USOO8247421B2

(12) United States Patent (10) Patent No.: US 8.247,421 B2 Mortimore et al. (45) Date of Patent: Aug. 21, 2012

(54) 5-CYANO-4-(PYRROLO 2.3B 7,507,826 B2 3/2009 Salituro et al. 7,767,816 B2 8, 2010 Farmer et al. PYRIDINE-3-YL)-PYRIMIDINE 7,795,259 B2 9/2010 Binch et al. DERVATIVES USEFUL AS PROTEINKNASE 2002fO147.189 A1 10, 2002 Cai et al. INHIBITORS 2003/0153560 A1 8/2003 Salituro et al. 2004/OOO9968 A1 1/2004 Binch et al.

(75) Inventors: Michael Mortimore, Oxfordshire (GB); 3.99. A. 58: MOme et al.al Stells E. E. 2004/0044203 A1 3/2004 Wittman et al...... 544/55 (GB); Simon Robert Lorrie Everitt, 2004/023611.0 A1* 11/2004 Ladouceur et al...... 546,277.4 Oxfordshire (GB); Ronald Knegtel, 2005, 01372O1 A1 6/2005 Aronov et al. Oxfordshire (GB); Joanne Louise 2005/0148603 A1 7/2005 Jimenez et al. Pinder, Oxfordshire (GB); Alistair 2006, OOO3968 A1 1/2006 Green et al. 2006, O122185 A1 6/2006 Green et al. t RECEIS, 2006/0122213 A1 6/2006 Pierard et al. teven Durrant, Oxfordshire (GB); Guy 2006/0183761 A1 8/2006 Ledeboer et al. Brenchley, Oxfordshire (GB); 2006/01839 11 A1 8/2006 Charrier et al. Jean-Damien Charrier, Oxfordshire 2006/0258662 A1 11/2006 Binch et al. (GB); Michael O'Donnell, Oxfordshire 2007.0043063 A1 2/2007 Salituro et al. (GB) s 2007/0135466 A1 6/2007 Ledeboer et al. 2007/0203142 A1 8, 2007 Farmer et al. 2007/02O7995 A1 9, 2007 Salituro et al. (73) Assignee: Vertex Pharmaceuticals Incorporated, 2007/0213327 A1 9, 2007 Collier et al. Cambridge, MA (US) 2009 OO48250 A1 2/2009 Aronov et al. 2009 OO88445 A1 4/2009 Ledeboer et al. (*) Notice: Subject to any disclaimer, the term of this 2009/0118278 Al 52009 Forster et al. tent is extended or adiusted under 35 2009/0176763 A1 T/2009 Salituro et al. pa 2009,029 1937 A1 11/2009 Jimenez et al. U.S.C. 154(b) by 296 days. 2010.0311743 A1 12/2010 Farmer et al. 2011/0224197 A1 9, 2011 Henkel et al. (21) Appl. No.: 12/448,489 FOREIGN PATENT DOCUMENTS (22) PCT Filed: Dec. 21, 2007 EP O557171 8, 1993 WO 88/O1997 3, 1988 (86). PCT No.: PCT/US2007/026,190 WO 95/33748 12/1995 WO 99.21859 5, 1999 S371 (c)(1), WO OOf 40581 T 2000 WO OOf43393 T 2000 (2), (4) Date: Nov. 10, 2009 WO O1/O1986 1, 2001 (87) PCT Pub. No.: WO2008/079346 WO O2/O51837 T 2002 (Continued) PCT Pub. Date: Jul. 3, 2008 OTHER PUBLICATIONS (65) Prior Publication Data Alvarez, Mercedes, et al., “Synthesis of 3-Aryl- and 3-Heteroaryl-7- US 2010/O189773 A1 Jul. 29, 2010 stols , Synthesis, Thieme Stuttgart, New York, No. 4, 1999, pp. O O Berge, Stephen M., et al., “Pharmaceutical Salts' Journal of Phar Related U.S. Application Data maceutical Sciences, vol. 66, No. 1, Jan. 1977, pp. 1-19. (60) Provisional application No. 60/876,307, filed on Dec. Chitaley, Kanchan, et al., “Antagonism of Rho-kinase stimulates rat 21, 2006 isional licati s N s 607922,291 penile erection via a nitric oxide-independent pathway'. Nature s provisional application, No. OU 94.491, Medicine, Nature Publishing Group, vol. 7, No. 1, Jan. 2001, pp. filed on Apr. 6, 2007, provisional application No. 119-122. 60/947,707, filed on Jul. 3, 2007, provisional Eto, Masato, et al., “Thrombin Suppresses Endothelial Nitric Oxide application No. 60/989,014, filed on Nov. 19, 2007. Synthase and Upregulates Endothelin-Converting Enzyme-l Expression by Distinct Pathways'. Circulation Research, vol. 89, (51) Int. C. 2001, pp. 583-590. AOIN 43/54 (2006.01) (Continued) (52) U.S. Cl...... 514/256; 544/333 Primary Examiner — James O Wilson (58) Field of Classification Search ...... Assistant Examiner — Alexander R Pagano (74) Attorney, Agent, or Firm — Honigman Miller Schwartz See application file for complete search history. and Cohn LLP: Jonathan P. O'Brien; Andrew N. Weber (56) References Cited (57) ABSTRACT The present invention relates to compounds useful as inhibi U.S. PATENT DOCUMENTS tors of protein kinase. The invention also provides pharma 5,304,121 A 4/1994 Sahatian ceutically acceptable compositions comprising said com 5,338,849 A 8, 1994 Festal et al. pounds and methods of using the compositions in the 5,886,026 A 3, 1999 Hunter et al. treatment of various disease, conditions, or disorders. The 6,099,562 A 8/2000 Ding et al. invention also provides processes for preparing compounds 6,265,403 B1 7/2001 Fraley et al. of the inventions. 6,900,201 B2 * 5/2005 Noe et al...... 514,227.8 7,135,550 B2 11/2006 Come et al. 79 Claims, No Drawings US 8,247.421 B2 Page 2

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Somlyo, Avril V... et al., Rho-Kinase Inhibitor Retards Migration and WO 2008/079346, International Search Report. in Vivo Dissemination of Human Prostate Cancer Cells, Biochemical WO 2008/079346, International Preliminary Reporton Patentability. and Biophysical Research Communications, vol. 269, No. 3, 2000, pp. 652-659. * cited by examiner US 8,247,421 B2 1. 2 5-CYANO-4-(PYRROLO 2.3B rophage-colony stimulating factor (GM-CSF), and fibroblast PYRIDINE-3-YL)-PYRIMIDINE growth factor (FGF)). An extracellular stimulus may affect DERVATIVESUSEFUL AS PROTEINKNASE one or more cellular responses related to cell growth, migra INHIBITORS tion, differentiation, secretion of hormones, activation of transcription factors, muscle contraction, glucose metabo CROSS-REFERENCE lism, control of protein synthesis, Survival and regulation of the cell cycle. This application is a national phase entry of PCT/US2007/ Many diseases are associated with abnormal cellular 026190 filed responses triggered by protein kinase-mediated events as Dec. 21, 2007 which claims the benefit of the U.S. Provi 10 described above. These diseases include, but are not limited sional Application No's 60/876,307, filed on Dec. 21, 2006: to, cancer, autoimmune diseases, inflammatory diseases, 60/922.291, filed on Apr. 6, 2007: 60/947,707, filed on Jul. 3, bone diseases, metabolic diseases, neurological and neurode 2007; and 60/989,014, filed on Nov. 19, 2007. The entire generative diseases, cardiovascular diseases, allergies and contents of the documents are incorporated herein in their entirety. 15 asthma, Alzheimer's disease and hormone related diseases. Incorporated herein by reference is a Sequence Listing Accordingly, there has been a Substantial effort in medicinal named “06-156 US Seq List ST25. txt” created on Nov. 3, chemistry to find protein kinase inhibitors that are effective as 2009, that is 1,886 bytes. The sequence listing does not therapeutic agents. include any new matter which goes beyond the disclosure of The Polo-like kinases (PLK) belong to a family of serine/ the application as filed. threonine kinases that are highly conserved across the spe cies, ranging from yeast to man (reviewed in Lowery, D. M. et TECHNICAL FIELD OF THE INVENTION al. Oncogene, 2005, 24; 248-259). The PLK kinases have multiple roles in cell cycle, including control of entry into and The present invention relates to compounds useful as progression through mitosis. inhibitors of protein kinases. The invention also provides 25 PLK1 is the best characterized of the PLK family mem pharmaceutically acceptable compositions comprising the bers. PLK1 is widely expressed and is most abundant in compounds of the invention and methods of using the com tissues with a high mitotic index. Protein levels of PLK1 rise positions in the treatment of various disorders. The invention and peak in mitosis (Hamanaka, R. et al., J. Biol. Chen., also provides processes for preparing the compounds of the 1995, 270, 21086-21091). The reported substrates of PLK1 invention. 30 are all molecules that are known to regulate entry and pro gression through mitosis, and include CDC25C, cyclin B, BACKGROUND OF THE INVENTION p53, APC, BRCA2 and the proteasome. PLK1 is upregulated in multiple cancer types and the expression levels correlate The search for new therapeutic agents has been greatly with severity of disease (Macmillan, J C et al., Ann. Surg. aided in recent years by a better understanding of the structure 35 Oncol., 2001, 8, 729-740). PLK1 is an oncogene and can of enzymes and other biomolecules associated with diseases. transform NIH-3T3 cells (Smith, M. R. et al., Biochem. Bio One important class of enzymes that has been the subject of phys. Res. Commun., 1997, 234,397-405). Depletion or inhi intensive study is protein kinases. bition of PLK1 by siRNA, antisense, microinjection of anti Protein kinases constitute a large family of structurally bodies, or transfection of a dominant negative construct of related enzymes that are responsible for the control of a 40 PLK1 into cells, reduces proliferation and viability of tumour variety of signal transduction processes within the cell (see cells in vitro (Guan, R. et al., Cancer Res., 2005, 65, 2698 Hardie, G. et al., The Protein Kinase Facts Book, I and II, 2704; Liu, X. et al., Proc. Natl. Acad. Sci. USA, 2003, 100, Academic Press, San Diego, Calif.: 1995). Protein kinases are 5789-5794; Fan, Y. et al., World J. Gastroenterol., 2005, 11, thought to have evolved from a common ancestral gene due to 4596-4599; Lane, H. A. et al., J. Cell Biol 1996, 135, 1701 the conservation of their structure and catalytic function. 45 1713; Wada, M. et al., Biochem. Biophys. Res. Commun., Almost all kinases contain a similar 250-300 amino acid 2007,357(2): 353-359; Rizki, A. et al., Cancer Res., 2007, 67 catalytic domain. The kinases may be categorized into fami (23): 11106-11100). Tumour cells that have been depleted of lies by the Substrates they phosphorylate (e.g., protein-ty PLK1 have activated spindle checkpoints and defects in rosine, protein-serine/threonine, lipids etc). Sequence motifs spindle formation, chromosome alignment and separation have been identified that generally correspond to each of 50 and cytokinesis. Loss in viability has been reported to be the these kinase families (see, e.g., Hanks, S. K., Hunter, T., result of an induction of apoptosis. In contrast, normal cells FASEB.J., 1995, 9, 576-596: Knighton et al., Science, 1991, have been reported to maintain viability on depletion of 253, 407-414; Hiles etal, Cell, 1992, 70,419-429; Kunzetal, PLK1. In vivo knock down of PLK1 by siRNA or the use of Cell, 1993, 73, 585-596; Garcia-Bustos et. al., EMBO J., dominant negative constructs leads to growth inhibition or 1994, 13, 2352-2361). 55 regression of tumours in Xenograft models. In general, protein kinases mediate intracellular signaling PLK2 is mainly expressed during the G1 phase of the cell by effecting a phosphoryl transfer from a nucleoside triphos cycle and is localized to the centroSome in interphase cells. phate to a protein acceptor that is involved in a signaling PLK2 knockout mice develop normally, are fertile and have pathway. These phosphorylation events act as molecular normal survival rates, but are around 20% smaller than wild on/off Switches that can modulate or regulate the target pro 60 type mice. Cells from knockout animals progress through the tein biological function. These phosphorylation events are cell cycle more slowly than in normal mice (Ma, S. et al., Mol. ultimately triggered in response to a variety of extracellular Cell Biol., 2003, 23, 6936-6943). Depletion of PLK2 by and other stimuli. Examples of Such stimuli include environ siRNA or transfection of kinase inactive mutants into cells mental and chemical stress signals (e.g., shock, heat shock, blocks centriole duplication. Downregulation of PLK2 also ultraviolet radiation, bacterial endotoxin, and H2O), cytok 65 sensitizes tumour cells to taxol and promotes mitotic catas ines (e.g., interleukin-1 (IL-1) and tumor necrosis factor trophe, in part by suppression of the p53 response (Burns, T. alpha (TNF-a), and growth factors (e.g., granulocyte mac F. et al., Mol. Cell Biol., 2003, 23, 5556-5571). US 8,247,421 B2 3 4 PLK3 is expressed throughout the cell cycle and increases Each R is independently H, Caliphatic, aryl, heteroaryl, from G1 to mitosis. Expression is upregulated in highly pro cycloalkyl, or 4-12 membered heterocyclic ring optionally liferating ovarian tumours and breast cancer and is associated with a worse prognosis (Weichert, W. et al., Br. J. Cancer, containing 1-3 groups selected from —N(R')—, -O-, or 2004,90, 815-821; Weichert, Wet al., Virchows. Arch., 2005, —S—, wherein each of the aliphatic, aryl, heteroaryl, 446, 442-450). In addition to regulation of mitosis, PLK3 is cycloalkyl, and heterocyclic ring are optionally Substituted believed to be involved in Golgi fragmentation during the cell with 1-3 of Q: cycle and in the DNA-damage response. Inhibition of PLK3 Each Q is independently selected from halogen, hydroxy, by dominant negative expression is reported to promote p53 C. alkyl, benzyl, oxo, —CF. W. —CN, NH, N(H) independent apoptosis after DNA damage and Suppresses W, N(W), N(H) SO W, S(O), N(H) W. colony formation by tumour cells (Li, Z. et al., J. Biol. Chem., 10 2005, 280, 16843-16850). —S(O), N(W), C(O) W. —C(O) N(W), PLK4 is structurally more diverse from the other PLK N(H) C(O) W, O C(O) W. C(O) O W, family members. Depletion of this kinase causes apoptosis in - SO W, SW or OW: cancer cells (Li, J. et. al., Neoplasia, 2005, 7,312-323). PLK4 Two Q can be linked together to form a 4- to 8-membered knockout mice arrest at E7.5 with a high fraction of cells in carbocyclic or heterocyclic ring optionally substituted with mitosis and partly segregated chromosomes (Hudson, J. W. 15 et. al., Current Biology, 2001, 11, 441-446). C. alkyl or CF; Molecules of the protein kinase family have been impli Each W is independently selected from —H, C alkyl, cated in tumour cell growth, proliferation and Survival. aralkyl, cycloalkyl or heterocyclic ring; each C alkyl, Accordingly, there is a great need to develop compounds aralkyl, cycloalkyl or heterocyclic ring is optionally Substi useful as inhibitors of protein kinases. The evidence impli tuted with 1-3 of halogen, OR, —CN, C- alkyl or cating the PLK kinases as essential for cell division is strong. NR'R'', or Blockade of the cell cycle is a clinically validated approach to inhibiting tumour cell proliferation and viability. It would One W, together with the nitrogen atom to which it is therefore be desirable to develop compounds that are useful attached and a carbon atom of R, form a 4- to 8-membered as inhibitors of the PLKfamily of protein kinases (e.g., PLK1, ring; or PLK2, PLK3 and PLK4), that would inhibit proliferation and 25 Two W, together with the same or different nitrogen atom reduce viability of tumour cells, particularly as there is a or carbon atom to which they are attached, form a 4- to strong medical need to develop new treatments for cancer. 8-membered heterocyclic ring: Each RandR' is independently hydrogen or C, alkyl: SUMMARY OF THE INVENTION O 30 R" and R', together with the nitrogenatom to which they Compounds of this invention are useful as inhibitors of are attached, form a 4- to 8-membered heterocyclic ring, PLK protein kinases and in some-embodiments, as inhibitors optionally substituted with C alkyl or CF; of PLK1 protein kinases. These compounds are as defined Two W can be linked together to form a 4- to 8-membered herein. cycloalkyl or heterocycloalkyl optionally substituted with These compounds, and pharmaceutically acceptable salts 35 thereof, are useful for treating or preventing a variety of C. alkyl or CF; diseases, disorders or conditions, including, but not limited R is NRR, OR, SR, or NR'R'': to, an autoimmune, inflammatory, proliferative, or hyperpro Each R is independently halogen, C. alkyl, aryl, or het liferative disease, a neurodegenerative disease, or an immu eroaryl; nologically-mediated disease. The compounds provided by 40 Each R" is independently —H or Caliphatic optionally this invention (and pharmaceutically acceptable salts and substituted with 1-3 R': pharmaceutically acceptable derivatives thereof) are also use Each R is independently Caliphatic optionally substi ful for the study of kinases in biological and pathological phenomena; the study of intracellular signal transduction tuted with 1-4 R' or a 4- to 8-membered monocyclic or 6- to pathways mediated by Such kinases; and the comparative 10-membered bicyclic ring optionally substituted with 1-4 45 R7, or evaluation of new kinase inhibitors. R" and Rican be joined together to form a monocyclic or DETAILED DESCRIPTION OF THE INVENTION bicyclic ring optionally substituted with 1-3 R’. Each R is independently H, C, alkyl, -L-aryl, or -L- Compounds of the Invention heteroaryl, wherein each of the C- alkyl, -L-aryl, or -L- This invention provides a compound of formula I: 50 heteroaryl is optionally and independently substituted with 1-3 R8; L is Co- alkyl, Each R7 is independently oxo, alkyl, halogen, —CN, —OR.—SR, N(R), Css cycloalkyl, aryl, heteroaryl or 55 a 4- to 8-membered heterocyclic ring containing 1-3 groups selected from N(R'7) , —O , or —S , wherein each alkyl, cycloalkyl, 4-8 membered heterocyclic monocyclic or 60 bicyclic ring, aryl, and heteroaryl is optionally and inde pendently substituted with 1-3 R', or Two R' on the same atom or adjacent atoms is joined to form a carbocyclic ring or a 4-8 membered heterocyclic ring or a pharmaceutically acceptable salt thereof, wherein: containing 1-3 groups selected from —N(R')—, -O-, or R" is -H, halogen, Caliphatic optionally substituted 65 —S—, wherein each of the carbocyclic ring and the 4- to with 1-3 R', O(Caliphatic) optionally substituted with 8-membered heterocyclic ring is optionally and indepen 1-3 R, or N(H)R; dently substituted with 1-3 R: US 8,247,421 B2 5 6 Each R is independently —R, -Q, R, OR, N In some embodiments, R is H or C, alkyl and R is a (R), halogen, or —CN: hydroxyalkyl, hydroxcycloalkyl oralkoxyalkyl selected from Each R is independently —H, N(R'), C. carbocy clic ring, C. heterocyclic ring, or Caliphatic, wherein C. carbocyclic ring, C. heterocyclic ring and C- ali 5 phatic are each optionally substituted with 1-3 Q; or Two R groups together with the Natom to which they are bound form a 4-8 membered ring additionally containing 1 or 2 groups each independently selected from N(R') , 10 —O—, or —S , wherein the 4- to 8-membered ring is optionally and independently substituted with 1-3 of W: Each R" is independently hydrogen or C. alkyl, or Two R' groups together with the Natom to which they are 15 bound form a 4- to 8-membered ring containing 1 or 2 groups selected from NR'7, O, or S: Each R'' is independently, hydrogen, Q, or Caliphatic A-l. - or cycloaliphatic, wherein each C aliphatic or cycloaliphatic is optionally substituted with 1-3 of Q; Q is C alkyl, benzyl, —SO. W. —S(O), N(H)—W. S(O), N(W), C(O) W. C(O) N(W), C(O) N(H) W, N(H) C(O) W. O. C(O) W. C(O) O W, or - SO W: R" is —H or C-C aliphatic optionally substituted with 25 1-3 R7; HO Each of R'' and R' is independently H or Caliphatic optionally substituted with 1-3 R'; or 30 R'' and R' can be joined togetherto formaring optionally substituted with 1-3 R'; or R'' and R' can bejoined togetherto formaring optionally substituted with 1-3 R’; and 35 Each R'' and R' is independently H, C alkyl, carbocy clic, or heterocyclic optionally substituted with 1-3 R'; or R'' and R' can bejoined togetherto formaring optionally substituted with 1-3 R. 40 In some embodiments, R is H or C. alkyl and R is selected from —CH,

45

OH

50

55

60

65 US 8,247,421 B2 7 8 -continued -continued o, O 5 NH2 or Cl, / s N H

o, 10 O OCH

H l OCH, OCH s 15 In some embodiments, R is H or C alkyl and R is a substituted alkyl selected from / s

CH3 >n-r-, N/ N(CH3)2, s 30 In some embodiments, R is H or C. alkyl, and R is an CH aryl or aralkyl selected from

35

X C 7-c. 40 Xo 1sO s OH, s F Xo s s 45 x5 Xr

Xol OCH, s 50

F, OCH3, Xr OCH x5 55 X C 60 Xc Br, OH, 65 OH US 8,247,421 B2 9 10 -continued -continued Xol1. 5 N OCH 1N

Xo-OCH Xo 10 N ~ 15 Null.

