Methods and compounds for restoring mutant p53 function

Abstract

Mutations in oncogenes and tumor suppressor genes contribute to the development and progression of cancer. The present disclosure describes compounds and methods for restoring wild-type function to p53 mutants. The compounds of the invention can bind to mutant p53 and restore the ability of p53 mutants to bind DNA and activate downstream effectors involved in tumor suppression. The disclosed compounds can be used to reduce the progression of cancers containing p53 mutations.

Description

[0001] This application is a divisional of Chinese Patent Application No. 201780013450.6, filed February 17, 2017, entitled "Methods and Compounds for Restoring Mutant p53 Function," which corresponds to PCT Application No. PCT / US2017 / 018511, filed February 17, 2017, of the same title.

[0002] CROSS-REFERENCE

[0003] This application claims the benefit of U.S. Provisional Application No. 62 / 297,450, filed February 19, 2016, which is incorporated by reference herein in its entirety.

[0004] SEQUENCE LIST

[0005] The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on February 17, 2017, is named 44727-702_601_SL.txt and is 2,540 bytes in size. BACKGROUND

[0006] Cancer is uncontrolled cell proliferation, a multi-factorial disease characterized by tumor formation, growth, and in some cases, metastasis. Cells bearing activated oncogenes, a damaged genome, or other pro-cancerous alterations can be prevented from replicating by a finely tuned tumor suppression network. A central component of this tumor suppression network is p53, one of the most potent tumor suppressors in the cell. Both wild-type and mutant conformations of p53 have been implicated in the progression of cancer.

[0007] INCORPORATION BY REFERENCE

[0008] Each patent, publication, and non-patent literature cited in this application is hereby incorporated by reference in its entirety as if each individual patent, publication, and non-patent literature was specifically and individually indicated to be incorporated by reference. SUMMARY

[0009] In some embodiments, the present application provides a compound of the formula:

[0010]

[0011] wherein:

[0012] - each is independently a single or double bond;

[0013] - X 1 is CR 5 , CR 5R 6 , N, NR 5 , O, S, C=O, C=S, or a carbon atom attached to Q 1 ;

[0014] -X 2 is CR 7 , CR 7 R 8 , N, NR 7 , O, S, C=O, C=S, or a carbon atom attached to Q 1 ;

[0015] -X 3 is CR 9 , CR 9 R 10 , N, NR 9 , O, S, C=O, C=S, or a carbon atom attached to Q 1 ;

[0016] -X 4 is CR 11 , CR 11 R 12 , N, NR 11 , O, S, C=O, C=S, or a carbon atom attached to Q 1 ;

[0017] -X 5 is CR 13 , N, or NR 13 ;

[0018] wherein at least one of X 1 , X 2 , X 3 , and X 4 is a carbon atom attached to Q 1 ;

[0019] -Q 1 is C=O, C=S, C=CR 14 R 15 , C=NR 14 , alkylene, alkenylene, or alkynylene, each of which is independently substituted or unsubstituted, or a bond;

[0020] -m is 1, 2, 3, or 4;

[0021] -Y is N, O, or absent;

[0022] -R 1 is -C(O)R 16 , -C(O)OR 16 , -C(O)NR 16 R 17、-OR 16 、-SR 16 、-

[0023] NR 16 R 17 、-NR 16 C(O)R 16 、-OC(O)R 16 、-SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or is hydrogen;

[0024] -Each R 3 and R 4 are independently -C(O)R 19 、-C(O)OR 19 、-

[0025] C(O)NR 19 R 20 、-SOR 19 、-SO2R 19 , alkyl, alkenyl, alkynyl, aromatic

[0026] alkyl, heteroaryl or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen, or R 3 and R 4 With R 3 and R 4 The nitrogen atoms to which they are bound together form a ring, wherein the ring is substituted or unsubstituted, or R 3 does not exist;

[0027] -Each R 2 、R 5 、R 6 、R 7 、R 8 、R 9 、R 10 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 and R 18 are independently -C(O)R 21 、-C(O)OR 21 、-C(O)NR 21 R 22 、-OR 21 、-SR 21 、-NR 21 R22 , -NR 21 C(O)R 22 , -OC(O)R 21 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen or halogen;

[0028] - each R 19 and R 20 is -C(O)R 23 , -C(O)OR 23 , -C(O)NR 23 R 24 , -

[0029] OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 , -OC(O)R 23 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen or halogen;

[0030] - each R 21 and R 22 is independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen;

[0031] and

[0032] - each R 23 and R 24 is independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen.

[0033] In some embodiments, the present application provides a method of increasing the activity of a p53 mutant in a cell, the method comprising contacting the cell with a therapeutically effective amount of a compound that binds the p53 mutant, wherein the compound increases the ability of the p53 mutant to bind DNA.

[0034] In some embodiments, the present application provides a method of inducing apoptosis in a cell, the method comprising contacting the cell with a therapeutically effective amount of a compound that binds a p53 mutant, wherein the compound increases the ability of the p53 mutant to bind DNA, wherein the cell expresses the p53 mutant.

[0035] In some embodiments, the present application provides a method of treating a condition, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound that binds to a p53 mutant in the subject, wherein binding of the compound to the p53 mutant increases the ability of the p53 mutant to bind DNA compared to the ability of the p53 mutant to bind DNA in the absence of the compound.

[0036] In some embodiments, the present application provides a method of determining the ability of a compound to activate the binding of a mutant p53 to DNA, the method comprising:

[0037] a) contacting the compound with a labeled mutant p53 moiety and an antibody conjugated to a fluorescent energy acceptor, the antibody directed against a label of the labeled mutant p53 moiety, in a test chamber;

[0038] b) contacting the labeled mutant p53 moiety with biotin-labeled DNA and streptavidin conjugated to a fluorescent energy donor, in the test chamber;

[0039] c) illuminating the test chamber with light that promotes fluorescence resonance energy transfer;

[0040] d) detecting the fluorescence resonance energy transfer;

[0041] e) determining the SC 150 value of the compound based on the fluorescence resonance energy transfer;

[0042] f) comparing the SC 150 value of the compound to the SC 150 value of a control sample, wherein the control sample comprises the labeled mutant p53 moiety, the antibody conjugated to a fluorescent energy acceptor directed against a label of the labeled mutant p53 moiety, the biotin-labeled DNA, and the streptavidin conjugated to a fluorescent energy donor, wherein the control sample does not comprise the compound; and

[0043] g) determining the level of activation of protein-DNA binding in the presence of the compound based on the comparison.

[0044] In some embodiments, the present application provides a method of increasing the activity of a p53 mutant in a cell, the method comprising contacting the cell with a therapeutically effective amount of a compound that binds to the p53 mutant, wherein the compound increases the ability of the p53 mutant to bind DNA in an assay by at least about 50%, wherein the assay comprises:

[0045] a) contacting the compound with a labeled mutant p53 moiety and an antibody conjugated to a fluorescent energy acceptor, the antibody directed against a label of the labeled mutant p53 moiety, in a test chamber;

[0046] b) contacting the labeled mutant p53 moiety with biotin-labeled DNA and streptavidin conjugated to a fluorescent energy donor in the test chamber;

[0047] c) illuminating the test chamber with light that promotes fluorescence resonance energy transfer;

[0048] d) detecting the fluorescence resonance energy transfer;

[0049] e) determining the SC 150 value of the compound based on the fluorescence resonance energy transfer;

[0050] f) comparing the SC 150 value of the compound to the SC 150 value of a control sample, wherein the control sample comprises the labeled mutant p53 moiety, the antibody against the tag of the labeled mutant p53 moiety conjugated to a fluorescent energy acceptor, the biotin-labeled DNA, and the streptavidin conjugated to a fluorescent energy donor, wherein the control sample does not comprise the compound; and

[0051] g) determining the level of activation of protein-DNA binding in the presence of the compound based on the comparison.

[0052] In some embodiments, the present application provides a method of inducing apoptosis in a cell, the method comprising contacting the cell with a therapeutically effective amount of a compound that binds to a p53 mutant, wherein the cell expresses the p53 mutant, wherein the compound increases the ability of the p53 mutant to bind DNA by at least about 50% in an assay, wherein the assay comprises:

[0053] a) contacting the compound with a labeled mutant p53 moiety and an antibody against a tag of the labeled mutant p53 moiety conjugated to a fluorescent energy acceptor in a test chamber;

[0054] b) contacting the labeled mutant p53 moiety with biotin-labeled DNA and streptavidin conjugated to a fluorescent energy donor in the test chamber;

[0055] c) illuminating the test chamber with light that promotes fluorescence resonance energy transfer;

[0056] d) detecting the fluorescence resonance energy transfer;

[0057] e) determining the SC 150 value of the compound based on the fluorescence resonance energy transfer;

[0058] f) comparing the SC 150 value of the compound to the SC 150value, wherein the control sample comprises the labeled mutant p53 portion, the antibody against the tag of the labeled mutant p53 portion conjugated to a fluorescent energy acceptor, the biotin-labeled DNA and the streptavidin conjugated to a fluorescent energy donor, wherein the control sample does not comprise the compound; and

[0059] g) determining the level of activation of protein-DNA binding in the presence of said compound based on said comparison.

[0060] In some embodiments, the present invention provides a method of treating a condition, comprising administering to a subject in need thereof a therapeutically effective amount of a compound that binds to a p53 mutant in the subject, wherein binding of the compound to the p53 mutant increases the ability of the p53 mutant to bind DNA by at least about 50% compared to the ability of the p53 mutant to bind DNA in the absence of the compound as determined by an assay comprising:

[0061] a) contacting the compound with a labeled mutant p53 portion and an antibody conjugated to a fluorescent energy acceptor in a test chamber, the antibody being directed against a tag of the labeled mutant p53 portion;

[0062] b) contacting the labeled mutant p53 portion with biotin-labeled DNA and streptavidin conjugated to a fluorescent energy donor in the test chamber;

[0063] c) illuminating the test chamber with light that promotes fluorescence resonance energy transfer;

[0064] d) detecting the fluorescence resonance energy transfer;

[0065] e) determining the SC of the compound based on the fluorescence resonance energy transfer 150 value;

[0066] f) Comparison of the SC of the compounds 150 The SC values ​​of the control samples were 150 value, wherein the control sample comprises the labeled mutant p53 portion, the antibody against the tag of the labeled mutant p53 portion conjugated to a fluorescent energy acceptor, the biotin-labeled DNA and the streptavidin conjugated to a fluorescent energy donor, wherein the control sample does not comprise the compound; and

[0067] g) determining the level of activation of protein-DNA binding in the presence of said compound based on said comparison. BRIEF DESCRIPTION OF THE DRAWINGS

[0068] FIG. 1 Shown is a protein DNA binding assay of mutant p53 in the presence of compounds of the present invention.

[0069] FIG. 2 Protein DNA binding assays for mutant p53 are shown in the presence of the compounds of the application to demonstrate the specificity of the compounds of the application. DETAILED DESCRIPTION

[0070] The present application provides compounds and methods for restoring wild-type function to mutant p53. The compounds of the application can bind to mutant p53 and restore the ability of the p53 mutant to bind DNA. Restoration of p53 mutant activity can allow activation of downstream effectors of p53, resulting in inhibition of cancer progression. The present application also provides methods of treating cancerous lesions or tumors harboring p53 mutations.

[0071] Cancer is a collection of related diseases characterized by uncontrolled cell proliferation with the ability to metastasize throughout the body. Cancer can be divided into five broad categories, including, for example: carcinomas, which can arise from cells covering the inside and outside of the body such as the lung, breast, and colon; sarcomas, which can arise from cells located in the bone, cartilage, fat, connective tissue, muscle, and other supportive tissues; lymphomas, which can arise in lymph nodes and immune system tissues; leukemias, which can arise in the bone marrow and accumulate in the bloodstream; and adenomas, which can arise in the thyroid, pituitary, adrenal, and other glandular tissues.

[0072] While different cancers can develop in almost any body tissue and contain unique characteristics, the underlying processes that lead to cancer can be similar in all forms of the disease. Cancer begins when cells break free from the normal restraints on cell division and begin to grow and divide uncontrollably. Genetic mutations in the cell can exclude the cell's ability to repair damaged DNA or initiate apoptosis, and can lead to uncontrolled cell growth and division.

[0073] The ability of a tumor cell population to expand depends not only on the rate of cell proliferation, but also on the rate of cell consumption. Programmed cell death or apoptosis represents the major mechanism of cell consumption. Cancer cells can evade apoptosis through a variety of strategies, for example, by inhibiting p53 function, thereby suppressing the expression of pro-apoptotic proteins.

[0074] Oncogenes and tumor suppressor genes can regulate cell proliferation. Genetic mutations can affect oncogenes and tumor suppressor genes, potentially aberrantly activating or inhibiting activity, further promoting uncontrolled cell division. Oncogenes contribute to cell growth, while tumor suppressor genes slow cell division by repairing damaged DNA and activating apoptosis. Cell oncogenes that can be mutated in cancer include, for example, Cdkl, Cdk2, Cdk3, Cdk4, Cdk6, EGFR, PDGFR, VEGF, HER2, Raf kinase, K-Ras, and myc. Tumor suppressor genes that can be mutated in cancer include, for example, BRCA1, BRCA2, cyclin-dependent kinase inhibitor 1C, retinoblastoma protein (pRb), PTEN, pl6, p27, p53, and p73.

[0075] Tumor suppressor p53

[0076] The tumor suppressor protein p53 is a 393 amino acid transcription factor that can regulate cell growth in response to cellular stress including, for example, UV radiation, hypoxia, oncogene activation, and DNA damage. p53 has multiple mechanisms to inhibit cancer progression including, for example, initiating apoptosis, maintaining genomic stability, cell cycle arrest, inducing senescence, and inhibiting angiogenesis. Due to the key role of p53 in tumor suppression, p53 is inactivated in almost all cancers either by direct mutation or by interference with related signaling pathways involved in tumor suppression. Homozygous loss of the p53 gene occurs in almost all types of cancer, including breast, colon, and lung cancer. The presence of certain p53 mutations in several types of human cancer can be associated with less favorable patient prognosis.

[0077] In the absence of a stress signal, p53 levels are maintained at low levels through the interaction of p53 with the E3 ubiquitin ligase Mdm2. In unstressed cells, Mdm2 can target p53 for degradation by the proteasome. Under stress conditions, the interaction between Mdm2 and p53 is disrupted, and p53 accumulates. The key event leading to p53 activation is the phosphorylation of the N-terminal domain of p53 by protein kinases, transducing the upstream stress signal. Phosphorylation of p53 leads to a conformational change that can promote DNA binding of p53 and allow transcription of downstream effectors. Activation of p53 can induce, for example, intrinsic apoptotic pathways, extrinsic apoptotic pathways, cell cycle arrest, senescence, and DNA repair. p53 can activate proteins involved in the above pathways, including, for example, Fas / Apo1, KILLER / DR5, Bax, Puma, Noxa, Bid, caspase-3, caspase-6, caspase-7, caspase-8, caspase-9, and p21 (WAF1). In addition, p53 can repress transcription of a variety of genes including, for example, c-MYC, cyclin B, VEGF, RAD51, and hTERT.

[0078] Each strand of the p53 tetramer is composed of several functional domains, including a transactivation domain (amino acids 1-100), a DNA binding domain (amino acids 101-306), and a tetramerization domain (amino acids 307-355), which are highly mobile and largely unstructured. Most p53 cancer mutations are located in the DNA binding core domain of the protein, which contains a central β- sandwich of antiparallel β-strands that serves as the basic scaffold for the DNA binding surface. The DNA binding surface is composed of two β-turn loops, L2 and L3, and a loop-helix-loop motif, stabilized by a zinc ion at, for example, Arg 175 and Arg 248. Together, these structural elements form an extended DNA binding surface that is rich in positively charged amino acids and makes specific contacts with various p53 response elements.

[0079] Reactivation of wild-type p53 function in cancer cells can be an effective therapy due to the prevalence of p53 mutations in almost all types of cancer. Mutations in p53 located at the protein's DNA binding domain or periphery of the DNA binding surface result in abnormal protein folding required for DNA recognition and binding. Mutations in p53 can occur, for example, at amino acids Val 143, His 168, Arg 175, Tyr 220, Gly 245, Arg 248, Arg 249, Phe 270, Arg 273, and Arg 282. p53 mutations that can abrogate p53 activity include, for example, R175H, Y220C, G245S, R248Q, R248W, R273H, and R282H. These p53 mutations can distort the structure of the DNA binding site or destabilize the folded protein thermodynamically at body temperature. Wild-type function of p53 mutants can be restored by binding of the p53 mutant to a compound that shifts the folding-unfolding equilibrium toward the folded state, thereby reducing the rate of unfolding and destabilization.

[0080] Non-limiting examples of amino acids include: alanine (A, Ala); arginine (R, Arg); asparagine (N, Asn); aspartic acid (D, Asp); cysteine (C, Cys); glutamic acid (E, Glu); glutamine (Q, Gin); glycine (G, Gly); histidine (H, His); isoleucine (I, He); leucine (L, Leu); lysine (K, Lys); methionine (M, Met); phenylalanine (F, Phe); proline (P, Pro); serine (S, Ser); threonine (T, Thr); tryptophan (W, Trp); tyrosine (Y, Tyr); and valine (V, Val).

[0081] Mechanism of the compounds of the invention

[0082] The compounds of the invention can selectively bind to p53 mutants and can restore wild-type activity of p53 mutants, including, for example, DNA binding function and activation of downstream targets involved in tumor suppression. In some embodiments, the compounds of the invention selectively bind to p53 Y220C mutants. The Y220C mutant is a temperature sensitive mutant that binds to DNA at lower temperatures and denatures at body temperature. The compounds of the invention can stabilize the Y220C mutant to reduce the likelihood of protein denaturation at body temperature.

[0083] The aromatic ring of Y220 is located at the periphery of the p53 β-sandwich structure connecting β-strands S7 and S8 and is an integral part of the hydrophobic core of the β-sandwich. The Y220C mutation is highly destabilizing due to the formation of an internal surface cavity. The compounds of the present invention can bind to and occupy this surface cleft, stabilizing the β-sandwich structure and thereby restoring wild-type p53 DNA binding activity.

[0084] To determine the ability of the compounds of the invention to bind to and stabilize mutant p53, assays can be used to detect, for example, conformational changes in p53 mutants or activation of wild-type p53 targets. Conformational changes in p53 can be measured, for example, by differential scanning fluorimetry (DSF), isothermal titration calorimetry (ITC), nuclear magnetic resonance spectroscopy (NMR), or X-ray crystallography. In addition, antibodies specific for wild-type mutant conformations of p53 can be used to detect conformational changes by, for example, immunoprecipitation (IP), immunofluorescence (IF), or immunoblotting.

[0085] Methods used to detect the ability of p53 mutants to bind DNA can include, for example, DNA affinity immunoblotting, modified enzyme-linked immunosorbent assay (ELISA), electrophoretic mobility shift assay (EMSA), fluorescence resonance energy transfer (FRET), homogeneous time-resolved fluorescence (HTRF), and chromatin immunoprecipitation (ChIP) assays.

[0086] In order to determine whether the compounds described herein can reactivate the transcriptional activity of p53, the activation of downstream targets in the p53 signaling cascade can be measured. The activation of p53 effector proteins can be detected by, for example, immunohistochemistry (IHC-P), reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. The activation of p53 can also be measured by inducing apoptosis via the caspase cascade and using methods including, for example, annexin V staining, TUNEL assays, procaspase and caspase levels and cytochrome c levels. Another consequence of p53 activation is aging, which can be measured using methods such as beta-galactosidase staining.

[0087] The p53 mutants that can be used to determine the effectiveness of the compounds of the present invention in enhancing the DNA binding ability of p53 mutants are p53 truncation mutants that contain only amino acids 94-312, comprising the DNA binding domain of p53. For example, the sequence of the p53Y220C mutant used to test the efficacy of the compounds can be:

[0088] SSSVPSQ KTYQGSYGFR LGFLHSGTAK SVTCTYSPAL

[0089] NKMFCQLAKT CPVQLWVDST PPPGTRVRAM

[0090] AIYKQSQHMT EVVRRCPHHE RCSDSDGLAP

[0091] PQHLIRVEGN LRVEYLDDRN TFRHSVVVPC

[0092] EPPEVGSDCT TIHYNYMCNS SCMGGMNRRP

[0093] ILTIITLEDS SGNLLGRNSF EVHVCACPGR

[0094] DRRTEEENLR KKGEPHHELP PGSTKRALSN NT (SEQ

[0095] ID NO. 1)

[0096] The compounds of the application can increase the ability of a p53 mutant to bind DNA by at least or up to about 0.1%, at least or up to about 0.2%, at least or up to about 0.3%, at least or up to about 0.4%, at least or up to about 0.5%, at least or up to about 0.6%, at least or up to about 0.7%, at least or up to about 0.8%, at least or up to about 0.9%, at least or up to about 1%, at least or up to about 2%, at least or up to about 3%, at least or up to about 4%, at least or up to about 5%, at least or up to about 6%, at least or up to about 7%, at least or up to about 8%, at least or up to about 9%, at least or up to about 10%, at least or up to about 11%, at least or up to about 12%, at least or up to about 13%, at least or up to about 14%, at least or up to about 15%, at least or up to about 16%, at least or up to about 17%, at least or up to about 18%, at least or up to about 19%, at least or up to about 20%, at least or up to about 21%, at least or up to about 22%, at least or up to about 23%, at least or up to about 24%, at least or up to about 25%, at least or up to about 26%, at least or up to about 27%, at least or up to about 28%, at least or up to about 29%, at least or up to about 30%, at least or up to about 31%, at least or up to about 32%, at least or up to about 33%, at least or up to about 34%, at least or up to about 35%, at least or up to about 36%, at least or up to about 37%, at least or up to about 38%, at least or up to about 39%, at least or up to about 40%, at least or up to about 41%, at least or up to about 42%, at least or up to about 43%, at least or up to about 44%, at least or up to about 45%, at least or up to about 46%, at least or up to about 47%, at least or up to about 48%, at least or up to about 49%, at least or up to about 50%, at least or up to about 51%, at least or up to about 52%, at least or up to about 53%, at least or up to about 54%, at least or up to about 55%, at least or up to about 56%, at least or up to about 57%, at least or up to about 58%, at least or up to about 59%, at least or up to about 60%, at least or up to about 61%, at least or up to about 62%, at least or up to about 63%, at least or up to about 64%, at least or up to about 65%, at least or up to about 66%, at least or up to about 67%, at least or up to about 68%, at least or up to about 69%, at least or up to about 70%, at least or up to about 71%, at least or up to about 72%, at least or up to about 73%, at least or up to about 74%, at least or up to about 75%, at least or up to about 76%, at least or up to about 77%, at least or up to about 78%, at least or up to about 79%, at least or up to about 80%, at least or up to about 81%, at least or up to about 82%, at least or up to about 83%, at least or up to about 84%, at least or up to about 85%,at least or at most about 86%, at least or at most about 87%, at least or at most about 88%, at least or at most about 89%, at least or at most about 90%, at least or at most about 91%, at least or at most about 92%, at least or at most about 93%, at least or at most about 94%, at least or at most about 95%, at least or at most about 96%, at least or at most about 97%, at least or at most about 98%, at least or at most about 99%, at least or at most about 100%, at least or at most about 125%, at least or at most about 150%, at least or at most about 175%, at least or at most about 200%, at least or at most about 225%, or at least or at most about 250%.

[0097] The compounds described herein can increase the activity of a p53 mutant, i.e., by at least or at most about 2-fold, at least or at most about 3-fold, at least or at most about 4-fold, at least or at most about 5-fold, at least or at most about 6-fold, at least or at most about 7-fold, at least or at most about 8-fold, at least or at most about 9-fold, at least or at most about 10-fold, at least or at most about 11-fold, at least or at most about 12-fold, at least or at most about 13-fold, at least or at most about 14-fold, at least or at most about 15-fold, at least or at most about 16-fold, at least or at most about 17-fold, at least or at most about 18-fold, at least or at most about 19-fold, at least or at most about 20-fold, at least or at most about 25-fold, at least or at most about 30-fold, at least or at most about 35-fold, at least or at most about 40-fold, at least or at most about 45-fold, at least or at most about 50-fold, at least or at most about 55-fold, at least or at most about 60-fold, at least or at most about 65-fold, at least or at most about 70-fold, at least or at most about 75-fold, at least or at most about 80-fold, at least or at most about 85-fold, at least or at most about 90-fold, at least or at most about 95-fold, at least or at most about 100-fold, at least or at most about 110-fold, at least or at most about 120-fold, at least or at most about 130-fold, at least or at most about 140-fold, at least or at most about 150-fold, at least or at most about 160-fold, at least or at most about 170-fold, at least or at most about 180-fold, at least or at most about 190-fold, at least or at most about 200-fold, at least or at most about 250-fold, at least or at most about 300-fold, at least or at most about 350-fold, at least or at most about 400-fold, at least or at most about 450-fold, at least or at most about 500-fold, at least or at most about 550-fold, at least or at most about 600-fold, at least or at most about 650-fold, at least or at most about 700-fold, at least or at most about 750-fold, at least or at most about 800-fold, at least or at most about 850-fold, at least or at most about 900-fold, at least or at most about 950-fold, at least or at most about 1,000-fold, at least or at most about 1,500-fold, at least or at most about 2,000-fold, at least or at most about 3,000-fold, at least or at most about 4,000-fold, at least or at most about 5,000-fold, at least or at most about 6,000-fold, at least or at most about 7,000-fold, at least or at most about 8,000-fold, at least or at most about 9,000-fold, or at least or at most about 10,000-fold, compared to the activity of the p53 mutant in the absence of the compound.

[0098] For example, the compounds of the application can be used to induce apoptosis, cell cycle arrest, or senescence in a cell. In some embodiments, the cell is a cancer cell. In some embodiments, the cell harbors a mutation in p53.

[0099] Compounds of the application

[0100] Non-limiting examples of compounds of the application include any of the following general formulae:

[0101]

[0102] In some embodiments, the compounds of the application are compounds of the formula:

[0103]

[0104] wherein each is independently a single or double bond; X 1 is CR 5 , CR 5 R 6 , N, NR 5 , O, S, C=O, C=S, or a carbon atom attached to Q 1 ; X 2 is CR 7 , CR 7 R 8 , N, NR 7 , O, S, C=O, C=S, or a carbon atom attached to Q 1 ; X 3 is CR 9 , CR 9 R 10 , N, NR 9 , O, S, C=O, C=S, or a carbon atom attached to Q 1 ; X 4 is CR 11 , CR 11 R 12 , N, NR 11 , O, S, C=O, C=S, or a carbon atom attached to Q 1 ; X 5 is CR 13 , N, or NR 13 ; wherein at least one of X 1 , X 2 , X 3 , and X 4 is a carbon atom attached to Q 1 ; Q 1 is C=O, C=S, C=CR 14 R 15 , C=NR 14 , alkylene, alkenylene, or alkynylene, each of which is independently substituted or unsubstituted, or a bond; m is 1, 2, 3, or 4; Y is N, O, or absent; R 1 is -C(O)R 16 , -C(O)OR 16 , -C(O)NR 16 R 17 , -OR16 -SR 16 -NR 16 R 17 -NR 16 C(O)R 16 -OC(O)R 16 -SiR 16 R 17 R 18 alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen; each R 3 and R 4 are independently -C(O)R 19 -C(O)OR 19 -C(O)NR 19 R 20 -SOR 19 -SO2R 19 alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen, or R 3 and R 4 together with the nitrogen atom to which they are bonded form a ring, wherein the ring is substituted or unsubstituted, or R 3 is absent; each R 4 , R 3 , R 2 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 and R 18 are independently -C(O)R 21 -C(O)OR 21 -C(O)NR 21 R 22 -OR 21 -SR 21 -NR 21 R 22 -NR 21 C(O)R 22 -OC(O)R 21 alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen or halogen; each R 19 and R 20-C(O)R 23 -C(O)OR 23 -C(O)NR 23 R 24 -OR 23 -SR 23 -NR 23 R 24 -NR 23 C(O)R 24 -OC(O)R 23 alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen or halogen; each R 21 and R 22 are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen; and each R 23 and R 24 are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen.

[0105] In some embodiments, the pattern of dashed bonds is selected to provide an aromatic system, such as indole, indolene, pyrrolopyridine, pyrrolopyrimidine, or pyrrolopyrazine.

[0106] In some embodiments, X 1 is CR 5 , CR 5 R 6 , or a carbon atom attached to Q 1 . In some embodiments, X 2 is CR 7 , CR 7 R 8 , or a carbon atom attached to Q 1 . In some embodiments, X 3 is CR 9 , CR 9 R 10 , or a carbon atom attached to Q 1 . In some embodiments, X 4 is CR 11 , CR 11 R 12 , or a carbon atom attached to Q 1 . In some embodiments, X 5 is CR 13 , N, or NR 13 . In some embodiments, X 1 is a carbon atom attached to Q 1 . In some embodiments, X 2is a carbon atom attached to Q 1 In some embodiments, X 3 is a carbon atom attached to Q 1 In some embodiments, X 4 is a carbon atom attached to Q 1 In some embodiments, X 5 is N.

[0107] In some embodiments, the compound is a compound of the formula:

[0108]

[0109] In some embodiments, the compound is a compound of the formula:

[0110]

[0111] wherein R 1 is -C(O)R 16 , -C(O)OR 16 , -C(O)NR 16 R 17 , -OR 16 , -SR 16 , -NR 16 R 17 , -NR 16 C(O)R 16 , -OC(O)R 16 , SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen.

[0112] In some embodiments, m is 1, 2, 3, or 4. In some embodiments, m is 1. In some embodiments, X 3 is a carbon atom attached to Q 1 , and m is 1. In some embodiments, the compound is a compound of the formula:

[0113]

[0114] wherein R 1 is -C(O)R 16 , -C(O)OR 16 , -C(O)NR 16 R 17 , -OR 16 , -SR 16 , -NR 16 R 17 , -NR16 C(O)R 16 , -OC(O)R 16 , SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen.

[0115] In some embodiments, R 1 is alkyl, alkenyl, -C(O)R 16 , -C(O)OR 16 , or -C(O)NR 16 R 17 . In some embodiments, R 1 is substituted alkyl. R 1 may be substituted with one or more substituents selected from the group consisting of hydroxyl, thiol, halogen, amino, nitro, nitroso, cyano, azido, sulfoxide, sulfone, sulfonamide, carboxyl, carboxaldehyde, imine, alkyl, halo-alkyl, cyclic alkyl, alkenyl, halo-alkenyl, alkynyl, halo-alkynyl, alkoxy, aryl, aryloxy, aralkyl, aralkoxy, heterocyclyl, acyl, acyloxy, carbamate, amide, carbamate, and ester. In some embodiments, R 1 is alkyl substituted with an amine group. In some embodiments, R 1 is alkyl substituted with NR 16 R 17 .

[0116] In some embodiments, Q 1 is C=O, C=S, C=CR 14 R 15 , C=NR 14 , alkylene, alkenylene, or alkynylene, each of which is independently substituted or unsubstituted, or a bond. In some embodiments, Q 1 is alkylene, alkenylene, or alkynylene. In some embodiments, Q 1 is C1-alkylene. In some embodiments, each R 16 and R 17 is independently alkyl, alkenyl, aryl, heteroaryl, heterocyclyl, or hydrogen. In some embodiments, Q 1 is C1-alkylene, R 16 is aryl, and R 17 is alkyl. In some embodiments, Q 1 is C1-alkylene, R 16 is aryl, and R 17 is hydrogen. In some embodiments, Q 1is C1-alkylene, R 16 is a heteroaryl group, and R 17 In some embodiments, Q 1 is C1-alkylene, R 16 is a heteroaryl group, and R 17 In some embodiments, Q 1 is C1-alkylene, R 16 is a substituted heteroaryl group, and R 17 In some embodiments, Q 1 is C1-alkylene, R 16 is a substituted alkyl group, and R 17 In some embodiments, R 17 is aryl, heteroaryl or heterocyclyl, each of which is independently substituted or unsubstituted by halogen, alkyl or hydroxy. In some embodiments, R 16 is hydrogen, and R 17 is aryl or heteroaryl, which may or may not be substituted by halogen or alkyl. 16 is an alkyl group, and R 17 is a heteroaryl group substituted with halogen or alkyl. 17 is aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted by alkyl. 17 is aryl or heteroaryl, each of which is independently substituted with alkyl, wherein the alkyl is optionally substituted with fluoro, chloro, bromo, iodo or cyano.

[0117] In some embodiments, R 2 In some embodiments, R 13 is alkyl, alkenyl, hydrogen or halogen. 2 is an alkyl group, and R 13 In some embodiments, R 2 is hydrogen, and R 13 In some embodiments, R 2 is methyl, ethyl, propyl, isopropyl, butyl or tert-butyl. 13 is methyl, ethyl, propyl, isopropyl, butyl or tert-butyl. 2 is hydrogen, and R 13 For hydrogen.

[0118] In some embodiments, R 3 -C(O)R 19 、-C(O)OR 19, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or is hydrogen, and R 4 -C(O)R 19 、-C(O)OR 19 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or is hydrogen. In some embodiments, R 3 is hydrogen and R 4 In some embodiments, R 3 is hydrogen and R 4 In some embodiments, R 3 is an alkyl group and R 4 In some embodiments, R 3 is an alkyl group and R 4 It is an aryl group.

[0119] In some embodiments, R 3 is hydrogen, and R 4 In some embodiments, the compound is a compound of the following formula:

[0120] In some embodiments, R 3 and R 4 With R 3 and R 4 The nitrogen atoms to which they are bound together form a ring, wherein the ring is substituted or unsubstituted. In some embodiments, R 3 and R 4 With R 3 and R 4 The nitrogen atoms to which they are attached together form a substituted heterocycle. 3 and R 4 With R 3 and R 4 The nitrogen atoms to which they are attached together form a heterocyclic ring substituted with hydroxy, halogen, amino or alkyl. 3 and R 4 With R 3 and R 4 The bound nitrogen atoms together form a heterocyclic ring, wherein the heterocyclic ring is substituted with a substituted or unsubstituted heterocyclic ring.

[0121] In some embodiments, R 3 and R 4 With R 3 and R 4 The bound nitrogen atoms together form a ring of the formula:

[0122] In some embodiments, R 16 is alkyl, alkenyl, aryl, heteroaryl, heterocyclyl, or hydrogen, and R 17 is aryl, heteroaryl, or heterocyclyl. In some embodiments, R 17 is phenyl, indolyl, piperidinyl, imidazolyl, thiazolyl, morpholinyl, pyrrolyl, or pyridyl.

[0123] In some embodiments, the compound is a compound of the formula:

[0124]

[0125] In some embodiments, the compound is a compound of the formula:

[0126]

[0127] In some embodiments, the compound is a compound of the formula:

[0128]

[0129] wherein each Z 1 and Z 2 is independently CR x or N; each R x is independently -C(O)R 21 , -C(O)OR 21 , -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -NR 21 R 22 , -NR 21 C(O)R 22 , -OC(O)R 21 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen or halogen; each R 25 and R 26 is independently -C(O)R 21 , -C(O)OR 21 , -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -SO2R 21 , -NR 21 R 22 , -NR 21 C(O)R 22, -OC(O)R 21 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen or halogen.

