd, J=3.0, 1.5 Hz, 1H), 5.37-5.29 (m, 1H), 5.19-5.11 (m, 1H), 4.44-4.31 (m, 1H), 4.11-3.97 (m, 1H), 3.09 (s, 3H), 2.46-2.32 (m, 2H). MS (m/z): 530.1 (M+1)+.
Compound 393: 1H NMR (400 MHz, CD3OD) δ 8.30 (s, 1H), 7.96 (s, 1H), 7.68-7.51 (m, 2H), 7.42-7.26 (m, 2H), 7.25 (br, 1H), 6.45 (br, 1H), 5.46-5.25 (m, 1H), 5.24-5.11 (m, 1H), 4.93 (m, 1H), 4.05-3.85 (m, 1H), 3.09 (s, 3H), 2.62-2.24 (m, 2H). MS (m/z): 530.1 (M+1)+.
According to the procedures described in Example 48 using the corresponding reagents and intermediates, 60c and 60c′ were given after purification by flash column chromatography from the reaction of 60b and NaCN in DMSO.
The solution of 60c (30 mg, 0.046 mmol) in TFA (5 mL) was stirred at 0° C. for 1 h, then concentrated, the resulting residue was dissolved in MeOH (5 mL), and treated with NH3.H2O (2 mL), the mixture was stirred at r.t for 1 h, then concentrated and purified by p-TLC to give Compound 394 as a yellow solid. 1H NMR (400 MHz, CD3OD) δ 8.20 (s, 1H), 7.86 (s, 1H), 7.63-7.41 (m, 5H), 7.29 (d, J=3.0 Hz, 1H), 6.49 (d, J=3.0 Hz, 1H), 5.24 (t, J=7.6 Hz, 1H), 4.28-4.13 (m, 2H), 3.28-3.22 (m, 1H), 3.06 (s, 3H), 2.54-2.47 (m, 2H). MS (m/z): 519.1 (M+1)+.
Compound 395 was prepared according to the procedure of Compound 394. 1H NMR (400 MHz, CD3OD) δ 8.14 (s, 1H), 7.99 (s, 1H), 7.61-7.51 (m, 2H), 7.44-7.38 (m, 2H), 7.36 (d, J=3.0 Hz, 1H), 7.30-7.26 (m, 1H), 6.50 (d, J=3.0 Hz, 1H), 5.38-5.36 (m, 1H), 4.47-4.45 (m, 1H), 4.17-4.15 (m, 1H), 3.27-3.20 (m, 1H), 3.12 (s, 3H), 2.65-2.46 (m, 2H). MS (m/z): 519.1 (M+1)+.
Under the HPLC analysis conditions below, the retention time of Compound 394 is 8.22 min, the retention time of Compound 395 is 8.24 min.
HPLC analysis conditions: Gilson system, Column: Daicel 4.6*250 mm IA; mobile phase: EtOH/DEA=100/0.1; flow rate, 0.5 mL/min; detector: UV 254 nm.
Compound 219 was resolved by p-TLC to produce the optically pure enantiomers Compound 396 and Compound 397 with at least 98% ee.
Under the HPLC analysis conditions below, the retention time of Compound 396 is 8.83 min, the retention time of Compound 397 is 8.50 min.
HPLC analysis conditions: Gilson system, Column: Daicel 4.6*250 mm IA; mobile phase: EtOH/DEA=100/0.1; flow rate, 0.5 mL/min; detector: UV 254 nm.
Compound 396: 1H NMR (400 MHz, DMSO-d6) δ 12.37 (brs, 1H), 8.25 (s, 1H), 7.73 (d, J=7.9 Hz, 1H), 7.67-7.54 (m, 5H), 7.26 (m, 1H), 6.413 (d, J=3.2 Hz, 1H), 4.79 (t, J=7.2 Hz, 1H), 3.84-3.80 (m, 2H), 2.93 (s, 3H), 2.11-2.05 (m, 2H), 2.01-1.96 (m, 1H), 1.81-1.76 (m, 1H). MS (m/z): 478.1 (M+1)+.