CF Xol O 2O OCH3, O

OH C F 30

O H,

OH, 35 S 40 ON” X O -COCH3, N 45

O, 50 O

55 O OCH, C 60

Xo, -SO3CH3. 65 Xo,Nuk US 8,247,421 B2 11 12 -continued -continued

lu O ux. 10

O.o 15 O

1

25

SOCH3, N v 30 NHCH,

35 Ol r 40

n

45

F

n u 50 Cl 8. 2

55 O NHCH2CH3, N 60

O, s 65 v US 8,247,421 B2 13 14 -continued -continued

to In some embodiments, R is H or C, alkyl and R is an optionally substituted heteroaralkyl selected from

NH2, 15 N O 2 NF N

Nr 25 Xr 2 N2 s s1 nu1 N N OCH O2 N 30 N s 2 s N C OH, 35 N

2 N r OH, 40 N YCH, O O H

45 N1

50 N. N NS 55 Xo O, NH2, O SO 60 N

2 1 NHCH3 or 65 N SO US 8,247,421 B2 15 16 -continued -continued

Z <%C) H C) H 10 ºA O 15 (T)Z

2O

25 OH

30

35

40

45

OH 50

55

60

65 US 8,247,421 B2 17 18 -continued -continued

N N

2 s N ^1

10 C N o / N 15 2 XCN N c H C

r % N 1-N-NH.

N 25 N

- 2 Clu)N N X H 30

1 r 35 N 4. ~ NS 40 XoXolXo4. N

2 N N 45 O (

N 50 2 N

55 C C

60

65 XolN ON US 8,247,421 B2 19 20 -continued -continued

10 In some embodiments, R is H and R is an optionally Substituted aralkyl or heteroaralkyl, which give compounds of formula Ic 15

Ic A-N-

NC | y 25 RI

N N. N) 30 wherein each X is independently CR or N and K is OR, —SR, or - N(R). In some embodiments, R is H or C alkyl and R is a 35 cycloalkyl or cycloalkyl(alkyl), each optionally Substituted, and selected from

40

45

50

55

60

65 US 8,247,421 B2 21 22 -continued In other embodiments, R is Hand R is an amino substi tuted cycloalkyl(alkyl), which give the compounds of for mula Ib OH OH, s 5 HO Ib OH, s NHR17.

10 N | Y-st NC leN OH 15 R rN\, 2 N N H o, ck 8, 2O In other embodiments, R is H or C. alkyl, and R is a NHSOCH, heterocycloalkyl or heterocycloalkyl (alkyl), each optionally substituted, and selected from

25 ..., O O N-CH o NHSOCH, 30 / \ s N O,

o, O 35

N 1sult >n-rr O O, s

H 40 N i. s -N S, c. Ol N M NHCOCH, 45 H N s Ol 4 C 55 %,

C 'NHCOCH, or N -4 O 60 CH

H 65 AC US 8,247,421 B2 23 24 -continued -continued

5

O

10

N

15 O O 20 O x 25

30 CH3 n 1 N

35 O N 1. 40 O

45

50 r N N ué O

55 " uC ls O 60

N OH, s-A 65 f 2 O US 8,247,421 B2 25 26 -continued -continued

OH 5 N s

O 10 XCAs OS 15 /\,

O 2O

O H,

25 gH N^- s

30

O H,

N H

N r N H2, 50 55 XO- NH2, N-1aO H,

r OH,

US 8,247,421 B2 31 32 In some embodiments, R is HandR is a heterocycloalkyl -continued (alkyl), which give the compounds of formula Ia

CF Ia 5 N1 3.

O

N 10 N1 No s

N OH, ry a NH 2 N N H 15 O

CH3, s O O

Or NH N1 No O represents a 4- to 8-membered nitrogen containing heterocy- 25 O clic ring. CF In one embodiment, Rand Rare joined together to form n-13

O 30 N In another embodiment, Ris-SR wherein R is selected from CH ,

35 NH2

In other embodiments, R is NR'R'' wherein R' is H Cl, -- )-OCH, and R'' is selected from 40

O H N

O s B C H N

60 s s O OCH H N CF3, N1 3 65 s s O US 8,247,421 B2 33 34 -continued —O—, or —S , wherein the 4- to 8-membered ring is optionally and independently substituted with 1-3 of W. OH or In still some embodiments of the compounds of this inven tion, R is optionally substituted at an ortho-position with fluoride. In some embodiments, the compounds of this inventions have the following features: In some embodiments, the compounds of this invention R is 5- to 7-membered heterocyclic ring optionally con have one of more of the following features: R is a 4- to taining 2 nitrogen atoms and optionally Substituted with 1-3 10-membered heterocyclic monocyclic or bicyclic ring 10 of Q; optionally substituted with 1-3 of R. R7 is a 4- to 6-mem Each R is independently —H. C. heterocyclic ring, or bered heterocyclic monocyclic ring optionally substituted Caliphatic, wherein C. heterocyclic ring and C-C ali with 1-3 of R: R is Q: Q is selected from C(O) W. phatic are each optionally substituted with 1-3 Q; or —C(O) N(W), C(O) O W, or -SO. W: Q is Two R groups together with the Natom to which they are C(O) W. 15 bound form a 6- to 7-membered ring containing an additional In some embodiments, the compounds of this invention nitrogenatom and optionally substituted with 1-3 of W. have one of more of the following features: R is a C-Cs Still some of the compounds of this invention have one or carbocycle optionally substituted with 1-3 R; one R is Q: Q more of the following features: R is piperazine optionally is selected from hydroxy, NH, N(H) W. N(W), substituted with 1-3 of Q: R is N(H) SO. W. —C(O) N(W), N(H) C(O). W. O O C(O) W: Q is N(H)–C(O) W. In some embodiments, the compounds of this invention have one of more of the following features: R' is halogen (e.g., —Cl); R' is Caliphatic (e.g., alkyl, alkenyl, or alky 25 nyl) optionally substituted with 1-3 of R, and each R is independently halo, aryl, or heteroaryl; R' is methyl option ally substituted with 1-3 R and each R is independently halo: R is CF; R is —CH; R is NHRandR is H. C. O aliphatic (e.g., alkyl), aryl, or Cs cycloalkyl. 30 In some embodiments, the compounds of this invention have one or more of the following features: R is NR'R' or NR'R'': R is NRR, wherein R is Hor Caliphatic optionally substituted with 1-3 R', and R is Caliphatic optionally substituted with 1-4 R7 or a 3- to 6-membered 35 monocyclic or 6- to 10-membered bicyclic ring optionally O substituted with 1-4 R'; R is H or Caliphatic, and R is Calkyl that is optionally substituted with 1-4 of R7: R is H. and R is C, alkyland optionally substituted with 1-4 R. R. 40 is ethyl substituted at the carbonatom attached to the nitrogen atom with R: R7 is an aryl or heteroaryl, and is optionally substituted with 1-3 of R: Risphenyl, pyridyl, or pyrimidyl, N and is optionally substituted with 1-3 of R. R7 is phenyl n CH3 optionally substituted with 1-3 of R: R is phenyl optionally 45 substituted at the ortho- or meta-position with R and also optionally substituted at the para-position with R (e.g., the optional substituent Rat an ortho- or meta-position, when present, is halo). In some embodiments of the compounds of this invention, 50 R’ is phenyl substituted at the para-position with -R or - N(R): O O r R is 4- to 8-membered heterocyclic ring optionally con taining 1-3 groups each independently selected from —N(R')-, -O-, or -S , and the heterocyclic ring is 55 optionally substituted with 1-3 of Q; Each Q is independently selected from halogen, hydroxy, C. alkyl, benzyl, —CF. W. —C(O)—W. —C(O)—N N (W), C(O) O W: N Each W is independently selected from —H, C alkyl, or 60 cycloalkyl; N- YCH: Each R is independently —H. C. heterocyclic ring, or Caliphatic, wherein C. heterocyclic ring and Cali R’ is phenyl substituted at the para-position with —OR. R. phatic are each optionally substituted with 1-3 Q; or is —H. C. heterocycloalkyl ring, or C- alkyl, wherein C Two R groups together with the Natom to which they are 65 heterocycloalkyl ring and C alkyl are each optionally Sub bound form a 4- to 8-membered ring containing additional 1 stituted with 1-3 Q; R is pyrrolidinyl or piperidinyl and or 2 groups each independently selected from N(R') , optionally substituted with 1-3 Q; R is US 8,247,421 B2 36 or different nitrogen atom or carbon atom to which they are N1 CH attached, form a 4- to 8-membered heterocyclic ring. N-CH or Some other embodiments of the compounds of this inven 5 tion have one or more of the following features: R is

R is CH3,

N1

10 O-ch. O-ch. O .. t 15 C YCH, CH3. N

r 2 N r N- N YCH, Still some of the compounds of this invention have one or more of the following features: R is pyrimidinyl optionally substituted with 1-3 of R: R is 5-pyrimidyl optionally sub 25 stituted at the 2-position with R: Ris-R, OR. -SR, —N(R), halogen, or—CN; Each R is independently —H. r 2 C. carbocyclic ring, C. heterocyclic ring, or Caliphatic, wherein C. carbocyclic ring, Cheterocyclic ring and C 30 N r aliphatic are each optionally substituted with 1-3 Q; R is N-l.

y H. O 35 r N Nals N -C) H \=N -(/ \)- 40 R’ is pyridinyl optionally substituted with 1-3 of R, R is N 3-pyridinyl optionally substituted with 1-3 of R: R is 3-py 45 ridinyl optionally substituted at the 6-position with R: R is -Q, R, OR, N(R), halogen, or —CN; each R is N n independently —H. C. heterocyclic ring, or Caliphatic, O C H3 wherein C. heterocyclic ring and Caliphatic are each 50 optionally substituted with 1-3 Q; or two R groups together with the Natom to which they are bound form a 4- to 8-mem bered hetercycloalkyl ring optionally containing an addi tional 1 or 2 groups selected from N(R'7)— —O—, or —S , wherein the 4- to 8-membered heterocycloalkyl ring 55 is optionally and independently substituted with 1-3 of W: each R'' is independently, hydrogen, or C, aliphatic, wherein each Caliphatic is optionally substituted with 1-3 of Q; each Q is independently selected from halogen, hydroxy, C alkyl, —CF, NH, N(H)—W, or —N 60 (W); each Wis independently selected from H. C. alkyl, aralkyl, cycloalkyl or heterocyclic ring; each Ce alkyl, O aralkyl, cycloalkyl or heterocyclic ring is optionally Substi tuted with 1-3 of halogen, OR, —CN, C- alkyl or —NR'R'; or one W group, together with the nitrogenatom 65 r to which it is attached and a carbon atom of R, form a 4- to 8-memberedg or two Wring; groups, together with the same US 8,247,421 B2 37 38 -continued Still some other compounds of this invention have one or