[0130] In some embodiments, Z 1 is N. In some embodiments, Z 1 and Z 2 are N. In some embodiments, each R 25 and R 26 is independently halogen. In some embodiments, R 25 is In some embodiments, R 25 is SO2R 21 . In some embodiments, R 25 is SO2R 21 , wherein R 21 is alkyl. In some embodiments, R 25 is SO2R 21 , wherein R 21 is methyl.

[0131] Non-limiting examples of compounds of the present application include the following:

[0132]

[0133]

[0134]

[0135]

[0136] or a pharmaceutically acceptable salt of any of the foregoing compounds.

[0137] Non-limiting examples of compounds of the present application include the following:

[0138]

[0139]

[0140]

[0141]

[0142] or a pharmaceutically acceptable salt of any of the foregoing compounds.

[0143] Non-limiting examples of compounds of the present application include the following:

[0144]

[0145]

[0146]

[0147]

[0148] or a pharmaceutically acceptable salt of any of the foregoing compounds.

[0149] In some embodiments, the compound is a compound of the formula:

[0150]

[0151] Each Q 1a and Q 1b are independently C=O, C=S, C=CR 14′ R 15′ 、C=NR 14′ , alkylene, alkenylene, or alkynylene, each of which is independently substituted or unsubstituted, or a bond; each R 1a and R 1b are independently -C(O)R 16′ 、-C(O)OR 16′ 、-C(O)NR 16′ R 17′ 、-OR 16′ 、-SR 16′ 、-NR 16′ R 17′ 、-NR 16′ C(O)R 16′ 、-OC(O)R 16′ 、-SiR 16′ R 17′ R 18′ , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or is hydrogen; each R 3a and R 3b is independently alkylene, alkenylene, alkynylene, arylene, heteroarylene, or heterocyclylene, each of which is independently substituted or unsubstituted, or is hydrogen; each R 4a and R 4b are independently absent, -C(O)R 19′ 、-C(O)OR 19′ 、-C(O)NR 19′ R 20′ 、-SOR 19′ 、-SO2R 19′, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or is hydrogen; each R 2a 、R 2b 、R 13a and R 13b are independently -C(O)R 21′ 、-C(O)OR 21′ 、-C(O)NR 21′ R 22′ 、-OR 21′ 、-SR 21′ 、-NR 21′ R 22′ 、-NR 21′ C(O)R 22′ 、-OC(O)R 21′ , alkyl, alkenyl, alkynyl, aryl, heteroaryl or heterocyclyl, each of which is independently substituted or unsubstituted, or is hydrogen or halogen; each R 19′ and R 20′ are independently -C(O)R 23′ 、-C(O)OR 23′ 、-C(O)NR 23′ R 24′ 、-OR 23′ 、-SR 23′ 、-NR 23′ R 24′ 、-NR 23′ C(O)R 24′ 、-OC(O)R 23′ , alkyl, alkenyl, alkynyl, aryl, heteroaryl or heterocyclyl, each of which is independently substituted or unsubstituted, or is hydrogen or halogen; each R 21′ and R 22′ is independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or is hydrogen; each R 23′ and R 24′ L is independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or is hydrogen; 1 is a linker portion; and L 2 For the connector part.

[0152] In some embodiments, each L 1 and L 2 L is independently an ester, ether, thioether, polyethylene glycol (PEG), alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocyclylene, arylene, heteroarylene, or heterocycloalkylene, any of which is substituted or unsubstituted. 1 and L 2independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, cycloarylene, or heterocycloalkylene.

[0153] In some embodiments, L 1 is alkylene and L 2 is ester.

[0154] The compounds herein can include all stereoisomers, enantiomers, diastereomers, mixtures, racemates, atropisomers, and tautomers thereof.

[0155] Non-limiting examples of optional substituents include hydroxyl, thiol, halogen, amino, nitro, nitroso, cyano, azido, sulfoxide, sulfone, sulfonamide, carboxyl, carboxaldehyde, imine, alkyl, halo-alkyl, alkenyl, halo-alkenyl, alkynyl, halo-alkynyl, alkoxy, aryl, aryloxy, aralkyl, aralkyloxy, heterocyclyl, acyl, acyloxy, carbamate, amide, urea, epoxy, and ester.

[0156] Non-limiting examples of alkyl and alkylene groups include straight chain, branched chain, and cyclic alkyl and alkylene groups. The alkyl or alkylene group can be, for example, substituted or unsubstituted C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , C 20 , C 21 , C 22 , C 23 , C 24 , C 25 , C 26 , C 27 , C 28 , C 29 , C 30 , C 31 , C 32 , C 33 , C 34 , C 35 , C 36 , C 37 , C 38 , C 39 , C 40 , C 41 , C 42 , C 43 , C 44, C 45 , C 46 , C 47 , C 48 , C 49 or C 50 group.

[0157] Non-limiting examples of straight chain alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, and decyl.

[0158] A branched alkyl group includes any straight chain alkyl group substituted with any number of alkyl groups. Non-limiting examples of branched alkyl groups include isopropyl, isobutyl, sec-butyl, and t-butyl.

[0159] Non-limiting examples of substituted alkyl groups include hydroxymethyl, chloromethyl, trifluoromethyl, aminomethyl, 1-chloroethyl, 2-hydroxyethyl, 1,2-difluoroethyl, and 3-carboxypropyl.

[0160] Non-limiting examples of cyclic alkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Cyclic alkyl groups also include fused, bridged, and spiro bicyclic and higher order fused, bridged, and spiro systems. Cyclic alkyl groups can be substituted with any number of straight chain, branched, or cyclic alkyl groups. Non-limiting examples of cyclic alkyl groups include cyclopropyl, 2-methyl-cycloprop-1-yl, cycloprop-2-en-1-yl, cyclobutyl, 2,3-dihydroxycyclobut-1-yl, cyclobut-2-en-1-yl, cyclopentyl, cyclopent-2-en-1-yl, cyclopent-2,4-dien-1-yl, cyclohexyl, cyclohex-2-en-1-yl, cycloheptyl, cyclooctyl, 2,5-dimethylcyclopent-1-yl, 3,5-dichlorocyclohex-1-yl, 4-hydroxycyclohex-1-yl, 3,3,5-trimethylcyclohex-1-yl, octahydro-pentenyl, octahydro-1H-indenyl, 3a,4,5,6,7,7a-hexahydro-3H-inden-4-yl, decahydroazulenyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[3.1.1]heptyl, 1,3-dimethyl[2.2.1]hept-2-yl, bicyclo[2.2.2]octyl, and bicyclo[3.3.3]undecyl.

[0161] Non-limiting examples of alkenyl and alkenylene groups include straight chain, branched, and cyclic alkenyl groups. One or more alkenes of the alkenyl group can be, for example, E, Z, cis, trans, terminal, or exo-methylene. The alkenyl or alkenylene group can be, for example, substituted or unsubstituted C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C17 、C 18 、C 19 、C 20 、C 21 、C 22 、C 23 、C 24 、C 25 、C 26 、C 27 、C 28 、C 29 、C 30 、C 31 、C 32 、C 33 、C 34 、C 35 、C 36 、C 37 、C 38 、C 39 、C 40 、C 41 、C 42 、C 43 、C 44 、C 45 、C 46 、C 47 、C 48 、C 49 or C 50 Non-limiting examples of alkenyl and alkenylene groups include vinyl, prop-1-en-1-yl, isopropenyl, but-1-en-4-yl; 2-chlorovinyl, 4-hydroxybuten-1-yl, 7-hydroxy-7-methyloct-4-en-2-yl, and 7-hydroxy-7-methyloct-3,5-dien-2-yl.

[0162] Non-limiting examples of alkynyl or alkynylene groups include straight chain, branched chain, and cyclic alkynyl groups. The triple bond of the alkynyl or alkynylene group can be internal or terminal. The alkynyl or alkynylene group can be, for example, substituted or unsubstituted C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 、C 18 、C 19 、C 20 、C 21 、C 22 、C 23 、C 24 、C 25 、C 26 、C 27 、C28 , C 29 , C 30 , C 31 , C 32 , C 33 , C 34 , C 35 , C 36 , C 37 , C 38 , C 39 , C 40 , C 41 , C 42 , C 43 , C 44 , C 45 , C 46 , C 47 , C 48 , C 49 or C 50 Non-limiting examples of alkynyl or alkynylene groups include ethynyl, prop-2-yn-1-yl, prop-1-yn-1-yl, and 2-methyl-hex-4-yn-1-yl; 5-hydroxy-5-methylhex-3-yn-1-yl, 6-hydroxy-6-methylhept-3-yn-2-yl, and 5-hydroxy-5-ethylhept-3-yn-1-yl.

[0163] Halo-alkyl can be any alkyl group substituted with any number of halogen atoms, such as fluorine, chlorine, bromine, and iodine atoms. Halo-alkenyl can be any alkenyl group substituted with any number of halogen atoms. Halo-alkynyl can be any alkynyl group substituted with any number of halogen atoms.

[0164] Alkoxy can be, for example, an oxygen atom substituted with any alkyl, alkenyl, or alkynyl group. Ether or ether groups include alkoxy groups. Non-limiting examples of alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, and isobutoxy.

[0165] Aryl groups can be heterocyclic or non-heterocyclic. Aryl groups can be monocyclic or polycyclic. Aryl groups can be substituted with any number of substituents described herein, such as hydrocarbyl groups, alkyl groups, alkoxy groups, and halogen atoms. Non-limiting examples of aryl groups include phenyl, toluyl, naphthyl, pyrrolyl, pyridyl, imidazolyl, thienyl, and furanyl. Non-limiting examples of substituted aryl groups include 3,4-dimethylphenyl, 4-tert-butylphenyl, 4-cyclopropylphenyl, 4-diethylaminophenyl, 4-(trifluoromethyl)phenyl, 4-(difluoromethoxy)-phenyl, 4-(trifluoromethoxy)phenyl, 3-chlorophenyl, 4-chlorophenyl, 3,4-dichlorophenyl, 2-fluorophenyl, 2-chlorophenyl, 2-iodophenyl, 3-iodophenyl, 4-iodophenyl, 2-methylphenyl, 3-fluorophenyl, 3-methylphenyl, 3-methoxyphenyl, 4-fluorophenyl, 4-methylphenyl, 4-methoxyphenyl, 2,3-difluorophenyl, 3,4-difluorophenyl, 3,5-difluorophenyl, 2,3-dichlorophenyl, 3,4-dichlorophenyl, 3,5-dichlorophenyl, 2-hydroxyphenyl, 3-hydroxyphenyl, 4-hydroxyphenyl, 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 2,3-dimethoxyphenyl, 3,4-dimethoxyphenyl, 3,5-dimethoxyphenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl, 2,3,4-trifluorophenyl, 2,3,5-trifluorophenyl, 2,3,6-trifluorophenyl, 2,4,5-trifluorophenyl, 2,4,6-trifluorophenyl, 2,4-dichlorophenyl, 2,5-dichlorophenyl, 2,6-dichlorophenyl, 3,4-dichlorophenyl, 2,3,4-trichlorophenyl, 2,3,5-trichlorophenyl, 2,3,6-trichlorophenyl, 2,4,5-trichlorophenyl, 3,4,5-trichlorophenyl, 2,4,6-trichlorophenyl, 2,3-dimethylphenyl, 2,4-dimethylphenyl, 2,5-dimethylphenyl, 2,6-dimethylphenyl, 2,3,4-trimethylphenyl, 2,3,5-trimethylphenyl, 2,3,6-trimethylphenyl, 2,4,5-trimethylphenyl, 2,4,6-trimethylphenyl, 2-ethylphenyl, 3-ethylphenyl, 4-ethylphenyl, 2,3-diethylphenyl, 2,4-diethylphenyl, 2,5-diethylphenyl, 2,6-diethylphenyl, 3,4-diethylphenyl, 2,3,4-triethylphenyl, 2,3,5-triethylphenyl, 2,3,6-triethylphenyl, 2,4,5-triethylphenyl, 2,4,6-triethylphenyl, 2-isopropylphenyl, 3-isopropylphenyl, and 4-isopropylphenyl.

[0166] Non-limiting examples of substituted aryl groups include 2-aminophenyl, 2-(N- methylamino)phenyl, 2-(N,N-dimethylamino)phenyl, 2-(N-ethylamino)phenyl, 2-(N,N- diethylamino)phenyl, 3-aminophenyl, 3-(N-methylamino)phenyl, 3-(N,N- dimethylamino)phenyl, 3-(N-ethylamino)phenyl, 3-(N,N-diethylamino)phenyl, 4- aminophenyl, 4-(N-methylamino)phenyl, 4-(N,N-dimethylamino)phenyl, 4-(N- ethylamino)phenyl, and 4-(N,N-diethylamino)phenyl.

[0167] Heterocycle can be any ring containing ring atoms that are not carbon, such as N, O, S, P, Si, B, or any other heteroatom. Heterocycle can be substituted with any number of substituents such as alkyl and halogen atoms. Heterocycle can be aromatic (heteroaryl) or non-aromatic. Non-limiting examples of heterocycle include pyrrole, pyrrolidine, pyridine, piperidine, succinamide, maleimide, morpholine, imidazole, thiophene, furan, tetrahydrofuran, pyran, and tetrahydropyran.

[0168] Non-limiting examples of heterocycle include: i) heterocyclic ring units having a single ring containing one or more heteroatoms, non-limiting examples of which include diaziridinyl, aziridinyl, azetidinyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolinyl, thiazolidinyl, isothiazolinyl, oxathiazolidinonyl, oxazolidinonyl, hydantoinyl, tetrahydrofuranyl, pyrrolidinyl, morpholinyl, piperazinyl, piperidinyl, dihydropyranyl, tetrahydropyranyl, piperidin-2- onyl, 2,3,4,5-tetrahydro-lH-azepinyl, 2,3-dihydro-lH-indolyl, and l,2,3,4-tetrahydroquinolinyl; and ii) heterocyclic ring units having 2 or more rings one of which is a heterocycle, non-limiting examples of which include hexahydro-lH-pyrrolidinyl, 3a,4,5,6,7,7a-hexahydro-lH- benzo[d]imidazolyl, 3a,4,5,6,7,7a-hexahydro-lH-indolyl, l,2,3,4-tetrahydroquinolinyl, and decahydro-lH-cycloocta[b]pyrrolyl.

[0169] ​Non-limiting examples of heteroaryl groups include: i) heteroaryl groups containing a monocyclic ring, non-limiting examples of which include 1,2,3,4-tetrazolyl, [1,2,3]triazolyl, [1,2,4]triazolyl, triazinyl, thiazolyl, 1H-imidazolyl, oxazolyl, isoxazolyl, isothiazolyl, furanyl, thienyl, pyrimidinyl, 2-phenylpyrimidinyl, pyridyl, 3-methylpyridyl, and 4-dimethylaminopyridyl; and ii) heteroaryl rings containing 2 or more fused rings of which one is a heteroaryl ring, non-limiting examples of which include 7H-purinyl, 9H-purinyl, 6-amino-9H-purinyl, 5H-pyrrolo[3,2-d]pyrimidinyl, 7H-pyrrolo[2,3-d]pyrimidinyl, pyrido[2,3-d]pyrimidinyl, 4,5,6,7-tetrahydro-1H-indolyl, quinoxalinyl, quinazolinyl, quinolinyl, 8-hydroxy-quinolinyl, and isoquinolinyl.

[0170] Any compound herein can be purified. A compound herein can be at least 1% pure, at least 2% pure, at least 3% pure, at least 4% pure, at least 5% pure, at least 6% pure, at least 7% pure, at least 8% pure, at least 9% pure, at least 10% pure, at least 11% pure, at least 12% pure, at least 13% pure, at least 14% pure, at least 15% pure, at least 16% pure, at least 17% pure, at least 18% pure, at least 19% pure, at least 20% pure, at least 21% pure, at least 22% pure, at least 23% pure, at least 24% pure, at least 25% pure, at least 26% pure, at least 27% pure, at least 28% pure, at least 29% pure, at least 30% pure, at least 31% pure, at least 32% pure, at least 33% pure, at least 34% pure, at least 35% pure, at least 36% pure, at least 37% pure, at least 38% pure, at least 39% pure, at least 40% pure, at least 41% pure, at least 42% pure, at least 43% pure, at least 44% pure, at least 45% pure, at least 46% pure, at least 47% pure, at least 48% pure, at least 49% pure, at least 50% pure, at least 51% pure, at least 52% pure, at least 53% pure, at least 54% pure, at least 55% pure, at least 56% pure, at least 57% pure, at least 58% pure, at least 59% pure, at least 60% pure, at least 61% pure, at least 62% pure, at least 63% pure, at least 64% pure, at least 65% pure, at least 66% pure, at least 67% pure, at least 68% pure, at least 69% pure, at least 70% pure, at least 71% pure, at least 72% pure, at least 73% pure, at least 74% pure, at least 75% pure, at least 76% pure, at least 77% pure, at least 78% pure, at least 79% pure, at least 80% pure, at least 81% pure, at least 82% pure, at least 83% pure, at least 84% pure, at least 85% pure, at least 86% pure, at least 87% pure, at least 88% pure, at least 89% pure, at least 90% pure, at least 91% pure, at least 92% pure, at least 93% pure, at least 94% pure, at least 95% pure, at least 96% pure, at least 97% pure, at least 98% pure, at least 99% pure, at least 99.1% pure, at least 99.2% pure, at least 99.3% pure, at least 99.4% pure, at least 99.5% pure, at least 99.6% pure, at least 99.7% pure, at least 99.8% pure, or at least 99.9% pure.

[0171] In some embodiments, the compounds of the present application can be used to treat cancer in a subject. For example, the compounds of the present application can slow the proliferation of a cancer cell line or kill a cancer cell. Non-limiting examples of cancers that can be treated by the compounds of the present application include: acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancers, AIDS-related lymphoma, anal cancer, appendix cancer, astrocytic tumor, basal cell carcinoma, biliary cancer, bladder cancer, bone cancer, brain tumors such as cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumors, visual pathway and hypothalamic glioma, breast cancer, bronchial adenomas, Burkitt lymphoma, cancer of unknown primary origin, central nervous system lymphoma, cerebellar astrocytoma, cervical cancer, childhood cancers, chronic lymphocytic leukemia, chronic myelogenous leukemia, chronic myeloproliferative disorders, colon cancer, cutaneous T-cell lymphoma, desmoplastic small round cell tumor, endometrial cancer, ependymoma, esophageal cancer, Ewing sarcoma, germ cell tumors, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, glioma, hairy cell leukemia, head and neck cancer, heart cancer, hepatocellular (liver) cancer, Hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell carcinoma, Kaposi sarcoma, kidney cancer, laryngeal cancer, lip and oral cavity cancer, liposarcoma, liver cancer, lung cancer such as non-small cell lung cancer and small cell lung cancer, lymphoma, leukemia, macroglobulinemia, malignant fibrous histiocytoma of bone / osteosarcoma, medulloblastoma, melanoma, mesothelioma, metastatic squamous neck cancer with occult primary, mouth cancer, multiple endocrine neoplasia syndrome, myelodysplastic syndromes, myeloid leukemia, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer, oral cancer, oropharyngeal cancer, osteosarcoma / malignant fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, pancreatic cancer, pancreatic islet cell carcinoma, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pineal astrocytic tumors, pineal germinoma, pituitary tumor, pleuropulmonary blastoma, plasmacytoma, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell cancer, renal pelvis and ureter transitional cell cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, skin cancer, Merkel cell skin cancer, small bowel cancer, soft tissue sarcoma, squamous cell carcinoma, stomach cancer, T-cell lymphoma, throat cancer, thymoma, thymic carcinoma, thyroid cancer, trophoblastic tumor (gestational), cancer of unknown primary site, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom macroglobulinemia, and Wilms tumor.

[0172] In some embodiments, the compounds of the present application exhibit non-lethal toxicity.

[0173] Pharmaceutically acceptable salts

[0174] The present application provides the use of a pharmaceutically acceptable salt of any of the therapeutic compounds described herein. Pharmaceutically acceptable salts include, for example, acid addition salts and base addition salts. The acid added to the compound to form the acid addition salt can be an organic or inorganic acid. The base added to the compound to form the base addition salt can be an organic or inorganic base. In some embodiments, the pharmaceutically acceptable salt is a metal salt. In some embodiments, the pharmaceutically acceptable salt is an ammonium salt.

[0175] Metal salts can be produced by adding an inorganic base to a compound of the present application. Inorganic bases consist of a metal cation paired with a basic counterion, such as hydroxide, carbonate, bicarbonate, or phosphate. The metal can be an alkali metal, an alkaline earth metal, a transition metal, or a main group metal. In some embodiments, the metal is lithium, sodium, potassium, cesium, cerium, magnesium, manganese, iron, calcium, strontium, cobalt, titanium, aluminum, copper, cadmium, or zinc.

[0176] In some embodiments, the metal salt is a lithium salt, a sodium salt, a potassium salt, a cesium salt, a cerium salt, a magnesium salt, a manganese salt, an iron salt, a calcium salt, a strontium salt, a cobalt salt, a titanium salt, an aluminum salt, a copper salt, a cadmium salt, or a zinc salt.

[0177] Ammonium salts can be produced by adding ammonia or an organic amine to a compound of the present application. In some embodiments, the organic amine is triethylamine, diisopropylethylamine, ethanolamine, diethanolamine, triethanolamine, morpholine, N-methylmorpholine, piperidine, N-methylpiperidine, N-ethylpiperidine, dibenzylamine, piperazine, pyridine, pyrrazole, pipyrrazole, imidazole, pyrazine, or pipyrazine.

[0178] In some embodiments, the ammonium salt is a triethylamine salt, a diisopropylethylamine salt, an ethanolamine salt, a diethanolamine salt, a triethanolamine salt, a morpholine salt, an N-methylmorpholine salt, a piperidine salt, an N-methylpiperidine salt, an N-ethylpiperidine salt, a dibenzylamine salt, a piperazine salt, a pyridine salt, a pyrrazole salt, a pipyrrazole salt, an imidazole salt, a pyrazine salt, or a pipyrazine salt.

[0179] Acid addition salts can be produced by adding an acid to a compound of the present application. In some embodiments, the acid is an organic acid. In some embodiments, the acid is an inorganic acid. In some embodiments, the acid is hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, nitrous acid, sulfuric acid, sulfurous acid, phosphoric acid, isonicotinic acid, lactic acid, salicylic acid, tartaric acid, ascorbic acid, gentisic acid, gluconic acid, glucaronic acid, saccaric acid, formic acid, benzoic acid, glutamic acid, pantothenic acid, acetic acid, propionic acid, butyric acid, fumaric acid, succinic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, oxalic acid, or maleic acid.

[0180] In some embodiments, the salt is a hydrochloride, hydrobromide, hydroiodide, nitrate, nitrite, sulfate, sulfite, phosphate, isonicotinate, lactate, salicylate, tartrate, ascorbate, gentisinate, gluconate, glucaronate, saccarate, formate, benzoate, glutamate, pantothenate, acetate, propionate, butyrate, fumarate, succinate, mesylate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, citrate, oxalate, or maleate.

[0181] Pharmaceutical compositions of the invention

[0182] For example, the pharmaceutical compositions of the invention can be used prior to, during, or after treatment of a subject with, for example, another agent.

[0183] The subject can be, for example, an elderly person, an adult, an adolescent, a pre-adolescent child, a child, an infant, a baby, a neonate, and a non-human animal. In some embodiments, the subject is a patient.

[0184] The pharmaceutical compositions of the invention can be a combination of any of the pharmaceutical compounds described herein with other chemical components, such as carriers, stabilizers, diluents, dispersing agents, suspending agents, thickening agents, and / or excipients. The pharmaceutical composition facilitates administration of the compound to an organism. Pharmaceutical compositions can be administered as a pharmaceutical composition in a therapeutically effective amount by a variety of forms and routes including, for example, intravenous, subcutaneous, intramuscular, oral, parenteral, ocular, subdermal, transdermal, nasal, vaginal, and topical administration.

[0185] The pharmaceutical composition can be administered in a local manner, for example, by direct injection of the compound into an organ, optionally in the form of a depot or sustained release formulation or implant. The pharmaceutical composition can be provided in the form of a fast release formulation, in the form of an extended release formulation, or in the form of an intermediate release formulation. Fast release forms can provide immediate release. Extended release formulations can provide controlled release or sustained, delayed release.

[0186] For oral administration, the pharmaceutical composition can be formulated by combining the active compound with a pharmaceutically acceptable carrier or excipient. Such a carrier can be used to formulate a liquid, gel, syrup, elixir, slurry, or suspension for oral ingestion by the subject. Non-limiting examples of solvents used in orally dissolvable formulations can include water, ethanol, isopropyl alcohol, saline, physiological saline, DMSO, dimethylformamide, potassium phosphate buffer, phosphate buffered saline (PBS), sodium phosphate buffer, 4-2-hydroxyethyl-l-piperazineethanesulfonic acid buffer (HEPES), 3-(N-morpholino)propanesulfonic acid buffer (MOPS), piperazine-N,N'-bis(2-ethanesulfonic acid) buffer (PIPES), and saline sodium citrate buffer (SSC). Non-limiting examples of co-solvents used in orally dissolvable formulations can include sucrose, urea, cremaphor, DMSO, and potassium phosphate buffer.

[0187] The pharmaceutical formulation can be prepared for intravenous administration. The pharmaceutical composition can be in a form suitable for parenteral injection as a sterile suspension, solution or emulsion in an oily or aqueous vehicle and can contain formulatory agents such as suspending, stabilizing and / or dispersing agents. Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Suspensions of the active compounds can be prepared as oil injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acids esters such as ethyl oleate or triglycerides, or liposomes. The suspension can also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for a highly concentrated solution. Alternatively, the active ingredient can be in powder form for reconstitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

[0188] The active compounds can be administered topically, and can be formulated into a wide variety of topically administrable compositions, such as solutions, suspensions, lotions, gels, pastes, medicated sticks, balms, creams, and ointments. Such pharmaceutical compositions can contain solubilizers, stabilizers, tonicity enhancing agents, buffers, and preservatives.

[0189] The compounds of the present application can be applied topically to the skin or body cavities of a subject, such as the oral cavity, vaginal cavity, bladder, cranial cavity, spinal column, thoracic cavity, or pelvic cavity. The compounds of the present application can be applied to accessible body cavities.

[0190] The compounds can also be formulated into rectal compositions, such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, jelly suppositories, or retention enemas, containing conventional suppository bases such as cocoa butter or other glycerides, as well as synthetic polymers such as polyvinylpyrrolidone and PEG. In suppository forms of the compositions, low-melting waxes such as a mixture of fatty acid glycerides optionally combined with cocoa butter can be melted, solidified, and then molded.

[0191] In practicing the methods of treatment or use provided herein, a therapeutically effective amount of a compound described herein is administered to a subject having a disease or condition to be treated in the form of a pharmaceutical composition. In some embodiments, the subject is a mammal, such as a human. The therapeutically effective amount can vary widely depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors. The compound can be used alone or in combination with one or more therapeutic agents as components of mixtures.

[0192] Pharmaceutical compositions can be formulated to release a compound described herein initially or at a predetermined rate or to continue to release the compound for a set period of time. The pharmaceutical compositions can be formulated to release the compound locally or systemically, and any suitable route of administration can be employed, for example, oral, transdermal, parenteral, subcutaneous, intravenous, intramuscular, intranasal, or rectal. Pharmaceutical compositions can be formulated using one or more physiologically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. Proper formulation is dependent upon the route of administration chosen.

[0193] The pharmaceutical compositions can comprise at least one pharmaceutically acceptable carrier, diluent or excipient and a compound described herein as a free base or a pharmaceutically acceptable salt form. Pharmaceutical compositions can contain solubilizers, stabilizers, tonicity enhancing agents, buffers, and preservatives.

[0194] Methods of preparing compositions comprising a compound described herein include formulating the compound with one or more inert, pharmaceutically acceptable excipients or carriers to form a solid, semi-solid, or liquid composition. Solid compositions include, for example, powders, tablets, dispersible granules, capsules, and cachets. Liquid compositions include, for example, solutions in which a compound is dissolved, emulsions containing a compound, or solutions containing liposomes, micelles, or nanoparticles comprising a compound as disclosed herein. Semi-solid compositions include, for example, gels, suspensions, and creams. The compositions can be liquid solutions or suspensions, solid forms suitable for dissolution or suspension in liquid prior to use, or as emulsions. These compositions also can contain minor amounts of nontoxic, auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, and other pharmaceutically acceptable additives.

[0195] Non-limiting examples of dosage forms suitable for use in the present application include liquids, powders, gels, nanosuspensions, nanoparticles, microgels, aqueous or oily suspensions, emulsions, and any combination thereof.

[0196] Non-limiting examples of pharmaceutically acceptable excipients suitable for use in the present application include binders, disintegrants, anti-tacking agents, anti-static agents, surfactants, antioxidants, coating agents, coloring agents, plasticizers, preservatives, suspending agents, emulsifying agents, antimicrobial agents, and spheronization agents, and any combination thereof.

[0197] The compositions of the present application can be, for example, immediate release or controlled release formulations. Immediate release formulations can be formulated to allow the compounds to act quickly. Non-limiting examples of immediate release formulations include readily dissolving formulations. Controlled release formulations can be drug formulations that have been adjusted so that the rate and pattern of release of the active agent can be matched with the physiological and time therapy requirements, or have been formulated to achieve release of the active agent at a programmed rate. Non-limiting examples of controlled release formulations include granules, delayed release granules, hydrogels (e.g., synthetic or naturally derived), other gelling agents (e.g., gel-forming dietary fibers), matrix-based formulations (e.g., formulations comprising polymeric materials with at least one active ingredient dispersed therein), granules within a matrix, polymeric mixtures, and particulate substances.

[0198] In some embodiments, the controlled release formulation is a delayed release form. The delayed release form can be formulated to delay the action of the compounds for an extended period of time. The delayed release form can be formulated to delay the release of an effective dose of one or more compounds, for example, for about 4, about 8, about 12, about 16, or about 24 hours.

[0199] The controlled release formulation can be a sustained release form. The sustained release form can be formulated to maintain, for example, the action of the compounds over an extended period of time. The sustained release form can be formulated to provide an effective dose of any of the compounds described herein (e.g., to provide a physiologically effective blood profile) over about 4, about 8, about 12, about 16, or about 24 hours.

[0200] Non-limiting examples of pharmaceutically acceptable excipients can be found, for example, in Remington: The Science and Practice of Pharmacy, 19th Ed. (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Ed. (Lippincott Williams & Wilkins 1999), each of which is incorporated herein by reference in its entirety.

[0201] The various therapeutic agents can be administered in any order or simultaneously. In some embodiments, the compound of the application is administered in conjunction with, prior to, or following administration of another therapeutic agent. If administered simultaneously, the various therapeutic agents can be provided in a single, unified form or in multiple forms, e.g., as multiple, separate pills. The agents can be packaged together in one package or packaged separately in multiple packages. One or all of the therapeutic agents can be administered in multiple doses. If not administered simultaneously, the timing between the multiple doses can vary up to about a month.

[0202] The therapeutic agents described herein can be administered before, during or after the occurrence of a disease or condition, and the timing of administration of the composition containing the therapeutic agent can vary. For example, the composition can be used as a prophylactic and can be administered continuously to a subject susceptible to a condition or disease to reduce the likelihood of the disease or condition occurring. The composition can be administered to the subject as soon as possible during or after the onset of symptoms. Administration of the therapeutic agent can be initiated within 48 hours prior to the onset of symptoms, within 24 hours prior to the onset of symptoms, within 6 hours prior to the onset of symptoms, or within 3 hours prior to the onset of symptoms. The initial administration can be made using any of the formulations described herein, via any practical route, such as by any of the routes described herein.

[0203] Upon detection or suspicion of the occurrence of a disease or condition, the compound can be administered as soon as it is available and for a length of time necessary to treat the disease, e.g., from about 1 month to about 3 months. In some embodiments, the length of time the compound can be administered can be about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 1 month, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 2 months, about 9 weeks, about 10 weeks, about 11 weeks, about 12 weeks, about 3 months, about 13 weeks, about 14 weeks, about 15 weeks, about 16 weeks, about 4 months, about 17 weeks, about 18 weeks, about 19 weeks, about 20 weeks, about 5 months, about 21 weeks, about 22 weeks, about 23 weeks, about 24 weeks, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 1 year, about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 19 months, about 20 months, about 21 months, about 22 months, about 23 months, about 2 years, about 2.5 years, about 3 years, about 3.5 years, about 4 years, about 4.5 years, about 5 years, about 6 years, about 7 years, about 8 years, about 9 years, or about 10 years. The length of treatment can vary from subject to subject.

[0204] The pharmaceutical compositions described herein can be in unit dosage form in a pack, suitable for single administration of precise dosage. In unit dosage form, the preparation is divided into unit doses containing appropriate quantities of one or more compounds. The unit dosage can be in the form of a package containing discrete quantities of the preparation. Non-limiting examples are packaged injections, vials, or ampoules. An aqueous suspension composition can be packaged in a single-dose container that is not reclosable. A multi-dose container that is reclosable can be used, for example, in combination with or without a preservative. The preparation for injection can be presented in unit dosage form, for example, in an ampoule or in a multi-dose container with a preservative.

[0205] The pharmaceutical compositions provided herein can be administered in combination with other therapies, such as chemotherapy, radiation therapy, surgery, anti-inflammatory agents, and selected vitamins. The other agents can be administered prior to, after, or simultaneously with the pharmaceutical compositions.

[0206] The pharmaceutical compositions can be in the form of solid, semi-solid, or liquid dosage forms, such as, for example, tablets, suppositories, pills, capsules, powders, liquids, suspensions, lotions, creams, or gels, for example, in unit dosage form suitable for single administration of precise dosages, depending on the intended mode of administration, according to the present application.

[0207] For a solid composition, non-toxic solid carriers include, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talcum, cellulose, glucose, sucrose, and magnesium carbonate.

[0208] Non-limiting examples of pharmaceutically active agents suitable for combination with the compositions of the present application include anti-infective agents, i.e., aminoglycosides, antiviral agents, antimicrobial agents, anticholinergics / antispasmodics, antidiabetic agents, antihypertensive agents, antineoplastic agents, cardiovascular agents, central nervous system agents, coagulation modulators, hormones, immunological agents, immunosuppressive agents, and ophthalmic agents.

[0209] The compounds can be delivered by liposome technology. The use of liposomes as drug carriers can increase the therapeutic index of the compounds. Liposomes are composed of natural phospholipids and can contain mixed lipid chains with surfactant properties (e.g., egg phosphatidyl ethanolamine). Liposome design can employ surface ligands to attach to unhealthy tissue. Non-limiting examples of liposomes include multilamellar vesicles (MLV), small unilamellar vesicles (SUV), and large unilamellar vesicles (LUV). Liposome physicochemical properties can be adjusted to optimize penetration through biological barriers and retention at the site of administration, and to reduce the likelihood of premature degradation and toxicity to non-target tissues. Optimal liposome properties depend on the route of administration: large size liposomes show good retention upon local injection, small size liposomes are better suited to achieve passive targeting. PEGylation reduces uptake of liposomes by the liver and spleen, and prolongs circulation time, resulting in increased localization at inflamed sites due to the enhanced permeability and retention (EPR) effect. Additionally, the liposome surface can be modified to achieve selective delivery of the encapsulated drug to specific target cells. Non-limiting examples of targeting ligands include monoclonal antibodies specific for receptors concentrated on the surface of cells associated with disease, vitamins, peptides, and polysaccharides.