Compound 397: 1H NMR (400 MHz, DMSO-d6) δ 12.40 (brs, 1H), 8.26 (s, 1H), 7.87 (s, 1H), 7.78-7.75 (m, 1H), 7.64-7.52 (m, 4H), 7.38-7.37 (m, 1H), 6.40 (d, J=3.2 Hz, 1H), 4.68-4.66 (m, 1H), 4.17-4.15 (m, 1H), 3.69-3.67 (m, 1H), 2.88 (s, 3H), 2.33-2.19 (m, 2H), 2.01-1.89 (m, 2H). MS (m/z): 478.1 (M+1)+.
60d (prepared according to the procedures described in Example 6 using the corresponding reagents and intermediates) was resolved chiral HPLC to produce the optically pure enantiomers 60e and 60e′. HPLC conditions: Gilson system, Column: CHIRALPAK Ia 20 mm I.D.×25 cm L; mobile phase: Hexane/EtOH/Et2NH=70/30/0.1; flow rate:10 mL/min; detector: UV 254 nm.
60e is the first eluent, 60e′ is the second eluent.
Compound 405 was prepared from 60e according to the procedures described in Example 6 using the corresponding reagents and intermediates. 1H NMR (400 MHz, CD3OD) δ 9.21 (d, J=7.0 Hz, 1H), 8.09 (d, J=0.9 Hz, 1H), 7.94 (s, 1H), 7.46-7.41 (m, 2H), 7.33 (d, J=7.9 Hz, 1H), 7.23-7.18 (m, 3H), 6.98 (t, J=7.7 Hz, 1H), 6.38-6.37 (m, 1H), 4.93-4.88 (m, 1H), 2.53 (s, 3H), 1.47 (d, J=6.7 Hz, 3H). MS (m/z): 431.1 (M+1)+.
Compound 406 was prepared from 60e′ according to the procedures described in Example 6 using the corresponding reagents and intermediates. 1H NMR (400 MHz, CD3OD) δ 9.21 (d, J=7.1 Hz, 1H), 8.09 (s, 1H), 7.94 (s, 1H), 7.46-7.41 (m, 2H), 7.33 (d, J=8.0 Hz, 1H), 7.23-7.18 (m, 3H), 6.97 (t, J=7.7 Hz, 1H), 6.398-6.38 (m, 1H), 4.93-4.88 (m, 1H), 2.53 (s, 3H), 1.47 (d, J=6.7 Hz, 3H). MS (m/z): 431.1 (M+1)+.
Compound 407 was prepared from 60e according to the procedures described in Example 1 using the corresponding reagents and intermediates. 1H NMR (400 MHz, DMSO-d6) δ 9.21 (d, J=7.6 Hz, 1H), 7.55-7.45 (m, 1H), 7.37-7.27 (m, 4H), 7.23-7.19 (m, 2H), 6.39-6.38 (m, 1H), 4.91-4.86 (m, 1H), 3.52-3.39 (m, 2H), 2.62-2.46 (m, 2H), 1.36 (d, J=6.8 Hz, 3H). MS (m/z): 434.1 (M+1)+.
Compound 449 was prepared from 60e according to the procedures described in Example 6 using the corresponding reagents and intermediates. 1H NMR (400 MHz, CD3OD) δ 7.99 (brs, 1H), 7.45 (t, J=6.9 Hz, 1H), 7.39 (brs, 1H), 7.29-7.20 (m, 5H), 6.39-6.38 (m, 1H), 5.07-5.02 (m, 1H), 1.39 (d, J=6.6 Hz, 3H). MS (m/z): 390.1 (M+1)+.