more of the following features: R is OR or - SR: R is optionally substituted phenyl. Specific examples of the compounds of this invention include compounds I-1 through I-304 or pharmaceutically acceptable salts thereof; compounds IA-0 through IA-13 or pharmaceutically acceptable salts thereof; compounds II-0 through II-17 or pharmaceutically acceptable salts thereof; and compounds EG4, EG5. EG6, EG7 and EG8 or pharma 10 ceutically acceptable salts thereof. The structures of these specific compounds are show hereinafter. one R is aryl, heteroaryl, C-Cs cycloalkyl, or 4- to 8-mem In another aspect, the invention features compounds of bered heterocyclic ring each optionally substituted with 1-3 formula Id of Q: R is a 4- to 8-membered heterocyclic ring optionally 15 substituted with 1-3 of Q; one R is a 5- to 6-membered Id heterocyclic ring optionally substituted with 1-3 of Q; one R is a piperazine ring optionally Substituted with 1-3 of Q; one R is Q: Q is NHW, NW, NH SOW, NH COW, CO. NHW, CO NW, SONHW, SO f Y - G)- NW, SW, OW, or W: Q is NHW, NW, or OW: NC eN W is C alkyl, aralkyl, cycloalkyl or heterocyclic ring; each RI C. alkyl, aralkyl, cycloalkyl or heterocyclic ring is option ally substituted with 1-3 of halogen, —OR, CN, C alkyl, Ce alkyl or NR'R'; W is C, alkyl or heterocy 25 clic ring; each C alkyl, aralkyl, cycloalkyl or heterocyclic ring is optionally substituted with 1-3 of halogen, OR, —CN, C-alkyl, C. alkyl or -NR'R'': one R is R. or a pharmaceutically acceptable salt thereof, wherein: —OR or N(R); R is independently H. C. carbocyclic A is phenyl, pyridinyl, or pyrmidinyl: 30 B is NR'R'"; ring, C. heterocyclic ring, or Caliphatic, wherein C R" is H or C alkyl: carbocyclic ring, C. heterocyclic ring and Caliphatic are R" is a Caliphatic substituted with NH or NHCH: each optionally substituted with 1-3 Q; or two R groups, O together with the Natom to which they are bound, form a 4 R" is an N-heterocycloaliphatic optionally substituted on N to 8-membered ring optionally containing additional 1 or 2 35 with CH; or groups selected from N(R)-, -O-, or—S , wherein R" and R" together with the N to which they are attached the 4- to 8-membered ring is optionally and independently form a 5- to 7-membered heterocycloaliphatic ring contain Substituted with 1-3 of W. ing two Natoms. Still some other compounds of this invention have one or Compounds of formula Id show selectivity for PLK1 over more of the following features: R is a 4- to 10-membered 40 PLK2 and PLK3. heterocyclic monocyclic or bicyclic ring optionally Substi In some embodiments, R' is trifluromethyl. tuted with 1-3 of R: R is a 4- to 6-membered heterocyclic In some embodiments, R' is Hor methyl and R" is 2-amino monocyclic ring optionally substituted with 1-3 of R: R is or 2-methylamino-ethane. Q: Q is selected from —C(O)—W. —C(O) N(W). C In other embodiments, B is piperazinyl or diazepanyl, (O) O Wor–SO. W: Q is selected from –C(O). W. 45 optionally substituted at the 4-position with methyl. Still some other compounds of this invention have one or In certain embodiments, R' is H or methyl, and R" is pyr more of the following features: R is a Cascarbocycle option rolidine or piperidine and is optionally substituted at N with ally substituted with 1-3 R; one R is Q, Q is selected from methyl. hydroxy, NH, -N (H) W. N(W), N(H) SO, In certain embodiments, R" is a Caliphatic substituted W. C(O) N(W), N(H)–C(O) W, or - O C(O) 50 with NH, or - NHCH. W; Q is selected from N(H) C(O) W: R is NR'R'': In certain embodiments, R" is cyclohexyl substituted with Ris HandR'' is C(R'R')C(=O)NR'R'': R is H: - NH or - NHCH R" is C alkyl; R'' is H; and R' is alkyl substituted with In certain embodiments, R" is piperazine or pyrrolidine. trifluoromethyl or hydroxy, or R' is cycloalkyl substituted The compounds of this invention include those described with hydroxy: R' is 55 herein, and are further illustrated by the classes, subclasses, and species disclosed herein. Another aspect of this invention provides a pharmaceutical composition containing one of the compounds described CF3, , or above, and a pharmaceutically acceptable carrier, adjuvant, or OH 60 vehicle. The composition may further include another thera HO peutic agent selected from the group consisting of synthetic small molecule VEGF receptor antagonists, small molecule growth factor receptor antagonists, inhibitors of the EGF receptor and/or VEGF receptor and/or integrin receptors or 65 any other protein tyrosine kinase receptors which are not classified under the synthetic small-molecules, inhibitors directed to EGF receptor and/or VEGF receptor and/or inte US 8,247,421 B2 39 40 grin receptors or any other protein tyrosine kinase receptors, (5-FU), pemetrexed, tegafurfuracil, uracil mustard, fludara which are fusion proteins, compounds which interact with bine, gemcitabine, capecitabine, mercaptopurine, cladribine, nucleic acids and which are classified as alkylating agents or thioguanine, methotrexate, pentostatin, hydroxyurea, or folic platinum compounds, compounds which interact with acid, a phleomycin, a bleomycin or a derivative or salt thereof, nucleic acids and which are classified as anthracyclines, as CHPP. BZPP, MTPP, BAPP, liblomycin, an acridine or a DNA intercalators or as DNA cross-linking agents, including derivative thereof, a rifamycin, an actinomycin, adramycin, a DNA minor-groove binding compounds, anti-metabolites, camptothecin Such as irinotecan or topotecan, an amsacrine naturally occurring, semi-synthetic or synthetic bleomycin or analogue thereof, a tricyclic carboxamide, an his type antibiotics, inhibitors of DNA transcribing enzymes, and tonedeacetylase inhibitor such as SAHA, MD-275, trichos especially the topoisomerase I or topoisomerase II inhibitors, 10 tatin A, CBHA, LAQ824, or valproic acid, an anti-cancer chromatin modifying agents, mitosis inhibitors, anti-mitotic drug from plants such as paclitaxel (taxol), docetaxel or taxo agents, cell-cycle inhibitors, proteasome inhibitors, enzymes, tere, a Vinca alkaloid such as navelbine, vinblastin, Vincristin, hormones, hormone antagonists, hormone inhibitors, inhibi vindesine or vinorelbine, a tropolone alkaloid Such as colchi tors of steroid biosynthesis, Steroids, cytokines, hypoxia-se cine or a derivative thereof, a macrollide Such as maytansine, lective cytotoxins, inhibitors of cytokines, lymphokines, anti 15 an ansamitocin or rhizoxin, an antimitotic peptide Such as bodies directed against cytokines, oral and parenteral phomopsin or dolastatin, an epipodophyllotoxin or a deriva tolerance induction agents, supportive agents, chemical tive of podophyllotoxin Such as etoposide or teniposide, a radiation sensitizers and protectors, photo-chemically acti Steganacin, an antimitotic carbamate derivative such as com vated drugs, synthetic poly- or oligonucleotides, optionally bretastatin oramphetinile, procarbazine, a proteasome inhibi modified or conjugated, non-steroidal anti-inflammatory tor Such as bortezomib, an enzyme Such as asparaginase, drugs, cytotoxic antibiotics, antibodies targeting growth fac pegylated asparaginase (pegaspargase) or a thymidine-phos tors or their receptors, antibodies targeting the Surface mol phorylase inhibitor, a gestagen or an estrogen Such as estra ecules of cancer cells, inhibitors of metalloproteinases, met mustine (T-66) or megestrol, an anti-androgen such as fluta als, inhibitors of oncogenes, inhibitors of gene transcription mide, casodex, anandron or cyproterone acetate, an or of RNA translation or protein expression, complexes of 25 aromatase inhibitor Such as aminogluthetimide, anastrozole, rare earth elements, compounds which reduces the transport formestan or letrozole, a GNrHanalogue Such as leuprorelin, of hyaluronan mediated by one or more ABC transporters, buserelin, goserelin or triptorelin, an anti-estrogen Such as and photo-chemotherapeutic agents. tamoxifen or its citrate salt, droloxifene, trioxifene, ralox Still, the pharmaceutical compositions of this invention ifene or Zindoxifene, a derivative of 17.beta-estradiol such as may further contains another therapeutic agent selected from 30 ICI 164.384 or ICI 182,780, aminoglutethimide, formestane, the group consisting of a small molecule VEGF receptor fadrozole, finasteride, ketoconazole, a LH-RH antagonist antagonist such as vatalanib (PTK-787/ZK222584), such as leuprolide, a steroid such as prednisone, predniso SU-5416, SU-6668, SU-11248, SU-14813, AZD-6474, lone, methylprednisolone, dexamethasone, budenoside, fluo AZD-2171, CP-547632, CEP-7055, AG-013736, IM-842 or cortolone or triamcinolone, an interferon Such as interferon GW-78.6034, a dual EGFR/HER2 antagonist such as gefi 35 ..beta., an interleukin such as IL-10 or IL-12, an anti-TNFal tinib, erlotinib, CI-1033 or GW-2016, an EGFR antagonist pha. antibody Suchasetanercept, an immunomodulatory drug such as iressa (ZD-1839), tarceva (OSI-774), PKI-166, EKB Such as thalidomide, its R- and S-enantiomers and its deriva 569, HKI-272 or herceptin, an antagonist of the mitogen tives, or revimid (CC-5013), a leukotrien antagonist, mito activated protein kinase such as BAY-43-9006 or BAY-57 mycin C, an aziridoquinone such as BMY-42355. AZQ or 9006, a quinazoline derivative such as 4-(3-chloro-4- 40 EO-9, a 2-nitroimidazole such as misonidazole, NLP-1 or fluorophenyl)amino-6-4-(N,N-dimethylamino)-1-oxo-2- NLA-1, a nitroacridine, a nitroquinoline, a nitropyrazoloacri bute-n-1-yl)amino-7-((S)-tetrahydrofuran-3-yloxy)- dine, a "dual-function’ nitro aromatic such as RSU-1069 or quinazoline or 4-(3-chloro-4-fluoro-phenyl)amino]-6-4- RB-6145, CB-1954, a N-oxide of nitrogen mustard such as (homomorpholin-4-yl)-1-oxo-2-bu-ten-1-yl)amino-7-(S)- nitromin, a metal complex of a nitrogen mustard, an anti-CD3 (tetrahydrofuran-3-yl)oxy-quinazoline, O a 45 oranti-CD25 antibody, a tolerance induction agent, a biphos pharmaceutically acceptable salt thereof, a protein kinase phonate or derivative thereof Such as minodronic acid or its receptor antagonist which is not classified under the synthetic derivatives (YM-529, Ono-5920, YH-529), Zoledronic acid Small molecules such as atrasentan, rituximab, cetuximab, monohydrate, ibandronate Sodium hydrate or clodronate AvastinTM (bevacizumab), IMC-1C11, erbitux (C-225), disodium, a nitroimidazole Such as metronidazole, misonida DC-101, EMD-72000, vitaxin, imatinib, a protein tyrosine 50 Zole, benznidazole or nimorazole, a nitroaryl compound Such kinase inhibitor which is a fusion protein such as VEGFtrap, as RSU-1069, a nitroxyl or N-oxide such as SR-4233, an an alkylating agent or a platinum compound Such as mel halogenated pyrimidine analogue Such as bromodeoxyuri phalan, cyclophosphamide, an oxazaphosphorine, cisplatin, dine, iododeoxyuridine, a thiophosphate such as WR-2721, a carboplatin, oxaliplatin, satraplatin, tetraplatin, iproplatin, photo-chemically activated drug such as porfimer, photofrin, mitomycin, streptozocin, carmustine (BCNU), lomustine 55 a benzoporphyrin derivative, a pheophorbide derivative, (CCNU), busulfan, ifosfamide, streptozocin, thiotepa, merocyanin 540 (MC-540) or tin etioporpurin, an ant-tem chlorambucil, a nitrogen mustard Such as mechlorethamine, plate or an anti-sense RNA or DNA such as oblimersen, a an ethyleneimine compound, an alkylsulphonate, daunorubi non-steroidal inflammatory drug such as acetylsalicyclic cin, doxorubicin (adriamycin), liposomal doxorubicin acid, mesalazin, ibuprofen, naproxen, flurbiprofen, fenopro (doxil), epirubicin, idarubicin, mitoxantrone, amsacrine, dac 60 fen, fenbufen, ketoprofen, indoprofen, pirprofen, carprofen, tinomycin, distamycin or a derivative thereof, metropsin, oxaprozin, pranoprofen, miroprofen, tioxaprofen, Suprofen, pibenzimol, mitomycin, CC-1065, a duocarmycin, mithra alminoprofen, tiaprofenic acid, fluprofen, indomethacin, mycin, chromomycin, olivomycin, aphtalanilide such as pro Sulindac, tolmetin, Zomepirac, nabumetone, diclofenac, fen pamidine or stilbamidine, an anthramycin, an aziridine, a clofenac, alclofenac, bromfenac, ibufenac, aceclofenac, nitrosourea or a derivative thereof, a pyrimidine or purine 65 acemetacin, fentiazac, clidanac, etodolac, oXpinac, mefe analogue or antagonist or an inhibitor of the nucleoside namic acid, meclofenamic acid, flufenamic acid, nifluminic diphosphate reductase Such as cytarabine, 5-fluorouracile acid, tolfenamic acid, diflunisal, flufenisal, piroxicam, US 8,247,421 B2 41 42 tenoxicam, lomoxicam, nimeSulide, meloxicam, celecoxib, fluorophenyl)amino-6-4-(N,N-dimethylamino)-1-oxo-2- rofecoxib, or a pharmaceutically acceptable salt of a non bute- n-1-yl)amino-7-((S)-tetrahydrofuran-3-yloxy)- steroidal inflammatory drug, a cytotoxic antibiotic, an anti quinazoline or 4-(3-chloro-4-fluoro-phenyl)amino]-6-4- body targeting the Surface molecules of cancer cells Such as (homomorpholin-4-yl)-1-oxo-2-bu-ten-1-yl)amino-7-(S)- apolizumab or 1D09C3, an inhibitor of metalloproteinases (tetrahydrofuran-3-yl)oxy-quinazoline, O a such as TIMP-1 or TIMP-2, Zinc, an inhibitor of oncogenes pharmaceutically acceptable salt thereof, an inhibitor of the such as P53 and Rb, a complex of rare earth elements such as transcription factor complex ESX/DRIP130/Sur-2, an inhibi the heterocyclic complexes of lanthanides, a photo-chemo tor of HER-2 expression, such as the heat shock protein therapeutic agent such as PUVA, an inhibitor of the transcrip HSP90 modulator geldanamycin and its derivative 17-ally tion factor complex ESX/DRIP130/Sur-2, an inhibitor of 10 HER-2 expression, such as the heat shock protein HSP90 laminogeldanamycin or 17-AAG, a protein kinase receptor modulator geldanamycin and its derivative 17-allylami antagonist which is not classified under the synthetic Small nogeldanamycin or 17-AAG, a compound which reduces the molecules such as atrasentan, rituximab, cetuximab, Avas transport of hyaluronan mediated by one or more ABC trans tinTM (bevacizumab), IMC-1C11, erbitux (C-225), DC-101, porters selected from a P-glycoprotein (P-gp) inhibitor mol 15 EMD-72000, vitaxin, imatinib, a P-glycoprotein (P-gp) ecule or inhibitor peptide, an MRP1 inhibitor, an antibody inhibitor molecule such as Zosuduidar (LY 335973), its salts directed againstand capable of blocking the ABC transporter, (especially the trichloride salt) and its polymorphs, an antisense oligomer, iRNA, siRNA or aptamer directed cyclosporin A, Verapamil or its R-isomer, tamoxifen, quini against one or more ABC transporters, or a therapeutic agent dine, d-alpha tocopheryl polyethylene glycol 1000 succinate, selected from IM-842, tetrathiomolybdate, squalamine, com VX-710, PSC833, phenothiazine, GF 120918 (II), SDZ PSC brestatin A4, TNP-470, marimastat, neovastat, bicalutamide, 833, TMBY, MS-073, S-9788, SDZ 280-446, XR(9051) and abarelix, oregovomab, mitumomab, TLK-286, alemtuzumab, functional derivatives, analogues and isomers of these, or an ibritumomab, temozolomide, denileukin diftitox, aldesleu antibody targeting the Surface molecules of cancer cells Such kin, dacarbazine, floxuridine, plicamycin, mitotane, pipobro as apolizumab or ID09C3. man, plicamycin, tamLOXifen and testolactone. 25 In some embodiments, the compositions of this invention The pharmaceutical composition of this invention may still may further include 4-(3-chloro-4-fluorophenyl)amino)-6- further contains another therapeutic agent selected from the 4-(N,N-dimethylamino)-1-oxo-2-bute-n-1-yl)amino-7- group consisting of an anti-cancer drug from plants such as ((S)-tetrahydrofuran-3-yloxy)-quinazoline, 4-(3 -chloro-4- paclitaxel (taxol), docetaxel or taxotere, a Vinca alkaloid Such fluorophenyl)amino-6-4-(N,N-dimethylamino)-1-oxo-2- as navelbine, vinblastin, Vincristin, vindesine or vinorelbine, 30 buten-1-yl)amino-7-(S)-tetrahydrofuran-3-yloxy)- a Vinca alkaloid such as navelbine, vinblastin, Vincristin, Vin quinazoline, 4-(3 -chloro-4-fluoro-phenyl)amino]-6-4- desine or vinorelbine, an alkylating agent or a platinum com (homomorpholin-4-yl)-1-oxo-2-buten-1-yl)amino)-7-(S)- pound Such as melphalan, cyclophosphamide, an oxazaphos (tetrahydrofuran-3-yl)oxy-quinazoline, 4-(3-chloro-4- phorine, cisplatin, carboplatin, oxaliplatin, satraplatin, fluoro-phenyl)amino]-6-4-(homomorpholin-4-yl)-1-oxo tetraplatin, iproplatin, mitomycin, Streptozocin, carmustine 35 2-bu-ten-1-yl)amino-7-(S)-(tetrahydrofuran-3-yl)oxy (BCNU), lomustine (CCNU), busulfan, ifosfamide, strepto quinazoline, 3-Z-1-(4-(N-(4-methyl-piperazin-1-yl)- Zocin, thiotepa, chlorambucil, a nitrogen mustard Such as methylcarbonyl)-N-methyl-amino)-anilino-)-1-phenyl mechlorethamine, an immunomodulatory drug such as thali methylene-6-methoxycarbonyl-2-indolinone, 3-Z-1-(4-(N- domide, its R- and S-enantiomers and its derivatives, or ((4-methyl-piperazin-1-yl)-methylcarbonyl)-N-methyl revimid (CC-5013)), an ethyleneimine compound, an alkyl 40 amino)-a-nilino)-1-phenyl-methylene-6-methoxycarbonyl Sulphonate, daunorubicin, doxorubicin (adriamycin), liposo 2-indolinone, 3-Z-1-(4-dimethylaminomethylanilino)-1-(4- mal doxorubicin (doxil), epirubicin, idarubicin, mitox (2-carboxyethyl)phenyl)methyle-ne-6-fluoro-2-indolinone, antrone, amsacrine, dactinomycin, distamycin or a derivative or a pharmaceutically acceptable salt thereof. thereof, netropsin, pibenzimol, mitomycin, CC-1065, a duo The compounds of this invention can be used to inhibit carmycin, mithramycin, chromomycin, olivomycin, a phta 45 PLK protein kinase activity in a biological sample. lanilide such as propamidine or stilbamidine, an anthramycin, Thus, the invention further provides a method of inhibiting an aziridine, a nitrosourea or a derivative thereof, a pyrimi PLK protein kinase activity in a patient, which includes dine or purine analogue or antagonist or an inhibitor of the administering to the patient one of the compounds described nucleoside diphosphate reductase such as cytarabine, 5-fluo above or one of the compositions described above. rouracile (5-FU), pemetrexed, tegafurfuracil, uracil mustard, 50 Another aspect of this invention provides a method of fludarabine, gemcitabine, capecitabine, mercaptopurine, treating a proliferative disorder, a neurodegenerative disor cladribine, thioguanine, methotrexate, pentostatin, hydrox der, an autoimmune disorder, an inflammatory disorder, oran yurea, or folic acid, an acridine or a derivative thereof, a immunologically mediated disorder in a patient. The method rifamycin, an actinomycin, adramycin, a camptothecin Such includes administering to the patient in need thereofathera as irinotecan or topotecan, an amsacrine or analogue thereof, 55 peutically effective amount of one of the compounds or com a tricyclic carboxamide, an histonedeacetylase inhibitor Such positions described above. This method may further include as SAHA, MD-275, trichostatin A, CBHA, LAQ824, or val administering to the patient an additional therapeutic agent proic acid, a proteasome inhibitor Such as bortezomib, a small selected from a chemotherapeutic or anti-proliferative agent, molecule VEGF receptor antagonist such as Vatalanib (PTK an anti-inflammatory agent, an immunomodulatory or immu 787/ZK222584), SU-5416, SU-6668, SU-11248, SU-14813, 60 nosuppressive agent, a neurotrophic factor, an agent for treat AZD-6474, AZD-2171, CP-547632, CEP-7055, AG-013736, ing cardiovascular disease, an agent for treating destructive IM-842 or GW-786034, an antagonist of the mitogen-acti bone disorders, an agent for treating liver disease, an anti vated protein kinase such as BAY-43-9006 or BAY-57-9006, viral agent, an agent for treating blood disorders, an agent for a dual EGFR/HER2 antagonist such as gefitinib, erlotinib, treating diabetes, or an agent for treating immunodeficiency CI-1033 or GW-2016, an EGFR antagonist such as iressa 65 disorders, wherein (1) said additional therapeutic agent is (ZD-1839), tarceva (OSI-774), PKI-166, EKB-569, HKI-272 appropriate for the disease being treated, and (2) said addi or herceptin, a quinazoline derivative such as 4-(3-chloro-4- tional therapeutic agent is administered together with said US 8,247,421 B2 43 44 composition as a single dosage form or separately from said (wherein each R" is independently C-C alkyl or two R' composition as part of a multiple dosage form. together with the atoms to which they are attached fom a 5 or Still in another aspect, the invention provides a method of 6 membered ring optionally substituted with 1 to 4 methyl treating an oncological disease in a patient, which includes groups), a pyrimidine of formula 7 administering to the patient in need thereofatherapeutically effective amount of a compound or a pharmaceutical compo sition of those described above. Examples of the oncological disease include, but are not limited to, refractory or relapsed multiple myeloma, acute or chronic myelogenous leukaemia, myelodysplastic syndrome, myeloproliferativesyndromes, 10 acute lymphoblastic leukaemia, Hodgkin’s and non NC Hodgkin’s lymphoma. C The invention further provides a method of treating solid tumor, melanoma, myeloma, leukemia, lymphoma, neuro 15 blastoma, or a cancer selected from colon, breast, gastric, and an alkali metal carbonate with a palladium catalyst in a ovarian, cervical, lung, central nervous system (CNS), renal, suitable solvent. An example of the suitable palladium cata prostate, bladder, or pancreatic, in a patient by administering lyst is bis-(tri-tert-butylphosphine)palladium(0) and an to said patient one of the compounds orpharmaceutical com example of the suitable solvent is dioxane and the alkali metal positions described above. carbonate is potassium carbonate. To practice this process, The invention still further provides a method of treating the pyrimidine can be 4-chloro-2-(methylthio)pyrimidine-5- cancer in a patient, which includes administering to the carbonitrile, and the boronate ester can be 5-Trifluoromethyl patient one of the compounds or pharmaceutical composi 3-(4,4,5,5-tetramethyl-1,3,2-dioxoborolan-2-yl)-1-tosyl tions described above. Examples of the cancer include uro 1H-pyrrolo2,3-bipyridine. genital cancer, lung cancer, gastrointestinal cancer, head and 25 neck cancer, malignant mesothelioma, breast cancer, malig In some embodiments of the process of this invention, R is nant melanoma, childhood cancer and bone or soft tissue —SCH. SaCOa. The process of this invention may further includes the steps The invention also relates to a method of treating a disease of: involving cell proliferation, migration or apoptosis of cancer 30 (a) oxidizing a compound of formula 26 cells, orangiogenesis, in a human or non-human mammalian body, by administering simultaneously, separately or sequen tially to the patient a therapeutically effective amount of one of the compounds or pharmaceutical compositions described N 26 above. Examples of Such disease include Coeliac disease, 35 diabetes mellitus type 1 (IDDM), systemic lupus erythema tosus (SLE), Sjögren's syndrome, multiple sclerosis (MS), | Y-ch Hashimoto's thyroiditis, Graves disease, idiopathic thromb NC leN ocytopenic purpura, and rheumatoid arthritis (RA). RI Also within the scope of the invention is a process for 40 preparing a compound of formula I rN\,2 N N Tos NC | y 45 leN RI (wherein R' is as as previously described) with chlorine in aqueous ethanol to provide a compound of formula 27 rN\, 50 as 27 N or a pharmaceutically acceptable salt thereof, wherein the | y-lost variables R' and R are as defined above. This process 55 NC leN includes contacting a mixture of a boronate ester of formula 8 RI rN\, B(OR)2 8 2 N 60 N V RI Tos