[0210] Non-limiting examples of dosage forms suitable for use in the present application include liquids, elixirs, nanosuspensions, aqueous or oily suspensions, drops, syrups, and any combination thereof. Non-limiting examples of pharmaceutically acceptable excipients suitable for use in the present application include granulating agents, binding agents, lubricating agents, disintegrating agents, sweetening agents, glidants, anti-sticking agents, anti-static agents, surfactants, antioxidants, gums, coating agents, coloring agents, flavoring agents, coating agents, plasticizers, preservatives, suspending agents, emulsifying agents, vegetable cellulose materials, and spheronization agents, and any combination thereof.

[0211] The compositions of the present application can be packaged as a kit. In some embodiments, the kit comprises written instructions for administration / use of the composition. The written material can be, for example, a label. The written material can suggest conditions and methods of administration. The instructions provide the best guidance to the subject and supervising physician regarding obtaining the best clinical results from administration of the therapy. The written material can be a label. In some embodiments, the label can be approved by a regulatory agency, such as the U.S. Food and Drug Administration (FDA), the European Medicines Agency (EMA), or other regulatory agency.

[0212] Dosing

[0213] The pharmaceutical compositions described herein can be in unit dosage form, in a form suitable for single administration of a precise dosage. In unit dosage form, the preparation is divided into unit doses containing appropriate quantities of one or more compounds. The unit dosage can be in the form of a package containing discrete quantities of the preparation. Non-limiting examples are a liquid in a vial or ampoule. An aqueous suspension composition can be packaged in a single-dose container that is not resealable. A multi-dose container that is resealable can be used, for example, in combination with a preservative. The preparation for parenteral injection can be presented in unit dosage form, for example, in an ampoule or in a multi-dose container with a preservative.

[0214] The compounds described herein can be present in a composition in a range of about 1 mg to about 2000 mg, about 100 mg to about 2000 mg, about 10 mg to about 2000 mg, about 5 mg to about 1000 mg, about 10 mg to about 500 mg, about 50 mg to about 250 mg, about 100 mg to about 200 mg, about 1 mg to about 50 mg, about 50 mg to about 100 mg, about 100 mg to about 150 mg, about 150 mg to about 200 mg, about 200 mg to about 250 mg, about 250 mg to about 300 mg, about 300 mg to about 350 mg, about 350 mg to about 400 mg, about 400 mg to about 450 mg, about 450 mg to about 500 mg, about 500 mg to about 550 mg, about 550 mg to about 600 mg, about 600 mg to about 650 mg, about 650 mg to about 700 mg, about 700 mg to about 750 mg, about 750 mg to about 800 mg, about 800 mg to about 850 mg, about 850 mg to about 900 mg, about 900 mg to about 950 mg, or about 950 mg to about 1000 mg.

[0215] The compounds described herein can be present in the composition in an amount of about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, about 1500 mg, about 1550 mg, about 1600 mg, about 1650 mg, about 1700 mg, about 1750 mg, about 1800 mg, about 1850 mg, about 1900 mg, about 1950 mg, or about 2000 mg.

[0216] In some embodiments, the dose can be expressed in terms of the amount of drug divided by the mass of the subject, e.g., milligrams of drug per kilogram of subject body weight. In some embodiments, the compound is administered in an amount of about 5 mg / kg to about 50 mg / kg, 250 mg / kg to about 2000 mg / kg, about 10 mg / kg to about 800 mg / kg, about 50 mg / kg to about 400 mg / kg, about 100 mg / kg to about 300 mg / kg, or about 150 mg / kg to about 200 mg / kg.

[0217] The present application provides, among other things, the following embodiments:

[0218] 1. A compound of the formula:

[0219]

[0220] wherein:

[0221] - each is independently a single or double bond;

[0222] - X 1 is CR 5 , CR 5 R 6 , N, NR 5 , O, S, C=O, C=S, or Q1 the attached carbon atom;

[0223] -X 2 CR 7 , CR 7 R 8 、N、NR 7 , O, S, C=O, C=S or with Q 1 the attached carbon atom;

[0224] -X 3 CR 9 , CR 9 R 10 、N、NR 9 , O, S, C=O, C=S or with Q 1 the attached carbon atom;

[0225] -X 4 CR 11 , CR 11 R 12 、N、NR 11 , O, S, C=O, C=S or with Q 1 the attached carbon atom;

[0226] -X 5 CR 13 , N or NR 13 ;

[0227] where X 1 、X 2 、X 3 and X 4 At least one of them is 1 the attached carbon atom;

[0228] -Q 1 C=O、C=S、C=CR 14 R 15 、C=NR 14 , alkylene, alkenylene, or alkynylene, each of which is independently substituted or unsubstituted, or a bond;

[0229] -m is 1, 2, 3, or 4;

[0230] -Y is N, O or absent;

[0231] -R 1 -C(O)R 16 、-C(O)OR 16 、-C(O)NR 16 R 17 、-OR 16 、-SR 16 、-

[0232] NR 16 R 17 、-NR 16 C(O)R 16 、-OC(O)R 16 、C=O、C=S、-CN、-

[0233] SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl or heterocyclic group,

[0234] each of which is independently substituted or unsubstituted, or is hydrogen;

[0235] -Each R 3 and R 4 are independently -C(O)R 19 、-C(O)OR 19 、-

[0236] C(O)NR 19 R 20 、-SOR 19 、-SO2R 19 , alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or is hydrogen, or R 3 and R 4 With R 3 and R 4 The nitrogen atoms to which they are bound together form a ring, wherein the ring is substituted or unsubstituted, or

[0237] R 3 does not exist;

[0238] -Each R 2 、R 5 、R 6 、R 7 、R 8 、R 9 、R 10 、R 11 、R 12 、R 13 、R 14 、R 15 、R 16 、R 17 and R 18 Independently for-

[0239] C(O)R 21 、-C(O)OR 21 、-C(O)NR 21 R 22 、-OR21 21

[0240] NR 21 R 22 21 22 21 alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen or halogen;

[0241] each R 19 and R 20 is independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen; 23 23 23 R 24

[0242] OR 23 23 23 R 24 23 24 23 alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen or halogen;

[0243] each R 21 and R 22 is independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen;

[0244] and

[0245] each R 23 and R 24 is independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen.

[0246] 2. The compound of embodiment 1, wherein X 3 is a carbon atom attached to Q 1 and Y is N or O.

[0247] 3. The compound of embodiment 1, wherein m is 1 and Y is N.

[0248] 4. The compound of embodiment 1, wherein the compound is of the formula:

[0249]

[0250] ​​​​​​​​​​​​​5. The compound according to embodiment 1, wherein R 1 Alkyl, alkenyl, -C(O)R 16 、-C(O)OR 16 or -C(O)NR 16 R 17 .

[0251] 6. The compound according to embodiment 1, wherein R 1 To be NR 16 R 17 Substituted alkyl.

[0252] 7. The compound according to embodiment 1, wherein the compound is a compound of the following formula:

[0253]

[0254] 8. The compound according to embodiment 1, wherein each R 16 and R 17 is independently alkyl, alkenyl, aryl, heteroaryl, heterocyclyl or hydrogen.

[0255] 9. The compound according to embodiment 1, wherein R 16 is hydrogen or alkyl.

[0256] 10. The compound according to embodiment 1, wherein R 17 is aryl, heteroaryl or heterocyclyl, each of which is independently substituted or unsubstituted by halogen, alkyl or hydroxy.

[0257] 11. The compound according to embodiment 1, wherein Q 1 C=O, C=NR 14 , a bond, an alkylene group or an alkenylene group.

[0258] 12. The compound of embodiment 1, wherein Q 1 It is a C1-alkylene group.

[0259] 13. The compound according to embodiment 1, wherein R 2 is hydrogen or alkyl.

[0260] 14. The compound of embodiment 13, wherein R 2 It is an alkyl group.

[0261] 15. The compound of embodiment 14, wherein R 2 It is a cycloalkyl group.

[0262] 16. The compound according to embodiment 1, wherein R 13 is alkyl, alkenyl, hydrogen or halogen.

[0263] 17. The compound according to embodiment 1, wherein R 13 For hydrogen.

[0264] 18. The compound according to embodiment 1, wherein R 3 and R 4 Each is independently alkyl, alkenyl, aryl, heteroaryl, heterocyclyl or hydrogen.

[0265] 19. The compound according to embodiment 1, wherein R 4 It is an alkyl group substituted by an aryl group.

[0266] 20. The compound according to embodiment 1, wherein R 3 and R 4 With R 3 and R 4 The bound nitrogen atoms together form a ring, wherein the ring is substituted or unsubstituted.

[0267] 21. The compound according to embodiment 1, wherein R 3 and R 4 With R 3 and R 4 The bound nitrogen atoms together form a heterocyclic ring, wherein the heterocyclic ring is substituted with at least one substituent.

[0268] 22. The compound according to embodiment 1, wherein X 1 For Q 1 Attached carbon atoms.

[0269] 23. The compound of embodiment 1, wherein m is 1.

[0270] 24. The compound according to embodiment 1, wherein the compound is a compound of the following formula:

[0271]

[0272] 25. The compound according to embodiment 1, wherein R 1 Alkyl, alkenyl, -C(O)R 16 、-C(O)OR 16 or -C(O)NR 16 R 17 .

[0273] 26. The compound according to embodiment 1, wherein R 1 To be NR 16 R 17 Substituted alkyl.

[0274] 27. The compound according to embodiment 1, wherein the compound is a compound of the following formula:

[0275]

[0276] 28. The compound of embodiment 1, wherein each R 16 and R 17 is independently alkyl, alkenyl, aryl, heteroaryl, heterocyclyl or hydrogen.

[0277] 29. The compound according to embodiment 1, wherein R 16 is hydrogen or alkyl.

[0278] 30. The compound according to embodiment 1, wherein R 17 is aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted with alkyl, each of which is independently substituted or unsubstituted with halogen, alkyl, or hydroxy.

[0279] 31. The compound of embodiment 1, wherein Q 1 C=O, C=NR 14 , a bond, an alkylene group or an alkenylene group.

[0280] 32. The compound of embodiment 1, wherein Q 1 It is a C1-alkylene group.

[0281] 33. The compound of embodiment 1, wherein R 2 is hydrogen or alkyl.

[0282] 34. The compound of embodiment 1, wherein R 13 is alkyl, alkenyl, hydrogen or halogen.

[0283] 35. The compound of embodiment 1, wherein R 13 For hydrogen.

[0284] 36. The compound of embodiment 1, wherein R 3 and R 4 Each is independently alkyl, alkenyl, aryl, heteroaryl, heterocyclyl or hydrogen.

[0285] 37. The compound of embodiment 1, wherein R 3 and R 4 With R 3 and R 4 The bound nitrogen atoms together form a ring, wherein the ring is substituted or unsubstituted.

[0286] 38. The compound of embodiment 1, wherein R 3 and R 4 With R 3 and R 4The bound nitrogen atoms together form a heterocyclic ring, wherein the heterocyclic ring is substituted with at least one substituent.

[0287] 39. The compound of embodiment 1, wherein R 3 and R 4 With R 3 and R 4 The bound nitrogen atoms together form a heterocyclic ring, wherein the heterocyclic ring is optionally substituted with a substituted or unsubstituted heterocyclic ring.

[0288] 40. A method of inducing apoptosis in a cell, the method comprising contacting the cell with a therapeutically effective amount of a compound that binds to a p53 mutant, wherein the compound increases the ability of the p53 mutant to bind DNA, wherein the cell expresses the p53 mutant, wherein the compound is a compound of the formula:

[0289]

[0290] in:

[0291] - Each are independently a single bond or a double bond;

[0292] -X 1 CR 5 , CR 5 R 6 、N、NR 5 , O, S, C=O, C=S or with Q 1 the attached carbon atom;

[0293] -X 2 CR 7 , CR 7 R 8 、N、NR 7 , O, S, C=O, C=S or with Q 1 the attached carbon atom;

[0294] -X 3 CR 9 , CR 9 R 10 、N、NR 9 , O, S, C=O, C=S or with Q 1 the attached carbon atom;

[0295] -X 4 CR 11 , CR 11 R 12 、N、NR 11 , O, S, C=O, C=S or with Q 1 the attached carbon atom;

[0296] -X 5 is CR 13 , N or NR 13 ;

[0297] wherein at least one of X 1 , X 2 , X 3 and X 4 is a carbon atom attached to Q 1 ;

[0298] -Q 1 is C=O, C=S, C=CR 14 R 15 , C=NR 14 , alkylene, alkenylene or alkynylene, each of which is independently substituted or unsubstituted, or a bond;

[0299] -m is 1, 2, 3 or 4;

[0300] -Y is N, O or absent;

[0301] -R 1 is -C(O)R 16 , -C(O)OR 16 , -C(O)NR 16 R 17 , -OR 16 , -SR 16 , -

[0302] NR 16 R 17 , -NR 16 C(O)R 16 , -OC(O)R 16 , C=O, C=S, -CN, -

[0303] SiR 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl or heterocyclyl,

[0304] each of which is independently substituted or unsubstituted, or hydrogen;

[0305] each R 3 and R 4 is independently -C(O)R 19 , -C(O)OR 19 , -

[0306] C(O)NR 19 R 20 , -SOR 19 , -SO2R 19, alkyl, alkylene, alkenyl, alkenylene, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen, or R 3 and R 4 together with the nitrogen atom to which they are bound form a ring, wherein the ring is substituted or unsubstituted, or 3 and R 4 together with the nitrogen atom to which they are bound form a ring, wherein the ring is substituted or unsubstituted, or

[0307] R 3 is absent;

[0308] - each R 2 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , and R 18 are independently

[0309] C(O)R 21 , -C(O)OR 21 , -C(O)NR 21 R 22 , -OR 21 , -SR 21 , -

[0310] NR 21 R 22 , -NR 21 C(O)R 22 , -OC(O)R 21 , alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen or halogen;

[0311] - each R 19 and R 20 is -C(O)R 23 , -C(O)OR 23 , -C(O)NR 23 R 24 , -

[0312] OR 23 , -SR 23 , -NR 23 R 24 , -NR 23 C(O)R 24 , -OC(O)R23 alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen, or halogen;

[0313] each R 21 and R 22 are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen;

[0314] and

[0315] each R 23 and R 24 are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or hydrogen.

[0316] 41. The method of embodiment 40, wherein the p53 mutant has a mutation at amino acid 220.

[0317] 42. The method of embodiment 41, wherein the p53 mutant is p53Y220C.

[0318] 43. The method of embodiment 40, wherein the compound induces a conformational change in the p53 mutant.

[0319] 44. The method of embodiment 40, wherein the compound selectively binds the p53 mutant as compared to wild-type p53.

[0320] 45. The method of embodiment 40, wherein the therapeutically effective amount is about 20 mg to about 2000 mg.

[0321] 46. A method of treating a condition, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound that binds to a p53 mutant in the subject, wherein the binding of the compound to the p53 mutant increases the ability of the p53 mutant to bind DNA by at least about 50% as compared to the ability of the p53 mutant to bind DNA in the absence of the compound, as determined by an assay, wherein the assay comprises:

[0322] a) contacting the compound with a labeled mutant p53 moiety and an antibody conjugated to a fluorescent energy acceptor in a test chamber, the antibody being directed against a tag of the labeled mutant p53 moiety;

[0323] b) contacting the labeled mutant p53 moiety with biotin-labeled DNA and streptavidin conjugated to a fluorescent energy donor in the test chamber;

[0324] c) illuminating the test chamber with light that promotes fluorescence resonance energy transfer;

[0325] d) detecting said fluorescence resonance energy transfer;

[0326] e) determining the SC 150 value of said compound based on said fluorescence resonance energy transfer;

[0327] f) comparing the SC 150 value of said compound to the SC 150 value of a control sample, wherein the control sample comprises said labeled mutant p53 moiety, said antibody against said labeled mutant p53 moiety conjugated to a fluorescent energy acceptor, said biotin-labeled DNA, and said streptavidin conjugated to a fluorescent energy donor, wherein the control sample does not comprise said compound; and

[0328] g) determining the level of activation of protein-DNA binding in the presence of said compound based on said comparison.

[0329] 47. The method of embodiment 46, wherein the p53 mutant has a mutation at amino acid 220.

[0330] 48. The method of embodiment 47, wherein the p53 mutant is p53Y220C.

[0331] 49. The method of embodiment 46, wherein the compound induces a conformational change in the p53 mutant.

[0332] 50. The method of embodiment 46, wherein the compound selectively binds to the p53 mutant compared to wild-type p53.

[0333] 51. The method of embodiment 46, wherein binding of the compound to the p53 mutant induces apoptosis in a cell.

[0334] 52. The method of embodiment 46, wherein binding of the compound to the p53 mutant induces cell cycle arrest in a cell.

[0335] 53. The method of embodiment 46, wherein the therapeutically effective amount is about 20 mg to about 2000 mg.

[0336] 54. The method of embodiment 46, wherein the condition is cancer.

[0337] 55. The method of embodiment 54, wherein the condition is ovarian cancer.

[0338] 56. The method of embodiment 54, wherein the condition is breast cancer.

[0339] 57. The method of embodiment 54, wherein the condition is lung cancer.

[0340] 58. The method of embodiment 46, wherein the administration is oral.

[0341] 59. The method of embodiment 46, wherein the administration is intravenous administration.

[0342] 60. The method of embodiment 46, wherein the administration is subcutaneous administration.

[0343] 61. The method of embodiment 46, wherein the administration is topical administration.

[0344] 62. The method of embodiment 46, wherein the subject is a human.

[0345] 63. The method of embodiment 46, wherein the compound enhances the stability of the biologically active conformation of the p53 mutant relative to the stability of the biologically active conformation of the p53 mutant in the absence of the compound.

[0346] 64. The method of embodiment 46, wherein the labeled p53 is His-labeled.

[0347] 65. The method of embodiment 64, wherein the labeled p53 comprises a Y220C mutation.

[0348] 66. The method of embodiment 65, wherein the labeled p53 comprises amino acids 94-312 of wild-type p53.

[0349] 67. The method of embodiment 46, wherein the antibody is conjugated to allophycocyanin (APC).

[0350] 68. The method of embodiment 46, wherein the DNA is a DNA duplex of SEQ ID NO.: 2.

[0351] 69. The method of embodiment 46, wherein the FRET assay is a homogeneous time-resolved fluorescence (HRTF) assay.

[0352] Example

[0353] Example 1 Preparation of 1-anilino-3-{1-ethyl-5-[(1-methyl-4-piperidinylamino)methyl]-1H- indol-2-yl}-2-propyne

[0354]

[0355] Preparation of 5-bromo-1 -(phenylsulfonyl)-1 H-indole

[0356] To a solution of 5-bromo-indole (60 g, 309 mmol) in tetrahydrofuran (600 mL) at 0°C was added NaH (18.5 g, 464 mmol, 60% in mineral oil). The mixture was stirred at 0°C for 1 h. Then PhS02CI (65 g, 370 mmol) in tetrahydrofuran (600 mL) was added dropwise at 0°C. The reaction mixture was stirred at 0°C for 1 h and allowed to warm to 25°C for 17 h. The residue was poured into a mixture of ice and saturated solution of ammonium chloride (2000 mL, w / w = 1 / 1) and stirred for 20 min. The aqueous phase was extracted with ethyl acetate (3 x 800 mL). The combined organic phases were washed with brine (3 x 800 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (silica gel; column height: 500 mm, diameter: 100 mm, 100-200 mesh silica gel, eluting with a gradient of petroleum ether / ethyl acetate 20 / 1 to 10 / 1) to give 5-bromo-1-(phenylsulfonyl)-1H-indole (74 g, 72% yield) as a yellow solid.

[0357] Preparation of 5-bromo-2-iodo-1 -(phenylsulfonyl)-1 H-indole

[0358] To a -70°C stirred solution of 5-bromo-1-(phenylsulfonyl)-1H-indole (28 g, 83.28 mmol) in anhydrous tetrahydrofuran (500 mL) was added dropwise a solution of LDA (2 M, 62.46 mL). After stirring the mixture at 0°C for 2 h, the progress of anion formation was checked by adding D20 to an aliquot. The solution was then cooled to -70°C and a solution of 12 (23.25 g, 91.61 mmol) in tetrahydrofuran (500 mL) was added. The reaction mixture was stirred at 0°C for 0.5 h and allowed to warm to 17°C for 15 h. The residue was poured into a saturated solution of ammonium chloride (1000 mL) and stirred for 3 min. The aqueous phase was extracted with ethyl acetate (3 x 500 mL). The combined organic phases were washed with brine (2 x 500 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography (silica gel, eluting with a gradient of petroleum ether / ethyl acetate 10 / 1 to 5 / 1) to give 5-bromo-2-iodo-1-(phenylsulfonyl)-1H-indole (20.00 g, 70% yield) as a yellow solid.

[0359] Preparation of 5-bromo-2-iodo-1 H-indole

[0360] A mixture of 5-bromo-2-iodo-l-(phenylsulfonyl)-lH-indole (30.00 g, 61.67 mmol) and potassium carbonate (2 M, 100 mL) in methanol (300 mL) was stirred at 90 °C for 2 h. After confirming the completion of the reaction by thin layer chromatography (petroleum ether / ethyl acetate 5 / 1), 80% of the solvent was removed and the reaction mixture was filtered to give crude 5-bromo-2-iodo-lH-indole (13 g, yellow solid). This crude product was used in the next reaction without further purification.

[0361] Preparation of 5-bromo-1 -ethyl-2-iodo-1 H-indole

[0362] To a mixture of 5-bromo-2-iodo-lH-indole (14.62 g, 36.34 mmol) and iodoethane (8.50 g, 54.51 mmol) in tetrahydrofuran (200 mL) was added NaH (2.91 g, 72.68 mmol, 60% in mineral oil) in one portion at 0 °C under nitrogen. The mixture was stirred at 0 °C for 30 min, then warmed to 25 °C and stirred for 11.5 h. Thin layer chromatography (petroleum ether / ethyl acetate, 5 / 1) showed the reaction was complete. The reaction mixture was poured into aqueous ammonium chloride solution (500 mL) and stirred for 5 min. The aqueous phase was extracted with ethyl acetate (2 x 200 mL). The combined organic phase was washed with brine (2 x 200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give crude 5-bromo-l-ethyl-2-iodo-lH-indole (10 g, yellow solid). This crude product was used directly without further purification.

[0363] 1 H NMR (400 MHz MeOD): δ 7.60-7.65 (dd, J = 1.92 Hz, 1H), 7.17-7.59 (m, 2H), 6.69 (s, 1H), 4.17-4.26 (m, 2H), 1.26-1.29 (t, 3H)

[0364] Preparation of [3-(5-bromo-1 -ethyl-1 H-indol-2-yl)-2-propynyl]aniline

[0365] A flask was charged with copper (I) iodide (544.2 mg, 2.86 mmol) and triethylamine (3.62 g, 35.73 mmol) and, under nitrogen, the crude 5-bromo-l-ethyl-2-iodo-lH-indole (5.00 g, 14.30 mmol) was added in portions. The mixture was stirred at 90 °C for 2 h. After confirming the completion of the reaction by thin layer chromatography (petroleum ether / ethyl acetate 5 / 1), the reaction mixture was filtered and the filtrate was concentrated in vacuo to give crude 5-bromo-l-ethyl-2-iodo-lH-indole (5.00 g, yellow solid). This crude product was used in the next reaction without further purification.

[0366] 14.29 mmol) and N-2-propynylaniline (2.25 g, 17.15 mmol) in tetrahydrofuran (50 mL) followed by tetrakis(triphenylphosphine)palladium(0) (825.4 mg, 714.50 pmol). The reaction mixture was stirred at 25 °C for 1 h. Thin layer chromatography (petroleum ether / ethyl acetate 5 / 1) showed the reaction to be complete. The reaction was diluted with 50 mL of ethyl acetate and 100 mL of 2 M EDTA and the biphasic mixture was stirred at 25 °C for 3 h. The reaction mixture was extracted with ethyl acetate (3 x 40 mL) and the combined organic extracts were washed with 150 mL of saturated brine, dried over sodium sulfate and concentrated in vacuo. The crude residue was purified by flash column chromatography (silica gel, column height: 20 mm, diameter: 10 mm, 100-200 mesh silica gel, eluted with a gradient of petroleum ether / ethyl acetate 30 / 1 to 20 / 1) to give [3-(5-bromo-1-ethyl-1H-indol-2-yl)-2- propynyl]phenylamine as a yellow solid (3.10 g, 49.13% yield).

[0367] 1 H NMR (400 MHz, MeOD) d 7.68 (s, 1H), 7.24-7.30 (m, 4H), 7.11-7.13 (d, J = 4.0 Hz, 1H), 6.78-6.83 (m, 3H), 6.62 (s, 1H), 4.27 (s, 1H), 4.08-4.13 (m, 2H), 4.02 (s, 1H), 1.21-1.24 (t, 3H).

[0368] Preparation of 2-(3-anilino-1 -propynyl)-1 -ethyl-1 H-indole-5-carbaldehyde

[0369] To a solution of [3-(5-bromo-1-ethyl-1H-indol-2-yl)-2-propynyl]phenylamine (500 mg, 1.42 mmol) in tetrahydrofuran (10.00 mL) cooled to -78 °C under nitrogen was added n-butyllithium (2.5 M, 3.41 mL) in one portion. The mixture was stirred at -78 °C for 30 min, then 4-morpholinecarboxaldehyde (1.63 g, 14.20 mmol) was added in one portion at -78 °C. The mixture was stirred for 1.5 h, thin layer chromatography (petroleum ether / ethyl acetate, 5 / 1) showed the reaction to be complete. The residue was poured into an aqueous ammonium chloride solution ice-water (20 mL, w / w = 1 / 1) and stirred for 3 min. The aqueous phase was extracted with ethyl acetate (2 x 10 mL). The combined organic phases were washed with brine (2 x 10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give crude 2-(3-phenylamino-1- propynyl)-1-ethyl-1H-indole-5-carbaldehyde (600 mg) as a yellow oil. This product was used directly in the next step without further purification.

[0370] Preparation of 1 -anilino-3-{1 -ethyl-5-[(1 -methyl-4-piperidinylamino)methyl]-1 H- indol-2-yl}-2-propynyl Example 2

[0371] To a mixture of 2-(3-anilino-1-propynyl)-1-ethyl-1H-indole-5-carbaldehyde (200 mg, 529.15 pmol, 1 eq) and 1-methylpiperidin-4-amine (53.5 mg, 529.15 pmol, 1 eq) in dichloromethane (10 mL) was added anhydrous magnesium sulfate (318.5 mg, 2.65 mmol, 5 eq) in one portion at 25 °C under nitrogen. The mixture was stirred at 25 °C for 60 min, then NaBH(OAc)3(336.5 mg, 1.59 mmol, 3 eq) was added and the reaction mixture was stirred for 5 h. LCMS showed the reaction was complete. The residue was poured into ice-water (10 mL, w / w = 1 / 1) and stirred for 3 min. The aqueous phase was extracted with dichloromethane (3 x 5 mL). The combined organic phase was washed with brine (3 x 5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude residue was purified by preparative HPLC (Gilson 281 semi-preparative HPLC system; column: Waters Xbridge 150 x 25, 5u; flow rate: 20 mL / min; eluted with a gradient of acetonitrile in water containing 0.04% HC1) to give 1-anilino-3-{1-ethyl-5-[(1-methyl-4-piperidinylamino)methyl]-1H-indol-2-yl}-2-propyne.

[0372] LC-MS (ES + ,m / z): 401.3 [(M+H) + ]

[0373] Example 3 Preparation of 1-anilino-3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}-2-propyne

[0374]

[0375] In a similar manner to that described in Example 1, 2-(3-anilino-1-propynyl)-1- ethyl-1H-indole-5-carbaldehyde was reacted with methylamine to give 1-anilino-3-{1- ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}-2-propyne.

[0376] LC-MS (ES + ,m / z): 287.2 [(M-NHMe) + ]

[0377] Example 4Preparation of 1 -anilino-3 -{ 1 -ethyl-5 - [(tetrahydro-2H-pyran-4-ylamino)methyl] - 1 H-indol-2-yl}-2-propyne

[0378]

[0379] In a similar manner to that described in Example 1, 2-(3-anilino-1 -propynyl)-1 - ethyl-1 H-indole-5-carboxaldehyde was reacted with 4-aminotetrahydropyran to give 1 - anilino-3-{1 -ethyl-5-[(tetrahydro-2H-pyran-4-ylamino)methyl]-1 H-indol-2-yl}-2- propyne. LC-MS (ES + ,m / z): 388.3 [(M+H) + ]

[0380] Example 5 Preparation of 1 -anilino-3 -{ 1 -ethyl-5 - [(tetrahydro-2H-pyran-4-ylamino)methyl] - 1 H-indol-2-yl}-2-propyne

[0381]

[0382] In a similar manner to that described in Example 1, 2-(3-anilino-1 -propynyl)-1 - ethyl-1 H-indole-5-carboxaldehyde was reacted with benzylamine to give 1 -anilino-3-[5- (benzylaminomethyl)-1 -ethyl-1 H-indol-2-yl]-2-propyne.

[0383] LC-MS (ES + ,m / z): 394.3 [(M+H) + ]

[0384] Preparation of [3-(5-bromo-1 -ethyl-1 H-indol-2-yl)-2-propynyl](p-fluorophenyl)amine Preparation of 3-{1 -ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}-1 -(p- fluorophenylamino)-2-propyne

[0385]

[0386] Preparation of 1 -ethyl-2-[3-(4-fluoroanilino)prop-1 -ynyl]indole-5-carbaldehyde

[0387] To a solution of 5-bromo-l-ethyl-2-iodo-indole (800 mg, 2.29 mmol, 1 eq) in tetrahydrofuran (5 mL) at 25 °C was added 4-fluoro-N-prop-2-ynyl- benzenamine (341 mg, 2.29 mmol, 1 eq), triethylamine (694 mg, 6.86 mmol, 951 μί, 3 eq), copper (I) iodide (44 mg, 228.58 μmol, 0.1 eq) and tetrakis(triphenylphosphine)palladium(0) (264 mg, 228.58 μmol, 0.1 eq). The mixture was stirred for 2 h, then poured into EDTA solution (2 M, 20 mL) and stirred for a further 2 h. The mixture was extracted with ethyl acetate (2 x 20 mL) and the combined organic phases were washed with water (20 mL) and brine (20 mL), dried over anhydrous sodium sulfate. The solids were filtered off and the solvent was removed under vacuum. The crude residue was purified by flash column chromatography (silica gel, eluting with petroleum ether / ethyl acetate 10 / 1) to give N-[3-(5-bromo-l-ethyl-indol-2-yl)prop-2-ynyl]-4-fluoro- benzenamine (750 mg, 88.2% yield) as a yellow solid.

[0388] Example 6

[0389] To a solution of N-[3-(5-bromo-l-ethyl-indol-2-yl)prop-2-ynyl]-4-fluoro- benzenamine (700 mg, 1.89 mmol, 1 eq) in tetrahydrofuran (5.00 mL) cooled to -78 °C was added n-butyllithium (2.5 M, 3.02 mL, 4 eq). After stirring the mixture at -78 °C for 0.5 h, morpholine-4-carboxaldehyde (1.09 g, 9.43 mmol, 944 μί, 5 eq) was added. After 2 h from the addition, thin layer chromatography showed the reaction to be complete. The reaction mixture was poured into aqueous ammonium chloride solution (5 mL) and extracted with ethyl acetate (5 mL). The combined organic phases were washed with water (5 mL) and brine (5 mL), dried over anhydrous sodium sulfate. The solids were filtered off and the filtrate was concentrated under vacuum. The crude residue was purified by flash column chromatography (silica gel, eluting with petroleum ether / ethyl acetate 1 / 1) to give l-ethyl-2-[3-(4-fluoroanilino)prop-l-ynyl]indole-5- carboxaldehyde (300 mg, 49.6% yield) as a yellow solid.

[0390] In a similar manner to that described in Example 1, l-ethyl-2-[3-(4-fluoroanilino)prop- 1-ynyl]indole-5-carboxaldehyde was reacted with methylamine to give 3-{l-ethyl-5-[(methylamino)methyl]-lH-indol-2-yl}-l-(p-fluorophenylamino)-2- propynyl.

[0391] LC-MS (ES +m / z): 305.2 [(M - NHMe) + ]

[0392] Example 7 Preparation of 3-{1 -ethyl-5-[(tetrahydro-2H-pyran-4-ylamino)methyl]-1 H- indol-2-yl}-1 -(p-fluorophenylamino)-2-propyne

[0393]

[0394] In a similar manner to that described in Example 1, 1 -ethyl-2-[3-(4- chloroanilino)prop-1 -ynyl]indole-5-carboxaldehyde was reacted with 1 -methylpiperidin-4- amine to give 1 -(4-chlorophenylamino)-3-{1 -ethyl-5-[(1 -methyl-4-piperidinylamino)methyl]- 1 H-indol-2-yl}-2-propyne.

[0395] LC-MS (ES + m / z): 406.3 [(M + H) + ]

[0396] Example 8 Preparation of 3-{1 -ethyl-5-[(tetrahydro-2H-pyran-4-ylamino)methyl]-1 H- indol-2-yl}-1 -(p-fluorophenylamino)-2-propyne

[0397]

[0398] In a similar manner to that described in Example 6 for the preparation of 1 -ethyl-2-[3-(4- chloroanilino)prop-1 -ynyl]indole-5-carboxaldehyde, 1 -ethyl-2-[3-(4-fluoroanilino)prop-1 - ynyl]indole-5-carboxaldehyde was prepared.

[0399] In a similar manner to that described in Example 1, 1 -ethyl-2-[3-(4- chloroanilino)prop-1 -ynyl]indole-5-carboxaldehyde was reacted with 1 -methylpiperidin-4- amine to give 1 -(4-chlorophenylamino)-3-{1 -ethyl-5-[(1 -methyl-4-piperidinylamino)methyl]- 1 H-indol-2-yl}-2-propyne.

[0400] LC-MS (ES + m / z): 435.3 [(M + H) + ]

[0401] Example 9 Preparation of 3-{1 -ethyl-5-[(tetrahydro-2H-pyran-4-ylamino)methyl]-1 H- indol-2-yl}-1 -(p-fluorophenylamino)-2-propyne

[0402]

[0403] 1 -Ethyl-2-{3-[(6-methylpyridin-3-yl)amino]prop-1 -yn-1 -yl}-1 H-indole-5- carboxaldehyde was prepared in a similar manner to that described in Example 6 for the preparation of 1 -ethyl-2-[3-(4-fluoroanilino)prop-1 -ynyl]indole-5- carboxaldehyde.