Compound 452 was prepared from 60e according to the procedures described in Example 6 using the corresponding reagents and intermediates. 1H NMR (400 MHz, CD3OD) δ 9.10 (d, J=7.5 Hz, 1H), 8.45 (s, 1H), 7.48-7.44 (m, 1H), 7.38-7.38 (m, 1H), 7.30-7.27 (m, 2H), 7.22-7.17 (m, 2H), 7.15-7.12 (m, 1H), 6.38 (d, J=3.1 Hz, 1H), 5.01-4.93 (m, 1H), 2.40 (s, 3H), 1.36 (d, J=6.8 Hz, 3H). MS (m/z): 407.1 (M+1)+.
60f (prepared according to the procedures described in Example 19 using the corresponding reagents and intermediates) was resolved chiral HPLC to produce the optically pure enantiomers 60g and 60g′. HPLC conditions: Gilson system, Column: CHIRALPAK Ia 20 mm I.D.×25 cm L; mobile phase: EtOH/Et2NH=100/0.1; flow rate: 8 mL/min; detector: UV 254 nm.
60g is the first eluent, 60g′ is the second eluent.
Compound 447 was prepared from 60g according to the procedures described in Example 38 using the corresponding reagents and intermediates. 1H NMR (400 MHz, DMSO-d6) δ 9.27 (d, J=7.6 Hz, 1H), 8.54 (s, 1H), 7.78-7.73 (m, 1H), 7.61-7.57 (m, 1H), 7.55-7.48 (m, 1H), 7.47-7.41 (m, 2H), 7.37 (s, 1H), 7.33-7.25 (m, 1H), 6.48 (s, 1H), 4.58-4.51 (m, 1H), 2.38 (s, 3H), 1.24 (d, J=6.8 Hz, 3H). MS (m/z): 424.2 (M+1)+.
Compound 448 was prepared from 60g′ according to the procedures described in Example 38 using the corresponding reagents and intermediates. 1H NMR (400 MHz, CD3OD) δ 8.51 (s, 1H), 7.57-7.53 (m, 1H), 7.50-7.46 (m, 2H), 7.44-7.38 (m, 2H), 7.25 (s, 1H), 6.61 (s, 1H), 4.88-4.83 (m, 1H), 2.43 (s, 3H), 1.37 (d, J=6.8 Hz, 3H). MS (m/z): 424.2 (M+1)+.
Compound 450 was prepared from 60g according to the procedures described in Example 1 using the corresponding reagents and intermediates. 1H NMR (400 MHz, CD3OD) δ 7.97 (s, 1H), 7.57 (d, J=8.0 Hz, 1H), 7.51-7.43 (m, 2H), 7.37-7.34 (m, 1H), 7.29-7.25 (m, 1H), 7.20 (d, J=1.2 Hz, 1H), 6.84 (d, J=2.8 Hz, 1H), 6.72 (s, 1H), 4.93-4.88 (m, 1H), 1.43 (d, J=6.8 Hz, 3H). MS (m/z): 424.1 (M+1)+.
Compound 451 was prepared from 60g′ according to the procedures described in Example 1 using the corresponding reagents and intermediates. 1H NMR (400 MHz, CD3OD) δ 8.00 (s, 1H), 7.61 (d, J=8.8 Hz, 1H), 7.54-7.46 (m, 2H), 7.40-7.37 (m, 1H), 7.31-7.28 (m, 1H), 7.23 (d, J=1.6 Hz, 1H), 6.87 (d, J=2.4 Hz, 1H), 6.75 (s, 1H), 4.96-4.41 (m, 1H), 1.65 (d, J=6.8 Hz, 3H). MS (m/z): 424.1 (M+1)+.
Compound 483 were resolved by chiral HPLC to produce the optically pure enantiomers Compound 484 and Compound 485. HPLC conditions: Gilson system, Column: CHIRALPAK Ia 20 mm I.D.×25 cm L; mobile phase: EtOH/DEA=100/0.1; flow rate, 8 mL/min; detector: UV 254 nm.