rN\,2 N N (b) reacting a compound of formula 27 with a compound of Tos 65 formula RH, wherein R is HNRR, HOR or HSRandR, R and R are as previously described, in the presence of microwave irradiation to provide a compound of formula 28 US 8,247,421 B2 45 46 atoms. In still other embodiments, aliphatic groups contain 28 1-6 aliphatic carbon atoms, and in yet other embodiments N aliphatic groups contain 1-4 aliphatic carbonatoms. Suitable f Y aliphatic groups include, but are not limited to, linear or NC leN branched, substituted or unsubstituted alkyl, alkenyl, or alky nyl groups. Specific examples include, but are not limited to, methyl, ethyl, isopropyl. n-propyl. Sec-butyl, vinyl. n-bute RI N N nyl, ethynyl, and tert-butyl. As used herein, the term "Coalkyl” refers to either a bond 4. N 10 or a C- alkyl. Other carbon number ranges containing Zero Tos have the same meaning. It should be understood that if the aliphatic is alkenyl or and alkynyl, then the aliphatic group has at least 2 carbon atoms. c) removing the ToS protecting group to provide com The term “oxo' for substitutent refers to the group “—O' 15 which replace, e.g., two hydrogen atoms on a carbon atom of pounds of formula I. a methylen group to form —C(O)— (equivalent to Definitions —C(=O)—), i.e., a carbonyl group. As used herein, the following definitions shall apply unless The term "cycloaliphatic' or “carbocyclic' refers to a otherwise indicated. For purposes of this invention, the monocyclic Cls hydrocarbon or bicyclic C-2 hydrocarbon chemical elements are identified in accordance with the Peri that is completely saturated or that contains one or more units odic Table of the Elements, CAS version, Handbook of of unsaturation, but which is not aromatic, that has a single Chemistry and Physics, 75th Ed. Additionally, general prin point of attachment to the rest of the molecule wherein any ciples of organic chemistry are described in "Organic Chem individual ring in said bicyclic ring system has 3-7 members. istry'. Thomas Sorrell, University Science Books, Sausalito: Suitable cycloaliphatic groups include, but are not limited to, 1999, and “March's Advanced Organic Chemistry, 5th Ed., 25 cycloalkyl, bicycloalkyl, cycloalkenyl and bicycloalkenyl Ed. Smith, M. B. and March, J., John Wiley & Sons, New groups. Specific examples include, but are not limited to, York: 2001, the entire contents of which are hereby incorpo cyclohexyl, bicycle 2.2.1]heptanyl, cyclopropenyl, and rated by reference. cyclobutyl. The term "heteroaliphatic', as used herein, means As described herein, a specified number range of atoms aliphatic groups wherein one or two carbon atoms are inde includes any integer therein. For example, a group having 30 pendently replaced by one or more of oxygen, Sulfur, nitro from 1-4 atoms could have 1, 2, 3, or 4 atoms. gen, phosphorus, or silicon. Heteroaliphatic groups may be As described herein, compounds of the invention may substituted or unsubstituted, branched or unbranched, cyclic optionally be substituted with one or more substituents, such or acyclic, and include "heterocycle”, “heterocyclyl”, “het as are illustrated generally above, or as exemplified by par erocycloaliphatic”, or "heterocyclic' groups. ticular classes, Subclasses, and species of the invention. It will 35 The term “heterocycle”, “heterocyclyl”, or "heterocyclic” be appreciated that the phrase “optionally substituted” is used as used herein means non-aromatic, monocyclic, bicyclic, or interchangeably with the phrase “substituted or unsubsti tricyclic ring systems in which one or more ring members are tuted.” In general, the term “substituted”, whether preceded an independently selected heteroatom. A heterocyclic ring by the term “optionally’ or not, refers to the replacement of includes heterocycloaliphatic, which in turn includes hetero hydrogen radicals in a given structure with the radical of a 40 cycloalkyl, heterocycloalkenyl, and heterocycloalkynyl. In specified Substituent. Unless otherwise indicated, an option some embodiments, the "heterocycle”, “heterocyclyl', or ally Substituted group may have a Substituent at each Substi "heterocyclic group has three to fourteen ring members in tutable position of the group, and when more than one posi which one or more ring members is a heteroatom indepen tion in any given structure may be substituted with more than dently selected from oxygen, Sulfur, nitrogen, orphosphorus, one Substituent selected from a specified group, the Substitu 45 and each ring in the system contains 3 to 7 ring members. ent may be either the same or different at every position. Suitable heterocycles include, but are not limited to, 3-1 H Combinations of substituents envisioned by this invention are benzimidazol-2-one, 3-(1-alkyl)-benzimidazol-2-one, 2-tet preferably those that result in the formation of stable or rahydrofuranyl, 3-tetrahydrofuranyl, 2-tetrahydrothiophe chemically feasible compounds. nyl, 3-tetrahydrothiophenyl, 2-morpholino, 3-morpholino, The term “stable', as used herein, refers to compounds that 50 4-morpholino, 2-thiomorpholino, 3-thiomorpholino, 4-thio are not substantially altered when subjected to conditions to morpholino. 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl, allow for their production, detection, recovery, purification, 1-tetrahydropiperazinyl, 2-tetrahydropiperazinyl, 3-tetrahy and use for one or more of the purposes disclosed herein. In dropiperazinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, Some embodiments, a stable compound or chemically fea 4-piperidinyl, 1-pyrazolinyl, 3-pyrazolinyl, 4-pyrazolinyl, sible compound is one that is not substantially altered when 55 2-thiazolidinyl, 3-thiazolidinyl, 4-thiazolidinyl, 1-imidazo kept at a temperature of 40° C. or less, in the absence of lidinyl, 2-imidazolidinyl, 4-imidazolidinyl, indolinyl, tet moisture or other chemically reactive conditions, for at least rahydroquinolinyl, tetrahydroisoquinolinyl, benzothiolane, a week. benzodithiane, and 1,3-dihydro-imidazol-2-one. The term “aliphatic' or “aliphatic group', as used herein, Cyclic groups, (e.g., cycloaliphatic and heterocycles), can means a straight-chain (i.e., unbranched), branched, or cyclic 60 be linearly fused, bridged, or spirocyclic. substituted or unsubstituted hydrocarbon chain that is com The term "heteroatom' means one or more of oxygen, pletely saturated or that contains one or more units of unsat Sulfur, nitrogen, or phosphorus, (including, any oxidized uration that has a single point of attachment to the rest of the form of nitrogen, Sulfur, or phosphorus; the quaternized form molecule. Unless otherwise specified, aliphatic groups con of any basic nitrogen or, a Substitutable nitrogen of a hetero tain 1-20 aliphatic carbon atoms. In some embodiments, ali 65 cyclic ring, for example N (as in 3,4-dihydro-2H-pyrrolyl), phatic groups contain 1-10 aliphatic carbon atoms. In other NH (as in pyrrolidinyl) or NR" (as in N-substituted pyrrolidi embodiments, aliphatic groups contain 1-8 aliphatic carbon nyl)). US 8,247,421 B2 47 48 The term "unsaturated’, as used herein, means that a moi The term “protecting group' and “protective group’ as ety has one or more units of unsaturation. used herein, are interchangeable and refer to an agent used to The term “nonaromatic', as used herein, describes rings temporarily block one or more desired reactive sites in a that are either saturated or partially unsaturated. multifunctional compound. In certain embodiments, a pro The term “aromatic’, as used herein, describes rings that tecting group has one or more, or preferably all, of the fol are fully unsaturated. lowing characteristics: a) is added selectively to a functional The term “alkoxy”, or “thioalkyl, as used herein, refers to group in good yield to give a protected Substrate that is b) stable to reactions occurring at one or more of the other an alkyl group, as previously defined, attached to the principal reactive sites; and c) is selectively removable in good yield by carbon chain through an oxygen (“alkoxy”) or sulfur (“thio 10 reagents that do not attack the regenerated, deprotected func alkyl) atom. tional group. Exemplary protecting groups are detailed in The terms “haloalkyl, "haloalkenyl', “haloaliphatic’, and Greene, T. W., Wuts, P. G in “Protective Groups in Organic "haloalkoxy' mean alkyl, alkenyl or alkoxy, as the case may Synthesis'. Third Edition, John Wiley & Sons, New York: be, substituted with one or more halogen atoms. The terms 15 1999 (and other editions of the book), the entire contents of “halogen”, “halo', and “hal' mean F, Cl, Br, or I. which are hereby incorporated by reference. The term "nitro The term “aryl used alone or as part of a larger moiety as gen protecting group', as used herein, refers to an agents used in “aralkyl”, “aralkoxy’, or “aryloxyalkyl, refers to mono to temporarily block one or more desired nitrogen reactive cyclic, bicyclic, and tricyclic ring systems having a total of sites in a multifunctional compound. Preferred nitrogen pro tecting groups also possess the characteristics exemplified five to fourteen ring members, wherein at least one ring in the above, and certain exemplary nitrogen protecting groups are system is aromatic and wherein each ring in the system con also detailed in Chapter 7 in Greene, T. W. Wuts, P. G in tains 3 to 7 ring members. The term “aryl' may be used “Protective Groups in Organic Synthesis'. Third Edition, interchangeably with the term “aryl ring. The term “aryl John Wiley & Sons, New York: 1999, the entire contents of also refers to heteroaryl ring systems as defined hereinbelow. 25 which are hereby incorporated by reference. The term "heteroaryl, used alone or as part of a larger In some embodiments, an alkyl or aliphatic chain can be moiety as in "heteroaralkyl or "heteroarylalkoxy’, refers to optionally interrupted with another atom or group. This monocyclic, bicyclic, and tricyclic ring systems having a total means that a methylene unit of the alkyl or aliphatic chain is of five to fourteen ring members, wherein at least one ring in optionally replaced with said other atom or group. Examples the system is aromatic, at least one ring in the system contains 30 of such atoms or groups would include, but are not limited to, one or more heteroatoms, and wherein each ring in the system NR— —O— —S , —CO. , —OC(O)— —C(O) contains 3 to 7 ring members. The term "heteroaryl' may be CO , C(O)—, C(O)NR , C(=N-CN) , used interchangeably with the term "heteroaryl ring” or the - NRCO -, - NRC(O)O-, -SO2NR-, - NRSO , term "heteroaromatic'. Suitable heteroaryl rings include, but 35 NRC(O)NR , OC(O)NR NRSONR , are not limited to, 2-furanyl, 3-furanyl, N-imidazolyl, 2-imi —SO , or - SO , wherein R is defined herein. Unless dazolyl, 4-imidazolyl, 5-imidazolyl, benzimidazolyl, 3-isox otherwise specified, the optional replacements form a chemi azolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, cally stable compound. Optional interruptions can occur both 5-oxazolyl, N-pyrrolyl 2-pyrrolyl, 3-pyrrolyl 2-pyridyl, within the chain and at either end of the chain; i.e., both at the 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrim 40 point of attachment and/or also at the terminal end. Two idinyl, pyridazinyl (e.g., 3-pyridazinyl), 2-thiazolyl, 4-thiaz optional replacements can also be adjacent to each other olyl, 5-thiazolyl, tetrazolyl (e.g., 5-tetrazolyl), triazolyl (e.g., within a chain so long as it results in a chemically stable 2-triazolyl and 5-triazolyl), 2-thienyl, 3-thienyl, benzofuryl, compound. The optional interruptions or replacements can also completely replace all of the carbon atoms in a chain. For benzothiophenyl, indolyl (e.g., 2-indolyl), pyrazolyl (e.g., 45 2-pyrazolyl), isothiazolyl, 1.2.3-oxadiazolyl, 1,2,5-oxadiaz example, a Caliphatic can be optionally interrupted or olyl, 1,2,4-oxadiazolyl, 1.2.3-triazolyl, 1.2.3-thiadiazolyl, replaced by —NR—, —C(O)—, and —NR— to form 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, purinyl, pyrazinyl, 1.3, - NRC(O)NR - (a urea). 5-triazinyl, quinolinyl (e.g., 2-quinolinyl, 3-quinolinyl, As used herein, the term “1-3 (or other similar numerical 4-quinolinyl), and isoquinolinyl (e.g., 1-isoquinolinyl, 3-iso 50 terms) refers to a range of 1 to 3. quinolinyl, or 4-isoquinolinyl). Unless otherwise specified, if the replacement or interrup tion occurs at the terminal end, the replacementatom is bound When the term “ortho”, “meta', or “para’ is used to iden to an H on the terminal end. For example, if CH2CHCH tify the position of a Substituent on a 6-member ring system, were optionally interrupted with —O—, the resulting com it is relative to the atom by which this ring system is attached 55 pound could be —OCHCH. —CHOCH or to the core of the copound of formula. For instance, a phenyl —CHCH-OH. substituted at the ortho-position with methyl, at the meta Unless otherwise stated, structures depicted herein are also position with fluoro, and at the para-position with isopropyl is meant to include all isomeric (e.g., enantiomeric, diastereo meric, and geometric (or conformational)) forms of the struc 60 ture; for example, the R and S configurations for each asym HC F metric center, (Z) and (E) double bond isomers, and (Z) and CH (E) conformational isomers. Therefore, single stereochemi cal isomers as well as enantiomeric, diastereomeric, and geo metric (or conformational) mixtures of the present com CH3. 65 pounds are within the scope of the invention. Unless otherwise stated, all tautomeric forms of the com pounds of the invention are within the scope of the invention. US 8,247,421 B2 49 50 Unless otherwise stated, a substituent can freely rotate around any rotatable bonds. For example, a Substituent drawn Scheme 1 aS NC-N/ S. y-sch, R N N O 1 10 N N 1 NC-N4 \ also represents S. Y-soci, 15 R N N R2-H 1 N N 2 NC-N/ 3. s2 R N sN R2 Additionally, unless otherwise stated, structures depicted 25 herein are also meant to include compounds that differ only in 1 the presence of one or more isotopically enriched atoms. For example, compounds having the present structures except for the replacement of hydrogen by deuterium or tritium, or the 30 replacement of a carbon by a C- or C-enriched carbon are Referring to Scheme 1, oxidation of compound 1 to pro within the scope of this invention. Such compounds are use vide compound 2 can be achieved with a known oxidizing ful, for example, as analytical tools or probes in biological agent such as oXone or m-CPBA or chlorine gas in a solvent assayS. Such as aqueous alcohol, acetonitrile or dichloromethane. The following abbreviations are used: 35 Displacement with R. H., wherein R H is HNRR, PG protecting group HOR or HSR, to give compound 3 can be achieved ther LG leaving group mally or using microwave heating in a solvent such as diox DCM dichloromethane ane, tetrahydrofuran, ethanol, isopropanol or butanol. Ac acetyl DMF dimethylformamide 40 An alternative method for preparing compounds of the EtOAc ethyl acetate invention is illustrated in Scheme 2. DMSO dimethylsulfoxide MeCN acetonitrile Scheme 2 TCA trichloroacetic acid ATPadenosine triphosphate 45 NC 21 NN EtOH ethanol Ph phenyl R N ls Hvorovsis Me methyl N N S(O)CH, SW Et ethyl 2 Bulbutyl 50 DEAD diethylazodicarboxylate HEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic 1 acid BSA bovine serum albumin 55 DTT dithiothreitol MOPS 4-morpholinepropanesulfonic acid NMR nuclear magnetic resonance HPLC high performance liquid chromatography NC 21 NN LCMS liquid chromatography-mass spectrometry 60 TLC thin layer chromatography R ls Rt retention time N s OH Chlorination Synthesis of the Compounds The compounds of this invention may, in general, be pre 2 pared by methods such as those depicted in the schemes 65 below. Unless otherwise indicated, all variables in the follow 2-2 ing schemes are as defined herein. US 8,247,421 B2 51 52 -continued -continued NC B(OR)2 21 NN 2 N R N N R N o 5 N N Cl st NC 2 N -- % N He 2 C 7 8 2-3 10 N NC 21 NN | y NC leN R N susN R2 15 R N N 2 N N H % N

Referring to Scheme 2, hydrolysis of compound 1 can be Referring to Scheme 3, oxidation of compound 4 can be achieved by water in the presence of a base such as triethy achieved by a range of standard oxidising agents such as lamine or ethyldiisopropylamine in a solvent such as aceto oxone or mGPBA in a solvent such as acetonitrile or dichlo nitrile, tetrahydrofuran or dioxane to provide compound 2-2. romethane. reaction of compound 5 with compound 6 may be Chlorination of compound 2-2 to provide compound 2-3 can 25 carried as described in Scheme I to provide compound 7. The be achieved by phosphorous oxychloride in a solvent Such as coupling reaction of compound 7 with compound 8, wherein toluene. Reaction of compound 2-3 with R2-H, wherein —B(OR") can be a boronic acid—B(OH)), a boronate ester R H is HNR'R'', HOR or HSR, can be achieved ther such as —B(OMe) or a cyclic boronate ester such as mally or using microwave heating in a solvent Such as diox ane, tetrahydrofuran, ethanol, isopropanol or butanol to pro 30 vide compound I. In some instances, the azaindole 1-nitrogen is optionally protected with an arylsulphonate, such as tosyl or benzene Sulphonate, a silyl group Such as tert-butyldimethylsilyl or 35 triisopropylsilyl, or is left unprotected. If the nitrogen is pro tected by a Sulphonate group then it can be removed in a provides compound I. Subsequent step by treatment with base Such as aqueous The azaindole nitrogen may be protected with an arylsul Sodium hydroxide, lithium hydroxide or potassium carbonate phonate, such as tosyl or benzene Sulphonate, a silyl group in a solvent Such as methanol, ethanol or isoproanol. If the 40 such as tert-butyldimethylsilyl or triisopropylsilyl, or is left nitrogen is protected by a silyl group then it can be removed unprotected. If the nitrogen is protected by a Sulphonate in a Subsequent step by treatment with aqueous acid Such as group then it can be removed in a Subsequent step by treat hydrochloric acid or Sulphuric acid in a solvent Such as ment with base such as aqueous Sodium hydroxide, lithium methanol, ethanol, isopropanol, tetrahydrofuran or dioxane. 45 hydroxide or potassium carbonate in a solvent Such as metha An alternative method for preparing compounds of the nol, ethanol or isoproanol. If the nitrogen is protected by a invention is shown in Scheme 3. silyl group then it can be removed in a Subsequent step by treatment with aqueous acid such as hydrochloric acid, Sul phuric acidina Solvent Such as methanol, ethanol, isoproanol, tetrahydrofuran or dioxane. Scheme 3 50 Compounds of the invention wherein R' is NHRandR is N S aryl may be prepared as illustrated in Scheme 4.

N NC 2 Scheme 4 55 C Br N N 4 O V % N N S CH 60 n1 \ 9 O R2H HN N 6 NC 2 rN\, 65 4. N US 8,247,421 B2 53 54 -continued -continued N Cbz 1 N N f Y He- 5 NC leN N N 11 HN N N He Br H Cbz -N N N 10 4. N He Ts 2 18 N N N 12 | y H B r 15 NC leN

Cbz -N N Y - R1 N N 2 N N Ts 2O 4. N 13 Ts 19 N O 25 | y Y - O NC leN H -N He Cbz N N R1 N N 4. N 30 2 V Ts NY N 14 N N SMe | 35 Referring to Scheme 4, reaction of the bromoazaindole NC leN compound 9 with ammonia provides the amine compound 10. H After protection of compound 10 as a Cbz, derivative com -N He Cbz N N pound 11, bromination provides the bromo compound 12. Protection of the azaindole 1-nitrogen as a tosylate derivative Na NN 40 provides compound 13 which is converted to the boronate V compound 14. Coupling of compound 14 with the chloropy Ts rimidine 4 in the presence of a palladium catalyst provides 15 compound 15. Oxidation and displacement as previously O 45 described provides compound 17. Removal of the Cbz group N and coupling with an arylboronate as described above pro | Y-i-Me vides compound 19. Removal of the tosyl group provides NC 2N 9 compounds of the invention I.

--- In one method, the azaindole boronate esters 8 may be Cbz1 N N 50 prepared as illustrated in Scheme 5. 2 N N Scheme 5 Ts 16 55 RI N RI N I

N Her Her 2 2 | Y N NH2 N NH2 NC leN 2O 21 60 TMS Cbz1 N N Hess RI 2 N 2 He N N 2 Ts 65 N NH2 17 22 US 8,247,421 B2 55 56 -continued -continued

RI He 24 Br R! RI N 10 N N Ts 2 28 N NV 4. N T S Ts 25 8 15

Referring to Scheme 5, an aminopyridine compound 20 is iodinated to provide the iodo compound 21. Reaction of com pound 21 with trimethylsilylacetylene provides compound 22 which can be cyclized to provide the azaindole compound 23. N2 C Protection of compound 23 as a tosylate followed by bromi RI nation provides the bromoazaindole compound 25. Reaction 21 N of compound 25 with a diboronate in the presence of a palla N dium catalyst provides boronate intermediates compound 8. 25 N Another method for preparing the compounds is illustrated I in Scheme 6. Referring to Scheme 6, coupling of compound 8 with the Scheme 6 30 chloropyrimidine compound 4 in the presence of a palladium B(OR) catalyst provides compound 26. Oxidation by m-CPBA or RI NC chlorine and displacement as previously described provides N NN compound 28. Compounds of formula I of this invention are 35 obtained by removal of the tosyl group. CIS4. N CDC N SMe In some embodiments, the compounds may be prepared as Ts illustrated in Scheme 7. 8 4

40 Scheme 7 ey-so 45 2-1N l RI -as

N 50 N N Ts 27

55 21Ney-ll l 60 RI He

N N N 65 Ts 29 US 8,247,421 B2 57 58 -continued -continued

N NPG N2 NC | -e-NS-C *N4 RI

s 10 rN\,2 N N 30 Ts 32

15 N2 N NH

*N4 NC | leNS-C N RI

rN\,2 28 N N 25 Ts 33 N2 N N 21 N 30 | y-C. S. NC leN N N RI I

35 rN\,2 Referring to Scheme 7, hydrolysis of compound 27 (n=2) N N furnishes compound 29 which is chlorinated under conditions Ts known in the art to provide compound 30. Displacement of 34 chloropyrimidine compound 30 with an appropriate R. H. as previously described, provides compound 28. Compounds 40 of formula I of this invention are obtained by removal of the tosyl group. | Y- H NR17 In other embodiments, the compounds may be prepared as NC leN illustrated in Scheme 8. 45 RI

rN\,2 Scheme 8 NY N N | y-lost 50 NC leN Referring to Scheme 8,

55 rN\,2 N N Ts 27 represents a 4 to 8 membered optionally Substituted nitrogen 60 heterocycle wherein PG is a protecting group. Reaction of compound 27 with amine compound 31 followed by depro tection provides compound 33. Further modifications of com HN pound 33 may be achieved by known reactions such as, for NPG example, acylation, alkylation, and reductive amination to 31 65 provide intermediates compound 34. Compounds of formula Ia of this invention are then obtained by removal of the tosyl group. US 8,247,421 B2 59 60 In certain embodiments, compounds of the invention are -continued prepared by modification of substituents as illustrated in NH2 Scheme 9.

N Scheme 9 | Y NC leN RI HN 10 He rN\,2 OH N N 35 Ts 39 15

HN 2 -- 2O OH NHR 36

N N 25 | y | Y NC -e-N NC leN RI RI N Her 30 rN\,a NN N V % N Ts V 40 Ts 30 35

OH 40 NHR N

| y N NC leN -> 45 | y RI rN\, NC leN 4. N R N N Ts 50 37 4. N Ib O Referring to Scheme 9, hydrogenation of compound 35 in N 55 the presence of PtC) furnishes compound 36. Reaction of | y compound 30 with amine compound 36 provides compound NC leN 37. Subsequent oxidation of alcohol compound 37 to com --- pound 38 followed by reductive amination of compound 38 RI 60 provides compound 39. Further modifications of compound 39 may beachieved by known reactions such as, for example, rN\, acylation, alkylation, and reductive amination to provide 4. N V intermediates compound 40. Compounds of formula Ib of this Ts invention are then obtained by removal of the tosyl group. 38 65 In other embodiments, compounds of the invention are prepared by modification of substituents as illustrated in Scheme 10. US 8,247,421 B2 62 Referring to Scheme 10, reaction of compound 27 with an amine compound 41, wherein Hal represents F, Cl, Br or I and each X is independently CR or N, provides compound 42. Reaction of compound 42 with H K wherein K is —OR, —SR, or N(R) can be achieved thermally using conven tional heating or microwave irradiation, in a solvent Such as as dioxane, tetrahydrofuran, ethanol, isopropanol or butanol. In the case where H K is HN(R), the reaction can be achieved via Buchwald coupling conditions. Compounds of formula Ic 10 of this invention are then obtained by removal of the tosyl V group of compound 43. The chiral version of compound 41 Ts can be prepared using known methods: Tetrahedron Asym 27 metry 2006, 17,3163-9. R8 15 An alternative method for preparing the compounds is illustrated in Scheme 11. HN | S$x --- 2 X Hal Scheme 11 41 2O R8 22. ry-so y-ll 21N l N RI --- | Ny-l 25 N NC -e-N -- N N Ts N N 30 27 % N Ts 42 35 H-K -> F F R8 F

F N N yl 40 | y NC leN He Ne- N 45 RI N-V 2 N N N N N Ts 50 Ts 43 44 R8 y N Null 2N R2 | Y NC leN Na N

R! 60 --- rN\, R 21 N % N s N Ic 65 Ts US 8,247,421 B2 63 64 -continued -continued 2N R2 2N R2

Ne- N 5 Ne- N R! RI 21 N 21 N N N N1 N 10 N1 N I I

Referring to Scheme 11, reaction of compound 27 with Referring to Scheme 12, reaction of compound 29 with pentafluorophenol provides compound 44, which can be dis 15 diethylchlorophosphate provides compound 45. Reaction of placed with a series of HNR, under similar conditions as compound 45 with HR under conditions similar to those described previously in Scheme 10. Compounds of formula I previously described provides the intermediate compound of this invention are obtained by removal of the tosyl group of 28. Compounds of formula I of this invention are then compound 28. obtained by removal of the tosyl group of compound 28. A further method for preparing the compounds is illus An alternative method for preparing compounds of fomula trated in Scheme 12. Ib is illustrated in Scheme 13.