[0404] Reaction of 1 -ethyl-2-{3-[(6-methylpyridin-3-yl)amino]prop-1 -yn-1 -yl}-1 H-indole-5- carboxaldehyde with 4-amino tetrahydropyran in a similar manner to that described in Example 1 gave 3-{1 -ethyl-5-[(tetrahydro-2H-pyran-4-ylamino)methyl]-1 H-indol-2-yl}-1 -(6-methyl-3-pyridinylamino)-2-propyne.

[0405] LC-MS (ES + , m / z): 403.3 [(M+H) + ]

[0406] Example 10 Preparation of 3-{1 -ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}-1 -(6-methyl-3- pyridinylamino)-2-propyne

[0407]

[0408] 1 -Ethyl-2-{3-[(6-methylpyridin-3-yl)amino]prop-1 -yn-1 -yl}-1 H-indole-5- carboxaldehyde was prepared in a similar manner to that described in Example 6 for the preparation of 1 -ethyl-2-[3-(4-fluoroanilino)prop-1 -ynyl]indole-5- carboxaldehyde.

[0409] Reaction of 1 -ethyl-2-{3-[(6-methylpyridin-3-yl)amino]prop-1 -yn-1 -yl}-1 H-indole-5- carboxaldehyde with methylamine in a similar manner to that described in Example 1 gave 3-{1 -ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}-1 -(6-methyl-3-pyridinylamino)-2- propyne.

[0410] LC-MS (ES + , m / z): 302.2 [(M-NHMe) + ]

[0411] Example 11Preparation of 3-{1 -ethyl-5-[(1 -methyl-4-piperidinylamino)methyl]-1 H- indol-2-yl}-1 -(2-methyl-4-pyridinylamino)-2-propyne

[0412]

[0413] 1 -Ethyl-2-{3-[(2-methylpyridin-4-yl)amino]prop-1 -yn-1 -yl}-1 H-indole-5- carboxaldehyde was prepared in a similar manner to that described in Example 6 for the preparation of 1 -ethyl-2-[3-(4-fluoroanilino)prop-1 -ynyl]indole-5- carboxaldehyde.

[0414] 1 -Ethyl-2-{3-[(2-methylpyridin-4-yl)amino]prop-1 -yn-1 -yl}-1 H-indole-5- carboxaldehyde was reacted with 1 -methylpiperidin-4-amine in a similar manner to that described in Example 1 to give 3-{1 -ethyl-5-[(1 -methyl-4- piperidinylamino)methyl]-1 H-indol-2-yl}-1 -(2-methyl-4-pyridinylamino)-2- propyne.

[0415] LC-MS (ES + ,m / z): 416.3 [(M+H) + ]

[0416] Example 12 Preparation of 3-{1 -ethyl-5-[(1 -methyl-4-piperidinylamino)methyl]-1 H- indol-2-yl}-1 -(2-methyl-4-pyridinylamino)-2-propyne

[0417]

[0418] 1 -Ethyl-2-{3-[(2-methylpyridin-4-yl)amino]prop-1 -yn-1 -yl}-1 H-indole-5- carboxaldehyde was prepared in a similar manner to that described in Example 6 for the preparation of 1 -ethyl-2-[3-(4-fluoroanilino)prop-1 -ynyl]indole-5- carboxaldehyde.

[0419] 1 -Ethyl-2-{3-[(2-methylpyridin-4-yl)amino]prop-1 -yn-1 -yl}-1 H-indole-5- carboxaldehyde was reacted with benzylamine in a similar manner to that described in Example 1 to give 3-[5-(benzylaminomethyl)-1 -ethyl-1 H-indol-2-yl]-1 -(2-methyl-4- pyridinylamino)-2-propyne.

[0420] LC-MS (ES + ,m / z): 409.1 [(M+H) + ]

[0421] Example 13 Preparation of N-(3-{5-[(diethylamino)methyl]-l-ethyl-lH-indol-2-yl}prop-2-yn-l- yl)aniline

[0422]

[0423] In a similar manner to that described in Example 1, 2-(3-anilino-l-propynyl)-l- ethyl-lH-indole-5-carboxaldehyde was reacted with diethylamine to give N-(3-{5- [(diethylamino)methyl]-l-ethyl-lH-indol-2-yl}prop-2-yn-l-yl)aniline.

[0424] LC-MS (ES + , m / z): 360.3 [(M+H) + ]

[0425] Example 14 Preparation of 4-chloro-N-(3-{5-[(diethylamino)methyl]-l-ethyl-lH-indol-2-yl}prop-2- yn-l-yl)aniline

[0426]

[0427] In a similar manner to that described in Example 1, l-ethyl-2-[3-(4-chloroanilino)prop- 1-ynyl]indole-5-carboxaldehyde was reacted with diethylamine to give 4-chloro-N-(3-{5- [(diethylamino)methyl]-l-ethyl-lH-indol-2-yl}prop-2-yn-l-yl)aniline.

[0428] LC-MS (ES + , m / z): 394.3 [(M+H) + ]

[0429] Example 15 Preparation of N-({l-ethyl-2-[3-(phenylamino)prop-l-y n-l-yl]-lH-indol-5-yl}methyl)oxetan- 3-amine

[0430]

[0431] In a similar manner to that described in Example 1, 2-(3-anilino-l-propynyl)-l- ethyl-lH-indole-5-carboxaldehyde was reacted with diethylamine to give N-(3-{5- [(diethylamino)methyl]-l-ethyl-lH-indol-2-yl}prop-2-yn-l-yl)aniline. + , m / z): 360.2 [(M+H)+ ]

[0432] Example 16 Preparation of N-[3-(1-ethyl-5-{[(2-methylpropyl)amino]methyl}-1H-indol-2- yl)prop-2-yn-1-yl]aniline

[0433]

[0434] In a similar manner to that described in Example 1, 2-(3-anilino-1-propynyl)-1- ethyl-1H-indole-5-carboxaldehyde was reacted with 2-methoxyethylamine to give N- [3-(1-ethyl-5-{[(2-methoxyethyl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1- yl]aniline. LC-MS (ES+) m / z): 362.2 [(M+H)+]. + + ]

[0435] Example 17 Preparation of N-[3-(1-ethyl-5-{[(2-methoxyethyl)amino]methyl}-1H-indol-2- yl)prop-2-yn-1-yl]aniline

[0436]

[0437] In a similar manner to that described in Example 1, 2-(3-anilino-1-propynyl)-1- ethyl-1H-indole-5-carboxaldehyde was reacted with 2-methoxyethylamine to give N- [3-(1-ethyl-5-{[(2-methoxyethyl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1- yl]aniline. LC-MS (ES+) m / z): 362.2 [(M+H)+]. + + ]

[0438] Example 18 Preparation of N-({1-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H-indol-5- yl}methyl)-1-methanesulfonylpiperidin-4-amine

[0439]

[0440] In a similar manner to that described in Example 1, 2-(3-anilino-1-propynyl)-1- ethyl-1H-indole-5-carboxaldehyde was reacted with 2-methoxyethylamine to give N- [3-(1-ethyl-5-{[(2-methoxyethyl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1- yl]aniline. LC-MS (ES+) m / z): 362.2 [(M+H)+].

[0441] ​​LC-MS (ES + m / z): 465.2 [(M+H) + ]

[0442] Example 19 Preparation of N-(3-{1-ethyl-5-[(ethylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1- yl)aniline

[0443]

[0444] N-(3-{1-ethyl-5-[(ethylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1-yl)aniline was obtained in a similar manner to that described in Example 1 by reacting 2-(3-anilino-1- propynyl)-1-ethyl-1 H-indole-5-carboxaldehyde with ethylamine.

[0445] LC-MS (ES + m / z): 332.2 [(M+H) + ]

[0446] Example 20 Preparation of N-{3-[5-({[2-(dimethylamino)ethyl]amino}methyl)-1-ethyl-1 H-indol-2- yl]prop-2-yn-1-yl}aniline

[0447]

[0448] N-{3-[5-({[2-(dimethylamino)ethyl]amino}methyl)-1-ethyl-1 H-indol-2-yl]prop-2-yn-1- yl}aniline was obtained in a similar manner to that described in Example 1 by reacting 2- (3-anilino-1-propynyl)-1-ethyl-1 H-indole-5-carboxaldehyde with 2-dimethylaminoethylamine.

[0449] LC-MS (ES + m / z): 375.3 [(M+H) + ]

[0450] Example 21 Preparation of 6-tert-butyl-N-[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1 H- indol-2-yl)prop-2-yn-1-yl]pyridin-3-amine

[0451]

[0452] 2-{3-[(6-tert-Butylpyridin-3-yl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indole-5- carboxaldehyde was prepared in a similar manner as described in Example 6 for the preparation of 1-ethyl-2-[3-(4-fluoroanilino)prop-1-ynyl]indole-5-carboxaldehyde.

[0453] Reaction of 2-{3-[(6-tert-butylpyridin-3-yl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indole-5- carboxaldehyde with 1-methylpiperidin-4-amine in a similar manner as described in Example 1 gave 6-tert-butyl-N-[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2- yl)prop-2-yn-1-yl]pyridin-3-amine.

[0454] LC-MS (ES + , m / z): 458.6 [(M+H) + ]

[0455] Example 22 Preparation of N-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5- yl)methyl]oxanyl-4-amine

[0456]

[0457] Reaction of 1-ethyl-2-[3-(4-chloroanilino)prop-1-ynyl]indole-5-carboxaldehyde with 4- aminotetrahydropyran in a similar manner as described in Example 1 gave N-[(2-{3-[(4- chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5-yl)methyl]oxanyl-4-amine.

[0458] LC-MS (ES + , m / z): 422.2 [(M+H) + ]

[0459] Example 23 Preparation of 6-tert-butyl-N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2- yn-1-yl)pyridin-3-amine

[0460] 2-{3-[(6-tert-Butylpyridin-3-yl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indole-5- carboxaldehyde was prepared in a similar manner as described in Example 6 for the preparation of 1-ethyl-2-[3-(4-fluoroanilino)prop-1-ynyl]indole-5-carboxaldehyde.

[0461] In a similar manner to that described in Example 1, 2-{3-[(6-tert-butylpyridin-3- yl)amino]prop-1 -yn-1 -yl}-1 -ethyl-1 H-indole-5-carbaldehyde was reacted with methylamine to give 6-tert-butyl-N-(3-{1 -ethyl-5-[(methylamino)methyl]-1 H-indol-2- yl}prop-2-yn-1 -yl)pyridin-3-amine.

[0462] LC-MS (ES + , m / z): 375.3 [(M+H) + ]

[0463] Example 24 Preparation of 4-[(3-{1 -ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1 - yl)amino]benzonitrile

[0464]

[0465] To a solution of N-[(1 -ethyl-2-iodo-1 H-indol-5-yl)methyl]amino 2,2-dimethylpropanamide methyl ester (1 equiv) in dimethyl sulfoxide (5.0 mL) at 25 °C was added p-(2- propynylamino)benzonitrile (1 equiv), copper (I) iodide (0.47 equiv), diisopropylamine (3 equiv) and tetrakis(triphenylphosphine)palladium (0) (0.1 equiv) and the mixture was stirred under a nitrogen atmosphere for 2 h. After LCMS showed the reaction was complete, the reaction mixture was poured into a solution of EDTA (20 mL) and stirred for 2 h. The mixture was extracted with ethyl acetate (20 mL) and the organic phase washed with water (50 mL) and brine (50 mL) and dried over anhydrous sodium sulphate. The solids were filtered off and the filtrate was concentrated in vacuo to give the crude product as a dark brown oil. The crude product was purified by flash column chromatography (silica gel, eluting with 2 / 1 petroleum ether / ethyl acetate) to give N-({2-[3-(p-cyanophenylamino)-1 -propynyl]-1 -ethyl-1 H-indol-5- yl}methyl)amino 2,2-dimethylpropanamide methyl ester as a yellow oil.

[0466] To a solution of N-({2-[3-(p-cyanophenylamino)-l-propynyl]-l-ethyl-lH- indol-5-yl}methyl)amino 2,2-dimethylpropionic acid methyl ester (1 equiv) in acetonitrile (2 mL) at 50 °C was added BiCl3(4 equiv) and the mixture was stirred under a nitrogen atmosphere for 1 h. LCMS showed the reaction to be complete. The reaction mixture was poured into a solution of EDTA (20 mL) and stirred for 2 h. The mixture was extracted with ethyl acetate (20 mL) and the organic phase was washed with water (50 mL) and brine (50 mL) and dried over anhydrous sodium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo to give the crude product as a yellow oil. The crude product was purified by flash column chromatography (silica gel, eluting with 10 / 1 dichloromethane / methanol) to give p-(3-{l-ethyl-5-[(methylamino)methyl]-lH-indol-2-yl}-2- propynylamino)benzonitrile as a white solid.

[0467] LC-MS (ES + ,m / z): 343.1 [(M+H) + ]

[0468] Example 25 Preparation of 4-tert-butyl-N-(3-{l-ethyl-5-[(methylamino)methyl]-lH-indol-2- yl}prop-2-yn-l-yl)benzamide

[0469]

[0470] 4-tert-Butyl-N-(3-{l-ethyl-5-[(methylamino)methyl]-lH-indol-2-yl}prop-2-yn-l- yl)benzamide was prepared in a similar manner to that described in Example 23 using (2- propynylamino)[p-(tert-butyl)phenyl]methanal.

[0471] LC-MS (ES + ,m / z): 371.1 [(M-NHMe) + ]

[0472] Example 26 Preparation of 4-chloro-N-(3-{l-ethyl-5-[(methylamino)methyl]-lH-indol-2-yl}prop- 2-yn-l-yl)-3-fluorobenzamide

[0473]

[0474] In a similar manner to that described in Example 23, using (4-chloro-3- fluorophenyl)(2-propynylamino)methan al, 4-chloro-N-(3-{1 -ethyl-5-[(methylamino)methyl]- 1 H-indol-2-yl}prop-2-yn-1 -yl)-3-fluorobenzamide was prepared.

[0475] LC-MS (ES + , m / z): 367.1 [(M-NHMe) + ]

[0476] Example 27 Preparation of 4-cyano-N-({1 -ethyl-2-[3-(phenylcarboxamido)prop-1 -yn-1 -yl]-1 H- indol-5-yl}methyl)-N-methylbenzamide

[0477]

[0478] In a similar manner to that described in Example 23, using phenyl(2-propynylamino)methan al, (3-{1 -ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}-2-propynylamino)phenyl- methan al was prepared.

[0479] To a solution of (3-{1 -ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}-2-propynylamino)phenyl- methan al (1 equiv) and triethylamine (3 equiv) in dichloromethane (5 mL) at 25 °C was added 4- cyanobenzoyl chloride (1 equiv) and the mixture was stirred under a nitrogen atmosphere for 5 h. LCMS showed the reaction to be complete. The reaction mixture was poured into a saturated solution of sodium bicarbonate (20 mL) and stirred for 5 min. The mixture was extracted with ethyl acetate (20 mL), the organic phase was washed with water (50 mL) and brine (50 mL) and dried over anhydrous sodium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo to give the crude product as a dark brown oil. The crude product was purified by flash column chromatography (silica gel, eluting with 10 / 1 dichloromethane / methanol) to give 4-cyano-N-({1 -ethyl-2-[3-(phenylcarboxamido)prop-1 -yn-1 -yl]-1 H-indol-5-yl}methyl)- N-methylbenzamide as a white solid.

[0480] LC-MS (ES + , m / z): 475.2 [(M+H) + ]

[0481] Example 28 Preparation of 3-(3-{1 -ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1 -yl)-1 - [4-(trifluoromethyl)phenyl]urea

[0482]

[0483] 3-(3-{1 -Ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1 -yl)-1 -[4- (trifluoromethyl)phenyl]urea was prepared in a similar manner to that described in Example 23 using 3-(2-propynyl)-1 -[p-(trifluoromethyl)phenyl]urea.

[0484] LC-MS (ES + , m / z): 398.1 [(M-NHMe) + ]

[0485] Example 29 Preparation of N-{[1 -(2-chloroethyl)-2-{3-[(4-chlorophenyl)amino]prop-1 -yn-1 -yl}-1 H- indol-5-yl]methyl}oxane-4-amine

[0486]

[0487] To a solution of 1 H-indole-5-carboxaldehyde (85 g, 585.56 mmol, 1 equiv) in dichloromethane (100 mL) was added potassium hydroxide (65.38 g, 1.17 mol, 2 equiv) and n-Bu4NSO4(198.82 g, 585.56 mmol, 1 equiv). The mixture was stirred at 25 °C for 30 min, then PhSO2CI (155.13 g, 878.34 mmol, 1 12.41 mL, 1.5 equiv) was added. The mixture was stirred at 25 °C for 1 h and thin layer chromatography showed the reaction was complete. The reaction mixture was poured into aqueous ammonium chloride solution (200 mL) and extracted with dichloromethane (3 x 200 mL). The combined organic layers were washed with brine (2 x 100 mL) and dried over anhydrous sodium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo. The crude residue was purified by flash column chromatography (silica gel, eluted with petroleum ether / dichloromethane: 15 / 1 ) to give 1 -(phenylsulfonyl)indole-5-carboxaldehyde (124 g, 71 % yield) as a white solid.

[0488] To a solution of 1-(phenylsulfonyl)indole-5-carboxaldehyde (4 g, 14.02 mmol, 1 eq) and tetrahydro-2H-pyran-4-ylamine (2.84 g, 28.04 mmol, 2 eq) in dichloromethane (80 mL) was added NaBH(OAc)3(5.94 g, 28.04 mmol, 2 eq) and anhydrous magnesium sulfate (16.9 g, 140.2 mmol, 10 eq). The reaction mixture was stirred at 25 °C for 12 h, thin layer chromatography showed the reaction was complete. The mixture was poured into aqueous sodium bicarbonate solution (100 mL) and extracted with dichloromethane (3 x 50 mL). The combined organic layers were washed with aqueous sodium bicarbonate solution (3 x 50 mL) and brine (50 mL) and dried over anhydrous sodium sulfate. The solid was filtered off, the filtrate was concentrated in vacuo to give N-[[1-(phenylsulfonyl)indol-5-yl]methyl]-tetrahydropyran-4-amine (4.6 g) as off-white gum. This crude product was used directly without further purification.

[0489] To a solution of N-[[1-(phenylsulfonyl)indol-5-yl]methyl]tetrahydropyran-4-amine (4.6 g, 12.42 mmol, 1 eq) in dioxane (60 mL) was added (Boc)20 (4.61 g, 21.11 mmol, 4.85 mL, 1.70 eq) and the reaction mixture was stirred at 90 °C for 1 h. Thin layer chromatography showed the reaction was complete. The reaction mixture was concentrated in vacuo, the crude was purified by flash column chromatography (silica gel, eluted with petroleum ether / ethyl acetate: 4 / 1) to give N-[[1-(phenylsulfonyl)indol-5-yl]methyl]-N-tetrahydropyran-4-yl-carbamic acid tert-butyl ester (4 g, 68% yield) as a white solid.

[0490] To a solution of N-[[1 -(phenylsulfonyl)indol-5-yl]methyl]-N- tetrahydropyran-4-yl-carbamic acid tert-butyl ester (500 mg, 1.05 mmol, 1 eq) in tetrahydrofuran (6 mL) was added LDA (2 M, 1.31 mL, 2.5 eq) in small portions and the reaction mixture was stirred at -78 °C for 1 h. Iodine (293.2 mg, 1.16 mmol, 1.1 eq) in 1 mL of tetrahydrofuran was added and the reaction mixture was stirred at -78 °C for another 1 h. LCMS showed about 20% conversion. The reaction mixture was poured into aqueous ammonium chloride solution (100 mL) and extracted with dichloromethane (3 x 30 mL). The combined organic layers were washed with water (3 x 30 mL) and brine (10 mL) and dried over anhydrous sodium sulfate. The solid was filtered off and the filtrate was concentrated in vacuo to give N-[[1 -(phenylsulfonyl)-2-iodo-indol-5-yl]methyl]-N- tetrahydropyran-4-yl-carbamic acid tert-butyl ester (2.5 g, 4.19 mmol) as a yellow oil. This crude product was used without further purification.

[0491] To a solution of N-[[1 -(phenylsulfonyl)-2-iodo-indol-5-yl]methyl]-N- tetrahydropyran-4-yl-carbamic acid tert-butyl ester (2.5 g, 4.19 mmol, 1 eq) in methanol (50 mL) was added potassium carbonate (2.9 g, 20.95 mmol, 5 eq) and the reaction mixture was stirred at 80 °C for 5 h. Thin layer chromatography showed the reaction was complete. The reaction mixture was poured into 100 mL of ice water and acidified with 37% HC1 to pH = 4. The mixture was extracted with dichloromethane (3 x 30 mL), the combined organic layers were washed with water (2 x 30 mL) and brine (2 x 30 mL) and dried over anhydrous sodium sulfate. The solid was filtered off and the filtrate was concentrated in vacuo. The crude residue was purified by flash column chromatography (silica gel, eluted with petroleum ether / ethyl acetate: 8 / 1) to give N-[(2-iodo-1 H-indol-5-yl)methyl]-N- tetrahydropyran-4-yl-carbamic acid tert-butyl ester (900 mg, 43% yield) as a white solid.

[0492] To a solution of N-[(2-iodo-lH-indol-5-yl)methyl]-N-tetrahydropyran-4-yl- carbamic acid tert-butyl ester (300 mg, 90% purity, 591.69 pmol, 1 eq) in dichloroethane (58.6 mg, 591.69 pmol, 1 eq) was added KOH (199.2 mg, 3.55 mmol, 6 eq), potassium carbonate (204.4 mg, 1.48 mmol, 2.5 eq) and TBAI (218.6 mg, 591.69 pmol, 1 eq). The reaction mixture was stirred at 25 °C for 6 h. Thin layer chromatography showed the reaction was complete. The reaction mixture was filtered and the filtrate was concentrated in vacuo. The crude residue was purified by preparative thin layer chromatography (silica gel, eluted with petroleum ether / ethyl acetate: 4 / 1) to give N-[[l-(2-chloroethyl)-2-iodo-indol-5-yl]methyl]-N- tetrahydropyran-4-yl-carbamic acid tert-butyl ester (260 mg, 83% yield) as a light yellow oil.

[0493] To a solution of 4-chloro-N-prop-2-ynyl-phenylamine (122 mg, 736.68 pmol, 1.5 eq) in dimethyl sulfoxide (5 mL) was added N-isopropylpropan-2-amine (149.09 mg, 1.47 mmol, 207.07 pL, 3.00 eq) and copper (I) iodide (28.06 mg, 147.34 pmol, 0.3 eq) followed by N-[[l-(2-chloroethyl)-2-iodo-indol-5-yl]methyl]-N- tetrahydropyran-4-yl-carbamic acid tert-butyl ester (260 mg, 491.12 pmol, 1 eq) and tetrakis(triphenylphosphine)palladium(0) (56.8 mg, 49.11 pmol, 0.1 eq). After the reaction mixture was stirred at 25 °C for 1 h, thin layer chromatography showed the reaction was complete. Aqueous EDTA (20 mL) was added and the mixture was stirred for 2 h. The mixture was extracted with dichloromethane (3 x 10 mL). The combined organic layers were washed with water (3 x 10 mL) and brine (10 mL) and dried over anhydrous sodium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo. The crude residue was purified by preparative thin layer chromatography (silica gel, eluted with petroleum ether / ethyl acetate: 3 / 1) to give N-[[2-[3-(4-chloroanilino)prop-l-ynyl]-l-(2-chloroethyl)indol-5-yl]methyl]-N- tetrahydropyran-4-yl-carbamic acid tert-butyl ester (200 mg, 71% yield) as a light yellow solid.

[0494] To a solution of N-[[2-[3-(4-chloroanilino)prop-1-ynyl]-1-(2- chloroethyl)indol-5-yl]methyl]-N-tetrahydropyran-4-yl-carbamic acid tert-butyl ester (100 mg, 161.72 μmol, 1 eq) in acetonitrile (2 mL) was added bismuthine (204 mg, 646.88 μmol, 42.94 μL, 4 eq) and the reaction mixture was stirred at 50 °C for 2 h. LCMS showed the reaction was complete. Aqueous EDTA (20 mL) was added and the mixture was stirred for 2 h. The mixture was extracted with dichloromethane (3 x 10 mL). The combined organic layers were washed with water (3 x 10 mL) and brine (10 mL) and dried over anhydrous sodium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo. The crude residue was purified first by preparative thin layer chromatography (silica gel, eluted with dichloromethane / methanol: 5 / 1) and then by preparative HPLC (C18 silica gel, eluted with acetonitrile and water) to give N-[[2-[3-(4-chloroanilino)prop-1-ynyl]-1-(2-chloroethyl)indol-5-yl]methyl]tetrahydropyran-4-amine (14.9 mg, 19% yield) as a yellow solid.

[0495] LC-MS (ES + ,m / z): 456.2 [(M+H) + ]

[0496] Example 30 Preparation of 2-(4-{[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2- yl)prop-2-yn-1-yl]amino}phenyl)-2-methylpropanenitrile

[0497]

[0498] To a solution of 2-methyl-2-[p-(2-propynylamino)phenyl]propionitrile (2 eq) and [(1- ethyl-2-iodo-1H-indol-5-yl)methyl](1-methyl-4-piperidinyl)amino 2,2-dimethylpropanoate (100 mg, 1 eq) in dimethyl sulfoxide (2 mL) was added N-isopropylpropan-2-amine (3 eq) and copper (I) iodide (0.2 eq) followed by tetrakis(triphenylphosphine)palladium(0) (0.1 eq). After the reaction mixture was stirred at 25 °C for 1 h, the mixture was poured into an aqueous EDTA solution (10 mL) and stirred at 25 °C for 1 h. The reaction mixture was extracted with ethyl acetate (2 x 20 mL) and the combined organic extracts were washed with water (10 mL) and brine (10 mL) and dried over anhydrous sodium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo to give a crude residue. The crude residue was purified by flash column chromatography (silica gel, eluted with 15 / 1 dichloromethane / methanol) to give ({1-ethyl-2-[3-(3-phenylureido)-1- propynyl]-1H-indol-5-yl}methyl)(1-methyl-4-piperidinyl)amino 2,2-dimethylpropanoate.

[0499] To a 50 °C solution of ({1-ethyl-2-[3-(3-phenylureido)-1-propynyl]-1H-indol-5- yl}methyl)(1-methyl-4-piperidinyl)amino 2,2-dimethylpropanoate (60 mg, 108.48 μmol, 1 eq) in acetonitrile (2 mL) was added trichlorobismuthane (4 eq). The reaction mixture was stirred at 50 °C for 30 min and then poured into an aqueous EDTA solution (15 mL). The biphasic mixture was stirred at 25 °C for 2 h and separated. The mixture was extracted with ethyl acetate (2 x 40 mL) and the organic phase was washed with water (10 mL) and brine (10 mL) and dried over anhydrous sodium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo. The crude residue was purified by reverse phase HPLC (C18, eluted with acetonitrile and water) to give 2-(4-{[3-(1-ethyl-5-{[(1-methylpiperidin-4- yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}phenyl)-2-methylpropanenitrile.

[0500] LC-MS (ES + ,m / z): 468.2 [(M+H) + ]

[0501] Example 31 Preparation of 4-cyano-N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1- yl)benzamide

[0502]

[0503] Prepared in a similar manner to that described in Example 23 using p-[(2- propynylamino)carbonyl]benzonitrile to give 4-cyano-N-(3-{1 -ethyl-5-[(methylamino)methyl]- 1 H-indol-2-yl}prop-2-yn-1 -yl)benzamide.

[0504] LC-MS (ES + , m / z): 330.2 [(M-NHMe) + ]

[0505] Example 32 Preparation of N-(3-{1 -ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1 -yl)-6- methylpyridine-3-carboxamide

[0506]

[0507] Prepared in a similar manner to that described in Example 23 using (6-methyl-3- pyridyl)(2-propynylamino)formaldehyde to give N-(3-{1 -ethyl-5-[(methylamino)methyl]- 1 H-indol-2-yl}prop-2-yn-1 -yl)-6-methylpyridine-3-carboxamide.

[0508] LC-MS (ES + , m / z): 330.2 [(M-NHMe) + ]

[0509] Example 33 Preparation of 3-[3-(1 -ethyl-5-{[(1 -methylpiperidin-4-yl)amino]methyl}-1 H-indol-2-yl)prop- 2-yn-1 -yl]-1 -phenylurea

[0510]

[0511] Prepared in a similar manner to that described in Example 29 using 3-phenyl-1 -(2- propynyl)urea to give 3-[3-(1 -ethyl-5-{[(1 -methylpiperidin-4-yl)amino]methyl}-1 H-indol-2- yl)prop-2-yn-1 -yl]-1 -phenylurea.

[0512] LC-MS (ES + , m / z): 444.3 [(M+H) + ]

[0513] Example 34 Preparation of N-[(2-{3-[(4-chloro-3-fluorophenyl)amino]prop-1 -yn-1 -yl}-1 -ethyl-1 H-indol- 5-yl)methyl]-1 -methylpiperidin-4-amine

[0514]

[0515] N-[(2-{3-[(4-chloro-3-fluorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5- yl)methyl]-1-methylpiperidin-4-amine was prepared in a similar manner to that described in Example 29 using N-2-propynyl(4-chloro-3-fluorophenyl)amine.

[0516] LC-MS (ES + , m / z): 453.3 [(M+H) + ]

[0517] Example 35 Preparation of 2-(5-{[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H- indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[0518]

[0519] To a solution of 2-methyl-2-[5-(2-propynylamino)-2-pyridyl]propanenitrile (2.5 eq) in dimethyl sulfoxide (2 mL) was added N-isopropylpropan-2-amine (3 eq) and copper (I) iodide (0.3 eq) followed by 1-ethyl-2-iodo-1H-indole-5-carbaldehyde (100 mg, 1 eq) and tetrakis(triphenylphosphine)palladium(0) (0.1 eq). After stirring the reaction mixture under a nitrogen atmosphere at 25 °C for 1 h, the mixture was poured into an aqueous EDTA solution (15 mL). The biphasic mixture was stirred at 25 °C for 2 h, diluted with water (10 mL) and extracted with ethyl acetate (2 x 40 mL). The organic phase was washed with brine (15 mL) and dried over anhydrous sodium sulphate. The solids were filtered off and the filtrate was concentrated in vacuo to give a crude residue. The crude product was purified by preparative thin layer chromatography (silica gel, eluting with 2 / 1 petroleum ether / ethyl acetate) to give 2-{5-[3-(1-ethyl-5-formyl-1H-indol-2-yl)-2-propynylamino]-2-pyridyl}-2- methylpropanenitrile.

[0520] To a solution of 2-{5-[3-(1-ethyl-5-formyl-1H-indol-2-yl)-2-propynylamino]-2- pyridyl}-2-methylpropanenitrile (1 eq) in dichloromethane / methanol (4 mL, 1 / 1 mixture) were added 1-methyl-4-piperidinylamine (4 eq) and anhydrous magnesium sulfate (15 eq) respectively. The mixture was stirred at 25 °C for 12 h, then sodium bicarbonate (1 eq) and NaBH3CN (4 eq) were added at 0 °C. The reaction mixture was stirred at 0 °C for 2 h, then quenched with a saturated solution of sodium bicarbonate (10 mL). The mixture was extracted with ethyl acetate (2 x 30 mL), the combined organic phases were washed with water (30 mL) and brine (30 mL) and dried over anhydrous sodium sulfate. The solids were filtered off, the filtrate was concentrated in vacuo to give a crude residue. The crude residue was purified by preparative HPLC (C18 silica gel eluted with acetonitrile and water) to give 2-(4-{[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}phenyl)-2-methylpropanenitrile.

[0521] LC-MS (ES + ,m / z): 469.4 [(M+H) + ]

[0522] Example 36 Preparation of N-{[1-(2-chloroethyl)-2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1H- indol-5-yl]methyl}-1-methylpiperidin-4-amine

[0523]

[0524] N-{[1-(2-chloroethyl)-2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1H-indol-5-yl]methyl}-1- methylpiperidin-4-amine was prepared in a similar manner to that described in Example 28 using 1- methylpiperidin-4-amine.

[0525] LC-MS (ES + ,m / z): 469.1 [(M+H) + ]

[0526] Example 37 Preparation of 6-tert-butyl-N-[3-(1-ethyl-5-{[(1-methanesulfonylpiperidin-4-yl)amino]methyl}- 1H-indol-2-yl)prop-2-yn-1-yl]pyridin-3-amine

[0527]

[0528] In a similar manner to that described in Example 34, using N-2-propynyl[6-(tert- butyl)-3-pyridyl]amine, 6-tert-butyl-N-[3-(1-ethyl-5-{[(1-methanesulfonylpiperidin-4- yl)amino]methyl}-1 H-indol-2-yl)prop-2-yn-1-yl]pyridin-3-amine was prepared.

[0529] LC-MS (ES + ,m / z): 522.4 [(M+H) + ]

[0530] Example 38 Preparation of 2-(4-{[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1 H- indol-2-yl)prop-2-yn-1-yl]amino}phenyl)-2-methylpropanoic acid

[0531]

[0532] In a similar manner to that described in Example 34, using 2-methyl-2-[p-(2- propynylamino)phenyl]propanoic acid ethyl ester, 2-(4-{[3-(1-ethyl-5-{[(1- methylpiperidin-4-yl)amino]methyl}-1 H-indol-2-yl)prop-2-yn-1-yl]amino}phenyl)-2- methylpropanoic acid ethyl ester was prepared. This ester was then saponified with sodium hydroxide in methanol and water to give 2-(4-{[3-(1-ethyl-5-{[(1- methylpiperidin-4-yl)amino]methyl}-1 H-indol-2-yl)prop-2-yn-1-yl]amino}phenyl)-2- methylpropanoic acid.

[0533] LC-MS (ES + ,m / z): 487.4 [(M+H) + ]

[0534] Example 39 Preparation of 3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1 H-indol-2-yl)- N-methylprop-2-ynoic amide

[0535]

[0536] To a solution of methylamine (1.23 g, 11.89 mmol, 30% in water, 1 eq) in methanol (20 mL) was added methyl prop-2-ynoate (1 g, 11.89 mmol, 990 μL, 1 eq) dropwise at -50 °C for 30 min and the reaction mixture was stirred at -50 °C for 1.5 h. LCMS showed the reaction to be complete. The reaction mixture was concentrated at 25 °C to give the crude product which was washed with petroleum ether (2 x 10 mL) to give N-methylprop-2-ynoic amide (460 mg, 4.98 mmol, 41.9% yield, 90% purity) as a white solid.

[0537] To a solution of 1-(methylamino)-2-propyn-1-one (3 eq) in dimethyl sulfoxide (4 mL) was added N-isopropylpropan-2-amine (3 eq) and copper (I) iodide (0.2 eq) followed by 1-ethyl-2-iodo-1H-indole-5-carbaldehyde (100 mg, 1 eq) and tetrakis(triphenylphosphine)palladium(0) (0.1 eq). After stirring the reaction mixture under nitrogen atmosphere at 20 °C for 12 h, the mixture was poured into an aqueous EDTA solution (10 mL). The biphasic mixture was stirred at 25 °C for 2 h and left standing for 4 h before extracting with ethyl acetate (20 mL). The organic phase was washed with water (20 mL) and brine (20 mL) and dried over anhydrous sodium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo to give a crude residue. The crude product was purified by flash column chromatography (silica gel, eluting with a gradient of 10 / 1 to 5 / 1 petroleum ether / ethyl acetate) to give a yellow oil. This oil was further purified by preparative HPLC (C18 silica gel, eluting with acetonitrile and water) to give 1-ethyl-2-[3-(methylamino)-3-oxo-1-propynyl]-1H-indole-5- carbaldehyde (50 mg, 16.3% yield) as a yellow solid.