Compound 484 is the first eluent with at least 98% ee. MS (m/z): 464.2 (M+1)+.
Compound 485 is the second eluent with at least 98% ee. MS (m/z): 464.2 (M+1)+.
A solution of 61a (300 mg, 0.674 mmol)(prepared according to the procedure of Example 1), NaI (404 mg, 2.646 mmol), trans-1,2-bis(methylamino)cyclohexane (96 mg, 0.674 mmol) and CuI (64 mg, 0.337 mmol) in dioxane (8 ml) was stirred at reflux for 3 days. After cooling to the r.t., the reaction mixture was filtered through celite and washed with ethyl acetate, the resulting filtrate was concentrated and purified by chromatography to give 61b as a yellow solid. MS (m/z): 492.9 (M+H)+.
Under N2 atmosphere 61b (200 mg, 0.4 mmol) and CuI (94 mg, 0.492 mmol) were dissolved in DMF (5 mL), to this mixture were added HMPA (0.35 mL, 2 mol) and methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (0.25 mL, 2 mmol), the resulting mixture was stirred at 80° C. for 24 h, then poured into abundant ice-water and extracted with ethyl acetate. The organic layer was washed with water and brine, then concentrated and purified by chromatography to give 61c as a white solid. MS (m/z): 456.9 (M+Na)+.
Compound 486 was prepared with 61c as the material according to the procedure of Example 1 from 1e to Compound 1. 1H NMR (400 MHz, DMSO-d6) δ 8.47 (s, 1H), 7.770-7.55 (m, 5H), 7.46-7.43 (m, 1H), 6.95 (d, J=2.9, 1H), 6.82 (brs, 2H), 4.90 (brs, 1H), 4.20-4.14 (m, 1H), 3.49 (brs, 1H), 2.47-2.43 (m, 1H), 2.27 (brs, 3H), 1.92 (brs, 1H). MS (m/z): 470.1 (M+H)+.
The following Compounds were prepared according to the procedure of Compound 486 using the corresponding reagents and intermediates under appropriate conditions that will be recognized by one skilled in the art:
1H NMR (400 MHz, DMSO-d6) δ 7.86 (d, J = 2.8, 1H), 7.64-7.56 (m, 4H), 7.47-7.46 (m, 1H), 6.98 (d, J = 2.9, 3H), 4.97 (brs, 1H), 4.08 (d, J = 13.2, 2H), 2.67-2.56 (m, 1H), 2.23 (s, 3H), 2.14- 2.04 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 8.16 (s, 1H), 7.87 (s, 1H), 7.68-7.66 (m, 1H), 7.56-7.48 (m, 3H), 7.37 (d, J = 2.8 Hz, 1H), 7.33- 7.30 (m, 1H), 6.76 (d, J = 2.9 Hz, 1H), 5.28-5.24 (m, 1H), 4.60-4.53 (m, 1H), 4.24-4.18 (m, 1H), 2.59-2.51 (m, 1H), 2.22-2.14 (m, 1H).
1H NMR (400 MHz, CD3OD) δ 8.36 (s, 1H), 7.66-7.51 (m, 4H), 7.33-7.29 (m, 2H), 6.76 (d, J = 2.9 Hz, 1H), 4.91 (brs, 1H), 4.34 (brs, 1H), 3.29 (brs, 1H), 2.50 (brs, 1H), 2.19 (s, 3H), 0.62 (d, J = 6.8 Hz, 3H).
1H NMR (400 MHz, CD3OD) δ 8.01 (s, 1H), 7.55-7.48 (m, 4H), 7.43 (d, J = 2.6 Hz, 1H), 7.32-7.30 (m, 1H), 6.78 (d, J = 2.9 Hz, 1H), 4.76 (brs, 1H), 4.34 (brs, 1H), 3.60 (brs, 1H), 2.65 (brs, 1H), 0.66 (d, J = 6.7 Hz, 3H).