25 Scheme 13

PG n N H Ho 30 OH 46

PG 35 n N H Her

OH 47

40 PG n N H He

OLG 45 48

PG n N H -- 50 N 49

PG 55 n N H Her

NH2 50 60 PG n N H --

NHW 65 28 51 US 8,247,421 B2 65 66 -continued -continued O

He Nx --- NHW 2 OH X 8 52 R8a Rsa N SOCH 55 f Y 10 NH NC leN

RI Ho Nx -- 2 OH rN\, 15 X 8 4. N R8a Rsa Ts 56 27 N

N | Y-ch NC leN f Y RI NC -e-N NHW RI 25 rN\, 2 N N V rN\, Ts % N 27 Ib X OH 30 R1 y–M X R8a R8a Referring to Scheme 13, hydrogenation of compound 46. s wherein PG is a Suitable protecting group (e.g. Boc), in the W Her presence of rhodium on alumina furnishes compound 47. N2 \ Activation of alcohol compound 47 with a suitable leaving 35 N group LG 1 (e.g., ToS or Mes) to provide compound 48 fol Ts lowed by a reaction with NaNs provides compound 49. After 57 reduction of the azide compound 49, amine compound 50 is X OH coupled with a range of W X by reactions known in the art N to give derivatives compound 51. Compounds of formula Ib 40 NC R Y NH) { XX-7( R8a R8a of this invention are obtained by reaction of deprotected Sr. amine compound 52 with a compound of e.g. formula 27. followed by removal of the tosyl group. Another method for preparing compounds of formula Ia is 45 shown in Scheme 14. N Ia

Scheme 14 50 Referring to Scheme 14, each X is independently CR or N and R is R or two R* together with the atom to which they O O are attached may form a 5- to 7-membered heterocyclic ring. Starting material 53 (see, e.g., Bioorg. Med. Chem., 2005, 13, n X --- 6763) is lithiated and reacted with ketones RC(O)R. Amines compound 56 are formed after deprotection of ketals als 55 compound 54, followed by reductive amination of interme X Br diates compound 55. Reaction of compound 56 with com 53 pound 27 provides intermediate compound 57. Compounds of formula Ia of this invention are obtained by removal of the O O tosyl group. 60 Schemes 1 through 14 above depict synthetic methods that n X may be used to provide compounds of this invention. Accord -- ingly, this invention also provides processes for preparing a 2 OH X compound of this invention according to Schemes 1 through 8 14. R8a Rsa 65 Use of the Compounds 54 The present invention provides compounds that are useful for the treatment of diseases, disorders, and conditions US 8,247,421 B2 67 68 including, but not limited to, autoimmune diseases, inflam of any basic nitrogen-containing groups of the compounds matory diseases, proliferative and hyperproliferative dis disclosed herein. Water or oil-soluble or dispersible products eases, immunologically-mediated diseases, bone diseases, may be obtained by Such quaternization. metabolic diseases, neurological and neurodegenerative dis Base addition salts can be prepared by first reacting the eases, cardiovascular diseases, hormone related diseases, purified compound in its acid form with a Suitable organic or allergies, asthma, and Alzheimer's disease. Another aspect of inorganic base, and then isolating the salt thus formed. Base this invention provides compounds that are inhibitors of pro addition salts include alkali or alkaline earth metal salts. tein kinases, and thus are useful for the treatment of the Representative alkali or alkaline earth metal salts include diseases, disorders, and conditions, along with other uses Sodium, lithium, potassium, calcium, magnesium, and the described herein. In another aspect of the present invention, 10 like. Further pharmaceutically acceptable salts include, when pharmaceutically acceptable compositions are provided, appropriate, nontoxic ammonium, quaternary ammonium, wherein these compositions comprise any of the compounds and amine cations formed using counterions such as halide, as described herein, and optionally comprise a pharmaceuti hydroxide, carboxylate, Sulfate, phosphate, nitrate, lower cally acceptable carrier, adjuvant or vehicle. In certain alkyl sulfonate and aryl sulfonate. Other acids and bases, embodiments, these compositions optionally further com 15 while not in themselves pharmaceutically acceptable, may be prise one or more additional therapeutic agents. employed in the preparation of salts useful as intermediates in It will also be appreciated that certain of the compounds of obtaining the compounds of the invention and their pharma present invention can exist in free form for treatment, or ceutically acceptable acid or base addition salts. where appropriate, as a pharmaceutically acceptable Salt or As described herein, the pharmaceutically acceptable com pharmaceutically acceptable derivative thereof. positions of the present invention additionally comprise a As used herein, a “pharmaceutically acceptable derivative' pharmaceutically acceptable carrier, adjuvant, or vehicle, is an adduct or derivative which, upon administration to a which, as used herein, includes any and all solvents, diluents, patient in need, is capable of providing, directly or indirectly, or other liquid vehicle, dispersion or Suspension aids, Surface a compound as otherwise described herein, or a metabolite or active agents, isotonic agents, thickening or emulsifying residue thereof. Examples of pharmaceutically acceptable 25 agents, preservatives, Solid binders, lubricants and the like, as derivatives include, but are not limited to, pharmaceutically Suited to the particular dosage form desired. Remington's acceptable prodrugs. Pharmaceutical Sciences, Sixteenth Edition, E. W. Martin As used herein, the term “pharmaceutically acceptable (Mack Publishing Co., Easton, Pa., 1980) discloses various salt” refers to salts of a compound which are, within the scope carriers used in formulating pharmaceutically acceptable of Sound medical judgment, Suitable for use in contact with 30 compositions and known techniques for the preparation the tissues of humans and lower animals without undue tox thereof. Except insofar as any conventional carrier medium is icity, irritation, allergic response and the like, and are com incompatible with the compounds of the invention, such as by mensurate with a reasonable benefit/risk ratio. producing any undesirable biological effect or otherwise Pharmaceutically acceptable salts are well-known in the interacting in a deleterious manner with any other art. For example, S. M. Berge et al., describe pharmaceuti 35 component(s) of the pharmaceutically acceptable composi cally acceptable salts in detail in J. Pharmaceutical Sciences, tion, its use is contemplated to be within the scope of this 1977, 66, 1-19, incorporated herein by reference. Pharmaceu invention. tically acceptable salts of the compounds of this invention Some examples of materials which can serve as pharma include those derived from Suitable inorganic and organic ceutically acceptable carriers include, but are not limited to, acids and bases. These salts can be prepared in situ during the 40 ion exchangers, alumina, aluminum Stearate, lecithin, serum final isolation and purification of the compounds. Acid addi proteins, such as human serum albumin, buffer Substances tion salts can be prepared by first reacting the purified com Such as phosphates, glycine, Sorbic acid, or potassium Sor pound in its free-based form with a suitable organic or inor bate, partial glyceride mixtures of Saturated vegetable fatty ganic acid, and then isolating the salt thus formed. acids, water, salts or electrolytes, such as protamine Sulfate, Examples of pharmaceutically acceptable, nontoxic acid 45 disodium hydrogen phosphate, potassium hydrogen phos addition salts are salts of an amino group formed with inor phate, sodium chloride, Zinc salts, colloidal silica, magne ganic acids such as hydrochloric acid, hydrobromic acid, sium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, phosphoric acid, Sulfuric acid and perchloric acid or with polyethylene-polyoxypropylene-block polymers, wool fat, organic acids such as acetic acid, oxalic acid, maleic acid, Sugars such as lactose, glucose and Sucrose; starches Such as tartaric acid, citric acid, Succinic acid or malonic acid or by 50 corn starch and potato starch, cellulose and its derivatives using other methods used in the art Such as ion exchange. Such as Sodium carboxymethyl cellulose, ethyl cellulose and Other pharmaceutically acceptable salts include adipate, algi cellulose acetate; powdered tragacanth; malt, gelatin; talc; nate, ascorbate, aspartate, benzenesulfonate, benzoate, bisul excipients such as cocoa butter and Suppository waxes; oils fate, borate, butyrate, camphorate, camphorsulfonate, citrate, Such as peanut oil, cottonseed oil; Safflower oil; sesame oil; cyclopentanepropionate, digluconate, dodecylsulfate, 55 olive oil; corn oil and soybean oil; glycols; Such a propylene ethanesulfonate, formate, fumarate, glucoheptonate, glycero glycol or polyethylene glycol, esters such as ethyl oleate and phosphate, glycolate, gluconate, hemisulfate, heptanoate, ethyl laurate; agar, buffering agents such as magnesium hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lacto hydroxide and aluminum hydroxide; alginic acid; pyrogen bionate, lactate, laurate, lauryl Sulfate, malate, maleate, mal free water, isotonic saline; Ringer's solution; ethyl alcohol, onate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, 60 and phosphate buffer solutions, as well as other non-toxic nitrate, oleate, oxalate, palmitate, palmoate, pectinate, per compatible lubricants such as sodium lauryl Sulfate and mag Sulfate, 3-phenylpropionate, phosphate, picrate, pivalate, nesium Stearate, as well as coloring agents, releasing agents, propionate, salicylate, Stearate. Succinate, Sulfate, tartrate, coating agents, Sweetening, flavoring and perfuming agents, thiocyanate, p-toluenesulfonate, undecanoate, Valerate salts, preservatives and antioxidants can also be present in the com and the like. Salts derived from appropriate bases include 65 position, according to the judgment of the formulator. alkali metal, alkaline earth metal, ammonium and N(C- One aspect of this invention provides a method for the 4alkyl) salts. This invention also envisions the quaternization treatment or lessening the severity of a disease selected from US 8,247,421 B2 69 70 an autoimmune disease, an inflammatory disease, a prolifera enzymatic activity by binding to the protein kinase. In some tive or hyperproliferative disease, Such as cancer, an immu embodiments, said protein kinase is PLK. nologically-mediated disease, a bone disease, a metabolic The activity of the compounds as protein kinase inhibitors disease, a neurological or neurodegenerative disease, a car may be assayed in vitro, in vivo or in a cell line. In vitro assays diovascular disease, allergies, asthma, Alzheimer's disease, include assays that determine inhibition of either the kinase or a hormone related disease, comprising administering an activity or ATPase activity of the activated kinase. Alternate in effective amount of a compound, or a pharmaceutically vitro assays quantitate the ability of the inhibitor to bind to the acceptable composition comprising a compound, to a subject protein kinase and may be measured either by radiolabelling in need thereof. The term "cancer includes, but is not limited the inhibitor prior to binding, isolating the inhibitor/kinase to the following cancers: breast; ovary; cervix: prostate; tes 10 complex and determining the amount of radiolabel bound, or tis, genitourinary tract; esophagus; larynx, glioblastoma; by running a competition experiment where new inhibitors neuroblastoma; stomach; skin, keratoacanthoma; lung, epi are incubated with the kinase bound to known radioligands. dermoid carcinoma, large cell carcinoma, Small cell carci The protein kinase inhibitors or pharmaceutical salts noma, lung adenocarcinoma; bone; colon, adenoma; pan thereof may be formulated into pharmaceutical compositions creas, adenocarcinoma; thyroid, follicular carcinoma, 15 for administration to animals or humans. These pharmaceu undifferentiated carcinoma, papillary carcinoma, seminoma; tical compositions, which comprise an amount of the protein melanoma; sarcoma; bladder carcinoma; liver carcinoma and inhibitor effective to treat or prevent a protein kinase-medi biliary passages; kidney carcinoma, myeloid disorders; lym ated condition and a pharmaceutically acceptable carrier, are phoid disorders, Hodgkin’s, hairy cells; buccal cavity and another embodiment of the present invention. In some pharynx (oral), lip, tongue, mouth, pharynx. Small intestine; embodiments, said protein kinase-mediated condition is a colon-rectum, large intestine, rectum; brain and central ner PLK-mediated condition. In some embodiments, a PLK1 Vous system; and leukemia. mediated condition. In certain embodiments, an “effective amount of the com The exact amount of compound required for treatment will pound or pharmaceutically acceptable composition is that vary from Subject to Subject, depending on the species, age, amount effective in order to treat said disease. The com 25 and general condition of the subject, the severity of the infec pounds and compositions, according to the method of the tion, the particular agent, its mode of administration, and the present invention, may be administered using any amount and like. The compounds of the invention are preferably formu any route of administration effective for treating or lessening lated in dosage unit form for ease of administration and uni the severity of said disease. In some embodiments, said dis formity of dosage. The expression “dosage unit form as used ease is selected from a proliferative disorder, a neurodegen 30 herein refers to a physically discrete unit of agent appropriate erative disorder, an autoimmune disorder, and inflammatory for the patient to be treated. It will be understood, however, disorder, and an immunologically-mediated disorder. In that the total daily usage of the compounds and compositions Some embodiments, said disease is a proliferative disorder. In of the present invention will be decided by the attending Some embodiments, said disease is cancer. physician within the scope of Sound medical judgment. The In other embodiments of this invention, said disease is a 35 specific effective dose level for any particular patient or protein-kinase mediated condition. In some embodiments, organism will depend upon a variety of factors including the said protein kinase in PLK. disorder being treated and the severity of the disorder; the The term “protein kinase-mediated condition', as used activity of the specific compound employed; the specific herein means any disease or other deleterious condition in composition employed; the age, body weight, general health, which a protein kinase plays a role. Such conditions include, 40 sex and diet of the patient; the time of administration, route of without limitation, autoimmune diseases, inflammatory dis administration, and rate of excretion of the specific com eases, proliferative and hyperproliferative diseases, immuno pound employed; the duration of the treatment; drugs used in logically-mediated diseases, bone diseases, metabolic dis combination or coincidental with the specific compound eases, neurological and neurodegenerative diseases, employed, and like factors well known in the medical arts. cardiovascular diseases, hormone related diseases, allergies, 45 The term "patient’, as used herein, means an animal, prefer asthma, and Alzheimer's disease. ably a mammal, and most preferably a human. The term "PLK-mediated condition', as used herein means The pharmaceutically acceptable compositions of this any disease or other deleterious condition in which PLK plays invention can be administered to humans and other animals a role. Such conditions include, without limitation, a prolif orally, rectally, parenterally, intracisternally, intravaginally, erative disorder, Such as cancer, a neurodegenerative disorder, 50 intraperitoneally, topically (as by powders, ointments, or an autoimmune disorder, and inflammatory disorder, and an drops), bucally, as an oral or nasal spray, or the like, depend immunologically-mediated disorder. ing on the severity of the infection being treated. In certain In some embodiments, the compounds and compositions embodiments, the compounds of the invention may be admin of the invention are inhibitors of protein kinases. As inhibitors istered orally or parenterally at dosage levels of about 0.01 of protein kinases, the compounds and compositions of this 55 mg/kg to about 50 mg/kg and preferably from about 1 mg/kg invention are particularly useful for treating or lessening the to about 25 mg/kg, of Subject body weight per day, one or severity of a disease, condition, or disorder where a protein more times a day, to obtain the desired therapeutic effect. kinase is implicated in the disease, condition, or disorder. In Liquid dosage forms for oral administration include, but one aspect, the present invention provides a method for treat are not limited to, pharmaceutically acceptable emulsions, ing or lessening the severity of a disease, condition, or disor 60 microemulsions, Solutions, Suspensions, syrups and elixirs. der where a protein kinase is implicated in the disease state. In In addition to the active compounds, the liquid dosage forms another aspect, the present invention provides a method for may contain inert diluents commonly used in the art Such as, treating or lessening the severity of a disease, condition, or for example, water or other solvents, solubilizing agents and disorder where inhibition of enzymatic activity is implicated emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl in the treatment of the disease. In another aspect, this inven 65 carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, tion provides a method for treating or lessening the severity of propylene glycol. 1,3-butylene glycol, dimethylformamide, a disease, condition, or disorder with compounds that inhibit oils (in particular, cottonseed, groundnut, corn, germ, olive, US 8,247,421 B2 71 72 castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, kaolin and bentonite clay, and i) lubricants such as talc, cal polyethylene glycols and fatty acid esters of Sorbitan, and cium Stearate, magnesium Stearate, Solid polyethylene gly mixtures thereof. Besides inert diluents, the oral composi cols, Sodium lauryl Sulfate, and mixtures thereof. In the case tions can also include adjuvants such as wetting agents, emul of capsules, tablets and pills, the dosage form may also com Sifying and Suspending agents, Sweetening, flavoring, and 5 prise buffering agents. perfuming agents. Solid compositions of a similar type may also be employed Injectable preparations, for example, Sterile injectable as fillers in Soft and hard-filled gelatin capsules using Such aqueous or oleaginous Suspensions may be formulated excipients as lactose or milk Sugar as well as high molecular according to the known art using Suitable dispersing or wet weight polyethylene glycols and the like. The Solid dosage ting agents and Suspending agents. The sterile injectable 10 forms of tablets, dragees, capsules, pills, and granules can be preparation may also be a sterile injectable solution, Suspen prepared with coatings and shells Such as enteric coatings and sion or emulsion in a nontoxic parenterally acceptable diluent other coatings well known in the pharmaceutical formulating or solvent, for example, as a solution in 1,3-butanediol. art. They may optionally contain opacifying agents and can Among the acceptable vehicles and solvents that may be also be of a composition that they release the active employed are water, Ringer's solution, U.S.P. and isotonic 15 ingredient(s) only, or preferentially, in a certain part of the sodium chloride solution. In addition, sterile, fixed oils are intestinal tract, optionally, in a delayed manner. Examples of conventionally employed as a solventor Suspending medium. embedding compositions that can be used include polymeric For this purpose any bland fixed oil can be employed includ Substances and waxes. Solid compositions of a similar type ing synthetic mono- or diglycerides. In addition, fatty acids may also be employed as fillers in soft and hard-filled gelatin Such as oleic acid are used in the preparation of injectables. capsules using Such excipients as lactose or milk Sugar as well The injectable formulations can be sterilized, for example, as high molecular weight polethylene glycols and the like. by filtration through a bacterial-retaining filter, or by incor The active compounds can also be in microencapsulated porating sterilizing agents in the form of sterile solid compo form with one or more excipients as noted above. The solid sitions which can be dissolved or dispersed in sterile water or dosage forms of tablets, dragees, capsules, pills, and granules other sterile injectable medium prior to use. 25 can be prepared with coatings and shells Such as enteric In order to prolong the effect of a compound of the present coatings, release controlling coatings and other coatings well invention, it is often desirable to slow the absorption of the known in the pharmaceutical formulating art. In Such solid compound from Subcutaneous or intramuscular injection. dosage forms the active compound may be admixed with at This may be accomplished by the use of a liquid Suspension of least one inert diluent such as Sucrose, lactose or starch. Such crystalline or amorphous material with poor water solubility. 30 dosage forms may also comprise, as is normal practice, addi The rate of absorption of the compound then depends upon its tional Substances other than inert diluents, e.g., tableting rate of dissolution that, in turn, may depend upon crystal size lubricants and other tableting aids such a magnesium stearate and crystalline form. Alternatively, delayed absorption of a and microcrystalline cellulose. In the case of capsules, tablets parenterally administered compound form is accomplished and pills, the dosage forms may also comprise buffering by dissolving or Suspending the compound in an oil vehicle. 35 agents. They may optionally contain opacifying agents and Injectable depot forms are made by forming microencapsule can also be of a composition that they release the active matrices of the compound in biodegradable polymers such as ingredient(s) only, or preferentially, in a certain part of the polylactide-polyglycolide. Depending upon the ratio of com intestinal tract, optionally, in a delayed manner. Examples of pound to polymer and the nature of the particular polymer embedding compositions that can be used include polymeric employed, the rate of compound release can be controlled. 40 Substances and waxes. Examples of other biodegradable polymers include poly Dosage forms for topical or transdermal administration of (orthoesters) and poly(anhydrides). Depot injectable formu a compound of this invention include ointments, pastes, lations are also prepared by entrapping the compound in creams, lotions, gels, powders, Solutions, sprays, inhalants or liposomes or microemulsions that are compatible with body patches. The active component is admixed under sterile con tissues. 45 ditions with a pharmaceutically acceptable carrier and any Compositions for rectal or vaginal administration are pref needed preservatives or buffers as may be required. Oph erably suppositories which can be prepared by mixing the thalmic formulation, eardrops, and eye drops are also con compounds of this invention with Suitable non-irritating templated as being within the scope of this invention. Addi excipients or carriers such as cocoa butter, polyethylene gly tionally, the present invention contemplates the use of color a Suppository wax which are solid at ambient tempera 50 transdermal patches, which have the added advantage of pro ture but liquid at body temperature and therefore melt in the viding controlled delivery of a compound to the body. Such rectum or vaginal cavity and release the active compound. dosage forms can be made by dissolving or dispensing the Solid dosage forms for oral administration include cap compound in the proper medium. Absorption enhancers can Sules, tablets, pills, powders, and granules. In Such solid dos also be used to increase the flux of the compound across the age forms, the active compound is mixed with at least one 55 skin. The rate can be controlled by either providing a rate inert, pharmaceutically acceptable excipient or carrier Such as controlling membrane or by dispersing the compound in a Sodium citrate or dicalcium phosphate and/or a) fillers or polymer matrix or gel. extenders such as starches, lactose, Sucrose, glucose, manni In addition to the compounds of this invention, pharma tol, and silicic acid, b) binders such as, for example, car ceutically acceptable derivatives or prodrugs of the com boxymethylcellulose, alginates, gelatin, polyvinylpyrrolidi 60 pounds of this invention may also be employed in composi none. Sucrose, and acacia, c) humectants such as glycerol. d) tions to treat or prevent the above-identified disorders. disintegrating agents such as agar-agar, calcium carbonate, A “pharmaceutically acceptable derivative or prodrug’ potato or tapioca starch, alginic acid, certain silicates, and means any pharmaceutically acceptable ester, salt of an ester Sodium carbonate, e) solution retarding agents such as paraf or other derivative of a compound of this invention which, fin, f) absorption accelerators such as quaternary ammonium 65 upon administration to a recipient, is capable of providing, compounds, g) wetting agents such as, for example, cetyl either directly or indirectly, a compound of this invention or alcohol and glycerol monostearate, h) absorbents such as an inhibitorily active metabolite or residue thereof. Particu US 8,247,421 B2 73 74 larly favoured derivatives or prodrugs are those that increase starch. When aqueous Suspensions are required for oral use, the bioavailability of the compounds of this invention when the active ingredient is combined with emulsifying and Sus Such compounds are administered to a patient (e.g., by allow pending agents. If desired, certain Sweetening, flavoring or ing an orally administered compound to be more readily coloring agents may also be added. absorbed into the blood) or which enhance delivery of the Alternatively, the pharmaceutical compositions of this parent compound to a biological compartment (e.g., the brain invention may be administered in the form of Suppositories or lymphatic system) relative to the parent species. for rectal administration. These can be prepared by mixing Pharmaceutically acceptable prodrugs of the compounds the agent with a Suitable non-irritating excipient which is of this invention include, without limitation, esters, amino Solid at room temperature but liquid at rectal temperature and acid esters, phosphate esters, metal salts and Sulfonate esters. 10 therefore will melt in the rectum to release the drug. Such Pharmaceutically acceptable carriers that may be used in materials include, but are not limited to, cocoa butter, bees these pharmaceutical compositions include, but are not lim wax and polyethylene glycols. ited to, ion exchangers, alumina, aluminum Stearate, lecithin, The pharmaceutical compositions of this invention may serum proteins, such as human serum albumin, buffer Sub also be administered topically, especially when the target of stances such as phosphates, glycine, Sorbic acid, potassium 15 treatment includes areas or organs readily accessible by topi Sorbate, partial glyceride mixtures of Saturated vegetable cal application, including diseases of the eye, the skin, or the fatty acids, water, salts or electrolytes, such as protamine lower intestinal tract. Suitable topical formulations are Sulfate, disodium hydrogen phosphate, potassium hydrogen readily prepared for each of these areas or organs. phosphate, Sodium chloride, Zinc salts, colloidal silica, mag Topical application for the lower intestinal tract can be nesium trisilicate, polyvinyl pyrrolidone, cellulose-based effected in a rectal suppository formulation (see above) or in Substances, polyethylene glycol, Sodium carboxymethylcel a Suitable enema formulation. Topically-transdermal patches lulose, polyacrylates, waxes, polyethylene-polyoxypropy may also be used. lene-block polymers, polyethylene glycol and wool fat. For topical applications, the pharmaceutical compositions The compositions of the present invention may be admin may be formulated in a suitable ointment containing the istered orally, parenterally, by inhalation spray, topically, rec 25 active component Suspended or dissolved in one or more tally, nasally, buccally, vaginally or via an implanted reser carriers. Carriers for topical administration of the compounds voir. The term “parenteral as used herein includes, but is not of this invention include, but are not limited to, mineral oil, limited to, Subcutaneous, intravenous, intramuscular, intra liquid petrolatum, white petrolatum, propylene glycol, poly articular, intra-synovial, intrasternal, intrathecal, intrahe oxyethylene, polyoxypropylene compound, emulsifying wax patic, intralesional and intracranial injection or infusion tech 30 and water. Alternatively, the pharmaceutical compositions niques. Preferably, the compositions are administered orally, can beformulated in a Suitable lotion or cream containing the intraperitoneally or intravenously. active components suspended or dissolved in one or more Sterile injectable forms of the compositions of this inven pharmaceutically acceptable carriers. Suitable carriers tion may be acqueous or oleaginous Suspension. These Sus include, but are not limited to, mineral oil, sorbitan pensions may be formulated according to techniques known 35 monostearate, polysorbate 60, cetyl esters wax, cetearyl alco in the art using Suitable dispersing or wetting agents and hol, 2-octyldodecanol, benzyl alcohol and water. Suspending agents. The sterile injectable preparation may For ophthalmic use, the pharmaceutical compositions may also be a sterile injectable solution or Suspension in a non be formulated as micronized suspensions in isotonic, pH toxic parenterally-acceptable diluent or solvent, for example adjusted Sterile Saline, or, preferably, as solutions in isotonic, as a solution in 1,3-butanediol. Among the acceptable 40 pH adjusted sterile saline, either with or without a preserva vehicles and solvents that may be employed are water, Ring tive such as benzylalkonium chloride. Alternatively, for oph er's solution and isotonic sodium chloride Solution. In addi thalmic uses, the pharmaceutical compositions may be for tion, sterile, fixed oils are conventionally employed as a sol mulated in an ointment such as petrolatum. Ventor Suspending medium. For this purpose, any bland fixed The pharmaceutical compositions of this invention may oil may be employed including synthetic mono- or di-glyc 45 also be administered by nasal aerosol or inhalation. Such erides. Fatty acids, such as oleic acid and its glyceride deriva compositions are prepared according to techniques well tives are useful in the preparation of injectables, as are natural known in the art of pharmaceutical formulation and may be pharmaceutically-acceptable oils, such as olive oil or castor prepared as Solutions in saline, employing benzyl alcohol or oil, especially in their polyoxyethylated versions. These oil other Suitable preservatives, absorption promoters to enhance Solutions or Suspensions may also contain a long-chain alco 50 bioavailability, fluorocarbons, and/or other conventional hol diluent or dispersant, such as carboxymethyl cellulose or solubilizing or dispersing agents. similar dispersing agents which are commonly used in the The amount of protein kinase inhibitor that may be com formulation of pharmaceutically acceptable dosage forms bined with the carrier materials to produce a single dosage including emulsions and Suspensions. Other commonly used form will vary depending upon the host treated, the particular Surfactants, such as Tweens, Spans and other emulsifying 55 mode of administration. Preferably, the compositions should agents or bioavailability enhancers which are commonly used be formulated so that a dosage of between 0.01-100 mg/kg in the manufacture of pharmaceutically acceptable solid, liq body weight/day of the inhibitor can be administered to a uid, or other dosage forms may also be used for the purposes patient receiving these compositions. of formulation. It should also be understood that a specific dosage and The pharmaceutical compositions of this invention may be 60 treatment regimen for any particular patient will depend upon orally administered in any orally acceptable dosage form a variety of factors, including the activity of the specific including, but not limited to, capsules, tablets, aqueous Sus compound employed, the age, body weight, general health, pensions or solutions. In the case of tablets for oral use, sex, diet, time of administration, rate of excretion, drug com carriers commonly used include, but are not limited to, lac bination, and the judgment of the treating physician and the tose and corn starch. Lubricating agents, such as magnesium 65 severity of the particular disease being treated. The amount of Stearate, are also typically added. For oral administration in a inhibitor will also depend upon the particular compound in capsule form, useful diluents include lactose and dried corn the composition. US 8,247,421 B2 75 76 According to another embodiment, the invention provides shown below are only examples, and are not meant to limit the methods for treating or preventing a protein kinase-mediated Scope of the conditions that can be used for making, analyZ condition (in Some embodiments, a PLK-mediated condition) ing, or testing the compounds of this invention. Instead, this comprising the step of administering to a patient one of the invention also includes conditions known to those skilled in above-described pharmaceutical compositions. The term that art for making, analyzing, and testing the compounds of "patient’, as used herein, means an animal, preferably a this invention. human. Preferably, that method is used to treat or prevent a condi All references cite in this documentare incorporated herein tion selected from cancers such as cancers of the breast, by reference. colon, prostate, skin, pancreas, brain, genitourinary tract, 10 lymphatic system, stomach, larynx and lung, including lung EXAMPLES adenocarcinoma and Small cell lung cancer, stroke, diabetes, myeloma, hepatomegaly, cardiomegaly, Alzheimer's disease, cystic fibrosis, and viral disease, or any specific disease Mass spec. Samples may be analyzed on a MicroMass described above. 15 Quattro Micro mass spectrometer operated in single MS Another aspect of the invention relates to inhibiting protein mode with electrospray ionization. Samples may be intro kinase activity in a patient, which method comprises admin duced into the mass spectrometer using chromatography. istering to the patient a compound of formula I or a compo 'H-NMR spectra may be recorded at 400 MHz using a sition comprising said compound. Bruker DPX 400 instrument. Depending upon the particular protein kinase-mediated conditions to be treated or prevented, additional drugs, which are normally administered to treat or prevent that condition, Preparation 1: 5-Trifluoromethyl-3-(4.4.5.5-tetram may be administered together with the inhibitors of this ethyl-1,3,2-dioxaborolan-2-yl)-1-tosyl-1H-pyrrolo2. invention. For example, chemotherapeutic agents or other 3-bipyridine anti-proliferative agents may be combined with the protein 25 kinase inhibitors of this invention to treat proliferative dis CaSCS. Those additional agents may be administered separately, as part of a multiple dosage regimen, from the protein kinase inhibitor-containing compound or composition. Alterna 30 tively, those agents may be part of a single dosage form, mixed together with the protein kinase inhibitor in a single composition. O In some embodiments, said protein kinase inhibitor is a Y l-O PLK kinase inhibitor. In other embodiments, said protein 35 kinase inhibitor is a PLK1 kinase inhibitor. FC N N This invention may also be used in methods other than 2 those involving administration to a patient. N N One aspect of the invention relates to inhibiting protein Tos kinase activity in a biological sample or a patient, which 40 method comprises contacting said biological sample with a compound of formula I or a composition comprising said compound. The term “biological sample, as used herein, means an in vitro or an ex vivo Sample, including, without limitation, cell cultures or extracts thereof; biopsied material 45 obtained from a mammal or extracts thereofandblood, Saliva, Step 1: 3-Iodo-5-Trifluoromethylpyridin-2-amine urine, feces, semen, tears, or other body fluids or extracts thereof Inhibition of protein kinase activity in a biological sample is useful for a variety of purposes that are known to one of skill 50 in the art. Examples of Such purposes include, but are not FC I limited to, blood transfusion, organ-transplantation, and bio N logical specimen storage. 2 Another aspect of this invention relates to the study of N NH2 protein kinases in biological and pathological phenomena; 55 the study of intracellular signal transduction pathways medi ated by Such protein kinases; and the comparative evaluation of new protein kinase inhibitors. Examples of Such uses To 2-amino 5-trifluoromethyl pyridine, (50g, 308 mmol) include, but are not limited to, biological assays such as was added acetic acid (500 mL), followed by pre-mixed water enzyme assays and cell-based assays. 60 The compounds of this invention may be prepared in gen (35 mL) and concentrated sulfuric acid (5 mL) and stirred eral by methods known to those skilled in the art. Those until completely solubilised. Periodic acid (13.3 g, 58 mmol) compounds may be analyzed by known methods, including was added followed by iodine (31.2g, 123 mmol) and then the but not limited to LCMS (liquid chromatography mass spec reaction mixture was heated to 85°C. for 18 hours. trometry) and NMR (nuclear magnetic resonance). Com 65 The reaction mixture was allowed to cool to room tempera pounds of this invention may be also tested according to these ture and then carefully basified with 4MNaOH solution to pH examples. It should be understood that the specific conditions 10-14. This gave a precipitate which was isolated by filtration US 8,247,421 B2 77 78 and stirred in water (1 litre), filtered and dried to give the filtration, to afford 5-Trifluoromethyl-1H-pyrrolo2,3-bpy product as a brown powder (61.38 g. 69%). ridine as a crystalline solid (26.38 g, 59% yield). Step 2: 5-Trifluoromethyl-3-((trimethylsilyl)ethynyl) Step 4: 5-Trifluoromethyl-1-tosyl-1H-pyrrolo pyridin-2-amine 2,3-bipyridine