[0538] To a solution of 1-ethyl-2-[3-(methylamino)-3-oxo-1-propynyl]-1H-indole-5- carbaldehyde (1 eq) in dichloromethane (4 mL) was added 1-methyl-4-piperidinylamine (7 eq) and anhydrous magnesium sulfate (15 eq) respectively. The mixture was stirred at 25 °C for 12 h before NaBH3CN (2 eq) was added at 0 °C. The reaction mixture was stirred at 25 °C for 1 h before being quenched with a saturated solution of sodium bicarbonate (10 mL). The mixture was concentrated in ethyl acetate (20 mL) and the organic phase was washed with water (50 mL) and brine (50 mL) and dried over anhydrous sodium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo to give a crude residue. The crude residue was purified by preparative HPLC (C18 silica gel, eluting with acetonitrile and water) to give 3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)-N- methylpropan-2-ynoic amide.

[0539] LC-MS (ES + ,m / z): 353.3 [(M+H) + ]

[0540] Example 40 Preparation of 2-(4-{[3-(1 -ethyl-5-{[(1 -methylpiperidin-4- yl)amino]methyl}-1 H-indol-2-yl)prop-2-yn-1 -yl]amino}phenyl)-2- methylpropanoic acid ethyl ester

[0541]

[0542] 2-(4-{[3-(1 -ethyl-5-{[(1 -methylpiperidin-4- yl)amino]methyl}-1 H-indol-2-yl)prop-2-yn-1 -yl]amino}phenyl)-2- methylpropanoic acid ethyl ester was prepared in a similar manner to that described in Example 34 using 2-methyl-2-[p-(2-propynylamino)phenyl]propanoic acid ethyl ester.

[0543] LC-MS (ES + ,m / z): 515.4 [(M+H) + ]

[0544] Example 41 Preparation of 2-(5-{[3-(1 -ethyl-5-{[(oxan-4- yl)amino]methyl}-1 H-indol-2-yl)prop-2-yn-1 -yl]amino}pyridin-2-yl)-2- methylpropanenitrile

[0545]

[0546] To a solution of 2-methyl-2-[5-(2-propynylamino)-2-pyridyl]propanenitrile (1 eq) in dimethyl sulfoxide (2 mL) was added N-isopropylpropan-2-amine (3 eq) and copper (I) iodide (0.47 eq) followed by 1 -ethyl-2-iodo-1 H-indole-5- carboxaldehyde (100 mg, 1 eq) and tetrakis(triphenylphosphine)palladium (0) (0.1 eq). After stirring the reaction mixture under a nitrogen atmosphere at 40 °C for 2 h, the mixture was poured into an aqueous EDTA solution (20 mL). The biphasic mixture was stirred at 25 °C for 2 h, then extracted with ethyl acetate (20 mL). The organic phase was washed with water (50 mL) and brine (50 mL) and dried over anhydrous sodium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo to give a crude residue. The crude product was purified by flash column chromatography (silica gel, eluting with 2 / 1 petroleum ether / ethyl acetate) to give 2-{5-[3-(1 -ethyl-5-formyl-1 H-indol-2-yl)-2- propynylamino]-2-pyridyl}-2-methylpropanenitrile.

[0547] To a solution of 2-{5-[3-(1-ethyl-5-formyl-1H-indol-2-yl)-2-propynylamino]-2- pyridyl}-2-methylpropanenitrile (1 eq) in dichloromethane (2 mL) were added tetrahydro- 2H-pyran-4-ylamine (8 eq) and anhydrous magnesium sulfate (10 eq) respectively. The mixture was stirred at 25 °C for 2 h, then NaBH3CN (10 eq) was added. The reaction mixture was stirred at 25 °C for 1 h, then quenched with a saturated solution of sodium bicarbonate (10 mL). The mixture was extracted with ethyl acetate (20 mL), and the organic phase was washed with water (50 mL) and brine (50 mL), and dried over anhydrous sodium sulfate. The solids were filtered off, and the filtrate was concentrated in vacuo to give a crude residue as a yellow oil. The crude residue was purified by preparative HPLC (C18 silica gel eluted with acetonitrile and water) to give 2-(5-{[3-(1-ethyl-5-{[(oxan-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile.

[0548] LC-MS (ES + ,m / z): 456.3 [(M+H) + ]

[0549] Example 42 Preparation of N-[(1-ethyl-2-{3-[(4-methylphenyl)amino]prop-1-yn-1-yl}-1H-indol-5-yl)methyl]-1-methylpiperidin-4-amine

[0550]

[0551] N-[(1-ethyl-2-{3-[(4-methylphenyl)amino]prop-1-yn-1-yl}-1H-indol-5-yl)methyl]-1- methylpiperidin-4-amine was prepared in a similar manner to that described in Example 34 using N-2-propynyl(p-tolyl)amine.

[0552] LC-MS (ES + ,m / z): 415.3 [(M+H) + ]

[0553] Example 43 Preparation of 4-{[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}benzonitrile

[0554]

[0555] In a similar manner to that described in Example 34, 4-{[3-(1-ethyl-5-{[(1- methylpiperidin-4-yl)amino]methyl}-1 H-indol-2-yl)prop-2-yn-1 -yl]amino}benzonitrile was prepared using p-(2-propynylamino)benzonitrile.

[0556] LC-MS (ES + ,m / z): 426.3 [(M+H) + ]

[0557] Example 44 Preparation of 3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1 H-indol-2- yl)-N-phenylprop-2-ynoic amide

[0558]

[0559] In a similar manner to that described in Example 38, 3-(1-ethyl-5-{[(1- methylpiperidin-4-yl)amino]methyl}-1 H-indol-2-yl)-N-phenylprop-2-ynoic amide was prepared. LC-MS (ES + ,m / z): 415.3 [(M+H) + ]

[0560] Example 45 Preparation of N-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1 H-indol-5- yl)methyl]-1-methanesulfonylpiperidin-4-amine

[0561]

[0562] A flask was charged with copper (I) iodide (0.3 eq) and triethylamine (2.5 eq), a solution of N-2-propynyl (p-chlorophenyl)amine (2 eq) and 5-bromo-1-ethyl-2-iodo-1H-indole (1 eq) in tetrahydrofuran (10 mL) was added under nitrogen followed by tetrakis(triphenylphosphine)palladium(0) (0.05 eq). The reaction mixture was stirred at 25 °C for 1 h. Thin layer chromatography (petroleum ether / ethyl acetate: 5 / 1) showed the reaction to be complete. The reaction was diluted with 50 mL ethyl acetate and 100 mL 2M EDTA and the biphasic mixture was stirred at 25 °C for 3 h. The phases were separated and the aqueous layer was extracted with ethyl acetate (3 x 40 mL). The combined organic phases were washed with saturated brine (150 mL) and dried over anhydrous sodium sulfate. The solids were filtered off and the filtrate was concentrated under reduced pressure. The crude residue was purified by flash column chromatography (silica gel, eluting with a gradient of 30 / 1 to 20 / 1 petroleum ether / ethyl acetate) to give [3-(5-bromo-1-ethyl-1H-indol-2-yl)-2-propynyl](p- chlorophenyl)amine as a yellow solid.

[0563] To a mixture of [3-(5-bromo-1-ethyl-1H-indol-2-yl)-2-propynyl](p- chlorophenyl)amine (1 eq) in tetrahydrofuran (10 mL) was added n-butyllithium (2.5 M, 6 eq) in one portion at -78 °C under nitrogen. The mixture was stirred at -78 °C for 30 min, then 4-morpholinecarboxaldehyde (10 eq) was added in one portion. The reaction mixture was then stirred at -78 °C for 1.5 h. Thin layer chromatography (petroleum ether / ethyl acetate: 5 / 1) showed the reaction to be complete. The reaction mixture was poured into a saturated ammonium chloride solution / ice mixture (20 mL, w / w = 1 / 1) and stirred for 3 min. The aqueous phase was extracted with ethyl acetate (2 x 10 mL). The combined organic phases were washed with brine (2 x 10 mL) and dried over anhydrous sodium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo to give crude 2-[3-(p-chlorophenylamino)-1-propynyl]-1-ethyl-1H-indole-5- carboxaldehyde as a yellow oil.

[0564] To a mixture of 2-[3-(p-chlorophenylamino)-1-propynyl]-1-ethyl-1H-indole-5- carboxaldehyde (1 eq) and 1-(methylsulfonyl)-4-piperidinylamine (2 eq) in dichloromethane (10 mL) at 25 °C under nitrogen was added anhydrous magnesium sulfate (5 eq) in one portion. The mixture was stirred at 25 °C for 60 min, then NaBH(OAc)3(3 eq) was added. The reaction mixture was stirred for 5 h, LCMS showed the reaction was complete. The reaction mixture was poured into ice-water (10 mL, w / w = 1 / 1), the biphasic mixture was stirred for 3 min, then separated. The aqueous phase was extracted with dichloromethane (3 x 5 mL). The combined organic phase was washed with brine (3 x 5 mL) and dried over anhydrous sodium sulfate. The solid was filtered off, the filtrate was concentrated in vacuo. The crude residue was purified by preparative HPLC (C18 silica gel, eluted with acetonitrile and water) to give N-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5-yl)methyl]-1- methanesulfonopyperidin-4-amine.

[0565] LC-MS (ES + ,m / z): 499.2 [(M+H) + ]

[0566] Example 46 Preparation of 1-(4-{[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5- yl)methyl]amino}piperidin-1-yl)ethan-1-one

[0567]

[0568] In a similar manner to that described in Example 44, 2-[3-(p-chlorophenylamino)-1- propynyl]-1-ethyl-1H-indole-5-carboxaldehyde was reacted with 1-(4-amino-1- piperidinyl)-1-ethanone to give 1-(4-{[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1- ethyl-1H-indol-5-yl)methyl]amino}piperidin-1-yl)ethan-1-one.

[0569] LC-MS (ES + ,m / z): 463.3 [(M+H) + ]

[0570] Example 47 Preparation of 6-tert-butyl-N-[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H- indol-2-yl)prop-2-yn-1-yl]pyridine-3-carboxamide

[0571]

[0572] In a similar manner to that described in Example 29, using (2-propynylamino)[6-(tert- butyl)-3-pyridyl]methanal, 6-tert-butyl-N-[3-(1-ethyl-5-{[(1-methylpiperidin-4- yl)amino]methyl}-1 H-indol-2-yl)prop-2-yn-1-yl]pyridine-3-carboxamide was prepared.

[0573] LC-MS (ES + , m / z): 486.4 [(M+H) + ]

[0574] Example 48 Preparation of N-(3-{1-ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1- yl)-4-(trifluoromethyl)aniline

[0575]

[0576] In a similar manner to that described in Example 23, using N-2-propynyl[p-(trifluoromethyl)phenyl]amine, N-(3-{1-ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1- yl)-4-(trifluoromethyl)aniline was prepared.

[0577] LC-MS (ES + , m / z): 355.2 [(M-NHMe) + ]

[0578] Example 49 Preparation of N-[(1-ethyl-2-{3-[(4-methylphenyl)amino]prop-1-yn-1-yl}-1 H-indol-5- yl)methyl]oxapane-4-amine

[0579]

[0580] In a similar manner to that described in Example 40, using N-2-propynyl(p-tolyl)amine, N-[(1-ethyl-2-{3-[(4-methylphenyl)amino]prop-1-yn-1-yl}-1 H-indol-5-yl)methyl]oxapane- 4-amine was prepared.

[0581] LC-MS (ES + , m / z): 402.3 [(M+H) + ]

[0582] Example 50 Preparation of N-(3-{1-ethyl-4-[(methylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1- yl)aniline

[0583]

[0584] A flask was charged with copper (I) iodide (-60 mg, 0.3 eq) and triethylamine (-100 mg, 2.5 eq) and a solution of N-(prop-2-yn-1 -yl)aniline (-200 mg, 2.0 eq) and 4-bromo-1 -ethyl-2-iodo-1 H-indole (-100 mg, 1.0 eq, prepared from 4-bromo-1 H-indole in a similar manner to that described in Example 1 ) in tetrahydrofuran (10 mL) was added under nitrogen followed by tetrakis(triphenylphosphine)palladium(0) (-20 mg, 0.05 eq). The reaction mixture was stirred at 25 °C for 1 h. TLC (petroleum ether: ethyl acetate = 5:1 ) showed the reaction to be complete. The reaction was diluted with ethyl acetate (50 mL) and EDTA (2 M, 100 mL) and the biphasic mixture stirred at 25 °C for 3 h. The reaction mixture was extracted with ethyl acetate (3 x 40 mL). The organic extracts were washed with saturated brine (150 mL) and dried over anhydrous sodium sulphate. The solids were filtered off and the filtrate was concentrated in vacuo. The crude residue was purified by flash column chromatography (silica gel, eluting with a gradient of petroleum ether / ethyl acetate 30 / 1 to 20 / 1 ) to give N-[3-(4-bromo-1 -ethyl-1 H-indol-2-yl)prop-2-yn-1 -yl]aniline as a yellow solid.

[0585] LC-MS (ES + ,m / z): 353.1 [(M+H) + ]

[0586] To a mixture of N-[3-(4-bromo-1 -ethyl-1 H-indol-2-yl)prop-2-yn-1 -yl]aniline (1 eq) in tetrahydrofuran (10 mL) was added n-butyllithium (2.5 M, 6 eq) in one portion at -78 °C under nitrogen. The mixture was stirred at -78 °C for 30 min and 4-morpholinecarboxaldehyde (10 eq) was added in one portion at -78 °C. The reaction mixture was stirred at -78 °C for 1.5 h. TLC (petroleum ether: ethyl acetate = 5:1 ) showed the reaction to be complete. The residue was poured into an aqueous solution of ammonium chloride ice-water (20 mL, w / w = 1 / 1 ) and stirred for 3 min. The aqueous phase was extracted with ethyl acetate (2 x 10 mL). The combined organic phases were washed with brine (2 x 10 mL), dried over anhydrous sodium sulphate, filtered and concentrated in vacuo to give crude 1 -ethyl-2-[3-(phenylamino)prop-1 -yn-1 -yl]-1 H-indole-4-carbaldehyde as a yellow oil. This product was used directly in the next step without further purification. LC-MS (ES + ,m / z): 275.1 [(M-CHO) + ]

[0587] To a mixture of 1 -ethyl-2-[3-(phenylamino)prop- 1 -yn- 1 -yl]- 1 H-indole-4- carbaldehyde (1 eq) and methylamine (1 eq) in dichloromethane (10 mL) at 25 °C under nitrogen was added anhydrous magnesium sulfate (5 eq) in one portion. The mixture was stirred at 25 °C for 60 min, then NaBH(OAc)3(3 eq) was added and the reaction mixture was stirred for 5 h. The residue was poured into ice-water (10 mL, w / w = 1 / 1) and stirred for 3 min. The aqueous phase was extracted with dichloromethane (3 x 5 mL). The combined organic phases were washed with brine (3 x 5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude residue was purified by preparative HPLC to give N-(3-{ 1 -ethyl-4-[(methylamino)methyl]- 1 H-indol-2-yl}prop-2-yn- 1 -yl)aniline.

[0588] LC-MS (ES + ,m / z): 318.2 [(M+H) + ]

[0589] Example 51 Preparation of N-[3-(l-ethyl-4-{[4-(pyrrolidin-l-yl)piperidin-l-yl]methyl}-lH-indol-2- yl)prop-2-yn-l-yl]aniline

[0590]

[0591] N-[3-(l-ethyl-4-{[4-(pyrrolidin-l-yl)piperidin-l-yl]methyl}-lH-indol-2- yl)prop-2-yn-l-yl]aniline was prepared in a similar manner as described in Example 49 using 4-(pyrrolidin-l-yl)piperidine.

[0592] LC-MS (ES + ,m / z): 441.3 [(M+H) + ]

[0593] Example 52 Preparation of N-({l-ethyl-2-[3-(phenylamino)prop-l-yn-l-yl]-lH-indol-4-yl}methyl)-l- methylpiperidin-4-amine

[0594]

[0595] N-({l-ethyl-2-[3-(phenylamino)prop-l-yn-l-yl]-lH-indol-4-yl}methyl)-l- methylpiperidin-4-amine was prepared in a similar manner as described in Example 49 using 1 -methylpiperidin-4-amine.

[0596] LC-MS (ES + , m / z): 401.3 [(M+H) + ]

[0597] Example 53 Preparation of 1-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H- indol-4-yl)methyl]piperidin-4-ol

[0598]

[0599] 1-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-4-yl)methyl]piperidin-4-ol was prepared in a similar manner to that described in Example 49 using 4-hydroxypiperidine and 4-chloro-N-(prop-2-yn-1-yl)aniline.

[0600] LC-MS (ES + , m / z): 422.1 [(M+H) + ]

[0601] Example 54 Preparation of 4-chloro-N-[3-(1-ethyl-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H- indol-2-yl)prop-2-yn-1-yl]aniline

[0602]

[0603] 4-chloro-N-[3-(1-ethyl-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]aniline was prepared in a similar manner to that described in Example 49 using 4-(pyrrolidin-1-yl)piperidine and 4-chloro-N-(prop-2-yn-1-yl)aniline.

[0604] LC-MS (ES + , m / z): 475.3 [(M+H) + ]

[0605] Example 55 Preparation of 1-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-4-yl)methyl]-N,N-dimethylpiperidin-4-amine

[0606]

[0607] 1-[(2-{3-[(4-Chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-4-yl)methyl]-N,N-dimethylpiperidin-4-amine was prepared in a manner similar to the method described in Example 49 using N,N-dimethylpiperidin-4-amine and 4-chloro-N-(prop-2-yn-1-yl)aniline.

[0608] LC-MS(ES + ,m / z):449.2[(M+H) + ]

[0609] Example 56 : Preparation of 4-chloro-N-(3-{1-ethyl-4-[(4-methylpiperazin-1-yl)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)aniline

[0610]

[0611] 4-Chloro-N-(3-{1-ethyl-4-[(4-methylpiperazin-1-yl)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)aniline was prepared in a manner analogous to the method described in Example 49 using 1-methylpiperazine and 4-chloro-N-(prop-2-yn-1-yl)aniline.

[0612] LC-MS(ES + ,m / z):421.3[(M+H) + ]

[0613] Example 57 : Preparation of 1-{1-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-4-yl)methyl]piperidin-4-yl}piperidin-4-ol

[0614]

[0615] 1-{1-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-4-yl)methyl]piperidin-4-yl}piperidin-4-ol was prepared in a manner similar to the method described in Example 49 using 1,4′-bipiperidin-4-ol and 4-chloro-N-(prop-2-yn-1-yl)aniline.

[0616] LC-MS(ES + ,m / z):505.1[(M+H) + ]

[0617] Example 58Preparation of 2-(5-{[3-(4-{[4-(4-aminopiperidin-1-yl)piperidin-1-yl]methyl}-1- ethyl-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[0618]

[0619] 2-(5-{[3-(4-{[4-(4-aminopiperidin-1-yl)piperidin-1-yl]methyl}-1- ethyl-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared in a similar manner to that described in Example 49 using 1,4'-bipiperidin-4-amine and 2-methyl-2-[5-(prop-2-yn-1-ylamino)pyridin-2-yl]propanenitrile.

[0620] LC-MS (ES + ,m / z): 538.3 [(M+H) + ]

[0621] Example 59 Preparation of 1-[(1-ethyl-2-{3-[(4-fluorophenyl)amino]prop-1-yn-1-yl}-1H-indol-5- yl)methyl]-N,N-dimethylpiperidin-4-amine

[0622]

[0623] 1-[(1-ethyl-2-{3-[(4-fluorophenyl)amino]prop-1-yn-1-yl}-1H-indol-5- yl)methyl]-N,N-dimethylpiperidin-4-amine was prepared in a similar manner to that described in Example 1.

[0624] LC-MS (ES + ,m / z): 433.3 [(M+H) + ]

[0625] Example 60 Preparation of 4-N-({1-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H-indol-5- yl}methyl)-1-N,1-N-dimethylcyclohexane-1,4-diamine

[0626]

[0627] 4-N-({1-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H-indol-5-yl}methyl)-1-N,1-N- dimethylcyclohexane-1,4-diamine was prepared in a similar manner to that described in Example 1.

[0628] LC-MS (ES + m / z): 429.3 [(M+H) + ]

[0629] Example 61 Preparation of 4-chloro-N-(3-{1-ethyl-5-[(methylamino)methyl]-1 H-indol-2- yl}prop-2-yn-1-yl)-3-fluoroaniline

[0630]

[0631] 4-chloro-N-(3-{1-ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1-yl)-3- fluoroaniline was prepared in a similar manner to that described in Example 1.

[0632] LC-MS (ES + m / z): 339.0 [(M-NHMe) + ] and 370.0 [(M+H) + ]

[0633] Example 62 Preparation of 6-tert-butyl-N-(3-{1-ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop- 2-yn-1-yl)pyridine-3-carboxamide

[0634]

[0635] 6-tert-butyl-N-(3-{1-ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1-yl)pyridine- 3-carboxamide was prepared in a similar manner to that described in Example 1.

[0636] LC-MS (ES + m / z): 372.2 [(M-NHMe) + ]

[0637] Example 63 Preparation of N-(3-{1-ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1-yl)benzamide

[0638] N-(3-{1-ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1-yl)benzamide was prepared in a similar manner to that described in Example 1.

[0639] LC-MS (ES +m / z): 315.1 [(M-NHMe) + ]

[0640] Example 64 Preparation of 3-(3-{1 -ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1 - yl)-1 -(4-methylphenyl)urea

[0641]

[0642] 3-(3-{1 -ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1 -yl)-1 -(4- methylphenyl)urea was prepared in a similar manner to that described in Example 1.

[0643] LC-MS (ES + ,m / z): 344.1 [(M-NHMe) + ]

[0644] Example 65 Preparation of 4-chloro-N-(3-{1 -ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop-2- yn-1 -yl)aniline

[0645]

[0646] 4-chloro-N-(3-{1 -ethyl-5-[(methylamino)methyl]-1 H-indol-2-yl}prop-2-yn-1 - yl)aniline was prepared in a similar manner to that described in Example 1.

[0647] LC-MS (ES + ,m / z): 321.0 [(M-NHMe) + ]

[0648] Example 66 Preparation of 4-{[3-(1 -ethyl-5-{[(oxan-4-yl)amino]methyl}-1 H-indol-2-yl)prop-2-yn-1 - yl]amino}benzonitrile

[0649]

[0650] 4-{[3-(1 -ethyl-5-{[(oxan-4-yl)amino]methyl}-1 H-indol-2-yl)prop-2-yn-1 -yl]amino} benzonitrile was prepared in a similar manner to that described in Example 28.

[0651] LC-MS (ES + ,m / z): 413.2 [(M+H) + ]

[0652] Example 67 Preparation of N-[(2-{3-[(4-chloro-3-fluorophenyl)amino]prop-1-yn-1-yl}-1- ethyl-1H-indol-5-yl)methyl]oxanyl-4-amine

[0653]

[0654] N-[(2-{3-[(4-chloro-3-fluorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5- yl)methyl]oxanyl-4-amine was prepared in a similar manner to that described in Example 28.

[0655] LC-MS (ES + , m / z): 440.1 [(M+H) + ]

[0656] Example 68 Preparation of 3-[3-(1-ethyl-5-{[(oxanyl-4-yl)amino]methyl}-1H-indol-2-yl)prop-2- yn-1-yl]-1-phenylurea

[0657]

[0658] 3-[3-(1-ethyl-5-{[(oxanyl-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]-1- phenylurea was prepared in a similar manner to that described in Example 28. LC-MS (ES + , m / z): 431.1 [(M+H) + ]

[0659] Example 69 Preparation of 6-tert-butyl-N-[3-(1-ethyl-5-{[(oxanyl-4-yl)amino]methyl}-1H-indol-2- yl)prop-2-yn-1-yl]pyridin-3-amine

[0660]

[0661] 6-tert-Butyl-N-[3-(1-ethyl-5-{[(oxanyl-4-yl)amino]methyl}-1H-indol-2-yl)prop-2- yn-1-yl]pyridin-3-amine was prepared in a similar manner to that described in Example 28.

[0662] LC-MS (ES + , m / z): 445.3 [(M+H) + ]

[0663] Example 70 : 4-{[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5- yl)methyl]amino}-1H-pyridin-2-one 6 Preparation of thiamethoxam

[0664]

[0665] In a similar manner to that described in Example 28, 4-{[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5-yl)methyl]amino}-1H-pyridin-2-one was prepared. 6 Preparation of thiamethoxam.

[0666] LC-MS (ES + , m / z): 492.2 [(M+Na) + ]

[0667] Example 71 Preparation of N-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5- yl)methyl]-1-(2-methanesulfonyl ethyl)piperidin-4-amine

[0668]

[0669] In a similar manner to that described in Example 28, N-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5-yl)methyl]-1-(2-methanesulfonyl ethyl)piperidin-4-amine was prepared.

[0670] LC-MS (ES + , m / z): 527.3 [(M+H) + ]

[0671] Example 72 Preparation of 1-(4-{[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5- yl)methyl]amino}piperidin-1-yl)-2-(dimethylamino)ethan-1-one

[0672]

[0673] In a similar manner to that described in Example 28, 1-(4-{[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5-yl)methyl]amino}piperidin-1-yl)-2-(dimethylamino)ethan-1-one was prepared.

[0674] LC-MS (ES + m / z): 506.3 [(M+H) + ]

[0675] Example 73 Preparation of 2-(4-{[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1- ethyl-1H-indol-5-yl)methyl]amino}piperidin-1-yl)-N,N-dimethylacetamide

[0676]

[0677] 2-(4-{[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5- yl)methyl]amino}piperidin-1-yl)-N,N-dimethylacetamide was prepared in a similar manner to that described in Example 28.

[0678] LC-MS (ES + m / z): 506.2 [(M+H) + ]

[0679] Example 74 Preparation of 2-tert-butyl-N-[3-(1-ethyl-5-{[(1-methylpiperidin-4- yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]pyrimidin-5-amine

[0680]

[0681] 2-tert-Butyl-N-[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H- indol-2-yl)prop-2-yn-1-yl]pyrimidin-5-amine was prepared in a similar manner to that described in Example 28.

[0682] LC-MS (ES + m / z): 459.4 [(M+H) + ]

[0683] Example 75 Preparation of 2-methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile

[0684]

[0685] In a similar manner to that described in Example 28, 2-methyl-2-(5-{[3-(5-{[(oxan-4- yl)amino]methyl}-l-(2,2,2-trifluoroethyl)-lH-indol-2-yl)prop-2-yn-l-yl]amino}pyridin-2- yl)propanenitrile was prepared.

[0686] LC-MS (ES + , m / z): 510.2 [(M+H) + ]

[0687] Example 76 Preparation of 2-[5-({3-[l-(2-fluoroethyl)-5-{[(oxan-4-yl)amino]methyl}-lH-indol-2- yl]prop-2-yn-l-yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[0688]

[0689] In a similar manner to that described in Example 28, 2-[5-({3-[l-(2-fluoroethyl)-5-{[(oxan-4- yl)amino]methyl}-lH-indol-2-yl]prop-2-yn-l-yl}amino)pyridin-2-yl]-2-methylpropanenitrile was prepared.

[0690] LC-MS (ES + , m / z): 474.2 [(M+H) + ]

[0691] Example 77 Preparation of 3-(l-ethyl-5-{[(l-methylpiperidin-4-yl)amino]methyl}-lH-indol-2- yl)prop-2-yn-l-ol

[0692]

[0693] In a similar manner to that described in Example 28, 3-(l-ethyl-5-{[(l-methylpiperidin-4- yl)amino]methyl}-lH-indol-2-yl)prop-2-yn-l-ol was prepared.

[0694] LC-MS (ES + , m / z): 326.2 [(M+H) + ]

[0695] Example 78 Preparation of 2-[5-({3-[l-(2-fluoroethyl)-5-{[(oxan-4-yl)amino]methyl}-lH-indol-2- yl]prop-2-yn-l-yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[0696]

[0697] In a similar manner to that described in Example 28, 2-[5-({3-[1-(2,2- difluoroethyl)-5-{[(oxan-4-yl)amino]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin- 2-yl]-2-methylpropanenitrile is prepared.

[0698] LC-MS (ES + ,m / z): 490.2 [(M+H) + ]

[0699] Example 79 Preparation of 2-[5-({3-[1-(2,2-difluoroethyl)-5-{[(oxan-4-yl)amino]methyl}-1H- indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[0700]

[0701] In a similar manner to that described in Example 28, 6-chloro-N-[3-(1-ethyl-5- {[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]pyridin-3-amine is prepared.

[0702] LC-MS (ES + ,m / z): 492.3 [(M+H) + ]

[0703] Example 80 Preparation of 6-chloro-N-[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}- 1H-indol-2-yl)prop-2-yn-1-yl]pyridin-3-amine

[0704]

[0705] In a similar manner to that described in Example 28, 6-chloro-N-[3-(1-ethyl-5- {[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]pyridin-3-amine is prepared.

[0706] LC-MS (ES + ,m / z): 436.1 [(M+H) + ]

[0707] Synthetic Scheme:Preparation of tert-butyl N-({3-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H- indol-6-yl}methyl)-N-(oxan-4-yl)carbamate

[0708]

[0709] Example 81

[0710]

[0711] To a 0 °C solution of 1H-indole-6-carboxaldehyde (1.5 g, 10.34 mmol) in dimethylformamide (30 mL) was added sodium hydride (455 mg, 11.3 mmol). The mixture was stirred at 25 °C for 30 min, then cooled to 0 °C before the addition of benzenesulfonyl chloride (1.39 mL, 1.92 g, 11.3 mmol). The mixture was stirred at 25 °C overnight. The reaction was monitored by thin layer chromatography. Upon completion of the reaction, the mixture was poured into ice / water (300 mL) and stirred for 1 h. The solid was filtered and washed with water (100 mL) and hexane (50 mL) and dried overnight to give 1-(phenylsulfonyl)-1H-indole-5-carboxaldehyde as a yellow solid (2.90 g, 96% yield).

[0712] To a suspension of 1-(phenylsulfonyl)-1H-indole-5-carboxaldehyde (1.7 g, 5.96 mmol) in methanol (30 mL) was added 4-amino-tetrahydropyran (0.74 mL, 0.72 g, 7.16 mmol) dropwise. The reaction mixture was stirred at 25 °C for 12 h, then cooled to 0 °C before the addition of sodium borohydride in small portions. The mixture was stirred at 0 °C for 10 min, then allowed to warm to 25 °C with an additional 10 min stirring. A 1 N sodium hydroxide solution (20 mL) was added slowly with 30 min stirring, then the mixture was concentrated in vacuo. The residue was diluted with brine (50 mL) and water (50 mL) and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (50 mL) and water (3 x 50 mL) and dried over anhydrous sodium sulfate. The solid was filtered off and the filtrate was concentrated in vacuo to give {[1-(phenylsulfonyl)-1H-indol-5-yl]methyl}-tetrahydro-2H-pyran-4-ylamine as a yellow gum (2.15 g, 96% crude yield). This crude product was used directly without further purification.

[0713] A dry flask (250 mL) was charged with dichloromethane (60 mL) and aluminum trichloride (2.27 g, 17.01 mmol) under nitrogen at 0 °C. {[l-(phenylsulfonyl)-lH-indol-5-yl]methyl}-tetrahydro-2H-pyran-4-ylamine (2.0 g, 4.86 mmol) in dichloromethane (15 mL) was added dropwise and the mixture was stirred for 10 min after addition. Acetyl chloride (0.36 mL, 0.39 g, 5.10 mmol) in dichloromethane (15 mL) was then added dropwise and the resulting mixture was stirred at 0 °C for 10 min. The mixture was then allowed to warm to 25 °C and stirring was continued for 3 h. The reaction mixture was decanted and the residue was treated with ice / water (30 mL) followed by the addition of IN sodium hydroxide solution (30 mL). The mixture was stirred for 30 min and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (50 mL) and water (3 x 50 mL) and dried over anhydrous sodium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo to give l-{l-(phenylsulfonyl)-5-[(tetrahydro-2H-pyran-4-ylamino)methyl]-lH-indol-3-yl}-l- ethanone (1.92 g, 87% crude yield) as a white foamy solid.

[0714] A dry flask (250 mL) was charged with trifluoroacetic acid (30 mL) under nitrogen at 0 °C and sodium borohydride (2.60 g, 69.01 mmol) was added portionwise with vigorous stirring. l-{l-(phenylsulfonyl)-5-[(tetrahydro-2H-pyran-4-ylamino)methyl]-lH-indol-3-yl}-l- ethanone (1.90 g, 4.60 mmol) in dichloromethane (30 mL) was added dropwise at 0 °C and the resulting mixture was allowed to warm to 25 °C. Stirring was continued for 5 h. The reaction mixture was quenched with ice / water (20 mL) and basified with IN sodium hydroxide (450 mL) while stirring over 30 min. The mixture was then extracted with dichloromethane (3 x 30 mL), the organic layers were washed with water (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give {[3-ethyl-l-(phenylsulfonyl)-lH-indol-5-yl]methyl}-tetrahydro-2H-pyran-4-ylamine (1.60 g, 86% crude yield) as a light brown foamy solid.

[0715] To a solution of { [3-ethyl-l-(phenylsulfonyl)-lH-indol-5-yl]methyl}- tetrahydro-2H-pyran-4-ylamine (0.96 g, 4.02 mmol) in dichloromethane (50 mL) was added dropwise Boc-anhydride (0.96 g, 4.42 mmol) followed by trimethylamine (0.83 mL, 6.03 mmol) dropwise at 25 °C. The reaction mixture was stirred at 25 °C for 3 h. The mixture was diluted with water (100 mL) and extracted with dichloromethane (3 x 40 mL). The combined organic layers were washed with water (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluted with hexane / ethyl acetate 4 / 1) to give tert-butyl (3-ethyl- 1 -(phenylsulfonyl)- 1 H-indol-6-yl)methyl(tetrahydro-2H-pyran-4-yl)carbamate (550 mg) as a white solid.