The following compounds may be made using the procedures described in previously
PI3K kinases including p110α/p85α, p110δ/p85α and p110γ are purchased from Invitrogen, and p110β/p85α is from Millipore.
Primary screening data and IC50 values are measured using Transcreener™ KINASE Assay (Bellbrook, Catalog #3003-10K). The assay can be carried out according to the procedures suggested by the manufacturer. It is a universal, homogenous, high throughput screening (HTS) technology using a far-red, competitive fluorescence polarization immunoassay based on the detection of ADP to monitor the activity of enzymes that catalyze group transfer reactions. Briefly, the Transcreener KINASE Assay is designed as a simple two-part, endpoint assay.
In the first step, the 25 ul kinase reaction is performed by preparing a reaction mixture containing 5 ul test compound (2% DMSO final concentration), 10 ul kinase, buffer (50 mM HEPES, 100 mM NaCl, 1 mM EGTA, 0.03% CHAPS, 3 mM MgCl2, and freshly supplemented 1 mM DTT), and 10 ul 30 uM PIP2/10 uM ATP. The plate is sealed and incubated for 80 min at room temperature. Next, 25 ul ADP detection mix is added per well. The plate is sealed again and incubated for 60 min at room temperature, and then measure fluorescence polarization by Tecan Infinite F500 Reader.
Data is analyzed and IC50s are generated using the add-in software for Microsoft Excel, Xlfit™ (version 2.0). IH %=(ADP amount under 2% DMSO-ADP amount under test compound)/ADP amount under 2% DMSO.
In vitro activity data:
Acumen® eX3 (A Multilaser Microplate Cytometer For Enhanced High Content Screening): TTP LabTech
3×104 Raw264.7 macrophage cells were seeded into 96-well plates with DMEM+10% heat-inactivated FBS at 2,700 cells/well, 90 ul/well, overnight. After starvation for 3 hr at 37° C. under 5% CO2, Raw264.7 cells were treated with 10 ul/well various concentrations of compound or 0.5% DMSO for 30 min, and then stimulated with 10 ul/well 10 nM C5a for 5 min.
1.) Cells were fixed 110 μL of 4% pre-warmed Paraformaldehyde (2% final), incubate for 45 min at room temperature.
2.) Remove paraformaldehyde solution. Add 100 μL of ice-cold 0.1% Triton X-100 in PBS and leave at 4° C. for 30 min.
3.) Wash once in 100 μL PBS.
4.) Incubate with 100 μL blocking buffer (1% BSA, in PBS) for 2 hours at room temperature.
5.) Wash once for 5 min with 100 ul PBS.
6.) Incubate with 40 μL 1:200 dilution of phospho AKT (Ser473) rabbit antibody in antibody dilution buffer (0.1% BSA, in PBS) overnight at 4° C.
7.) Wash for 3 times for 10 min with 100 ul PBS.
8.) Incubate for 90 min at room temperature with 50 μL of goat anti-rabbit Alex488 antibody at a 1:1,000 dilution in antibody dilution buffer (0.1% BSA, in PBS). Cover plate in foil to keep out of light.
9.) Wash for 3 times for 10 min with 100 μL PBS.
10.) Add 50 μL of 1.5 μM Propidium Iodide solution to each well to determine cell number at a 1:1,000 dilution in PBS (stock: 1.5 mM).
11.) Incubate at room temperature for 30 min.
12.) Seal the plate with a black cover-seal (supplied with plate).
13.) Load the plate into the Acumen Explorer and scan with the appropriate instrument settings.
| Number | Date | Country | Kind |
|---|---|---|---|
| PCT/CN2012/079290 | Jul 2012 | CN | national |
| PCT/CN2013/072686 | Mar 2013 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2013/080195 | 7/26/2013 | WO | 00 |