58 TMS 10 2 FC N N 2 N NH2 15 N )N To a stirred solution of 3-Iodo-5-trifluoromethylpyridin-2- Ts amine (61.38 g. 213 mmol) in anhydrous THF (200 mL) under a nitrogen atmosphere was added Pd(PPh)C1 (1.5 g. 2.1 mmol), and copper iodide (0.406 g, 2.1 mmol) and the reaction mixture degassed under nitrogen. The reaction mix ture was cooled in an ice bath and TMS acetylene (36 mL. 256 mmol) was added dropwise over 30 minutes. Upon complete To a stirred solution of sodium hydride (6.2g, 156 mmol. addition the reaction was allowed to warm to room tempera 25 60% dispersion), tosyl chloride (27 g. 141.8 mmol), in anhy ture and-monitored until the reaction was complete. The mix drous THF (200 mL) under a nitrogen atmosphere was added ture was diluted with ethyl acetate (200 mL) and filtered a solution of 5-trifluoromethyl-1H-pyrrolo2,3-bipyridine through silica washed through with 4x150 mL ethyl acetate. (26.38 g. 141.8 mmol) in THF (150 mL) dropwise at a rate The filtrates were concentrated and the solid was triturated which maintained the internal temperature of the reaction from heptane (250 mL) to give 56 g of product. 30 below 40°C. The reaction mixture was stirred overnight at room temperature and analysis by LCMS showed no starting material remained after this time. The reaction mixture was Step 3: 5-Trifluoromethyl concentrated to remove most of THF and diluted with DCM -1H-pyrrolo2,3-bipyridine and water. The organic layer was separated and the aqueous 35 layer further extracted with DCM. The organic layers are combined and washed with brine, dried over magnesium Sulphate and concentrated to a Volume which was appropriate to load onto the top of a silica plug (300 mL) pre-wet with DCM, and the silica was washed with 2.5 L DCM. The filtrate FC 40 was concentrated to give the product as a slightly brown solid Oy (44.1 g, 92% yield). 2 N N H Step 5:3-Bromo-5-Trifluoromethyl-1-tosyl-1H-pyr 45 rolo2,3-bipyridine

Br To a stirred solution of potassium tert-butoxide (53.6 g. 479 50 mmol) in anhydrous NMP (200 mL) under a nitrogen atmo FC N N sphere at 80°C. was added a solution of 5-trifluoromethyl 3-((trimethylsilyl)ethynyl)pyridin-2-amine (62g, 239 mmol) % N in NMP (100 mL) at a rate to maintain the internal tempera Tos ture below 100° C. After complete addition the reaction was 55 deemed complete by LCMS. The reaction mixture was allowed to cool to room temperature and poured carefully into To a stirred solution of 5-trifluoromethyl-1-tosyl-1H-pyr saturated brine (700 mL) cooled in an ice bath to 10°C. The rolo2,3-bipyridine (53.7 g. 158 mmol) in DCM (250 mL) precipitate that formed was aged for 45 min at this tempera was added a solution of bromine (16 mL, 316 mmol) in DCM ture and isolated by filtration through buchner funnel. The 60 (50 mL) dropwise. No significant exotherm was observed. collected precipitate was stirred in ethyl acetate (1 litre) and After an hour at room temperature, the reaction mixture had water then filtered through celite to remove insoluble poly thickened and stirring ceased. The reaction was deemed com meric material and organic layer was separated. The aqueous plete by tic and was diluted with 1.2 litres of DCM and 50:50 layer was further extracted with ethyl acetate and the com saturated bicarbonate: Saturated Sodium thiosulfate aqueous bined organic layers dried over magnesium sulfate and con 65 Solution. The organic layer was separated and further washed centrated to give a brown solid which was triturated by stir with the 50:50 aqueous mixture, dried over magnesium sul ring for an hour in hot heptane and then cooling before fate and evaporated. The crude yellow solid was triturated US 8,247,421 B2 79 80 with diethyl ether and solid isolated and dried to give the After sonication for a further 5 minutes the reaction was product as a yellow powder (59 g, 59% yield) heated under microwave conditions at 130°C. for 15 minutes. The reaction was diluted with EtOAc and washed with brine. Step 6: 5-Trifluoromethyl-3-(4,4,5,5-tetramethyl-1,3, The organic phase was dried over MgSO and concentrated in 2-dioxaborolan-2-yl)-1-tosyl-1H-pyrrolo2,3-bipyri 5 vacuo. The crude product was purified by column chroma dine tography (ISCO CompanionTM, 80 g column, 0-100% EtOAc/Petroleum Ether). The product was seen to precipitate out in the collection tubes and was filtered off Further product was obtained by concentration of the fractions and trituration 10 with EtO to give the title compound as an off-white solid (195 mg, 40%). O Y l-O Preparation 2a: 2-(methylthio)-4-(1-tosyl-5-(trifluo FC 15 romethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimi 3 N N dine-5-carbonitrile; alternate procedure 2 N N 5-Trifluoromethyl-3-(4.4.5.5-tetramethyl-1,3,2-diox Tos oborolan-2-yl)-1-tosyl-1H-pyrrolo2,3-bipyridine (50 g, 107.2 mmol) and 4-chloro-2-(methylthio)pyrimidine-5-car To a mixture of 3-Bromo-5-trifluoromethyl-1-tosyl-1H bonitrile (39.8g. 214.4 mmol) were dissolved in dioxane (900 pyrrolo2,3-bipyridine (5 g, 11.9 mmol), bis-(pinacolato)di mL). Potassium carbonate (44.45 g, 321.6 mmol) was added boron (40 g, 95.5 mmol), potassium acetate (28.1 g, 266.7 followed by water (135 mL). The mixture was degassed 3 mmol) was added 1,4-dioxane (800 mL). The reaction mix 25 times with vacuum/nitrogen cycles, then bis-(tri-tert-bu ture was stirred vigorously while nitrogen gas was purged tylphosphine)palladium(0) (Strem, 5.0 g, 9.78 mmol) was through the solution for 2 hours. After this time tetrakis(triph added. After 30 minutes an additional 50 mL of water was enylphosphine)palladium (1Omol %) was added in one por added. A thick precipitate formed. The mixture was filtered tion and the reaction mixture brought to reflux after an hour. and the product washed with water. The product was dried in After 4 hours more tetrakis(triphenylphosphine)palladium 30 a vacuum oven at 60° C. for 18 hours. This gave 2-(meth (10 mol %) added and the reaction mixture refluxed under ylthio)-4-(1-tosyl-5-(trifluoromethyl)-1H-pyrrolo2,3-bpy nitrogen overnight. Heating continued until TLC confirmed no starting material remained. The reaction mixture was ridin-3-yl)pyrimidine-5-carbonitrile as an off-white solid allowed to cool then diluted with an equivalent volume of (48.1 g, 91%). petroleum ether. Solid potassium acetate was removed by 35 H NMR (DMSO-d 400 MHz) & 2.07 (3H, s), 2.65 (3H, filtration through celite. The filtrate was evaporated to dry s), 7.49 (2H, d), 8.12 (2H, d), 8.93 (1H, s), 9.03 (1H, s), 9.12 ness and the residue suspended in 20% ethyl acetate 80% (1H, s), 9.17 (1H, s); MS (ES") 490, (ES) 488. petroleum ether and filtered through a plug of Florisil, eluting with 3 volumes followed-by one volume 30% ethyl acetate Preparation 3: 2-(methylsulfonyl)-4-(1-tosyl-5-(trif 70% petroleum ether. The mixture was concentrated and 40 luoromethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimi triturated with heptane to give an off-white powder (35.72 g, 80% yield) dine-5-carbonitrile Preparation 2: 2-(methylthio)-4-(1-tosyl-5-(trifluo romethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimi 45 dine-5-carbonitrile

A mixture of 5-trifluoro-3-(4,4,5,5-tetramethyl-1,3,2-di 60 oxaborolan-2-yl)-1-tosyl-1H-pyrrolo2,3-bipyridine (415 mg, 1.08 mmol. 1.0 Eq.) and 4-chloro-2-(methylthio)pyrimi 2-(methylthio)-4-(1-tosyl-5-(trifluoromethyl)-1H-pyrrolo dine-5-carbonitrile (146 mg, 1.08 mmol. 1.0 Eq.) was sus 2,3-bipyridin-3-yl)pyrimidine-5-carbonitrile (50 mg, 0.1 pended in toluene (20 mL) and EtOH (5 mL) and treated with mmol) in DCM (10 mL) was treated with MCPBA (38 mg, 2M KCO (1.62 mL, 3.24 mmol. 3.0 Eq.). The mixture was 65 0.22 mmol. 2.2 Eq) and allowed to stir overnight at room Sonicated under an atmosphere of nitrogen for 20 min and temperature. The mixture was concentrated in vacuo and then treated with Pd(PPh) (75 mg, 0.06 mmol, 0.1 eq.). treated with EtOAc. The insoluble material was filtered off US 8,247,421 B2 81 82 and the resultant filtrate was concentrated to give the title -continued compound which was taken forward (33 mg, 63%). I-129

Example 1

2-(benzylamino)-4-(5-(trifluoromethyl)-1H-pyrrolo 2,3-bipyridin-3-yl)pyrimidine-5-carbonitrile

10

I-1

15

Step 1: 2-(methylsulfonyl)-4-(1-tosyl-5-(trifluorom ethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimidine-5- carbonitrile 2-(methylsulfonyl)-4-(1-tosyl-5-(trifluoromethyl)-1H pyrrolo2,3-bipyridin-3-yl)pyrimidine-5-carbonitrile (50 mg, 0.096 mmol) was heated with benzylamine (21 uL, 0.19 25 mmol. 2.0 Eq.) and DIPEA (50 uL, 0.29 mmol. 3.0 Eq.) in THF (2 mL) under microwave conditions at 100° C. for 10 minutes. The reaction was concentrated and the residue dis solved in THF (5 mL). LiOH.HO (20 mg, 0.48 mmol. 5.0 30 Eq.) in water (1 mL) was added and the reaction stirred for 1 hour. The reaction mixture was concentrated and the residue treated with MeOH. The obtained white precipitate was iso lated by filtration and dried under vacuum (13 mg, 36% over 2 steps). 35 MS (ES") m/e=395. H NMR (DMSO)4.67 (2H, d), 4.74 (2H, d), 7.21-7.38 (10H, m), 8.68 (1H, s), 8.72-8.74 (5H, m), 8.86 (1H, t), 8.93 (1H, s), 9.03 (1H, t),9.27(1H,s), 12.77 (2H, vbrs). 1:1 Mixture of rotamers 40 Example 2

2-((R)-1-(6-(4-methylpiperazin-1-yl)pyridin-3-yl) ethylamino)-4-(5-(trifluoromethyl)-1H-pyrrolo2,3- 45 bipyridin-3-yl)pyrimidine-5-carbonitrile I-147 and 2-((S)-1-(6-(4-methylpiperazin-1-yl)pyridin-3-yl) ethylamino)-4-(5-(trifluoromethyl)-1H-pyrrolo2,3- bipyridin-3-yl)pyrimidine-5-carbonitrile (I-129) 50

I-147 55 2-(Methylsulfonyl)-4-(1-tosyl-5-(trifluoromethyl)-1H pyrrolo2,3-bipyridin-3-yl)pyrimidine-5-carbonitrile (1.4g, 2.86 mmol) was suspended in ethanol (60 mL) and water (4 mL). The mixture was cooled in an ice bath, and chlroine gas 60 was slowly bubbled through the solution for 20 minutes. After addition, the suspension was stirred for another 30 minutes at 0° C. The reaction mixture was allowed to vent for 15 min utes, then, ether (100 mL) was added and the solid was filtered off. The solid was dried under vacuo to give a light pink solid (1.3g, 87% yield). MS (ES) 522. US 8,247,421 B2 83 84 Step 2: 2-((R)-(1-(6-chloropyridin-3-yl)ethylamino)- -continued 4-(1-tosyl-5-(trifluoromethyl)-1H-pyrrolo2,3-b I-129 pyridin-3-yl)pyrimidine-5-carbonitrile