[0716] To a solution of tert-butyl (3-ethyl-2-iodo-l-(phenylsulfonyl)-lH-indol-6- yl)methyl(tetrahydro-2H-pyran-4-yl)carbamate (270 mg, 0.40 mmol) in tetrahydrofuran (10 mL) at -78 °C under nitrogen was added dropwise lithium diisopropylamide (1 M, 1.18 mL, 3 eq) while stirring at -78 °C for 1 h. A solution of iodine (105 mg, 0.413 mmol, 1.05 eq) in 1 mL of tetrahydrofuran was added dropwise and the reaction mixture was stirred at -78 °C for an additional 5 min after addition. The reaction mixture was poured into aqueous ammonium chloride (5 mL) and extracted with ethyl acetate (3 x 30 mL). The combined organic layers were washed with water (3 x 30 mL) and brine (10 mL) and dried over anhydrous sodium sulfate. The solid was filtered off and the filtrate was concentrated in vacuo to give tert-butyl (3-ethyl-2-iodo-l-(phenylsulfonyl)-lH-indol-6- yl)methyl(tetrahydro-2H-pyran-4-yl)carbamate (270 mg) as a yellow oil. The crude product was used without further purification.

[0717] To a solution of tert-butyl (3-ethyl-2-iodo-1-(phenylsulfonyl)-1H-indol-6- yl)methyl(tetrahydro-2H-pyran-4-yl)carbamate (248 mg, 0.4 mmol, 1 equiv) in methanol (6.8 mL) was added 2 M potassium carbonate solution (3.7 mL, 6.4 mmol) and the reaction mixture was stirred at 80 °C for 16 h. The reaction mixture was concentrated in vacuo, diluted with water (10 mL) and extracted with ethyl acetate (3 x 30 mL). The combined organic layers were washed with water (2 x 30 mL) and brine (2 x 30 mL) and dried over anhydrous magnesium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo to give crude tert-butyl (3-ethyl-2-iodo-1H-indol-6-yl)methyl(tetrahydro-2H-pyran-4-yl)carbamate (200 mg) as an orange oil. This crude product was used directly without further purification.

[0718] To a solution of N-prop-2-yn-1-yl-aniline (20 mg, 150 μmol) in DMSO (1 mL) was added N-isopropylpropan-2-amine (53 μL, 375 μmol) and copper (I) iodide (5 mg, 25 μmol) at room temperature under nitrogen while stirring for 5 min. Then a solution of tert-butyl (3-ethyl-2-iodo-1H-indol-6-yl)methyl(tetrahydro-2H-pyran-4-yl)carbamate (60 mg, 125 μmol, 1 equiv) in DMSO (0.5 mL) and tetrakis(triphenylphosphine)palladium(0) (15 mg, 12.5 μmol, 0.1 equiv) was added and the reaction mixture was stirred at 25 °C for 1 h. The reaction was quenched with ice / water (10 mL). Ethyl acetate (30 mL) and aqueous EDTA (20 mL) were added and the mixture was stirred for 2 h. The mixture was extracted with ethyl acetate (3 x 30 mL) and the combined organic layers were washed with water (3 x 10 mL) and brine (10 mL) and dried over anhydrous magnesium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo. The crude residue was purified by preparative HPLC to give tert-butyl N-({3-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H-indol-6-yl}methyl)-N-(oxan-4- yl)carbamate (40 mg) as a light yellow solid.

[0719] LC-MS (ES + , m / z): 532.3 [(M+HCO2H) + ]

[0720] Example 82 Preparation of 6-chloro-N-[3-(1-ethyl-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H- indol-2-yl)prop-2-yn-1-yl]pyridin-3-amine

[0721]

[0722] In a similar manner to that described in Example 28, 6-chloro-N-[3-(1-ethyl-4-{[4- (pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]pyridin-3- amine was prepared.

[0723] LC-MS (ES + ,m / z): 476.2 [(M+H) + ]

[0724] Example 83 Preparation of 3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2- yl)prop-2-yn-1-yl benzoate

[0725]

[0726] In a similar manner to that described in Example 28, 3-(1-ethyl-5-{[(1- methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl benzoate was prepared. LC-MS (ES + ,m / z): 430.3 [(M+H) + ]

[0727] Example 84 Preparation of 2-[5-({3-[1-(2-chloroethyl)-5-{[(1-methylpiperidin-4-yl)amino]methyl}- 1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[0728]

[0729] In a similar manner to that described in Example 28, 2-[5-({3-[1-(2-chloroethyl)-5-{[(1- methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2- yl]-2-methylpropanenitrile was prepared.

[0730] LC-MS (ES + ,m / z): 503.2 [(M+H) + ]

[0731] Example 85 Preparation of N-(6-chloropyridin-3-yl)-3-(1-ethyl-5-{[(1-methylpiperidin-4- yl)amino]methyl}-1H-indol-2-yl)prop-2-ynamide

[0732]

[0733] N-(6-chloropyridin-3-yl)-3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1 H- indol-2-yl)prop-2-ynoic amide was prepared in a similar manner to that described in Example 28.

[0734] LC-MS (ES + ,m / z): 450.2 [(M+H) + ]

[0735] Example 86 Preparation of N-[6-(1-cyano-1-methylethyl)pyridin-3-yl]-3-(1-ethyl-5-{[(1- methylpiperidin-4-yl)amino]methyl}-1 H-indol-2-yl)prop-2-ynoic amide

[0736]

[0737] N-[6-(1-cyano-1-methylethyl)pyridin-3-yl]-3-(1-ethyl-5-{[(1-methylpiperidin-4- yl)amino]methyl}-1 H-indol-2-yl)prop-2-ynoic amide was prepared in a similar manner to that described in Example 28.

[0738] LC-MS (ES + ,m / z): 483.3 [(M+H) + ]

[0739] Example 87 Preparation of N-({3-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1 H-indol-6-yl}methyl)oxane- 4-amine

[0740]

[0741] To a solution of tert-butyl N-({3-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H- indol-6-yl}methyl)-N-(oxan-4-yl)carbamate (11 mg, 22.5 pmol, prepared as described in Example 80) in acetonitrile (1 mL) was added bismuthine (50 mg, 158.7 pmol, 42.94 pL) and the reaction mixture was stirred at 50 °C for 25 min. Aqueous EDTA (20 mL) and ethyl acetate (5 mL) were added and the mixture was stirred for 2 h. The reaction mixture was extracted with ethyl acetate (3 x 10 mL), the combined organic layers were washed with water (3 x 10 mL) and brine (10 mL) and dried over anhydrous magnesium sulfate. The solids were filtered off and the filtrate was concentrated in vacuo. The crude residue was immediately purified by preparative HPLC (C18 silica gel eluted with acetonitrile and water) to give N-({3-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H-indol-6- yl}methyl)oxan-4-amine as a yellow solid (5 mg, 57% yield).

[0742] LC-MS (ES + ,m / z): 388.2 [(M+H) + ]

[0743] Example 88 Preparation of 2-[5-({3-[1-(2-chloroethyl)-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}- 1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[0744]

[0745] 2-[5-({3-[1-(2-chloroethyl)-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2- yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile was prepared in a similar manner to that described in Example 1, using 4-bromo-indole instead of 5-bromo-indole.

[0746] LC-MS (ES + ,m / z): 543.3 [(M+H) + ]

[0747] Example 89 Preparation of 2-(5-{[3-(5-{[(1-methanesulfonylpiperidin-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[0748]

[0749] 2-(5-{[3-(5-{[(1-methanesulfonylpiperidin-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 -yl]amino}pyridin-2-yl)-2- methylpropanenitrile was prepared in a similar manner to that described in Example 28.

[0750] LC-MS (ES + ,m / z): 587.3 [(M+H) + ]

[0751] Example 90 Preparation of 2-[5-({3-[5-({[1-(2-methanesulfonyl ethyl)piperidin-4-yl]amino}methyl)- 1-(2,2,2-trifluoroethyl)-1 H-indol-2-yl]prop-2-yn-1 -yl}amino)pyridin-2-yl]-2- methylpropanenitrile

[0752]

[0753] 2-[5-({3-[5-({[1-(2-methanesulfonyl ethyl)piperidin-4-yl]amino}methyl)-1-(2,2,2- trifluoroethyl)-1 H-indol-2-yl]prop-2-yn-1 -yl}amino)pyridin-2-yl]-2- methylpropanenitrile was prepared in a similar manner to that described in Example 28.

[0754] LC-MS (ES + ,m / z): 615.3 [(M+H) + ]

[0755] Example 91 Preparation of 2-(5-{[3-(5-{[(1-acetylpiperidin-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 -yl]amino}pyridin-2-yl)-2- methylpropanenitrile

[0756]

[0757] 2-(5-{[3-(5-{[(1-acetylpiperidin-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1 H- indol-2-yl)prop-2-yn-1 -yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared in a similar manner to that described in Example 28.

[0758] LC-MS (ES + ,m / z): 551.3 [(M+H) +]

[0759] Example 92 Preparation of 2-methyl-2-(5-{[3-(5-{[(1-methylpiperidin-4-yl)amino]methyl}-1- (2,2,2-trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 -yl]amino}pyridin-2-yl)propanenitrile

[0760]

[0761] In a similar manner to that described in Example 28, 2-methyl-2-(5-{[3-(5-{[(1- methylpiperidin-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 - yl]amino}pyridin-2-yl)propanenitrile was prepared.

[0762] LC-MS (ES + ,m / z): 523.3 [(M+H) + ]

[0763] Example 93 Preparation of 2-methyl-2-(5-{[3-(5-{[(1-methylpiperidin-4-yl)amino]methyl}-1- (2,2,2-trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 -yl]amino}pyridin-2-yl)propanenitrile

[0764]

[0765] In a similar manner to that described in Example 28, 2-methyl-2-(5-{[3-(5-{[(1- methylpiperidin-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 - yl]amino}pyridin-2-yl)propanenitrile was prepared.

[0766] LC-MS (ES + ,m / z): 523.3 [(M+H) + ]

[0767] Example 94 Preparation of 2-methyl-2-(5-{[3-(5-{[(1-methylpiperidin-4-yl)amino]methyl}-1- (2,2,2-trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 -yl]amino}pyridin-2-yl)propanenitrile

[0768]

[0769] In a similar manner to that described in example 28, 6-chloro-N-[3-(5-{[(1- methylpiperidin-4-yl)amino]methyl}-1 -(2,2,2-trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 - yl]pyridin-3-amine was prepared.

[0770] LC-MS (ES + , m / z): 440.3 [(M+H) + ]

[0771] Example 95 Preparation of 6-chloro-N-[3-(5-{[(oxan-4-yl)amino]methyl}-1 -(2,2,2- trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 -yl]pyridin-3-amine

[0772]

[0773] In a similar manner to that described in example 28, 6-chloro-N-[3-(5-{[(1- methylpiperidin-4-yl)amino]methyl}-1 -(2,2,2-trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 - yl]pyridin-3-amine was prepared.

[0774] LC-MS (ES + , m / z): 490.1 [(M+H) + ]

[0775] Example 96 Preparation of 6-chloro-N-[3-(5-{[(oxan-4-yl)amino]methyl}-1 -(2,2,2- trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 -yl]pyridin-3-amine

[0776]

[0777] In a similar manner to that described in example 28, 6-chloro-N-[3-(5-{[(1- methylpiperidin-4-yl)amino]methyl}-1 -(2,2,2-trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 - yl]pyridin-3-amine was prepared.

[0778] LC-MS (ES + , m / z): 490.1 [(M+H) + ]

[0779] Example 97Preparation of 2-[5-({3-[1-(cyclopropylmethyl)-5-{[(oxan-4-yl)amino]methyl}-1H- indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[0780]

[0781] 2-[5-({3-[1-(cyclopropylmethyl)-5-{[(oxan-4-yl)amino]methyl}-1H-indol-2-yl]prop-2- yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile was prepared in a similar manner to that described in Example 28.

[0782] LC-MS (ES + ,m / z): 482.3 [(M+H) + ]

[0783] Example 98 Preparation of 2-(5-{[3-(4-{[4-(diethylamino)piperidin-1-yl]methyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[0784]

[0785] 2-(5-{[3-(4-{[4-(diethylamino)piperidin-1-yl]methyl}-1-(2,2,2-trifluoroethyl)-1H- indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared in a similar manner to that described in Example 1 using 4-bromo-indole.

[0786] LC-MS (ES + ,m / z): 565.2 [(M+H) + ]

[0787] Example 99 Preparation of 2-methyl-2-{5-[(3-{4-[(4-methylpiperazin-1-yl)methyl]-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}propanenitrile

[0788]

[0789] In a similar manner to that described in Example 1, using 4-bromo-7-fluoro-indole, 2-(5-{[3-(1-ethyl-7-fluoro-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared.

[0790] LC-MS (ES + , m / z): 509.1 [(M+H) + ]

[0791] Example 100 Preparation of 2-(5-{[3-(1-ethyl-7-fluoro-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[0792]

[0793] In a similar manner to that described in Example 1, using 4-bromo-7-fluoro-indole, 2-(5-{[3-(1-ethyl-7-fluoro-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared.

[0794] LC-MS (ES + , m / z): 527.2 [(M+H) + ]

[0795] Example 101 Preparation of 2-(5-{[3-(1-ethyl-7-fluoro-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[0796]

[0797] In a similar manner to that described in Example 1, using 4-bromo-7-fluoro-indole, 2-(5-{[3-(1-ethyl-7-fluoro-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared.

[0798] LC-MS (ES + , m / z): 563.3 [(M+H) + ]

[0799] Example 102 Preparation of 2-(5-{[3-(4-{[4-(4-hydroxypiperidin-l-yl)piperidin-l-yl]methyl}- l-(2,2,2-trifluoroethyl)-lH-indol-2-yl)prop-2-yn-l-yl]amino}pyridin-2-yl)-2- methylpropanenitrile

[0800]

[0801] 2-(5-{[3-(4-{[4-(4-hydroxypiperidin-l-yl)piperidin-l-yl]methyl}-l-(2,2,2- trifluoroethyl)-lH-indol-2-yl)prop-2-yn-l-yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared in a similar manner to that described in Example 1 using 4-bromo-indole.

[0802] LC-MS (ES + ,m / z): 593.2 [(M+H) + ]

[0803] Example 103 Preparation of N-(6-cyanopyridin-3-yl)-3-(l-ethyl-5-{[(l-methylpiperidin-4- yl)amino]methyl}-lH-indol-2-yl)prop-2-ynoic amide

[0804]

[0805] N-(6-cyanopyridin-3-yl)-3-(l-ethyl-5-{[(l-methylpiperidin-4-yl)amino]methyl}- lH-indol-2-yl)prop-2-ynoic amide was prepared in a similar manner to that described in Example 28.

[0806] LC-MS (ES + ,m / z): 441.3 [(M+H) + ]

[0807] Example 104 Preparation of N-[6-(l-cyano-l-methylethyl)pyridin-3-yl]-3-(5-{[(oxan-4- yl)amino]methyl}-l-(2,2,2-trifluoroethyl)-lH-indol-2-yl)prop-2-ynoic amide

[0808]

[0809] N-[6-(1-cyano-1-methylethyl)pyridin-3-yl]-3-(5-{[(oxan-4-yl)amino]methyl}-1- (2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-ynoic amide was prepared in a similar manner to that described in Example 28.

[0810] LC-MS (ES + , m / z): 524.2 [(M+H) + ]

[0811] Example 105 Preparation of N-[6-(1-cyano-1-methylethyl)pyridin-3-yl]-3-(5-{[(1-methylpiperidin-4- yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-ynoic amide

[0812]

[0813] N-[6-(1-cyano-1-methylethyl)pyridin-3-yl]-3-(5-{[(1-methylpiperidin-4-yl)amino]methyl}- 1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-ynoic amide was prepared in a similar manner to that described in Example 28.

[0814] LC-MS (ES + , m / z): 537.2 [(M+H) + ]

[0815] Example 106 Preparation of 2-methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(oxan-2-ylmethyl)-1H- indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile

[0816]

[0817] 2-methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(oxan-2-ylmethyl)-1H-indol-2-yl)prop- 2-yn-1-yl]amino}pyridin-2-yl)propanenitrile was prepared in a similar manner to that described in Example 28.

[0818] LC-MS (ES + , m / z): 484.4 [(M+H) + ]

[0819] Example 107: Preparation of 2-(5-{[3-(5-{[(2-methoxyethyl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile

[0820]

[0821] In a manner similar to that described in Example 28, 2-(5-{[3-(5-

[0822] {[(2-methoxyethyl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile.

[0823] LC-MS(ES + ,m / z):484.3[(M+H) + ]

[0824] Example 108 : Preparation of 2-methyl-2-[5-({3-[5-({[2-(morpholin-4-yl)ethyl]amino}methyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propionitrile

[0825]

[0826] Following the procedure described in Example 28, 2-methyl-2-[5-({3-[5-({[2-(morpholin-4-yl)ethyl]amino}methyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propanenitrile was prepared.

[0827] LC-MS(ES + ,m / z):539.3[(M+H) + ]

[0828] Example 109 : Preparation of 2-methyl-2-(5-{[3-(4-{[(1-methylpiperidin-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propionitrile

[0829]

[0830] In a similar manner to that described in Example 1, using 4-bromo-indole, 2-methyl-2-(5-{[3-(4-{[(1-methylpiperidin-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile was prepared.

[0831] LC-MS (ES + , m / z): 523.4 [(M+H) + ]

[0832] Example 110 Preparation of 2-methyl-2-(5-{[3-(4-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile

[0833]

[0834] In a similar manner to that described in Example 1, using 4-bromo-indole, 2-methyl-2-(5-{[3-(4-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile was prepared.

[0835] LC-MS (ES + , m / z): 510.3 [(M+H) + ]

[0836] Example 111 Preparation of 2-[5-({3-[5-({[2-(dimethylamino)ethyl]amino}methyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[0837]

[0838] In a similar manner to that described in Example 28, 2-[5-({3-[5-({[2-(dimethylamino)ethyl]amino}methyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile was prepared.

[0839] LC-MS (ES + , m / z): 497.4 [(M+H) + ]

[0840] Example 112Preparation of 2-(5-{[3-(7-fluoro-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1- (2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[0841]

[0842] In a similar manner to that described in Example 1, using 4-bromo-7-fluoro-indole, 2-(5-{[3-(7-fluoro-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1-(2,2,2-trifluoroethyl)-1H- indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared. LC-MS (ES + ,m / z): 581.4 [(M+H) + ]

[0843] Example 113 Preparation of 2-methyl-2-[5-({3-[1-(2,2,2-trifluoroethyl)-5-{[(2,2,2-trifluoroethyl)amino]methyl}- 1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propanenitrile

[0844]

[0845] In a similar manner to that described in Example 28, 2-methyl-2-[5-({3-[1-(2,2,2-trifluoroethyl)-5-{[(2,2,2-trifluoroethyl)amino]methyl}-1H-indol-2-yl]prop-2-yn-1- yl}amino)pyridin-2-yl]propanenitrile was prepared.

[0846] LC-MS (ES + ,m / z): 508.2 [(M+H) + ]

[0847] Example 114 Preparation of 2-[5-({3-[5-({[1-(2-hydroxyethyl)piperidin-4-yl]amino}methyl)-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[0848]

[0849] In a similar manner to that described in Example 28, 2-[5-({3-[5-({[1 -(2- methoxyethyl)piperidin-4-yl]amino}methyl)-1 -(2,2,2-trifluoroethyl)-1 H-indol-2- yl]prop-2-yn-1 -yl}amino)pyridin-2-yl]-2-methylpropanenitrile was prepared.

[0850] LC-MS (ES + , m / z): 553.4 [(M+H) + ]

[0851] Example 115 Preparation of 2-[5-({3-[5-({[1 -(2-methoxyethyl)piperidin-4-yl]amino}methyl)-1 -(2,2,2- trifluoroethyl)-1 H-indol-2-yl]prop-2-yn-1 -yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[0852]

[0853] In a similar manner to that described in Example 28, 2-[5-({3-[5-({[1 -(2- methoxyethyl)piperidin-4-yl]amino}methyl)-1 -(2,2,2-trifluoroethyl)-1 H-indol-2- yl]prop-2-yn-1 -yl}amino)pyridin-2-yl]-2-methylpropanenitrile was prepared.

[0854] LC-MS (ES + , m / z): 567.4 [(M+H) + ]

[0855] Example 116 Preparation of 2-[5-({3-[5-({[4-(dimethylamino)cyclohexyl]amino}methyl)-1 -(2,2,2- trifluoroethyl)-1 H-indol-2-yl]prop-2-yn-1 -yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[0856]

[0857] In a similar manner to that described in Example 28, 2-[5-({3-[5-({[4-(dimethylamino)cyclohexyl]amino}methyl)-1 -(2,2,2-trifluoroethyl)-1 H-indol-2- yl]prop-2-yn-1 -yl}amino)pyridin-2-yl]-2-methylpropanenitrile was prepared.

[0858] LC-MS (ES + , m / z): 551.6 [(M+H) + ]

[0859] Example 117 Preparation of 2-methyl-2-{5-[(3-{5-[({1-[2-(morpholin-4-yl)acetyl]piperidin-4- yl}amino)methyl]-1-(2,2,2-trifluoroethyl)-1 H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin- 2-yl}propionitrile

[0860]

[0861] 2-methyl-2-{5-[(3-{5-[({1-[2-(morpholin-4-yl)acetyl]piperidin-4-yl}amino)methyl]-1- (2,2,2-trifluoroethyl)-1 H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile was prepared in a similar manner to that described in Example 28.

[0862] LC-MS (ES + ,m / z): 636.4 [(M+H) + ]

[0863] Example 118 Preparation of 2-(5-{[3-(4-{[(2-methoxyethyl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile

[0864]

[0865] 2-(5-{[3-(4-{[(2-methoxyethyl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1 H-indol-2- yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile was prepared in a similar manner to that described in Example 1 using 4-bromo-indole.

[0866] LC-MS (ES + ,m / z): 484.3 [(M+H) + ]

[0867] Example 119 Preparation of 2-methyl-2-{5-[(3-{4-[(methylamino)methyl]-1-(2,2,2- trifluoroethyl)-1 H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile

[0868]

[0869] In a similar manner to that described in Example 1, using 4-bromo-indole, 2-methyl-2-{5-[(3-{4-[(methylamino)methyl]-1 -(2,2,2-trifluoroethyl)-1 H-indol-2-yl}prop-2-yn-1 - yl)amino]pyridin-2-yl}propionitrile was prepared.

[0870] LC-MS (ES + ,m / z): 440.3 [(M+H) + ]

[0871] Example 120 Preparation of 2-{5-[(3-{4-[(4-acetylpiperazin-1 -yl)methyl]-1 -(2,2,2- trifluoroethyl)-1 H-indol-2-yl}prop-2-yn-1 -yl)amino]pyridin-2-yl}-2-methylpropionitrile

[0872]

[0873] In a similar manner to that described in Example 1, using 4-bromo-indole, 2-{5-[(3-{4-[(4-acetylpiperazin-1 -yl)methyl]-1 -(2,2,2-trifluoroethyl)-1 H-indol-2- yl}prop-2-yn-1 -yl)amino]pyridin-2-yl}-2-methylpropionitrile was prepared.

[0874] LC-MS (ES + ,m / z): 537.3 [(M+H) + ]

[0875] Example 121 Preparation of 2-methyl-2-[5-({3-[4-(morpholin-4-ylmethyl)-1 -(2,2,2- trifluoroethyl)-1 H-indol-2-yl]prop-2-yn-1 -yl}amino)pyridin-2-yl]propionitrile

[0876]

[0877] In a similar manner to that described in Example 1, using 4-bromo-indole, 2-methyl-2-[5-({3-[4-(morpholin-4-ylmethyl)-1 -(2,2,2-trifluoroethyl)-1 H-indol-2- yl]prop-2-yn-1 -yl}amino)pyridin-2-yl]propionitrile was prepared.

[0878] LC-MS (ES + ,m / z): 496.2 [(M+H) + ]

[0879] Example 122Preparation of 2-(5-{[3-(4-{[4-(dimethylamino)piperidin-1-yl]methyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[0880]

[0881] 2-(5-{[3-(4-{[4-(dimethylamino)piperidin-1-yl]methyl}-1-(2,2,2-trifluoroethyl)-1H- indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared in a similar manner as described in Example 1 using 4-bromo-indole.

[0882] LC-MS (ES + ,m / z): 537.3 [(M+H) + ]

[0883] Example 123 Preparation of 2-[5-({3-[4-(hydroxymethyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2- yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[0884]

[0885] 2-[5-({3-[4-(hydroxymethyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino) pyridin-2-yl]-2-methylpropanenitrile was prepared in a similar manner as described in Example 1 using 4-bromo-indole.

[0886] LC-MS (ES + ,m / z): 409.0 [(M-OH) + ]

[0887] Example 124 Preparation of 2-methyl-2-[5-({3-[4-({4-[2-(morpholin-4-yl)-2-oxoethyl]piperazin-1-yl}methyl)- 1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propanenitrile

[0888]

[0889] In a similar manner to that described in Example 1, using 4-bromo-indole, 2-methyl-2-[5-({3-[4-({4-[2-(morpholin-4-yl)-2-oxoethyl]piperazin-1-yl}methyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propionitrile was prepared. LC-MS (ES + m / z): 622.4 [(M+H) + ]

[0890] Example 125 Preparation of 2-(5-{[3-(3-ethyl-7-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile

[0891]

[0892] In a similar manner to that described in Examples 81 and 86, 2-(5-{[3-(3-ethyl-7-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile was prepared.

[0893] LC-MS (ES + m / z): 509.4 [(M+H) + ]

[0894] Example 126 Preparation of methyl 5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridine-2-carboxylate

[0895]

[0896] In a similar manner to that described in Example 28, methyl 5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridine-2-carboxylate was prepared.

[0897] LC-MS (ES + m / z): 501.4 [(M+H) + ]

[0898] Example 127Preparation of N-methyl-5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridine-2-carboxamide

[0899]

[0900] N-methyl-5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2- yl)prop-2-yn-1-yl]amino}pyridine-2-carboxamide was prepared in a similar manner as described in Example 28.

[0901] LC-MS (ES + ,m / z): 530.3 [(M+H) + ]

[0902] Example 128 Preparation of N-(2-hydroxyethyl)-5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridine-2-carboxamide

[0903]

[0904] N-(2-hydroxyethyl)-5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H- indol-2-yl)prop-2-yn-1-yl]amino}pyridine-2-carboxamide was prepared in a similar manner as described in Example 28.

[0905] LC-MS (ES + ,m / z): 530.3 [(M+H) + ]

[0906] Example 129 Preparation of N-(2-methoxyethyl)-5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridine-2-carboxamide

[0907]

[0908] N-(2-methoxyethyl)-5-{[3-(5-{[(oxan-4-yl)amino]methyl}-l-(2,2,2- trifluoroethyl)-lH-indol-2-yl)prop-2-yn-l-yl]amino}pyridine-2-carboxamide was prepared in a similar manner to that described in Example 28.

[0909] LC-MS (ES + ,m / z): 544.4 [(M+H) + ]

[0910] Example 130 Preparation of 2-[(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-l-(2,2,2- trifluoroethyl)-lH-indol-2-yl)prop-2-yn-l-yl]amino}pyridin-2-yl)formamido]acetic acid

[0911]

[0912] 2-[(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-l-(2,2,2-trifluoroethyl)-lH-indol-2- yl)prop-2-yn-l-yl]amino}pyridin-2-yl)formamido]acetic acid was prepared in a similar manner to that described in Example 28.

[0913] LC-MS (ES + ,m / z): 544.3 [(M+H) + ]

[0914] Example 131 Preparation of 5-{[3-(5-{[(oxan-4-yl)amino]methyl}-l-(2,2,2-trifluoroethyl)-lH- indol-2-yl)prop-2-yn-l-yl]amino}pyridine-2-carboxylic acid

[0915]

[0916] 5-{[3-(5-{[(oxan-4-yl)amino]methyl}-l-(2,2,2-trifluoroethyl)-lH-indol-2- yl)prop-2-yn-l-yl]amino}pyridine-2-carboxylic acid was prepared in a similar manner to that described in Example 28.

[0917] LC-MS (ES + ,m / z): 487.4 [(M+H) + ]

[0918] Example 132Preparation of N-(2-methanesulfonyl ethyl)-5-{[3-(5-{[(oxan-4-yl)amino]methyl}- 1 -(2,2,2-trifluoroethyl)- 1 H-indol-2-yl)prop-2-yn- 1 -yl]amino}pyridine-2-carboxamide

[0919]

[0920] N-(2-methanesulfonyl ethyl)-5-{[3-(5-{[(oxan-4-yl)amino]methyl}- 1 -(2,2,2- trifluoroethyl)- 1 H-indol-2-yl)prop-2-yn- 1 -yl]amino}pyridine-2-carboxamide was prepared in a similar manner to that described in Example 28.

[0921] LC-MS (ES + ,m / z): 592.3 [(M+H) + ]

[0922] Example 133 Preparation of 2-[5-({3-[l-(cyanomethyl)-5-{[(oxan-4-yl)amino]methyl}-lH-indol-2- yl]prop-2-yn- 1 -yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[0923]

[0924] 2-[5-({3-[l-(cyanomethyl)-5-{[(oxan-4-yl)amino]methyl}-lH-indol-2-yl]prop-2-yn- 1 -yl}amino)pyridin-2-yl]-2-methylpropanenitrile was prepared in a similar manner to that described in Example 28.

[0925] LC-MS (ES + ,m / z): 467.3 [(M+H) + ]

[0926] Example 134 Preparation of 2-methyl-2-[5-({3-[l-(2-methylpropyl)-5-{[(oxan-4-yl)amino]methyl}- 1 H-indol-2-yl]prop-2-yn- 1 -yl}amino)pyridin-2-yl]propanenitrile

[0927]

[0928] Following the procedure described in Example 28, 2-methyl-2-[5-({3-[1-(2-methylpropyl)-5-{[(oxan-4-yl)amino]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propanenitrile was prepared.

[0929] LC-MS(ES + ,m / z):484.2[(M+H) + ]

[0930] ​ : Preparation of 2-methyl-2-{5-[(3-{4-[(oxacyclohexane-4-yl)amino]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile

[0931]

[0932] 2-Methyl-2-{5-[(3-{4-[(oxan-4-yl)amino]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile was prepared in a manner similar to the method described in Example 1 using 4-bromo-indole.

[0933] LC-MS(ES + ,m / z):496.3[(M+H) + ]

[0934] Example 135 : Preparation of 5-{[3-(5-{[(oxacyclohexane-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridine-2-carbonitrile

[0935]

[0936] Following the procedure described in Example 28, 5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridine-2-carbonitrile was prepared.

[0937] LC-MS(ES + ,m / z):468.3[(M+H) + ]

[0938] Example 136Preparation of N,N-dimethyl-5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridine-2-carboxamide

[0939]

[0940] N,N-dimethyl-5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H- indol-2-yl)prop-2-yn-1-yl]amino}pyridine-2-carboxamide was prepared in a similar manner as described in Example 28.

[0941] LC-MS (ES + ,m / z): 514.3 [(M+H) + ]

[0942] Example 137 Preparation of N-(oxan-4-yl)-5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridine-2-carboxamide

[0943]

[0944] N-(oxan-4-yl)-5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H- indol-2-yl)prop-2-yn-1-yl]amino}pyridine-2-carboxamide was prepared in a similar manner as described in Example 28.

[0945] LC-MS (ES + ,m / z): 570.3 [(M+H) + ]

[0946] Example 138 Preparation of 2-tert-butyl-N-[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]pyrimidin-5-amine

[0947]

[0948] In a similar manner to that described in Example 28, N-(1 -methylpiperidin-4-yl)-5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1 -(2,2,2-trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 - yl]amino}pyridine-2-carboxamide was prepared.

[0949] LC-MS (ES + , m / z): 500.3 [(M+H) + ]

[0950] Example 139 Preparation of N-(1 -methylpiperidin-4-yl)-5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1 -(2,2,2-trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 - yl]amino}pyridine-2-carboxamide

[0951]

[0952] In a similar manner to that described in Example 28, N-(1 -methylpiperidin-4-yl)-5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1 -(2,2,2-trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 - yl]amino}pyridine-2-carboxamide was prepared.

[0953] LC-MS (ES + , m / z): 583.5 [(M+H) + ]

[0954] Example 140 Preparation of N-[6-(1 -cyano-1 -methyl ethyl)pyridin-3-yl]-3-[1 -(2-fluoroethyl)-5-{[(oxan-4-yl)amino]methyl}-1 H-indol-2-yl]prop-2-ynoic amide

[0955]

[0956] In a similar manner to that described in Example 28, N-[6-(1 -cyano-1 -methyl ethyl)pyridin-3-yl]-3-[1 -(2-fluoroethyl)-5-{[(oxan-4-yl)amino]methyl}-1 H-indol-2-yl]prop-2-ynoic amide was prepared.

[0957] LC-MS (ES + , m / z): 488.3 [(M+H) + ]

[0958] Example 141Preparation of 2-(5-{[3-(7-chloro-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1- (2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2- methylpropanenitrile

[0959]

[0960] Preparation of 2-(5-{[3-(7-chloro-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1- (2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2- methylpropanenitrile was prepared in a similar manner to that described in Example 1 using 4-bromo-6-chloro-indole. LC-MS (ES+) + ,m / z): 597.5 [(M+H) + ]

[0961] Example 142: Preparation of 2-(5-{[3-(7-chloro-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1- (2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2- methylpropanenitrile was prepared in a similar manner to that described in Example 1 using 4-bromo-6-chloro-indole. LC-MS (ES+)

[0962]

[0963] Synthesis scheme:

[0964]

[0965] To a solution of 4-bromo-6-fluoro-indole (2 g, 9.38 mmol) in anhydrous tetrahydrofuran (30 mL) cooled in an ice bath was added sodium hydride (0.452 g, 11.3 mmol, 60% in mineral oil). The reaction mixture was stirred under nitrogen for 10 min before adding benzenesulfonyl chloride (1.44 mL, 11.3 mmol). The black solution was allowed to warm to room temperature over 4 h. TLC and LCMS indicated the reaction was complete. A saturated aqueous solution of ammonium chloride was added slowly and the resulting solution was extracted with ethyl acetate (2x). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude solid was triturated with ethanol. The resulting beige solid was collected by filtration to give 4-bromo-6-fluoro-1-(phenylsulfonyl)-1H-indole (2.6 g, 79% yield).

[0966] To a solution of 4-bromo-6-fluoro-l-(phenylsulfonyl)-lH-indole (0.6 g, 1.7 mmol) in anhydrous tetrahydrofuran (10 mL) cooled at -78 °C under a nitrogen atmosphere was added a 1M solution of diisopropylamino lithium in tetrahydrofuran / hexanes (2.7 mL, 2.7 mmol). The reaction mixture was kept at -78 °C for 90 min. Then a solution of iodine (0.427 mg, 1.7 mmol) in tetrahydrofuran (10 mL) was added slowly (over 2 min). The reaction mixture was allowed to warm to room temperature over 14 h. LCMS indicated the reaction was complete. A saturated aqueous solution of ammonium chloride was added slowly and the resulting solution was extracted with ethyl acetate (2x). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated to give 4-bromo-7-fluoro-2-iodo-l-(phenylsulfonyl)-lH-indole. The crude solid was used in the next step without further purification.