N

2 10 N -> NS

15 A mixture of 2-(1-(6-chloropyridin-3-yl)ethylamino)-4- (1-tosyl-5-(trifluoromethyl)-1H-pyrrolo2,3-bipyridin-3-yl) pyrimidine-5-carbonitrile (157 mg, 0.26 mmol) and N-meth ylpiperazine (2 mL) was heated under microwave irradiation at 180° C. for 40 minutes. The reaction mixture was parti tioned between EtOAc and water. The organic phase was dried over sodium sulphate, filtered and evaporated to dry ness. The residue was purified on silica gel by flash column chromatography to afford the title compound as an off-white solid (52 mg, 39% yield). 25 The two enantiomers were separated using a Berger Mini gram SFC system 250x4.6 mm Daicel Chiralcel OD-H col umn; 4 mLS/min flowrate; co-solvent was 32% Methanol (with 0.25% TEA (Tri-ethylamine) modifier added); column 30 temperature: 35° C.I. Data for I-147 (First Eluted): Diisopropylethylamine (730 ul, 4.2 mmol) was added to a "H NMR (DMSO-d 400 MHz) & 1.54 (3H, m), 2.00 (3H, mixture of 2-(methylsulfonyl)-4-(1-tosyl-5-(trifluorom m), 2.32-2.37 (4H, m), 3.38 (4H, m), 5.19-5.26 (1H, m), ethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimidine-5-carboni 6.77-6.84 (1H, m), 7.56-7.62 (1H, m), 8.08-8.16 (1H, m), trile (330 mg, 2.1 mmol) and 1-(6-chloropyridin-3-yl)etha 35 8.70-8.77 (4H, m), 9.01-9.19 (1H, m), 12.98 (1H, m) mixture namine (described in the literature: EP375,907 (B1))(500mg, of rotamers; MS (ES") 508, (ES) 506. 1 mmol) intetrahydrofuran (3 mL). The reaction mixture was Data for I-129 (Second Eluted): heated under microwave irradiation at 110°C. for 15 minutes. H NMR (DMSO-de, 400 MHz) & 1.54 (3H, m), 2.18 (3H, The reaction mixture was partitioned between EtOAc and m), 2.33-2.37 (4H, m), 3.38-3.43 (4H, m), 5.18-5.27 (1H, m), water. The organic phase was dried over magnesium Sulphate, 40 6.77-6.84 (1H, m), 7.56-7.62 (1H, m), 8.08-8.16 (1H, m), filtered and evaporated to dryness. The residue was purified 8.70-8.77 (4H, m), 9.04-9.18 (1H, m), 12.98 (1H, m) mixture on silica gel by flash column chromatography to afford the of rotamers; MS (ES") 508, (ES) 506. title compound as a wax (345 mg, 58% yield). Example 3 MS (ES") 598. 45 Step 3: 2-((R)-1-(6-(4-methylpiperazin-1-yl)pyridin N-((1R,4r)-4-((1R)-1-(5-cyano-4-(5-(trifluorom 3-yDethylamino)-4-(5-(trifluoromethyl)-1H-pyrrolo ethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimidin-2- 2,3-bipyridin-3-yl)pyrimidine-5-carbonitrile I-147 ylamino)ethyl)cyclohexyl)methanesulfonamide and 2-((S)-1-(6-(4-methylpiperazin-1-yl)pyridin-3- I-202 and N-(1S.4s)-4-(1R)-1-(5-cyano-4-(5-(trif yl)ethylamino)-4-(5-(trifluoromethyl)-1H-pyrrolo2. 50 luoromethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimi 3-bipyridin-3-yl)pyrimidine-5-carbonitrile I-129 din-2-ylamino)ethyl)cyclohexyl)methanesulfona mide I-203

55 I-147

60

NH s O. O

65 US 8,247,421 B2 85 86 -continued 2-hydroxy-4-(1-tosyl-5-(trifluoromethyl)-1H-pyrrolo2, I-2O3 3-bipyridin-3-yl)pyrimidine-5-carbonitrile (880.2 mg, 1.916 mmol) was cooled down with an ice bath. Phosphorus oxy chloride (2.938 g, 1.786 mL, 19.16 mmol) was added drop wise and the reaction mixture was stirred for 30 minutes at 0° C. and then allowed to warm up to rt and stirred at room temperature for 18 hours. The reaction mixture was quenched by slow addition onto ice. After 1 hour of stirring, the mixture O. O was extracted into ethyl acetate. The organic layer was dried 10 N1\/ N over magnesium sulfate, filtered and concentrated in vacuo to H afford a pale orange solid. The solid was triturated further in EtOAc and filtered off (715.4 mg, 78% yield) MS (ES") 478. Step 1: 2-hydroxy-4-(1-tosyl-5-(trifluoromethyl)-1H 15 pyrrolo2,3-bipyridin-3-yl)pyrimidine-5-carbonitrile Step 3: (R)-4-(1-aminoethyl)cyclohexanol

25 HN

OH

30

35 A slurry of platinum oxide (285.5 mg, 1.257 mmol) in water (2.9 mL) was added to 4-(1R)-1-Aminoethylphenol DIPEA (7.421 g, 10.00 mL, 57.42 mmol) was added to a suspension of 2-(methylsulfonyl)-4-(1-tosyl-5-(trifluorom (5.025 g, 36.63 mmol) in methanol (100 mL). Acetic acid (2.9 ethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimidine-5-carboni mL) was added to the reaction mixture. The reaction mixture trile (1 g, 1.918 mmol) in acetonitrile (20 mL). The reaction was hydrogenated in a Parr hydrogenator at 60 psi for 18 mixture was stirred for 1 hour then water (5 mL) was added. 40 hours. The platinum oxide was filtered off and the mother The mixture was allowed to stir at room temperature for 18 liquors were concentrated in vacuo leaving a colourless oil hours. The reaction mixture was concentrated in vacuo to give (8.44 g, crude: predominantly product contamined with some the desired product (880 mg, quantitative yield). (R)-1-cyclohexylethanamine and some starting material). MS (ES") 460, (ES) 458. The crude mixture was dissolved in THF (35 mL) and 45 MeCN (7 mL). NaOH (73.32 mL of 0.5M, 36.66 mmol) was Step 2: 2-chloro-4-(1-tosyl-5-(trifluoromethyl)-1H added followed by di-tert-butyl dicarbonate (12.00 g, 54.99 pyrrolo2,3-bipyridin-3-yl)pyrimidine-5-carbonitrile mmol) in portions. The reaction mixture was allowed to stir at ambient temperature for 18 hours. The reaction mixture was 50 extracted with EtOAc (3 times). The combined organic

extracts were washed twice with 1M HCl and brine, dried over magnesium Sulfate, filtered and concentrated in vacuo. The residue was purified on silica gel by flash column chro matography to afford tert-butyl (R)-1-(4-hydroxycyclohexyl) ethylcarbamate as a white solid (4.49 g, 50% yield). 55 Tert-butyl (R)-1-(4-hydroxycyclohexyl)ethylcarbamate (4.49 g, 18.45 mmol) was dissolved in dichloromethane (25 mL) and cooled to 0°C. under nitrogen. HCl 2M in Ether (27.68 mL of 2 M., 55.35 mmol) was added and the reaction 60 allowed to warm slowly to ambient temperature for 18 hours. The reaction mixture was concentrated in vacuo. The crude mixture was dissolved in MeOH (25 mL). MP-Carbonate was added (11.75 g, 36.90 mmol) and the reaction mixture was stirred for 3 hours. The resin was filtered and the mother 65 liquors were concentrated to dryness to afford the title com pound as an off white solid (2.39 g, 90% yield). MS (ES") 144. US 8,247,421 B2 87 88 Step 4: 2-((R)-1-(4-hydroxycyclohexyl)ethylamino)- dine-5-carbonitrile (442 mg 0.7561 mmol) was dissolved in 4-(1-tosyl-5-(trifluoromethyl)-1H-pyrrolo2,3-b dichloromethane (20 mL) and cooled down to 0°C. Dess pyridin-3-yl)pyrimidine-5-carbonitrile Martin periodinane (384.8 mg, 0.9073 mmol) was added and the reaction was allowed to warm up to room temperature. The mixture was stirred for 18 hours. The reaction mixture was partitioned between dichloromethane and a 1:1 saturated Solution of Sodium hydrosulfite and sodium hydrogen car 12. OH bonate. The aqueous phase was further extracted with dichlo romethane. The combined organic phases were dried over magnesium sulfate, filtered and concentrated to dryness. The N 10 residue was purified on silica gel by flash column chromatog e y raphy (ISCO Companion, 40 g column, 0-20% EtOAc/ F N2 N N DCM) to afford the title compound as a white solid (411 mg, F 93% yield). MS (ES) 583, (ES) 581. F 21 N 15 Step 6: 2-((R)-1-(4-aminocyclohexyl)ethylamino)-4- Sa N (1-tosyl-5-(trifluoromethyl)-1H-pyrrolo2,3-bipyri N \e din-3-yl)pyrimidine-5-carbonitrile s

4-((R)-1-aminoethyl)cyclohexanol (309.4 mg, 2.16 mmol) 25 was dissolved in THF (10 mL) and DMF (4 mL) and stirred over molecular sieves for 10 minutes. Diisopropylethylamine (376.3 uL., 2.160 mmol) was added, followed by 2-chloro-4- (1-tosyl-5-(trifluoromethyl)-1H-pyrrolo2,3-bipyridin-3-yl) 2. NH2 pyrimidine-5-carbonitrile (516.1 mg, 1.080 mmol). The reac 30 tion mixture was stirred under nitrogen at ambient temperature for 18 hours. The molecular sieves was removed N by filtration and washed with EtOAc. The filtrate was washed ry-st with water, dried over magnesium Sulfate, filtered and con 35 F N21 Nu centrated in vacuo. The residue was purified on silica gel by F flash column chromatography (ISCO Companion, 80 g col umn, 0-100% EtOAC/DCM) to afford the title compound as a F r N yelow solid (558 mg, 88% yield). N N MS (ES") 585, (ES) 583. 40 N \2Os3 Step 5: 2-((R)-1-(4-oxocyclohexyl)ethylamino)-4-(1- So tosyl-5-(trifluoromethyl)-1H-pyrrolo2,3-bipyridin 3-yl)pyrimidine-5-carbonitrile 45

O % 50 N e y-l F N2 N N F 2-((R)-1-(4-oxocyclohexyl)ethylamino)-4-(1-tosyl-5-(tri 55 fluoromethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimidine-5- F 21 N carbonitrile (105 mg, 0.1802 mmol) was disolved in metha nol (5 mL) and dichloromethane (5 mL). Ammonium acetate N N (138.9 mg, 118.7 uL. 1.802 mmol), followed by sodium tri N \e acetoxyborohydride (38.19 mg, 0.1802 mmol) were added s 60 and the reaction mixture was stirred for 4 hours at room temperature. The mixture was partitioned between dichlo romethane and a saturated Solution of sodium bicarbonate. The aqueous phase was extracted with dichloromethane. The organic layer was dried over magnesium Sulfate, filtered and 65 concentrated to dryness to afford the desired product as a 2-((R)-1-(4-hydroxycyclohexyl)ethylamino)-4-(1-tosyl yellow solid (95 mg, 90% yield). 5-(trifluoromethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimi MS (ES) 584, (ES) 582. US 8,247,421 B2 89 90 Step 7: N-((1R,4r)-4-((1R)-1-(5-cyano-4-(5-(trifluo s), 4.46-4.58 (1H, m), 5.49-5.64 (1H, m), 8.48-8.58 (1H, m), romethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimidin 8.71-8.88 (2H, m), 9.10-9.23 (1H, m), 9.56-10.10 (1H, m): 2-ylamino)ethyl)cyclohexyl)methanesulfonamide MS (ES") 508, (ES) 506. I-202 and N-(1S.4s)-4-((1R)-1-(5-cyano-4-(5-(trif luoromethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimi- 5 Example 4 din-2-ylamino)ethyl)cyclohexyl)methanesulfona mide I-203 2-((R)-1-(1-(2-hydroxyacetyl)piperidin-4-yl)ethy lamino)-4-(5-(trifluoromethyl)-1H-pyrrolo 2,3-b pyridin-3-yl)pyrimidine-5-carbonitrile I-205 10 I-2O2

15

25

Step 1: tert-butyl 2-((R)-1-(4-aminocyclohexyl)ethylamino)-4-(1-tosyl-5- 4-(1-aminoethyl)piperidine-1-carboxylate (trifluoromethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimi 35 dine-5-carbonitrile (95 mg, 0.1628 mmol) was dissolved in THF (5 mL) and diisopropylethylamine (25.25 mg. 34.03uL. 0.1954 mmol) was added. The reaction mixture was cooled down to 0°C. under nitrogen and methanesulfonylchloride (15.12 uL, 0.1954 mmol) was added. The reaction mixture 40 was allowed to warm up to ambient temperature and was stirred at that temperature for 1 hour. An aqueous solution of lithium hydroxide (651.2 LL of 1 M, 0.6512 mmol) was added HN and the reaction stirred at room temperature for 3 hours. The mixture was partitioned between ethyl acetate and water. The 45 aqueous phase was acidified to pH1 with 1M HCl. The aque ous phase was extracted with ethyl acetate. The organic layer was dried over magnesium sulfate, filtered and concentrated to dryness. The cis and trans isomers were separated by 50 reverse phase preparative HPLC Waters Sunfire C18, 10uM, 100 A column, gradient 10%-95% B (solvent A: 0.05%TFA in water; solvent B: CHCN) over 16 minutes at 25 mL/min and the resulting fractions were basified by passing them A mixture of tert-butyl 4-acetylpiperidine-1-carboxylate through a bicarbonate cartridge. Concentration under vacuo 55 (145g, 0.64 mol) and ammonium acetate (225g, 2.9 mol) in afforded the desired products (I-202: 12 mg, 27% yield; methanol (2 L) was stirred at room temperature for 1.5 hours. I-203: 16 mg, 37% yield). Sodium cyanoborohydride (30 g, 0.48 mol) was added in one portion and the stirring was continued overnight at room Data for I-202 (First Eluted): temperature. The reaction mixture was poured in 2N aq. "H NMR (DMSO-de, 400 MHz) & 1.19-127 (7H, m), 60 NaOH (2 L) and extracted with dichloromethane (2x1 L). The 1.85-1.97 (2H, m), 2.06-2.10 (2H, m), 2.91 (3H, s), 3.16-3.22 combined extracts were washed with water (1 L), brine (0.3 (1H, m), 4.08-4.17 (2H, m), 5.47-5.55 (1H, m), 8.43-8.50 L), and evaporated to dryness. The residue was dissolved in (1H, m), 8.65-8.80 (2H, m), 9.02-9.14 (1H, m), 10.07 (1H, br TBME (1.5 L), dried over sodium sulfate, filtered, and evapo d); MS (ES) 508, (ES) 506. rated to dryness under reduced pressure. The residue was Data for I-203 (Second Eluted): 65 purified by bulb to bulb distillation at 180° C. and 0.1 mbar to H NMR (DMSO-d 400 MHz) & 1.39-1.86 (11H, m), afford the title compound as a colorless oil that partly solidi 2.97& 3.00 (3H rotamers), 3.70-3.74 (1H, m), 4.27-4.32(1H, fied upon standing (131 g, 90% yield). US 8,247,421 B2 91 92 "H NMR (CDC1, 300 MHz) & 4.21-4.08 (2H, m), 2.75 short to 50-60 degrees. 1 mL sample was used. Chiracel 2.58 (3H, m), 1.78-1.60 (2H, m), 1.45 (9H, s), 1.44-1.05 (5H, OJ-H, eluent Heptane/EtOH/EtNH (85/15/02), Rf 11.8 and m), 1.04 (3H, d). 13.0 min. Step 3: 2-(perfluorophenoxy)-4-(1-tosyl-5-(trifluo Step 2: (R)- and (S)-tert-butyl 4-(1-aminoethyl)pip romethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimi eridine-1-carboxylate dine-5-carbonitrile

10

HN

15

25

2-(Methylthio)-4-(1-tosyl-5-(trifluoromethyl)-1H-pyrrolo 2,3-bipyridin-3-yl)pyrimidine-5-carbonitrile (150 mg, 0.31 HN mmol) was dissolved in dichloromethane at room tempera 30 ture. Meta-chloroperbenzoic acid (110 mg, 0.64 mmol) was added in one portion and the reaction was stirred for 5 min riskO utes. Pentafluorophenol (285 mg, 1.55 mmol) was added and the solution was stirred at room temperature for 3 hours. The 35 crude mixture was washed with a diluted solution of sodium A Suspension of tert-butyl 4-(1-aminoethyl)piperidine-1- bicarbonate and brine. The organic phase was dried over carboxylate (40 g, 175 mmol) and (S)-(+)-mandelic acid (27 magnesium sulfate, filtered and concentrated in vacuo. The g, 177 mmol) in acetone (400 mL) was heated until a clear residue was purified on silica gel by flash column chromatog solution was obtained. The solution was allowed to cool down raphy to afford the title compound as a solid (60 mg, 31% to room temperature during 5 hours. The newly formed pre 40 yield). cipitate was isolated and rinsed with some acetone (30 mL). H NMR (CDC1,400 MHz): 8 2.44 (3H, s), 7.38 (2H, d), The material was recrystallized twice from 300 mL acetone 8.21 (2H, d), 8.47 (1H, s), 8.78 (1H, s), 8.95 (1H, s), 9.32(1H, and some ethanol (around 25 mL). After drying, 18 g of salt s): MS (ES") 626. was obtained which was treated in an acid base separation, 100 mL dichloromethane and 100 mL 1 Naq. NaOH. The two 45 Step 4: tert-butyl 4-((1R)-1-(5-cyano-4-(1-tosyl-5- layers were separated and the organic layer washed with 40 (trifluoromethyl)-1H-pyrrolo2,3-bipyridin-3-yl) mL 1N aq. NaOH and water, dried over sodium sulfate, fil pyrimidin-2-ylamino)ethyl)piperidine-1-carboxylate tered and concentrated in vacuo to afford (S)-tert-butyl 4-(1- aminoethyl)piperidine-1-carboxylate (9.0g,99+% ee. 22.5% 50 yield). From the mother liquor of the above mentioned first crys tallization tert-butyl 4-(1-aminoethyl)piperidine-1-carboxy late (20g, 88 mmol) could be isolated (enriched in the other enantiomer). To the material was added (R)-(-)-mandelic 55 acid (14g, 92 mmol) and acetone (300 mL). The mixture was heated until a clear solution was obtained and allowed to cool to room temperature while stirring. The obtained precipitate was collected, rinsed with some acetone and recrystallized once as described above. After acid base separation, (R)-tert 60 butyl 4-(1-aminoethyl)piperidine-1-carboxylate WaS obtained (7.5g, 99.4% ee. 19% yield). E.e. was determined as followed: sample (one drop) of the amine and one drop 1-naphthyl isocyanate in 1 mL dichlo romethane was mixed and left for 1 day at room temperature 65 (most of the dichloromethane was evaporated). The sample (R)-tert-butyl 4-(1-aminoethyl)piperidine-1-carboxylate was concentrated and 5 mL ethanol was added and heated (43.82 mg, 0.19 mmol) was added to a solution of 2-(perfluo US 8,247,421 B2 93 94 rophenoxy)-4-(1-tosyl-5-(trifluoromethyl)-1H-pyrrolo2,3- H NMR (DMSO-d 400 MHz) & 1.19-128 (6H, m), 2.33 bipyridin-3-yl)pyrimidine-5-carbonitrile (100 mg, 0.16 (3H, s), 4.17-4.27 (4H, m), 7.51 (2H, d), 8.15 (2H, d), 8.94 mmol) and the reaction mixture was stirred at room tempera (1H, s), 9.24 (1H, s),9.41 (1H, s), 9.48 (1H, s); MS (ES") 596. ture for 2 hours. The crude mixture was concentrated to dryness and the resulting residue was purified on silica gel by 5 Step 4B: tert-butyl 4-((1R)-1-(5-cyano-4-(1-tosyl-5- flash column chromatography to afford the title compound (trifluoromethyl)-1H-pyrrolo2,3-bipyridin-3-yl) (70 mg, 65% yield). pyrimidin-2-ylamino)ethyl)piperidine-1-carboxylate MS (ES) 668. This intermediate could also be prepared by step 3B and 4B. 10 Step 3B: 5-cyano-4-(1-tosyl-5-(trifluoromethyl)-1H pyrrolo 2,3-bipyridin-3-yl)pyrimidin-2-yl diethyl phosphate 15

25 5-cyano-4-(1-tosyl-5-(trifluoromethyl)-1H-pyrrolo2,3-b pyridin-3-yl)pyrimidin-2-yl diethyl phosphate (200 mg. 0.3359 mmol) was treated with a solution of (R)-tert-butyl 4-(1-aminoethyl)piperidine-1-carboxylate (76.70 mg. 30 0.3359 mmol) in THF (2 mL), and diisopropylethylamine (70.22 uL, 0.4031 mmol). The reaction mixture was stirred at room temperature for 24 hours. The crude mixture was diluted with ethylacetate and washed with aqueous NaHCO. as The organic layer was dried over magnesium sulfate and concentrated under reduced pressure. The residue was puri fied on silica gel by flash column chromatography to afford the title compound as a colourless glass (188 mg, 84% yield). MS (ES) 668. 40 Step 5: 2-((R)-1-(piperidin-4-yl)ethylamino)-4-(1- tosyl-5-(trifluoromethyl)-1H-pyrrolo2,3-bipyridin 3-yl)pyrimidine-5-carbonitrile 2.2.2-trifluoroacetate