[0967] To a suspension of crude 4-bromo-7-fluoro-2-iodo-l-(phenylsulfonyl)-lH-indole (1.7 mmol) in methanol (30 mL) was added a 2M aqueous solution of K2CO3(10 mL, 20 mmol). The reaction mixture was heated at 60 °C for 18 h. The methanol was removed in vacuo. The residual aqueous phase was diluted with a saturated aqueous solution of ammonium chloride and extracted with ethyl acetate (2x). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated. The crude residue was purified by flash column chromatography (ISCO 24 g, eluting with a gradient of ethyl acetate and hexanes). The desired product was eluted with 20% ethyl acetate in hexanes. Fractions were combined to give 4-bromo-6-fluoro-2-iodo-lH-indole as an oil (310 mg, 54% yield).

[0968] To a solution of 4-bromo-6-fluoro-2-iodo-lH-indole (200 mg, 0.59 mmol) in anhydrous tetrahydrofuran (4 mL) was added sodium hydride (48 mg, 1.2 mmol, 60% in mineral oil). The reaction mixture was stirred under a nitrogen atmosphere for 10 min. Then, trifluoro-methanesulfonic acid 2,2,2-trifluoroethyl ester (0.170 mL, 1.2 mmol) was added. The reaction mixture was stirred at room temperature for 14 h. A saturated aqueous solution of ammonium chloride was added slowly and the resulting solution was extracted with ethyl acetate (2x). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated. The crude solid was dried in vacuo to give 4-bromo-7-fluoro-2-iodo-l-(2,2,2-trifluoroethyl)-lH-indole. The crude product was used in the next step without further purification.

[0969] To a scintillation vial containing crude 4-bromo-7-fluoro-2-iodo-l-(2,2,2- trifluoroethyl)-lH-indole (0.59 mmol) was added 2-methyl-2-[5-(2-propynylamino)-2- pyridyl]propanenitrile (207 mg, 0.89 mmol), CuI (22 mg, 0.12 mmol), Pd(PPh3)4(65 mg, 0.006 mmol). Anhydrous tetrahydrofuran (5 mL) was added followed by TEA (0.206 mL, 1.5 mmol) and the reaction mixture was heated at 40 °C for 1 h under a nitrogen atmosphere. The reaction mixture was cooled to room temperature and diluted with 0.5 M EDTA (5 mL) and ethyl acetate (5 mL). The reaction mixture was stirred at room temperature for 30 min and the organic phase was extracted with ethyl acetate (2x). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude material was purified by flash column chromatography (ISCO 20 g, eluting with a gradient of ethyl acetate and hexanes). The desired fractions were eluted with 50% ethyl acetate in hexanes. The fractions were combined to give 2-(5-{3-[4-bromo-7-fluoro-l-(2,2,2-trifluoroethyl)-lH-indol-2-yl]-2- propynylamino}-2-pyridyl)-2-methylpropanenitrile (120 mg, 41% yield).

[0970] To a solution of 2-(5-{3-[4-bromo-7-fluoro-l-(2,2,2-trifluoroethyl)-lH-indol-2-yl]-2- propynylamino}-2-pyridyl)-2-methylpropanenitrile (120 mg, 0.24 mmol) in anhydrous dimethylformamide (2 mL) in a scintillation vial was added Johnphos (4 mg, 0.012 mmol), sodium carbonate (25 mg, 0.24 mmol), palladium acetate (3 mg, 0.012 mmol), triethylsilane (0.114 mL, 0.72 mmol), and tert-butyl isocyanide (0.055 mL, 0.49 mmol). The reaction mixture was heated at 65 °C for 10 h under a nitrogen atmosphere. To the reaction mixture was added 0.5 M EDTA (4 mL) and ethyl acetate (10 mL) and it was stirred at room temperature for 1 h. The organic phase was extracted with ethyl acetate (2x). The combined organic phases were washed with brine (3x), dried over anhydrous sodium sulfate, and then concentrated. The crude material was purified by flash column chromatography (ISCO 12 g, eluting with a gradient of ethyl acetate and hexanes) to give recovered starting material (60 mg) and the desired 2-(5-{3-[7-fluoro-4-formyl-l-(2,2,2-trifluoroethyl)-lH-indol-2-yl]-2- propynylamino}-2-pyridyl)-2-methylpropanenitrile (40 mg, 75% yield based on recovery of starting material) as the more polar compound.

[0971] To a solution of 2-(5-{3-[7-fluoro-4-formyl-1-(2,2,2-trifluoroethyl)-1H- indol-2-yl]-2-propynylamino}-2-pyridyl)-2-methylpropanenitrile (20 mg, 0.045 mmol) in dichloromethane (1 mL) was added 4-pyrrolidin-1-yl-piperidine (14 mg, 0.09 mmol) and sodium triacetoxyborohydride (28 mg, 0.14 mmol). The reaction mixture was stirred at room temperature for 3 h. The reaction mixture was diluted with ethyl acetate and washed with aqueous sodium bicarbonate solution. The organic phase was extracted with ethyl acetate (2x). The combined organic phases were washed with brine (3x), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude material was purified by preparative HPLC (Combiflash™ EZ-prep system eluting with a gradient of water (0.1% formic acid) and acetonitrile (0.1% formic acid)) to give 2-(5-{[3-(6-fluoro-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile (5 mg, 19% yield). LC-MS (ES+, m / z): 581.1 [(M+H) + ]

[0972] Example 143 Preparation of 2-(5-{[3-(1-ethyl-5-{[(oxan-4-yl)amino]methyl}-1H-pyrrolo[2,3- c]pyridin-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[0973]

[0974] 2-(5-{[3-(1-ethyl-5-{[(oxan-4-yl)amino]methyl}-1H-pyrrolo[2,3-c]pyridin-2-yl)prop-2- yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared in a similar manner to that described in Example 28 using 1,6-diaza-1H-indene-5-carbaldehyde as the starting material.

[0975] LC-MS (ES+, m / z): 511.1 [(M+H) + ]

[0976] Example 144 Preparation of 2-methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-pyrrolo[2,3-c]pyridin-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile

[0977]

[0978] 2-methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-l-(2,2,2- trifluoroethyl)-lH-pyrrolo[2,3-c]pyridin-2-yl)prop-2-yn-l-yl]amino}pyridin-2- yl)propanenitrile was prepared in a similar manner to that described in Example 28 using l,6-diaza-lH-indene-5-carbaldehyde as the starting material.

[0979] LC-MS (ES + ,m / z): 457.3 [(M+H) + ]

[0980] Example 145 Preparation of 2-(5-{[3-(5-{[4-(dimethylamino)piperidin-l-yl]methyl}-l-(2,2,2- trifluoroethyl)-lH-pyrrolo[2,3-c]pyridin-2-yl)prop-2-yn-l-yl]amino}pyridin-2- yl)-2-methylpropanenitrile

[0981]

[0982] 2-(5-{[3-(5-{[4-(dimethylamino)piperidin-l-yl]methyl}-l-(2,2,2- trifluoroethyl)-lH-pyrrolo[2,3-c]pyridin-2-yl)prop-2-yn-l-yl]amino}pyridin-2- yl)-2-methylpropanenitrile was prepared in a similar manner to that described in Example 28 using l,6-diaza-lH-indene-5-carbaldehyde as the starting material.

[0983] LC-MS (ES + ,m / z): 538.1 [(M+H) + ]

[0984] Example 146 Preparation of 2-(5-{[3-(7-chloro-4-{[4-(pyrrolidin-l-yl)piperidin-l- yl]methyl}-l-(2,2,2-trifluoroethyl)-lH-pyrrolo[2,3-c]pyridin-2-yl)prop-2-yn-l- yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[0985]

[0986] Synthesis scheme:

[0987]

[0988] Dissolve 5-bromo-2-chloro-3-nitro-pyridine (20.2 g, 85.07 mmol, 1 eq) in dry tetrahydrofuran (100 mL) under nitrogen atmosphere and cool the solution to -78 °C. To this solution, add bromo(vinyl)magnesium (1 M, 340 mL, 4 eq) and stir the reaction mixture at -78 °C for 4 h. LCMS analysis shows the reaction to be complete. Quench the reaction with saturated ammonium chloride (150 mL). Extract the aqueous phase with ethyl acetate (3 x 150 mL) and dry the combined organic layers over magnesium sulfate, filter and concentrate. Purify the crude product by flash column chromatography (silica gel, eluting with a 1 / 1 mixture of ethyl acetate and petroleum ether) to obtain 4-bromo-7-chloro-1H-pyrrolo[2,3-c]pyridine as a yellow solid (4.12 g, 21% yield).

[0989] To a mixture of sodium hydride (1.16 g, 28.98 mmol, 60% purity, 3.15 eq) stirred in tetrahydrofuran (20 mL) at 0 °C, add dropwise 4-bromo-7-chloro-1H-pyrrolo[2,3- c]pyridine (2.13 g, 9.20 mmol, 1 eq) dissolved in tetrahydrofuran (10 mL). After the addition, stir the mixture at 0 °C for 1 h and add dropwise benzene sulfonyl chloride (2.06 g, 11.68 mmol, 1.5 mL, 1.27 eq) dissolved in tetrahydrofuran (10 mL) at 0 °C. Stir the resulting mixture at 0 °C for 2 h and quench by adding saturated ammonium chloride (50 mL) at 0 °C.

[0990] Extract the resulting reaction mixture with ethyl acetate (3 x 50 mL). Wash the combined organic layers with water (50 mL), dry over anhydrous sodium sulfate, filter and concentrate in vacuo. Purify the crude product by flash column chromatography (silica gel, eluting with dichloromethane / petroleum ether: 1 / 1) to obtain 1-(benzenesulfonyl)-4-bromo-7-chloro-pyrrolo[2,3-c]pyridine as a light yellow solid (2.03 g, 59% yield).

[0991] To a solution of 1-(phenylsulfonyl)-4-bromo-7-chloro-pyrrolo[2,3-c]pyridine (400 mg, 1.08 mmol) in tetrahydrofuran (20 mL) cooled to -78 °C was added dropwise diisopropylamino lithium (2 M, 1.62 mL). After addition, the mixture was stirred at -78 °C temperature for 1 h, then iodine (312 mg, 1.23 mmol) in tetrahydrofuran (12 mL) was added dropwise at -78 °C. The resulting mixture was stirred at -78 °C for 1 h, before partitioning between saturated ammonium chloride (40 mL) and ethyl acetate (40 mL). The reaction mixture was then extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with water (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluted with dichloromethane / petroleum ether: 1 / 1) to give 1-(phenylsulfonyl)-4-bromo-7-chloro-2-iodo-pyrrolo[2,3-c]pyridine (240 mg, 45% yield) as a light yellow solid.

[0992] A mixture of 1-(phenylsulfonyl)-4-bromo-7-chloro-2-iodo-pyrrolo[2,3-c]pyridine (600 mg, 1.21 mmol) and potassium carbonate (500 mg, 3.62 mmol) dissolved in methanol (20 mL) was stirred at 80 °C for 4 h under nitrogen atmosphere. The solution was concentrated under reduced pressure to give a crude product. The crude product was purified by preparative thin layer chromatography (dichloromethane: petroleum ether 1:1) to give 4-bromo-7-chloro-2-iodo-1H-pyrrolo[2,3-c]pyridine (350 mg, 81% yield) as a white solid.

[0993] 4-Bromo-7-chloro-2-iodo-1H-pyrrolo[2,3-c]pyridine was used in the methods described in examples 1 and 49 to prepare 2-(5-{[3-(7-chloro-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1-(2,2,2- trifluoroethyl)-1H-pyrrolo[2,3-c]pyridin-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2- methylpropanenitrile.

[0994] LC-MS (ES + ,m / z): 598.2 [(M+H) + ]

[0995] The following compounds were prepared in a similar manner to that described in example 142:

[0996]

[0997]

[0998] Starting from 2-iodo-1H-indole-4-carboxylic acid methyl ester and using similar procedures as described in Examples 1 and 49, the following compounds were prepared:

[0999]

[1000]

[1001] Example 166 Preparation of 2-methyl-2-[5-({3-[1-(oxan-2-ylmethyl)-4-{[4-(pyrrolidin-1- yl)piperidin-1-yl]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propanenitrile

[1002] In a similar manner to that described in Examples 1 and 49, 2-iodo-1H-indole-4- carboxaldehyde was reacted with 2-(bromomethyl)oxirane, followed by coupling with 2-methyl-2-{5-[(prop-2-yn-1-yl)amino]pyridin-2-yl}propanenitrile. Reductive amination of the resulting intermediate with 4-(pyrrolidin-1-yl)piperidine gave 2-methyl-2-[5-({3-[1-(oxan-2-ylmethyl)-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propanenitrile.

[1003] LC-MS (ES + ,m / z): 537.4 [(M+H) + ]

[1004] Example 167 Preparation of 2-(5-{[3-(3-ethyl-6-{[(oxan-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1- yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[1005]

[1006] In a similar manner to that described in Examples 80 and 86, 2-(5-{[3-(3-ethyl-6-{[(oxan-4- yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared.

[1007] LC-MS (ES + ,m / z): 456.2 [(M+H) + ]

[1008] Example 168 : Preparation of 2-methyl-2-(5-{[3-(6-{[(oxan-4-yl)amino]methyl}-3-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propionitrile

[1009]

[1010] Following the procedures described in Examples 80 and 86, 2-methyl-2-(5-{[3-(6-{[(oxan-4-yl)amino]methyl}-3-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile was prepared.

[1011] LC-MS(ES + ,m / z):510.3[(M+H) + ]

[1012] Example 169 : Preparation of 2-(5-{[3-(1-acetyl-3-ethyl-6-{[(oxacyclohexane-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile

[1013]

[1014] Following the procedures described in Examples 80 and 86, 2-(5-{[3-(1-acetyl-3-ethyl-6-{[(oxan-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile was prepared.

[1015] LC-MS(ES + ,m / z):498.3[(M+H) + ]

[1016] Example 170 : Preparation of 2-(5-{[3-(3-ethyl-6-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile

[1017]

[1018] In a similar manner to that described in Examples 80 and 86, 2-(5-{[3-(3-ethyl-6-{[(1- methylpiperidin-4-yl)amino]methyl}-1 H-indol-2-yl)prop-2-yn-1 -yl]amino}pyridin-2-yl)-2- methylpropanenitrile was prepared.

[1019] LC-MS (ES + , m / z): 469.3 [(M+H) + ]

[1020] Example 171 Preparation of 2-{5-[(3-{6-chloro-4-[(1-methylpiperidin-4-yl)amino]-1-(2,2,2- trifluoroethyl)-1 H-indol-2-yl}prop-2-yn-1 -yl)amino]pyridin-2-yl}-2-methylpropanenitrile

[1021]

[1022] In a similar manner to that described in Examples 80 and 86, 2-methyl-2-(5-{[3-(6-{[(1- methylpiperidin-4-yl)amino]methyl}-3-(2,2,2-trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 - yl]amino}pyridin-2-yl)propanenitrile was prepared.

[1023] LC-MS (ES + , m / z): 523.4 [(M+H) + ]

[1024] Example 172 Preparation of 2-{5-[(3-{6-chloro-4-[(1-methylpiperidin-4-yl)amino]-1-(2,2,2- trifluoroethyl)-1 H-indol-2-yl}prop-2-yn-1 -yl)amino]pyridin-2-yl}-2-methylpropanenitrile

[1025]

[1026] Synthesis scheme:

[1027]

[1028] Sodium hydride (1.39 g, 34.70 mmol, in mineral oil 60%) was added to a 0°C solution of 4-bromo-6-chloro-indole (4 g, 17.35 mmol) in dimethylformamide (60 mL). The reaction was stirred at 0°C for 1 h and to the reaction mixture was added 2- (trimethylsilyl)ethoxymethyl chloride (SEMCl, 4.34 g, 26.03 mmol). The reaction was stirred at 20°C for 1 h and a saturated solution of ammonium chloride (50.0 mL) was added. The reaction mixture was extracted with dichloromethane (3 x 100 mL). The combined organic layers were washed with water (3 x 100 mL), brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluting with petroleum ether / ethyl acetate: 300 / 1 to 150 / 1) to give 4-bromo-6-chloro-1-{[2- (trimethylsilyl)ethoxy]methyl}-1H-indole as a yellow oil (90% purity).

[1029] To a solution of 4-bromo-6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-indole (1.50 g, 3.74 mmol, 90% purity) and 1-methylpiperidin-4-amine (1.5 g, 13.13 mmol) dissolved in toluene (30 mL) was added t-BuONa (720 mg, 7.48 mmol), JohnPhos (302 mg, 1.01 mmol) and Pd2(dba)3(308 mg, 336.60 μmol). The reaction was then stirred at 80°C in a sealed tube for 12 h. The reaction mixture was treated with water (10 mL) and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluting with dichloromethane / methanol: 50 / 1) to give 6-chloro-N-(1-methylpiperidin-4-yl)-1-{[2- (trimethylsilyl)ethoxy]methyl}-1H-indol-4-amine as a yellow oil (1.6 g, 33% yield, 90% purity).

[1030] To a solution of 6-chloro-N-(1-methylpiperidin-4-yl)-1-{[2-(trimethylsilyl)-ethoxy]methyl}-1H-indol-4-amine (1.55 g, 3.55 mmol, 90% purity) in t-BuOH (21 mL) was added di-tert-butyl dicarboxylate (19 g, 87.06 mmol). The reaction was stirred at 80 °C for 24 h, cooled to room temperature and concentrated. The crude product was purified by flash column chromatography (silica gel eluting with dichloromethane / methanol: 100 / 1) to give tert-butyl N-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-indol-4-yl)-N-(1- methylpiperidin-4-yl)carbamate (1.20 g, 2.31 mmol, 65% yield, 95% purity) as a yellow oil.

[1031] A solution of tert-butyl N-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-indol-4-yl)-N-(1- methylpiperidin-4-yl)carbamate (470 mg, 665.80 μmol) dissolved in TBAF (1M in tetrahydrofuran, 8 mL) was stirred at 100 °C for 3 h, cooled to room temperature and concentrated. The crude residue was dissolved in dichloromethane (60 mL) and washed with water (20 mL), brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give 470 mg of crude tert-butyl N-(6-chloro-1H-indol-4-yl)-N-(1-methylpiperidin-4-yl)carbamate which was used without further purification.

[1032] To a 0 °C solution of crude tert-butyl N-(6-chloro-1H-indol-4-yl)-N-(1-methylpiperidin-4-yl)carbamate (470 mg, 1.29 mmol) in tetrahydrofuran (10 mL) was added sodium hydride (113 mg, 2.82 mmol, 60% in mineral oil). The resulting mixture was stirred at 0 °C for 30 min. To the reaction mixture was added benzenesulfonyl chloride (345 mg, 1.95 mmol). The ice bath was removed, the reaction mixture was allowed to warm to room temperature and the reaction mixture was stirred at room temperature for 30 min. The reaction was cooled to 0 °C and quenched with water (10 mL). The mixture was extracted with ethyl acetate (2 x 20 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel eluting with dichloromethane / methanol: 50 / 1) to give tert-butyl N-[1-(benzenesulfonyl)-6-chloro-1H-indol-4-yl]-N-(1-methylpiperidin-4-yl)- carbamate (800 mg, 88% yield, 72% purity) as a yellow oil.

[1033] To a -65 °C solution of N-[1-(phenylsulfonyl)-6-chloro-1H-indol-4-yl]-N-(1- methylpiperidin-4-yl)-carbamic acid tert-butyl ester (260 mg, 371.40 pmol, 72% purity) in tetrahydrofuran (6 mL) was added lithium diisopropylamide (2 M in tetrahydrofuran, 520 pL). The reaction was stirred at -65 °C for 1.5 h and to the reaction was added iodine solution (144 mg in 1.5 mL tetrahydrofuran, 567.35 pmol) dropwise. The reaction was stirred at -65 °C for 0.5 h and saturated solution of ammonium chloride (1.0 mL) was added. The reaction mixture was warmed to 20 °C and extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give crude N-[1-(phenylsulfonyl)-6-chloro-2-iodo-1H-indol-4-yl]-N-(1- methylpiperidin-4-yl)-carbamic acid tert-butyl ester (700 mg) as a yellow solid. This crude product was used directly in the next step without further purification.

[1034] To a solution of N-[1-(phenylsulfonyl)-6-chloro-2-iodo-1H-indol-4-yl]-N-(1- methylpiperidin-4-yl)-carbamic acid tert-butyl ester (900 mg) in methanol (15 mL) was added aqueous potassium carbonate solution (0.5 mL, 2 M). The reaction was then stirred at 80 °C for 30 min, cooled to room temperature, and concentrated. The residue was extracted with ethyl acetate (3 x 10 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative thin layer chromatography (silica gel, eluted with dichloromethane / methanol: 10 / 1 containing 0.1% triethylamine) to give N-(6-chloro-2-iodo-1H-indol-4-yl)-N-(1- methylpiperidin-4-yl)-carbamic acid tert-butyl ester (250 mg, 41% yield, 80% purity) as a yellow solid.

[1035] 2-{5-[(3-{6-chloro-4-[(1-methylpiperidin-4-yl)amino]-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}-2- methylpropanenitrile was prepared in a similar manner as described in Example 28 using N-(6-chloro-2-iodo-1H-indol-4-yl)-N-(1-methylpiperidin-4-yl)carbamic acid tert-butyl ester.

[1036] LC-MS (ES + ,m / z): 543.3 [(M+H) + ]

[1037] Example 173Preparation of 2-(3-{[6-(1-cyano-1-methylethyl)pyridin-3-yl]amino}prop-1-yn-1-yl)-6- fluoro-N-(1-methylpiperidin-4-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-4-carboxamide

[1038]

[1039] To a solution of 2-[5-({3-[6-fluoro-4-formyl-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2- yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile (250 mg, 553.78 μmol, Example 142) in tetrahydrofuran (4 mL) and water (1 mL) was added 2-methylbut-2-ene (388 mg, 5.54 mmol), NaH2PO4(133 mg, 1.11 mmol) and NaClO2(400 mg, 4.43 mmol, added in small portions). The resulting reaction mixture was stirred at 25 °C for 1 h, poured into 30 mL of aqueous ammonium chloride and extracted with ethyl acetate (3 x 10 mL). The combined organic layers were washed with water (3 x 10 mL), brine (10 mL), dried over anhydrous sodium sulfate and concentrated. The residue was washed with 2 mL of 1:1 dichloromethane and petroleum ether to give 2-(3-{[6-(1-cyano-1-methylethyl)pyridin-3-yl]amino}prop-1-yn-1-yl)-6-fluoro-1-(2,2,2- trifluoroethyl)-1H-indole-4-carboxylic acid (120 mg, 38% yield, 80% purity) as a yellow solid.

[1040] To a solution of 2-(3-{[6-(1-cyano-1-methylethyl)pyridin-3-yl]amino}prop-1-yn-1-yl)-6-fluoro-1-(2,2,2-trifluoroethyl)-1H-indole-4-carboxylic acid (50 mg, 87.26 pmol, 80% purity) in dichloromethane (3 mL) was added diisopropylethylamine (56 mg, 436.29 pmol) and HATU (50 mg, 130.89 pmol). Subsequently, to the reaction mixture was added 1-methylpiperidin-4-amine (20 mg, 174.52 pmol). The resulting reaction mixture was stirred at 25 °C for 30 min, poured into 30 mL of aqueous ammonium chloride solution and extracted with ethyl acetate (3 x 10 mL). The combined organic layers were washed with water (3 x 10 mL), brine (10 mL), dried over anhydrous sodium sulfate and concentrated. The crude residue was purified by reverse phase preparative high performance liquid chromatography (HPLC) to give 2-(3-{[6-(1-cyano-1-methylethyl)pyridin-3-yl]amino}prop-1-yn-1-yl)-6-fluoro-N-(1-methylpiperidin-4-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-4-carboxamide (13.7 mg, 26% yield) as a white solid.

[1041] LC-MS (ES + ,m / z): 555.2 [(M+H) + ]

[1042] Example 174 Preparation of 2-[5-({3-[6-fluoro-4-(4-methylpiperazine-1-carbonyl)-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[1043]

[1044] 2-[5-({3-[6-fluoro-4-(4-methylpiperazine-1-carbonyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2- yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile was prepared in a similar manner as described in the method described in Example 173.

[1045] LC-MS (ES + ,m / z): 541.2 [(M+H) + ]

[1046] Example 175Preparation of 6-fluoro-2-{3-[(6-methanesulfonylpyridin-3-yl)amino]prop-1-yn-1-yl}-N-(1- methylpiperidin-4-yl)-1-(2,2,2-trifluoroethyl)-1 H-indol-4-amine

[1047]

[1048] Using 4-bromo-6-fluoro-indole, 6-fluoro-2-{3-[(6-methanesulfonylpyridin-3-yl)amino]prop-1-yn-1-yl}-N-(1- methylpiperidin-4-yl)-1-(2,2,2-trifluoroethyl)-1 H-indol-4-amine was prepared in a similar manner as described in Example 172.

[1049] LC-MS (ES + ,m / z): 538.2 [(M+H) + ]

[1050] Example 176 Preparation of 2-{5-[(3-{6-fluoro-4-[(1-methylpiperidin-4-yl)amino]-1-(2,2,2-trifluoroethyl)-1 H-indol-2- yl}prop-2-yn-1-yl)amino]pyridin-2-yl}-2-methylpropanenitrile

[1051]

[1052] Using 4-bromo-6-fluoro-indole, 2-{5-[(3-{6-fluoro-4-[(1-methylpiperidin-4-yl)amino]-1-(2,2,2-trifluoroethyl)-1 H-indol-2- yl}prop-2-yn-1-yl)amino]pyridin-2-yl}-2-methylpropanenitrile was prepared in a similar manner as described in Example 172.

[1053] LC-MS (ES + ,m / z): 527.2 [(M+H) + ]

[1054] Example 177 Preparation of 5-[(3-{6-fluoro-4-[(1-methylpiperidin-4-yl)amino]-1-(2,2,2-trifluoroethyl)-1 H-indol-2-yl}prop-2-yn-1-yl)amino]-N-(pyridin-3-yl)pyridine-2-carboxamide

[1055]

[1056] Using 4-bromo-6-fluoro-indole, 5-[(3-{6-fluoro-4-[(1-methylpiperidin-4-yl)amino]-1- (2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]-N-(pyridin-3-yl)pyridine-2- carboxamide was prepared in a similar manner to that described in Example 172.

[1057] LC-MS (ES+, m / z): 511 [(M+H)+] + , m / z): 580.3 [(M+H) + ]

[1058] Example 178 Preparation of 2-methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile

[1059]

[1060] 2-methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H- pyrrolo[2,3-b]pyridin-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile was prepared in a similar manner to that described in Example 142.

[1061] LC-MS (ES+, m / z): 511 [(M+H)+]

[1062] Example 179 Preparation of 2-methyl-2-{5-[(3-{4-[(1-methylpiperidin-4-yl)amino]-1-(2,2,2- trifluoroethyl)-1H-pyrrolo[2,3-b]pyridin-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}propanenitrile

[1063]

[1064] Synthesis scheme:

[1065]

[1066] To a 0 °C mixture of sodium hydride (4.41 g, 110.34 mmol, 60% in mineral oil) in tetrahydrofuran (20 mL) was added a solution of 4-nitro-lH-pyrrolo[2,3-b]pyridine (6 g, 36.78 mmol) in tetrahydrofuran (5 mL). The mixture was stirred at 0 °C for 1 h, then benzenesulfonyl chloride (9.74 g, 55.17 mmol, 7.06 mL) was added. The mixture was stirred at 0 °C for 1 h, then quenched by the addition of saturated aqueous ammonium chloride solution. The reaction mixture was extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluting with petroleum ether / ethyl acetate: 30 / 1 to 5:1) to give l-(benzenesulfonyl)-4-nitro-lH-pyrrolo[2,3-b]pyridine (4.90 g, 44% yield) as a yellow solid.

[1067] A solution of l-(benzenesulfonyl)-4-nitro-pyrrolo[2,3-b]pyridine (1 g, 3.30 mmol) in tetrahydrofuran (10 mL) was added to a -78 °C solution of lithium 2,2,6,6-tetramethylpiperidide (1.70 g, 11.55 mmol) in tetrahydrofuran (27 mL). The mixture was stirred at -78 °C for 1 h, then iodine solution (1.84 g, 7.26 mmol, dissolved in 1.46 mL tetrahydrofuran) was added. After stirring at -78 °C for 1 h, the reaction mixture was quenched by the addition of saturated aqueous ammonium chloride solution. The resulting mixture was extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give crude l-(benzenesulfonyl)-2-iodo-4-nitro-lH-pyrrolo[2,3-b]pyridine (600 mg), which was used without further purification.

[1068] To a solution of l-(benzenesulfonyl)-2-iodo-4-nitro-lH-pyrrolo[2,3-b]pyridine (2 g, 4.66 mmol) in methanol (20 mL) was added potassium carbonate (2.58 g, 18.64 mmol), and the resulting reaction mixture was stirred at 80 °C for 1 h. The solids were filtered off, and the resulting filtrate was concentrated. The crude product was purified by flash column chromatography (silica gel, eluting with petroleum ether / ethyl acetate: 1 / 1) to give the product 2-iodo-4-nitro-lH-pyrrolo[2,3-b]pyridine (435 mg, 1.51 mmol, 32% yield) as a yellow solid.

[1069] To a 0 °C mixture of sodium hydride (187 mg, 4.68 mmol, 60% in mineral oil) in tetrahydrofuran (20 mL) was added 2,2,2-trifluoroethyl trifluoromethanesulfonate (1.81 g, 7.80 mmol). The resulting reaction mixture was stirred at 0 °C for 1 h and to the reaction mixture was added 2-iodo-4-nitro-1H-pyrrolo[2,3-b]pyridine (451 mg, 1.56 mmol). The mixture was stirred at 0 °C for 1 h and quenched by the addition of saturated aqueous ammonium chloride solution. The resulting mixture was extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluting with petroleum ether / ethyl acetate: 30 / 1 to 5 / 1) to give 2-iodo-4-nitro-1-(2,2,2-trifluoroethyl)-1H-pyrrolo[2,3-b]pyridine (260 mg, 45%) as a yellow oil.

[1070] To a solution of 2-iodo-4-nitro-1-(2,2,2-trifluoroethyl)-1H-pyrrolo[2,3-b]pyridine (235 mg, 633.34 pmol) in acetic acid (5 mL) was added Fe (142 mg, 2.53 mmol) and the mixture was stirred at 50 °C for 2 h. The reaction mixture was then poured into water and extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluting with petroleum ether / ethyl acetate: 30 / 1 to 5 / 1) to give 2-iodo-1-(2,2,2-trifluoroethyl)-1H-pyrrolo[2,3-b]pyridin-4-amine (100 mg, 46% yield) as a yellow oil.

[1071] To a solution of 2-methyl-2-{5-[(prop-2-yn-1 -yl)amino]pyridin-2-yl}propanenitrile (180 mg, 762.30 pmol) in dimethyl sulfoxide (2 mL) was added N-isopropylpropan-2-amine (536 pL, 3.81 mmol). Then, to the reaction mixture was added 2-iodo-1 -(2,2,2-trifluoroethyl)pyrrolo[2,3- b]pyridin-4-amine (130 mg, 381.15 pmol), CuI (7 mg, 38.11 pmol) and tetrakis(triphenylphosphine)palladium(0) (44 mg, 38.11 pmol). The reaction was stirred at 25 °C under nitrogen atmosphere for 2 h and poured into water (5 mL). The reaction mixture was extracted with ethyl acetate (3 x 20 mL) and the combined organic layers were stirred with a saturated ethylenediaminetetraacetic acid (EDTA) solution (-20 mL) for 1 h. The combined organic layers were washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by preparative thin layer chromatography (silica gel, eluting with petroleum ether / ethyl acetate: 1 / 2) to afford 2-[5-[3-[4-amino-1 -(2,2,2-trifluoroethyl)pyrrolo[2,3- b]pyridin-2-yl]prop-2-ynylamino]-2-pyridyl]-2-methyl-propanenitrile (130 mg, 83% yield) as a yellow solid.

[1072] To a solution of 1 -methylpiperidin-4-one (11 mg, 97 pmol, 11 pL) in ethanol (2 mL) and acetic acid (7.3 mg, 121.25 pmol, 7 pL) was added 2-[5-[3-[4-amino-1 -(2,2,2-trifluoroethyl)pyrrolo[2,3- b]pyridin-2-yl]prop-2-ynylamino]-2-pyridyl]-2-methyl-propanenitrile (10 mg, 24.25 pmol) and molecular sieves (30 mg). The mixture was stirred at 50 °C for 12 h and sodium cyanoborohydride (15 mg, 242.50 pmol) was added. The resulting reaction mixture was stirred at 50 °C for 2 h and then diluted with water (-10 mL). The reaction mixture was extracted with ethyl acetate (3 x 10 mL) and the combined organic layers were washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by preparative HPLC to afford 2-methyl-2-{5-[(3-{4-[(1 - methylpiperidin-4-yl)amino]-1 -(2,2,2-trifluoroethyl)-1 H-pyrrolo[2,3-b]pyridin-2-yl}prop-2-yn-1 - yl)amino]pyridin-2-yl}propanenitrile (3 mg, 22% yield). LC-MS (ES+, m / z): 510.3 [(M+H)+]

[1073] Example 180Preparation of 2-(5-{[3-(7-chloro-1-ethyl-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H- indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[1074]

[1075] 2-(5-{[3-(7-chloro-1-ethyl-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2-yl)prop-2- yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared in a similar manner to that described in Example 141.

[1076] LC-MS (ES+, m / z): 543.4 [(M+H)+]

[1077] Example 181 Preparation of 2-(5-{[3-(7-chloro-5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2- yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[1078]

[1079] 2-(5-{[3-(7-chloro-5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2- yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared from 5-bromo-7-chloro-1H- indole using a similar method to that described in Example 1.

[1080] LC-MS (ES+, m / z): 544.3 [(M+H)+]

[1081] Example 182 Preparation of 2-(5-{[3-(7-chloro-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[1082]

[1083] 2-(5-{[3-(7-chloro-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared starting from 5-bromo-7-chloro-1H-indole using a similar procedure as described in Example 1.

[1084] LC-MS (ES+, m / z): 557.3 [(M+H)+]

[1085] Example 183 Preparation of 2-{5-[(3-{7-fluoro-4-[(1-methylpiperidin-4-yl)amino]-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}-2-methylpropanenitrile

[1086]

[1087] 2-{5-[(3-{7-fluoro-4-[(1-methylpiperidin-4-yl)amino]-1-(2,2,2-trifluoroethyl)- 1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}-2-methylpropanenitrile was prepared starting from 4-bromo-7-fluoro-1H-indole following a similar procedure as described in Example 172.

[1088] LC-MS (ES+, m / z): 527.3 [(M+H)+]

[1089] Example 184 Preparation of 2-(5-{[3-(7-fluoro-5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile

[1090]

[1091] 2-(5-{[3-(7-fluoro-5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H- indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropanenitrile was prepared starting from 5-bromo-7-fluoro-1H-indole using a similar procedure as described in Example 1.