45

2-hydroxy-4-(1-tosyl-5-(trifluoromethyl)-1H-pyrrolo2. 3-bipyridin-3-yl)pyrimidine-5-carbonitrile (87% pure, 1.23 50 s g, 2.329 mmol) in dry THF (24.60 mL) was stirred at -25°C. and treated with potassium t-butoxide (2.795 mL of 1.0 M, 2.795 mmol) in THF. The reaction mixture was warmed up to 0° C. for 20 minutes, cooled to -50° C. and treated with NHTFA diethylchlorophosphate (482.3 mg, 403.9 uL., 2.795 mmol). 55 The reaction mixture was stirred at -40°C. for 20 minutes and at room temperature for 2 hours. The reaction mixture was diluted with ice cold ethyl acetate (60 mL), cooled in ice and shaken quickly with ice cold water containing 1M HCl (2.8 mL, 2.8 mmol), basified with aqueous NaHCO and extracted 60 into ethyl acetate. The organic extracts were dried over mag nesium sulfate and concentrated to give a pale yellow solid which was triturated with ether and filtered to give a pale Trifluoroacetic acid (2 mL, 25.96 mmol) was added to a yellow solid (810 mg). The mother liquors were concentrated solution of tert-butyl 4-((1R)-1-(5-cyano-4-(1-tosyl-5-(trif in vacuo and the residue purified on silica gel by flash column 65 luoromethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimidin-2- chromatography to afford a colourless Solid (219 mg) (total: ylamino)ethyl)piperidine-1-carboxylate (1.2 g, 1.79 mmol) 1.029 g, 74% yield). in dichloromethane (25 mL) and the reaction mixture was US 8,247,421 B2 95 96 stirred at room temperature for 1 hour. The crude mixture was acetate. The organic phase was washed with water, dried over concentrated to dryness to give the title compound as a white magnesium sulfate, filtered and concentrated in vacuo. The solid (1.021 g, 98% yield). residue was purified on silica gel by flash column chromatog MS (ES) 570, (ES) 568. raphy (ISCO Companion) to afford the title compound as a 5 white solid (30 mg, 50% yield). Step 6: 2-((R)-1-(1-(2-hydroxyacetyl)piperidin-4-yl) ethylamino)-4-(1-tosyl-5-(trifluoromethyl)-1H-pyr H NMR (DMSO-de, 400 MHz) & 1.05-1.2 (5H,m), 1.75 rolo2,3-bipyridin-3-yl)pyrimidine-5-carbonitrile 1.95 (5H,m), 2.85-2.95 (1H,m), 3.63-3.68 (2H,m), 3.97-4.03 (2H.m), 4.4-4.5 (1H.m), 8.23-8.28 (1H,m), 8.7-8.8 (3H.m), 10 9.1-9.2 (1H,m), 13.1-13.2 (1H,m); MS (ES")474, (ES) 472. Example 5

15 Trans-(2R)-N-(4-((1R)-1-(5-cyano-4-(5-(trifluorom ethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimidin-2- ylamino)ethyl)cyclohexyl)-2-hydroxypropanamide I-265

25

Diisopropylethylamine (179 ul, 1.025 mmol) was added to a mixture of 2-((R)-1-(piperidin-4-yl)ethylamino)-4-(1-to OH syl-5-(trifluoromethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyri 30 midine-5-carbonitrile 2.2.2-trifluoroacetate (140 mg, 0.205 mmol), TBTU (131.7 mg, 0.14 mmol) and glycolic acid (15.6 mg, 0.205 mmol) in tetrahydrofuran (3 mL). The reaction mixture was stirred at room temperature for 1 hour. The crude Step 1: (R)-tert-butyl mixture was concentrated to dryness and redissolved in ethyl 35 1-(4-hydroxycyclohexyl)ethylcarbamate acetate. The organic phase was washed with a saturated Solu tion of sodium bicarbonate and brine, dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified on silica gel by flash column chromatography to afford the title compound (60 mg. 47% yield). Also obtained was 20 mg (21% yield) of the detosylated compound. > 1, MS (ES) 628, (ES) 626. Step 7: 2-((R)-1-(1-(2-hydroxyacetyl)piperidin-4-yl) OH ethylamino)-4-(5-(trifluoromethyl)-1H-pyrrolo2,3- 45 bipyridin-3-yl)pyrimidine-5-carbonitrile (R)-tert-butyl 1-(4-hydroxyphenyl)ethylcarbamate (de scribed in the literature: WO 2007035154 (A1)) (100 mg, 0.4214 mmol) was dissolved in methanol (5 mL). A methan

olic (5 mL) solution of rhodium on alumina (10 Wt 96, 10 mg) 50 was added. The reaction mixture was place under a hydrogen atmosphere and stir at room temperature for 18 hours. The rhodium catalyst was filtered off and the mother liquors were concentrated in vacuo. The residue was purified on silica gel by flash column chromatography to afford the title compound 55 (62 mg, 61% yield). Data for the cis isomer (26 mg as a white solid): R(30% EtOAc/Petrol)0.40; "H NMR (DMSO-d 400 MHz) & 1.04 (3H, d), 1.19 (1H, 60 m), 1.30-1.40 (6H, m), 1.37 (9H, s), 1.52-1.62 (2H, m), 3.29 1M solution of lithium hydroxide (382.5 ul, 0.38 mmol) (1H, m), 3.73 (1H, brs), 4.21 (1H, d), 6.59 (1H, d). was added to a solution of 2-((R)-1-(1-(2-hydroxyacetyl)pi peridin-4-yl)ethylamino)-4-(1-tosyl-5-(trifluoromethyl)- Data for the trans isomer (36 mg as a colourless oil): 1H-pyrrolo2,3-bipyridin-3-yl)pyrimidine-5-carbonitrile R(30% EtOAc/Petrol) 0.33; (80 mg, 0.128 mmol) in tetrahydrofuran (2 mL). The reaction 65 H NMR (DMSO-d 400 MHz) & 0.85-121 (5H, m), 1.03 mixture was stirred at room temperature for 1 hour. The crude (3H, d), 1.37 (9H, s), 1.60- 1.67 (2H, m), 1.78-1.82 (2H, m), mixture was concentrated to dryness and redissolved in ethyl 3.20-3.30 (2H, m), 4.41 (1H, br s), 6.62 (1H, d). US 8,247,421 B2 97 98 Step 2: Cis-(R)-4-(1-(tert-butoxycarbonylamino) H NMR (CDC1, 400 MHz) & 1.02-1.32 (8H, m), 1.45 ethyl)cyclohexyl methanesulfonate (9H, s), 1.62-1.84 (4H, m), 1.87-196 (2H, m), 2.60 (1H, m), 3.53 (1H, m), 4.39 (1H, m). Step 4: Trans-tert-butyl (R)-1-(4-((R)-2-hydroxypro > O panamido)cyclohexyl)ethylcarbamate O ul 1%., H 10

Cis-(R)-tert-butyl 1-(4-hydroxycyclohexyl)ethylcarbam ate (575 mg, 2.363 mmol) was dissolved in dichloromethane (10 mL) and cooled to 0°C. under nitrogen. Triethylamine 15 (478.2 mg, 658.7 uL, 4.726 mmol) followed by methane us o, sulfonylchloride (284.2 mg, 192.0 LL, 2.481 mmol) were O N1 O added dropwise. The reaction mixture was stirred at room temperature for 30 minutes. The crude mixture was washed N OH with 1M HCl, water and a saturated aqueous solution of H sodium bicarbonate. The organic extracts were dried over magnesium Sulfate, filtered and concentrated in vacuo to give the compound as a white solid (706 mg, 93% yield). "H NMR (CDC1, 400 MHz) & 1.11 (3H, d), 1.38-1.52 (12H, m), 1.54-1.70 (4H, m), 2.06-2.20 (2H, m), 3.03 (3H, s), 25 3.60 (1H, brs), 4.39 (1H, d), 5.00 (1H, s). Trans-tert-butyl (R)-1-(4-aminocyclohexyl)ethylcarbam Step 3: Trans-(R)-tert-butyl ate (125 mg, 0.52 mmol) was dissolved in DMF (3 mL). 1-(4-aminocyclohexyl)ethylcarbamate D-Lactic Acid (49.52 mg, 0.52 mmol), diisopropylethy 30 lamine (166.6 mg, 224.5 uL. 1.29 mmol) and TBTU (165.6 mg, 0.5158 mmol) were successively added. The reaction mixture was allowed to stir at ambient temperature for 3 hours. The crude mixture was partitioned between ethyl acetate and brine. The organic extracts were dried over mag 35 nesium sulfate, filtered and concentrated in vacuo. The resi due was purified on silica gel by column chromatography (ISCO Companion), 12g column, 0-10% MeOH/DCM) to afford the title compound (66.1 mg, 41% yield). 'HNMR (CDC1,400 MHz) & 1.10(3H,d), 1.12-1.38 (5H, > O 40 m), 1.44 (3H, d), 1.46 (9H, s), 1.73-1.88 (2H, m), 1.97-2.08 O 1%.o, H (2H, m), 2.40 (1H, m), 3.58 (1H, m), 3.72 (1H, m), 4.22 (1H, q), 4.37 (1H, m), 6.24 (1H, m). Step 5: Trans-(R)-N-(4-((R)-1-aminoethyl)cyclo 45 hexyl)-2-hydroxypropanamide

50 HN1 , O Cis-(R)-4-(1-(tert-butoxycarbonylamino)ethyl)cyclo hexylmethanesulfonate (706 mg, 2.196 mmol) was dissolved N aOH H in DMF (10 mL). Sodium azide (713.8 mg, 10.98 mmol) was 55 added and the reaction was heated to 80°C. for 18 hours. The reaction mixture was cooled down to ambient temperature and partitioned between EtOAc (50 mL) and water (50 mL). Trans-tert-butyl (R)-1-(4-((R)-2-hydroxypropanamido) The organic layer was separated, dried over magnesium Sul cyclohexyl)ethylcarbamate (66 mg, 0.21 mmol) was dis fate and filtered. The ethyl acetate solution was taken into 60 solved in dichloromethane (10 mL). A solution of HCl in next step without further manipulation. dioxane (209.9 uL of 4M, 0.84 mmol) was added. The reac The above solution was diluted with MeOH (50 mL) and tion mixture was stirred at ambient temperature for 18 hours. 10% Pd on carbon (Degussa) (120 mg) was added. The reac The crude mixture was concentrated in vacuo and coevapo tion mixture was place under a hydrogen atmosphere and stir rated three times with diethylether. The residue was dissolved at room temperature for 18 hours. The catalyst was filtered off 65 in methanol (10 mL), passed through a sodium bicarbonate and the mother liquors were concentrated in vacuo to afford cartridge and concentrated in vacuo to give the desired com the title compound as an oil (558.4 mg., quantitative yield). pound as a white Solid (45 mg, quantitative yield). US 8,247,421 B2 99 100 "H NMR (DMSO-de, 400 MHz) & 0.80-1.10 (6H, m), d), 8.86 (0.67H, d), 9.09 (0.33H, s), 9.23 (0.67H, s), 9.64 (1H, 1.12-1.40(6H, m), 1.50-1.82 (5H, m), 2.54 (1H, m), 3.45 (1H, brs); MS (ES") 502, (ES) 500. m), 3.91 (1H, quint), 5.40 (1H, d), 7.35 (1H, d). Example 6

luoromethyl)-1H-pyrrolo2,3-bipyridin-3-yl)pyrimi 2-(1-(6-(4-hydroxy-1-methylpiperidin-4-yl)pyridin din-2-ylamino)ethyl)cyclohexyl)-2-hydroxypropana 3-yl)ethylamino)-4-(5-(trifluoromethyl)-1H-pyrrolo 2,3-bipyridin-3-yl)pyrimidine-5-carbonitrile I-266 mide I-265 10

15

25

OH

30

35 Step 1: 1-methyl-4-(5-(2-methyl-1,3-dioxolan-2-yl) pyridin-2-yl)piperidin-4-ol

Trans-(R)-N-(4-((R)-1-aminoethyl)cyclohexyl)-2-hy 40 droxypropanamide (45 mg, 0.21 mmol) was dissolved in THF (6 mL) and DMF (2 mL) and stirred over 4A molecular sieves for 10 minutes. 4-(5-(trifluoromethyl)-1-tosyl-1H-pyrrolo2, 3-bipyridin-3-yl)-2-(methylsulfonyl)pyrimidine-5-carboni trile (131.9 mg, 0.2100 mmol) was added. The reaction mix 45 ture was stirred under nitrogen at ambient temperature for 18 hours. The molecular sieves was removed by filtration and washed with EtOAc. The filtrate was washed with water, dried over magnesium sulfate, filtered and concentrated in 50 vacuo. The residue dissolved in tetrahydrofuran (6 mL). Lithium hydroxide (840.0 uL of 1 M, 0.8400 mmol) was added and the reaction mixture was stirred at room tempera ture for 18 hours. The crude mixture was partitioned between 2-bromo-5-(2-methyl-1,3-dioxolan-2-yl)pyridine (500 ethyl acetate and brine. The organic extracts were dried over 55 magnesium sulfate, filtered and concentrated in vacuo. The mg, 2.048 mmol) (described in the literature: Bioorg. Med. residue was purified by reverse phase preparative HPLC Wa Chem., 2005, 13, 6763) was dissolved in anhydrous tetrahy ters Sunfire C18, 10uM, 100 A column, gradient 10%-95% B drofuran (10 mL) and cooled to -78° C. under nitrogen. (solvent A: 0.05%TFA in water; solvent B: CH3CN) over 16 n-BuLi (901.2 LL of 2.5M, 2.253 mmol) was added dropwise 60 and the reaction mixture was stirred at -78°C. for 30 minutes. minutes at 25 mL/min. The fractions were collected, passed 1-Methyl-4-piperidone (602.6 mg, 655.0 u, 5.325 mmol) through a sodium bicarbonate cartridge and freeze-dried to was added and the reaction was allowed to stir at -78°C. for give the title compound as a white solid (19 mg, 18% yield). 20 minutes then warmed slowly to ambient temperature. The "H NMR (CDC1, 400 MHz) & 120-135 (8H, m), 1.44 reaction mixture was stirred at room temperature for 18 hours (3H, d), 1.90-2.09 (4H, m), 2.34 (1H, d), 3.69-3.81 (1H, m), 65 and concentrated in vacuo. The residue was purified on silica 4.18-4.28 (2H, m), 5.52 (0.33H, d), 5.58 (0.67H,d), 6.27(1H, gel by flash column chromatography to afford the title com d), 8.50 (0.66H, s), 8.58 (0.33H, s), 8.72 (1H, s), 8.77 (0.33H, pound as a sticky orange Solid (483.4 mg. 85% yield). US 8,247,421 B2 101 102 "H NMR (DMSO-de, 400 MHz) & 120-1.70 (5H, m), Lithium hydroxide (1.020 mL of 1 M, 1.020 mmol) was 2.03-2.40 (9H, m), 3.74 (2H, t), 4.00 (2H, t), 5.03 (1H, m), added and the reaction mixture was stirred at room tempera 7.65 (1H, d), 7.77 (1H, dd), 8.54 (1H, s); MS (ES") 279. ture for 4 hours. The crude mixture was partitioned between ethyl acetate and brine. The organic extracts were dried over Step 2: 4-(5-(1-aminoethyl)pyridin-2-yl)-1-methylpi 5 magnesium sulfate, filtered and concentrated in vacuo. The peridin-4-ol residue was purified by reverse phase preparative HPLC Wa ters Sunfire C18, 10uM, 100 A column, gradient 10%-95% B (solvent A: 0.05%TFA in water; solvent B: CHCN) over 16 minutes at 25 mL/min. The fractions were collected, passed NH2 10 through a sodium bicarbonate cartridge and freeze-dried to give the title compound as a white solid (12.5 mg.9% yield). 21 OH "H NMR (DMSO-d 400 MHz) & 1.40-1.49 (2H, m), 1.59 N (3H, d), 2.05-2.21 (2H, m), 2.16 (1.5H, s), 2.16 (1.5H, s), 15 2.25-2.33 (2H, m), 2.50-2.55 (2H, m), 4.96 (1H, d), 5.31-5.41 (1H, m), 7.58-7.65 (1H, m), 7.78-7.87 (1H, m), 8.51 (0.5H,s), Nn 8.57 (0.5H, s), 8.70-8.72 (3H, m), 8.52-8.56 (1H, m), 8.97 (0.5H, s), 9.20 (0.5H, s), 13.03 (1H, brs); MS (ES") 523, 1-Methyl-4-(5-(2-methyl-1,3-dioxolan-2-yl)pyridin-2-yl) (ES") 521 piperidin-4-ol (483 mg, 1.735 mmol) was suspended in tet Compounds prepared using the methods of Examples 1-6 rahydrofuran (10 mL). 1M HCl (6.97 mL, 6.97 mmol) was to give the title compounds I-1 to I-299 are shown in Table I. added. The reaction mixture was stirred at room temperature The characterization data for these compounds is Summa for 48 hours and concentrated in vacuo. rized in Table II-A below and includes LC/MS (observed) and "H NMR data. The residue was dissolved in MeCH (20 mL). Ammonium 25 acetate (1.337 g. 17.35 mmol), acetic acid (1.5 mL, 26.38 mmol) and MP-CNBH (2.871 g, 6.947 mmol) were succes TABLE 1 sively added. The reaction mixture was stirred at room tem perature for 18 hours then at 40°C. for a further 7 hours. The reaction mixture was allowed to cool down to room tempera 30 ture. The resin was filtered off and the mother liquors were concentrated in vacuo. The residue was dissolved in methanol (20 mL) and MP-Carbonate (2.763 g, 8.675 mmol) was added. The reaction mixture was stirred for 6 hours. The resin was filtered off and the mother liquors were concentrated in 35 vacuo. The residue was purified on silica gel by flash column chromatography to afford the title compound as a yellow solid (159 mg, 39% yield). "H NMR (DMSO-de, 400 MHz) & 1.27 (3H, d), 1.44-1.48 (2H, m), 2.08-2.60 (9H, m), 4.02(1H, q), 4.12 (2H, brs), 4.97 40 (1H, s), 7.58 (1H, d), 7.75 (1H, dd), 8.46 (1H, d); MS (ES") 236. Step 3: 2-(1-(6-(4-hydroxy-1-methylpiperidin-4-yl) pyridin-3-yl)ethylamino)-4-(5-(trifluoromethyl)-1H 45 pyrrolo2,3-bipyridin-3-yl)pyrimidine-5-carbonitrile I-266

50

N es N 55 H OH 2 N

NS 60 2-(Methylsulfonyl)-4-(1-tosyl-5-(trifluoromethyl)-1H pyrrolo2,3-bipyridin-3-yl)pyrimidine-5-carbonitrile (160.2 mg, 0.255 mmol) was added to a solution of 4-(5-(1-amino ethyl)pyridin-2-yl)-1-methylpiperidin-4-ol (66.01 mg, 0.28 65 mmol) in tetrahydrofuran (8 mL). The reaction mixture was stirred under nitrogen at room temperature for 18 hours. 1M US 8,247,421 B2 103 104 TABLE 1-continued TABLE 1-continued

10 I-11

15

25

30

I-13

35

40 N 2 N I-14

45

50 O

55 I-15

60

I-10 65 I-16 US 8,247,421 B2 105 106 TABLE 1-continued TABLE 1-continued

F F N F Šs

10 I-17 I-23

15

I-24 I-18

25

30

I-19 I-2S

35

40

O I-26

I-20 45

50

I-27 I-21 55

US 8,247,421 B2 119 120 TABLE 1-continued TABLE 1-continued

5

e NH OH N 10 O

I-106

15

y 2O O I-107 I-101 25

30

N I-108

35 N I-102

40

OH I-109 45

I-103

OH 50 On

O I-110 55

I-104

OH 60 OH

O 65 I-111

I-105

US 8,247,421 B2 123 124 TABLE 1-continued TABLE 1-continued

21 10 s N ls I-121

15

I-125 C 2O

OH

25

I-122

30

35 I-126

40

45

I-123

50

I-127

55

60

I-124 65 I-128 US 8,247,421 B2 125 126 TABLE 1-continued TABLE 1-continued

10

I-133 15

I-129

25

I-134

30

35 N I-130 40 I-135

I-136

55

60

I-132 65 US 8,247,421 B2 127 128 TABLE 1-continued TABLE 1-continued

5

10

15 I-138

20 I-142

25

I-139

35 I-143

40

45

I-140 50 I-144

55

60

I-141 65 I-145

US 8,247,421 B2 131 132 TABLE 1-continued TABLE 1-continued

CF CF NC NC 21 NN 5 an E

NS N N NS N N

HN NS 10 HN YCH, O O I-157 I-163

CF NC Z 21 NN \ N ls N NS | N N H H HN Nch, O I-158 I-164

CF NC 25 CF, Z a NN NCN4 N

NS y l N N NS() N. J N. HN | N A 30 HN | H Nns O O I-165 I-159

CF 35 CF 3. NC NC 21 NN

Sa NS N N NS l N N 40 | H N CN

HN s's HN I-166 N1 O I-160 45 CF3 NC

21 NN

NS N N N 50 | HN N Nui^ I-167 55

CF NC 21 60 Y N ls NS N N

S HN | O N O H

I-162 65 I-168

US 8,247,421 B2 135 136 TABLE 1-continued TABLE 1-continued

CF NC Z an \ N ls NS l N NH NH HN

I-181 I-187

N

I-188

I-183 I-189

I-184

2

I-191

N HN OH

I-186 65 I-192

US 8,247,421 B2 139 140 TABLE 1-continued TABLE 1-continued

I-212

35

OH

O

40

45 I-213

50

I-208

55 I-214

NN : 60 N1 .. O H 'N ls H I-209 65 N N1 No

US 8,247,421 B2 143 144 TABLE 1-continued TABLE 1-continued

I-226 I-231 15

2O

NH

25 I-232

I-228 I-233 40

I-234

55

60

65 I-235 US 8,247,421 B2 145 146 TABLE 1-continued TABLE 1-continued

I-237

O I-238 I-243 40

45

I-239

55

1. "/NN-N ~ 65 I-245 US 8,247,421 B2 147 148 TABLE 1-continued TABLE 1-continued

10

O I-246 15

s 2O N H N NH2 O 1 a 25 O I-252 I-247

I-248 40 I-253

I-254

55

NH NH2 60 N NH2

I-2SO 65 I-255 US 8,247,421 B2 149 150 TABLE 1-continued TABLE 1-continued

10 OH

I-256 I-261 15

25

I-262

I-258 40 I-263

65 I-265

US 8,247,421 B2 153 154 TABLE 1-continued TABLE 1-continued

I-283

I-278

25 I-284

30

35

I-285

40

45

50

I-281 I-286 55

60

I-282 65 I-287