[1092] LC-MS (ES + ,m / z): 528.3 [(M+H) + ]

[1093] Example 185Preparation of 2-methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-1,3-benzimidazol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile

[1094]

[1095] 2-methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-1,3- benzimidazol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile was prepared starting from 5-bromo-7-fluoro-1H-indole and using a similar procedure as described in Example 1.

[1096] LC-MS (ES + ,m / z): 541.3 [(M+H) + ]

[1097] Example 186 Preparation of 2-methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-1,3- benzimidazol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile

[1098]

[1099] Synthesis scheme

[1100]

[1101] To a solution of 4-fluoro-3-nitro-benzaldehyde (6 g, 35.48 mmol) in dimethylformamide (10 mL) was added Cs2CO3(23.12 g, 70.96 mmol). Then, to the reaction mixture was added 2,2,2-trifluoroethylamine (3.87 g, 39.03 mmol, 3.07 mL). The reaction mixture was stirred at 100 °C for 1 h, cooled to room temperature and concentrated. To the mixture was added water (500 mL) and the resulting mixture was extracted with ethyl acetate (3 x 500 mL). The combined organic layers were washed with water (3 x 300 mL), brine (300 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel eluted with petroleum ether / tetrahydrofuran: 100 / 1). The obtained yellow solid was further washed with petroleum ether / dichloromethane / methanol: 30 / 10 / 2 mL and filtered to give 3-nitro-4-[(2,2,2-trifluoroethyl)amino]benzaldehyde (2.50 g, 80% purity).

[1102] A solution of 3-nitro-4-[(2,2,2-trifluoroethyl)amino]benzaldehyde (2.80 g, 9.03 mmol, 80% purity) and 4-amino-pyran (2.74 g, 27.09 mmol) in tetrahydrofuran (30 mL) and methanol (30 mL) was stirred at 20 °C for 2 h. To the solution was added NaBH4(1.02 g, 27.09 mmol). The reaction mixture was stirred at 20 °C for 1 h, poured into water (20 mL) and concentrated. The residue was extracted with dichloromethane / methanol (5 x 40 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude compound was purified by flash column chromatography (silica gel, eluting with dichloromethane / methanol: 150 / 1 to 50 / 1) to give N-({3-nitro-4-[(2,2,2-trifluoroethyl)amino]phenyl}methyl)oxane-4-amine (3.01 g, 94% yield) as a yellow solid.

[1103] To a solution of N-({3-nitro-4-[(2,2,2-trifluoroethyl)amino]phenyl}methyl)oxane-4- amine (3 g, 8.46 mmol) in dichloromethane (70 mL) was added triethylamine (3.52 mL, 25.38 mmol and di-tert-butyl dicarboxylate (2.03 g, 9.31 mmol). The reaction mixture was stirred at 20 °C for 24 h, water (20 mL) was added to the reaction mixture and the resulting mixture was extracted with dichloromethane (3 x 50 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluting with dichloromethane / methanol: 200 / 1) to give tert-butyl N-({3-nitro-4-[(2,2,2-trifluoroethyl)amino]phenyl}methyl)-N-(oxan-4- yl)carbamate (3.90 g, 96% yield) as a yellow solid.

[1104] To a solution of tert-butyl N-({3-nitro-4-[(2,2,2-trifluoroethyl)amino]phenyl}methyl)-N- (oxan-4-yl)carbamate (3.50 g, 7.27 mmol) in t-BuOH (80 mL) and tetrahydrofuran (18 mL) was added 10% palladium on carbon (500 mg, 7.27 mmol). The mixture was stirred at 20 °C under hydrogen atmosphere (15 psi) for 30 min. The reaction was filtered and the filtrate was concentrated to give tert-butyl N-({3-amino-4-[(2,2,2-trifluoroethyl)amino]phenyl}methyl)-N- (oxan-4-yl)carbamate (3 g) as a white solid. The crude product was used in the next step without further purification.

[1105] To a solution of tert-butyl N-({3-amino-4-[(2,2,2-trifluoroethyl)amino]phenyl}methyl)- N-(oxan-4-yl)carbamate (1 g, 2.23 mmol) in ethanol (30 mL) was added potassium hydroxide (626 mg, 11.15 mmol). To the reaction was then added carbon disulfide (6.30 g, 82.74 mmol, 5 mL) and the reaction mixture was stirred at 80 °C for 1 h. After concentration, the pH of the crude mixture was adjusted to 9 with 12 M HC1. The reaction mixture was extracted with dichloromethane (3 x 50 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluted with dichloromethane) to give tert-butyl N-(oxan-4-yl)-N-{[2-mercapto-1-(2,2,2- trifluoroethyl)-1H-1,3-benzimidazol-5-yl]methyl}carbamate (1 g, 99% yield).

[1106] To a 20 °C solution of tert-butyl N-(oxan-4-yl)-N-{[2-mercapto-1-(2,2,2- trifluoroethyl)-1H-1,3-benzimidazol-5-yl]methyl}carbamate (150 mg, 329.97 pmol) in acetic acid (1.70 mL) and HBr (69 pL, 442.16 pmol, 35% solution in acetic acid) was added dropwise bromine (1.32 mmol, 68 pL) in acetic acid (0.5 mL). To the reaction was then added HBr (154 pL, 35% in acetic acid) in acetic acid (1 mL) and the reaction mixture was stirred at 50 °C for 2 h. After adjusting the pH to 10 with 20% sodium hydroxide (10 mL), the mixture was extracted with dichloromethane (3 x 20 mL). The combined organic layers were washed with water (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative thin layer chromatography (silica gel, eluted with dichloromethane / methanol: 10 / 1) to give N-{[2-bromo-1-(2,2,2-trifluoroethyl)-1H-1,3-benzimidazol-5-yl]methyl}oxan-4-amine (25 mg, 9% yield).

[1107] A solution of N-{[2-bromo-l-(2,2,2-trifluoroethyl)-lH-l,3-benzimidazol-5-yl]methyl}oxane-4- amine (30 mg, 68.84 pmol) and di-tert-butyl dicarboxylate (35 mg, 158.33 pmol) in dioxane (3 mL) was stirred at 70 °C for 2 h. After the reaction mixture was cooled to room temperature and concentrated, the residue was purified by preparative thin layer chromatography (eluted with dichloromethane / methanol: 10 / 1) to give tert-butyl N-{[2-bromo-l-(2,2,2-trifluoroethyl)-lH-l,3-benzimidazol-5-yl]methyl}-N- (oxan-4-yl)carbamate (30 mg, 80% yield) as a white solid.

[1108] To a solution of 2-methyl-2-[5-(prop-2-ynylamino)-2-pyridyl]propanenitrile (50 mg, 212.41 pmol), diisopropylamine (1.42 mmol, 200 pL) and CuI (1.2 mg, 6.40 pmol) in dimethyl sulfoxide (2.5 mL) was added tert-butyl N-{[2-bromo-l-(2,2,2-trifluoroethyl)-lH-l,3-benzimidazol-5-yl]methyl}-N- (oxan-4-yl)carbamate (35 mg, 63.98 pmol) and tetrakis(triphenylphosphine)palladium(0) (7 mg, 6.40 pmol, 0.10 equiv). The reaction mixture was stirred at 80 °C under nitrogen atmosphere for 1 h, then treated with a saturated EDTA solution (5 mL). The biphasic mixture was stirred at 20 °C for 1 h, then extracted with dichloromethane (3 x 10 mL). The combined organic layers were washed with water (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified twice by preparative thin layer chromatography (silica gel, eluted with dichloromethane / methanol: 10 / 1, ethyl acetate / methanol: 30 / 3, respectively) to give tert-butyl N-{[2-(3-{[6-(l-cyano-l-methylethyl)pyridin-3-yl]amino}prop-l-yn-l-yl)-l-(2,2,2- trifluoroethyl)-lH-l,3-benzimidazol-5-yl]methyl}-N-(oxan-4-yl)carbamate (41 mg, 84% yield, 80% purity) as a black solid.

[1109] To a solution of tert-butyl N-{[2-(3-{[6-(1-cyano-1-methylethyl)pyridin-3-yl]amino}prop-1-yn-1-yl)-1-(2,2,2-trifluoroethyl)-1H-1,3-benzimidazol-5-yl]methyl}-N-(oxan-4-yl)carbamate (35 mg, 45.85 μmol) in acetonitrile (3 mL) was added bismuth trichloride (443.83 μmol, 30 μL). The reaction mixture was stirred at 60 °C for 1 h and treated with a saturated EDTA solution (10 mL). The biphasic mixture was stirred at 20 °C for 1 h and extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with brine (2 x 10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC to give 2-methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-1,3-benzimidazol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile (6.5 mg, 25% yield, 98% purity) as a white solid.

[1110] LC-MS (ES + ,m / z): 511.3 [(M+H) + ]

[1111] Example 187 Preparation of N-{[2-(2-phenylethynyl)-1-(2,2,2-trifluoroethyl)-1H-1,3-benzimidazol-5-yl]methyl}oxan-4-amine

[1112]

[1113] To a solution of tert-butyl N-({3-amino-4-[(2,2,2-trifluoroethyl)amino]phenyl}methyl)- N-(oxan-4-yl)carbamate (200 mg, 446.16 pmol) in dimethylformamide (5 mL) was added 3- phenylprop-2-ynal (174 mg, 1.34 mmol, 163 pL). Then potassium hydrogen sulfate (Oxone) (274 mg, 446.16 pmol) was added and the reaction mixture was stirred at 20 °C for 2 h. The reaction mixture was then treated with saturated sodium bicarbonate solution (5 mL) and saturated Na2SO3solution (5 mL) and the mixture was stirred at 20 °C for 1 h. The mixture was then extracted with ethyl acetate (3 x 10 mL). The combined organic layers were washed with water (3 x 10 mL), brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative thin layer chromatography (silica gel eluting with ethyl acetate / petroleum ether: 1 / 1) to give tert-butyl N-(oxan-4-yl)-N-{[2-(2-phenylethynyl)-1-(2,2,2- trifluoroethyl)-1H-1,3-benzimidazol-5-yl]methyl}carbamate (80 mg, 31% yield) as a yellow oil.

[1114] Deprotection of tert-butyl N-(oxan-4-yl)-N-{[2-(2-phenylethynyl)-1-(2,2,2- trifluoroethyl)-1H-1,3-benzimidazol-5-yl]methyl}carbamate (60 mg, 105.2 mmol) was performed using the method described in Example 186 with bismuth(III) chloride to give N-{[2-(2- phenylethynyl)-1-(2,2,2-trifluoroethyl)-1H-1,3-benzimidazol-5-yl]methyl}oxan-4-amine (35 mg, yellowish solid).

[1115] LC-MS (ES + ,m / z): 414.3 [(M+H) + ]

[1116] Example 188 Preparation of 2-methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2- trifluoroethyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile

[1117]

[1118] To a 0 °C solution of methyl 1H-pyrrolo[3,2-b]pyridine-5-carboxylate (3.01 g, 17.05 mmol) in dimethylformamide (30 mL) was added sodium hydride (890 mg, 22.17 mmol, 60% in mineral oil). The mixture was stirred at 0 °C for 30 min and PhS02CI (2.2 mL, 17.05 mmol) was added. The ice bath was then removed and the mixture was allowed to warm to room temperature and stirred at room temperature overnight. The reaction mixture was poured into ice / water (300 mL) and stirred for 1 h. The solid was filtered, washed with water (100 mL) and hexanes (50 mL) and the resulting solution was dried overnight to give methyl 1-(phenylsulfonyl)-1H-pyrrolo[3,2-b]pyridine-5-carboxylate (3.50 g, 66% yield) as an off-white solid.

[1119] Methyl 1-(phenylsulfonyl)-1H-pyrrolo[3,2-b]pyridine-5-carboxylate (500 mg, 1.58 mmol) was dissolved in anhydrous tetrahydrofuran (50 mL) and cooled to -78 °C under a nitrogen atmosphere. A solution of DIBAL (1.8 M, 5.3 mL, 9.48 mmol) was added dropwise and the reaction mixture was stirred at -78 °C for 20 min. The mixture was then poured into a mixture of water (20 mL), 1 N sodium hydroxide (20 mL) and brine (20 mL) and stirred for 10 min. The mixture was then extracted with ethyl acetate (3 x 30 mL). The combined organic layers were washed with water, dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel eluted with dichloromethane / ethyl acetate: 4 / 1) to give 1-(phenylsulfonyl)-1H-pyrrolo[3,2-b]pyridine-5-carbaldehyde (320 mg, 70% yield) as a white solid.

[1120] 2-Methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-pyrrolo[3,2- b]pyridin-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile was prepared in a similar manner as described in Examples 1 and 49 using 1-(phenylsulfonyl)-1H-pyrrolo[3,2-b]pyridine-5- carbaldehyde and 2-methyl-2-{5-[(prop-2-yn-1-yl)amino]pyridin-2-yl}propanenitrile.

[1121] LC-MS (ES + ,m / z): 511.1 [(M+H) + ]

[1122] Example 189: Preparation of 2-{3-[(4-methanesulfonylphenyl)amino]prop-1-yn-1-yl}-5-methyl-N-(1-methylpiperidin-4-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine

[1123]

[1124] 2-{3-[(4-Methanesulfonylphenyl)-amino]prop-1-yn-1-yl}-5-methyl-N-(1-methylpiperidin-4-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine was prepared in a manner analogous to the method described in Example 179 using 5-methyl-4-nitro-1H-indole and 4-methanesulfonyl-N-(prop-2-yn-1-yl)aniline.

[1125] LC-MS(ES + ,m / z):533.1[(M+H) + ]

[1126] Example 190 :4-[(2-{3-[(4-methanesulfonylphenyl)amino]prop-1-yn-1-yl}-5-methyl-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino]-1λ 6 -Preparation of 1,1-thiol-1,2-dione

[1127]

[1128] 4-[(2-{3-[(4-methanesulfonylphenyl)-amino]prop-1-yn-1-yl}-5-methyl-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino]-1λ was prepared in a manner similar to that described in Example 179 using 5-methyl-4-nitro-1H-indole and 4-methanesulfonyl-N-(prop-2-yn-1-yl)aniline. 6 -thiazole-1,1-dione.

[1129] LC-MS(ES + ,m / z):568.2[(M+H) + ]

[1130] Example 191 : Preparation of 2-methyl-2-{5-[(3-{5-methyl-4-[(1-methylpiperidin-4-yl)amino]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile

[1131]

[1132] N-[l-(2-methanesulfonyl ethyl)piperidin-4-yl]-2-{3-[(4-methanesulfonylphenyl) amino]prop- 1 -yn- 1 -yl} -5 -methyl- 1 -(2,2,2-trifluoroethyl)- 1 H-indol-4-amine was prepared in a similar manner to that described in Example 179 using 5-methyl-4-nitro-lH-indole and 4- methanesulfonyl-N-(prop-2-yn-l-yl)aniline.

[1133] LC-MS (ES + , m / z): 523.0 [(M+H) + ]

[1134] Example 192 Preparation of N-[l-(2-methanesulfonyl ethyl)piperidin-4-yl]-2-{3-[(4- methanesulfonylphenyl)amino]prop- 1 -yn- 1 -yl} -5 -methyl- 1 -(2,2,2-trifluoroethyl)- 1 H- indol-4-amine

[1135]

[1136] N-[l-(2-methanesulfonyl ethyl)piperidin-4-yl]-2-{3-[(4-methanesulfonylphenyl) amino]prop- 1 -yn- 1 -yl} -5 -methyl- 1 -(2,2,2-trifluoroethyl)- 1 H-indol-4-amine was prepared in a similar manner to that described in Example 179 using 5-methyl-4-nitro-lH-indole and 4- methanesulfonyl-N-(prop-2-yn-l-yl)aniline.

[1137] LC-MS (ES + , m / z): 625.3 [(M+H) + ]

[1138] Example 193 Preparation of 4-[(3-{5-methyl-4-[(piperidin-4-yl)amino]-l-(2,2,2- trifluoroethyl)-lH-indol-2-yl}prop-2-yn-l-yl)amino]benzene-l-sulfonamide

[1139]

[1140] 4-[(3-{5-methyl-4-[(piperidin-4-yl)amino]-l-(2,2,2- trifluoroethyl)-lH-indol-2-yl}prop-2-yn-l-yl)amino]benzene-l-sulfonamide was prepared in a similar manner to that described in Example 179 using 5-methyl-4-nitro-lH-indole and 4- [(prop-2-yn-l-yl)amino]benzene-l-sulfonamide.

[1141] LC-MS(ES + ,m / z):519.9[(M+H) + ]

[1142] Example 194 : Preparation of 2-{3-[(4-methanesulfonylphenyl)amino]prop-1-yn-1-yl}-5-methyl-N-[1-(oxacyclohexane-4-yl)piperidin-4-yl]-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine

[1143]

[1144] 2-{3-[(4-Methanesulfonylphenyl)-amino]prop-1-yn-1-yl}-5-methyl-N-[1-(oxan-4-yl)piperidin-4-yl]-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine was prepared in a manner analogous to the method described in Example 179 using 5-methyl-4-nitro-1H-indole and 4-methanesulfonyl-N-(prop-2-yn-1-yl)aniline.

[1145] LC-MS(ES + ,m / z):603.3[(M+H) + ]

[1146] Example 195 : Preparation of 2-{4-[(2-{3-[(4-methanesulfonylphenyl)amino]prop-1-yn-1-yl}-5-methyl-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino]piperidin-1-yl}ethan-1-ol

[1147]

[1148] 2-{4-[(2-{3-[(4-Methanesulfonyl-phenyl)-amino]prop-1-yn-1-yl}-5-methyl-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino]piperidin-1-yl}ethan-1-ol was prepared in a manner analogous to the method described in Example 179 using 5-methyl-4-nitro-1H-indole and 4-methanesulfonyl-N-(prop-2-yn-1-yl)aniline.

[1149] LC-MS(ES + ,m / z):563.3[(M+H) + ]

[1150] Example 196Preparation of 2-[5-({3-[4-(methoxymethyl)-l-(2,2,2-trifluoroethyl)-lH-indol-2- yl]prop-2-yn-l-yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[1151]

[1152] 2-[5-({3-[4-(methoxymethyl)-l-(2,2,2-trifluoroethyl)-lH-indol-2- yl]prop-2-yn-l-yl}amino)pyridin-2-yl]-2-methylpropanenitrile was prepared in a similar manner to that described in Examples 1 and 49 using 2-iodo-lH-indole-4- carboxaldehyde (prepared from methyl lH-indole-4-carboxylate) and 2-methyl-2-{5- [(prop-2-yn-l-yl)amino]pyridin-2-yl}propanenitrile.

[1153] LC-MS (ES + ,m / z): 441.2 [(M+H) + ]

[1154] Example 197 Preparation of 2-[5-({3-[4-(cyanomethyl)-l-(2,2,2-trifluoroethyl)-lH-indol-2- yl]prop-2-yn-l-yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[1155]

[1156] 2-[5-({3-[4-(cyanomethyl)-l-(2,2,2-trifluoroethyl)-lH-indol-2- yl]prop-2-yn-l-yl}amino)pyridin-2-yl]-2-methylpropanenitrile was prepared in a similar manner to that described in Examples 1 and 49 using 2-iodo-lH-indole-4- carboxaldehyde (prepared from methyl lH-indole-4-carboxylate) and 2-methyl-2-{5- [(prop-2-yn-l-yl)amino]pyridin-2-yl}propanenitrile.

[1157] LC-MS (ES + ,m / z): 436.2 [(M+H) + ]

[1158] Example 198 Preparation of 2-methyl-2-[5-({3-[5-(morpholine-4-carbonyl)-l-(2,2,2- trifluoroethyl)-lH-indol-2-yl]prop-2-yn-l-yl}amino)pyridin-2-yl]propanenitrile

[1159]

[1160] To a solution of methyl 2-[3-[[6-(1-cyano-1-methyl-ethyl)-3-pyridinyl]amino]prop-1-ynyl]-1-(2,2,2-trifluoroethyl)-1H-indole-5-carboxylate (300 mg, 594.14 pmol) in methanol (4.5 mL) was added sodium hydroxide (5 M, 810 pL) and the resulting reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was then poured into aqueous ammonium chloride (10 mL) and the pH was adjusted to 3 using 1 N HC1. The reaction mixture was then extracted with ethyl acetate (3 x 10 mL). The combined organic layers were washed with water (2 x 10 mL), brine (2 x 10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give crude 2-[3-[[6-(1-cyano-1-methyl-ethyl)-3-pyridinyl]amino]prop-1-ynyl]-1-(2,2,2-trifluoroethyl)-1H-indole-5-carboxylic acid (brown solid) which was used without further purification.

[1161] 2-[3-[[6-(1-cyano-1-methyl-ethyl)-3-pyridinyl]amino]prop-1-ynyl]-1-(2,2,2-trifluoroethyl)-1H-indole-5-carboxylic acid (1 eq) was coupled with morpholine (2 eq) using HATU (2 eq) and diisopropylethylamine (3 eq) in dichloromethane at 25 °C for 1 h to give 2-methyl-2-[5-({3-[5-(morpholine-4-carbonyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propanenitrile after purification by preparative HPLC.

[1162] LC-MS (ES + ,m / z): 510.3 [(M+H) + ]

[1163] Example 199 Preparation of 2-methyl-2-[5-({3-[5-(4-methylpiperazine-1-carbonyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propanenitrile

[1164]

[1165] 2-methyl-2-[5-({3-[5-(4-methylpiperazine-1-carbonyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propanenitrile was prepared in a similar manner as described in the method described in Example 198.

[1166] LC-MS (ES + ,m / z): 523.3 [(M+H)+ ]

[1167] Example 200 Preparation of 2-(3-{[6-(1-cyano-1-methylethyl)pyridin-3-yl]amino}prop-1-yn-1-yl)-N-{1-[2-(dimethylamino)acetyl]piperidin-4-yl}-1-(2,2,2-trifluoroethyl)-1H-indole-5- carboxamide

[1168]

[1169] 2-(3-{[6-(1-cyano-1-methylethyl)pyridin-3-yl]amino}prop-1-yn-1-yl)-N-{1-[2- (dimethylamino)acetyl]piperidin-4-yl}-1-(2,2,2-trifluoroethyl)-1H-indole-5-carboxamide was prepared in a similar manner as described in Example 198.

[1170] LC-MS (ES + ,m / z): 608.4 [(M+H) + ]

[1171] Example 201 Preparation of 2-(3-{[6-(1-cyano-1-methylethyl)pyridin-3-yl]amino}prop-1-yn-1-yl)-N-{1-[2- (dimethylamino)acetyl]piperidin-4-yl}-1-(2,2,2-trifluoroethyl)-1H-indole-5-carboxamide

[1172]

[1173] 2-(3-{[6-(1-cyano-1-methylethyl)pyridin-3-yl]amino}prop-1-yn-1-yl)-N-{1-[2- (dimethylamino)acetyl]piperidin-4-yl}-1-(2,2,2-trifluoroethyl)-1H-indole-5-carboxamide was prepared in a similar manner as described in Example 198.

[1174] LC-MS (ES + ,m / z): 608.4 [(M+H) + ]

[1175] Example 202 Preparation of 2-(3-{[6-(1-cyano-1-methylethyl)pyridin-3-yl]amino}prop-1-yn-1-yl)-N-{1-[2- (dimethylamino)acetyl]piperidin-4-yl}-1-(2,2,2-trifluoroethyl)-1H-indole-5-carboxamide

[1176]

[1177] In a similar manner to that described in Example 198, 2-(3-{[6-(1-cyano-1- methylethyl)pyridin-3-yl]amino}prop-1-yn-1-yl)-N-(oxan-4-yl)-1-(2,2,2- trifluoroethyl)-1H-indole-5-carboxamide was prepared.

[1178] LC-MS (ES + ,m / z): 524.3 [(M+H) + ]

[1179] Example 203 Preparation of 2-methyl-2-(5-{[3-(5-{1-[(oxan-4-yl)amino]ethyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile

[1180]

[1181] In a similar manner to that described in Examples 1 and 49, using 1-(1H-indol-5- yl)ethan-1-one and 2-methyl-2-{5-[(prop-2-yn-1-yl)amino]pyridin-2-yl}propanenitrile, 2-methyl-2-(5-{[3-(5-{1-[(oxan-4-yl)amino]ethyl}-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile was prepared.

[1182] LC-MS (ES + ,m / z): 524.4 [(M+H) + ]

[1183] Example 204 Preparation of 2-methyl-2-{5-[(3-{4-[(1-methylpiperidin-4-yl)amino]-1-(2,2,2- trifluoroethyl)-1H-pyrrolo[3,2-c]pyridin-2-yl}prop-2-yn-1-yl)amino]pyridin-2- yl}propanenitrile

[1184]

[1185] To a solution of 2-(5-amino pyridin-2-yl)-2-methyl propionitrile (4.0 g, 24.81 mmol) in dioxane (100 mL) was added di-tert-butyl dicarboxylate (10.83 g, 49.62 mmol) and the reaction mixture was stirred at 100 °C for 6 h. The reaction mixture was then concentrated to give the crude product which was washed with petroleum ether (2 x 6 mL) and filtered to give tert-butyl N-[6-(1-cyano-1-methylethyl)-pyridin-3-yl]carbamate (5.80 g, 89% yield) as a white solid.

[1186] To a solution of tert-butyl N-[6-(1-cyano-1-methylethyl)-pyridin-3-yl]carbamate (5.80 g, 21.97 mmol) in dimethylformamide (100 mL) was added sodium hydride (2.64 g, 65.91 mmol, 60% in mineral oil). The reaction mixture was stirred at 0 °C for 30 min and to the reaction mixture was added propargyl bromide (7.5 mL, 87.44 mmol). The resulting reaction mixture was stirred at 25 °C for a further 30 min, poured into ice water (100 mL) and extracted with ethyl acetate (3 x 30 mL). The combined organic layers were washed with water (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel eluting with petroleum ether / ethyl acetate: 6 / 1) to give tert-butyl N-[6-(1-cyano-1-methylethyl)pyridin-3-yl]-N-(prop-2-yn-1-yl)carbamate (6.5 g, 98% yield) as a colourless oil.

[1187] A solution of tert-butyl N-[6-(1-cyano-1-methylethyl)pyridin-3-yl]-N-(prop-2-yn-1-yl)carbamate (2 g, 6.65 mmol) in HCl / ethyl acetate (4 M, 10 mL) was stirred at 25 °C for 3 h, then quenched with aqueous sodium bicarbonate solution. The resulting mixture was concentrated and the residue washed with petroleum ether (2 x 5 mL) to give 2-methyl-2-{5-[(prop-2-yn-1-yl)amino]pyridin-2-yl}propanenitrile (1.50 g, 94% yield, hydrochloride salt) as an off-white solid.

[1188] 2-methyl-2-{5-[(3-{4-[(1-methylpiperidin-4-yl)amino]-1-(2,2,2-trifluoroethyl)-1H-pyrrolo[3,2- c]pyridin-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}propanenitrile was prepared in a similar manner to that described in Example 172 using 4-chloro-1H-pyrrolo[3,2-c]pyridine and 2-methyl-2-{5-[(prop-2-yn-1-yl)amino]pyridin-2-yl}propanenitrile.

[1189] LC-MS (ES + m / z): 510.3 [(M+H) + ]

[1190] Example 205 Preparation of 2-methyl-2-[5-({3-[5-(morpholin-4-ylmethyl)-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propanenitrile

[1191]

[1192] Synthesis scheme:

[1193]

[1194] To a solution of 1H-indole-5-carboxylic acid (150 g, 930.75 mmol) in dimethylformamide (1.50 L) was added sodium bicarbonate (312.77 g, 3.72 mol) and iodomethane (528.44 g, 3.72 mol, 232 mL). The mixture was stirred at 30 °C for 2 h, quenched by the addition of water (3000 mL) at 30 °C, and extracted with ethyl acetate (3 x 500 mL). The combined organic layers were washed with brine (500 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give 1H-indole-5-carboxylic acid methyl ester (300 g, crude) as a yellow solid.

[1195] To a solution of 1H-indole-5-carboxylic acid methyl ester (80 g, 456.67 mmol) in tetrahydrofuran (800 mL) was added sodium hydride (27.4 g, 685.01 mmol, 60% in mineral oil) and benzenesulfonyl chloride (80.66 g, 456.67 mmol, 58.5 mL). The mixture was stirred at 0 °C for 2 h, quenched by the addition of saturated ammonium chloride (1000 mL) at 0 °C, and extracted with ethyl acetate (3 x 1000 mL). The combined organic layers were washed with brine (1000 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give crude 1-(benzenesulfonyl)-1H-indole-5-carboxylic acid methyl ester (400 g) as a yellow solid.

[1196] To 1- (benzenesulfonyl) -1H- indole -5- methyl formate (70g, 221.98mmol) in toluene (1L) -78 DEG C of stirring solution, add DIBAL-H solution (1M, 888mL). The mixture is stirred at -78 DEG C for 50min. It is quenched by adding saturated ammonium chloride (1000mL) at 0 DEG C and extracted with ethyl acetate (3x 1000mL). The combined organic layer is washed with brine (2000mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product is purified by flash column chromatography (silica gel, eluted with petroleum ether / ethyl acetate: 30 / 1 to 5 / 1) to obtain [1- (benzenesulfonyl) -1H- indole -5- base] methanol (180g, 94% yield) as a yellow solid.

[1197] To a solution of [1-(phenylsulfonyl)-1H-indol-5-yl]methanol (34 g, 118.33 mmol) in dichloromethane (500 mL) was added imidazole (24.17 g, 354.99 mmol) and TBSCl (26.75 g, 177.50 mmol). The mixture was stirred at 30 ° C for 1 h, quenched by adding water (500 mL), and extracted with ethyl acetate (3 x 500 mL). The combined organic layers were washed with brine (300 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give crude 1-(phenylsulfonyl)-5-[(tert-butyldimethylsilyl)oxy]-1H-indole (150 g).

[1198] To a stirred solution of 1-(phenylsulfonyl)-5-[(tert-butyldimethylsilyl)oxy]-1H-indole (30 g, 74.70 mmol) in anhydrous tetrahydrofuran (18 mL) at -78°C was added a solution of lithium diisopropylamide (2 M, 112 mL). The mixture was stirred at -78°C for 10 min, then an iodine solution (56.88 g, 224.11 mmol in 45 mL of tetrahydrofuran) was added. The mixture was stirred at -78°C for 50 min, quenched with saturated ammonium chloride (1000 mL), and extracted with ethyl acetate (3 x 1000 mL). The organic layer was washed with aqueous Na2S2O3 (1000 mL) and brine (1000 mL), dried over anhydrous sodium sulfate and concentrated to give crude 1-(benzenesulfonyl)-5-[(tert-butyldimethylsilyl)oxy]-2-iodo-1H-indole (200 g) as a yellow solid.

[1199] To a solution of 1-(phenylsulfonyl)-5-[(tert-butyldimethylsilyl)oxy]-2-iodo-1H- indole (20.0 g, 37.92 mmol) in methanol / water (2.40 L) was added potassium carbonate (5.24 g, 37.92 mmol) in methanol / water (2.40 L). The mixture was stirred at 120 °C under nitrogen atmosphere for 5 h. The solids were filtered off and washed with dichloromethane. The filtrate was washed with water (2.4 L), brine (2.4 L), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluted with petroleum ether / ethyl acetate: 1 / 1) to give (2-iodo-1H-indol-5-yl)methanol (19 g, 28% yield, 60% purity) as a yellow solid.

[1200] A mixture of (2-iodo-1H-indol-5-yl)methanol (15.83 g, 34.79 mmol) and MnO2(24.20 g, 278.32 mmol) in dichloromethane (150 mL) was stirred at 30 °C under nitrogen atmosphere for 5 h. The reaction mixture was filtered using celite and washed with dichloromethane (2 x 150 mL). The filtrate was washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluted with petroleum ether / ethyl acetate: 10 / 1) to give 2-iodo-1H-indole-5-carbaldehyde (8 g, 66.17% yield, 78% purity) as a yellow solid. This crude product was used without further purification.

[1201] To a solution of 2-iodo-1H-indole-5-carbaldehyde (3 g, 8.63 mmol) in tetrahydrofuran (40 mL) cooled to 0 °C was added sodium hydride (1.04 g, 25.90 mmol, 60% in mineral oil). The mixture was stirred at 0 °C under nitrogen atmosphere for 30 min and to the reaction mixture was added 2,2,2-trifluoroethyl trifluoromethanesulfonate (5.01 g, 21.58 mmol) dropwise. The ice bath was then removed and the reaction mixture was stirred under nitrogen atmosphere for 1 h, quenched with saturated ammonium chloride (100 mL), and extracted with ethyl acetate (2 x 150 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluted with petroleum ether / ethyl acetate: 10 / 1) to give 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-5-carbaldehyde (3 g, 98% yield) as a red solid.

[1202] To a solution of 2-methyl-2-[5-(prop-2-ynylamino)-2-pyridyl]propanenitrile (2.40 g, 9.18 mmol) in dimethyl sulfoxide (18 mL) was added N-isopropylpropan-2-amine (3.10 g, 30.60 mmol, 4.3 mL) and Cul (291 mg, 1.53 mmol). Then, to the reaction mixture was added 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-5- carboxaldehyde (1.80 g, 5.10 mmol) and tetrakis(triphenylphosphine)palladium(0) (589 mg, 510 μmol). The mixture was stirred at room temperature under nitrogen atmosphere for 2 h. The reaction mixture was quenched with EDTA (50 mL) and extracted with ethyl acetate (2 x 25 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative thin layer chromatography (silica gel, eluted with ethyl acetate / petroleum ether: 1 / 2) to give 2-[5-({3-[5-formyl-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile (2 g, 92.40% yield) as a yellow oil.

[1203] To a solution of 2-[5-({3-[5-formyl-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile (1 eq) and morpholine (1 eq) in dichloromethane (2 mL) and methanol (2 mL) was added magnesium sulfate (15 eq). After the mixture was stirred at room temperature for 12 h, sodium bicarbonate (1 eq) and sodium cyanoborohydride (5 eq) were added to the reaction mixture and the reaction was stirred at room temperature for 1 h, then the mixture was quenched by the addition of saturated sodium bicarbonate (10 mL). The mixture was extracted with ethyl acetate (2 x 30 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC to give 2-methyl-2-[5-({3-[5-(morpholin-4-ylmethyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propanenitrile.

[1204] LC-MS (ES + ,m / z): 496.3 [(M+H) + ]

[1205] The following compounds were prepared in a similar manner as described in Example 205:

[1206]

[1207]

[1208]

[1209]

[1210]

[1211]

[1212]

[1213] Example 266 Preparation of N-[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H- indol-2-yl)prop-2-yn-1-yl]-6-(pyrrolidine-1-carbonyl)pyridin-3-amine

[1214]

[1215] To a solution of 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-5-carbaldehyde (1 eq) in dichloromethane (20 mL) was added tetrahydro-2H-pyran-4-amine (8 eq) and magnesium sulfate (10 eq). The reaction mixture was then stirred at room temperature under nitrogen atmosphere for 2 h. Sodium cyanoborohydride (4 eq) was then added to the reaction and the mixture was stirred under nitrogen atmosphere for 1 h. The reaction mixture was poured into aqueous sodium bicarbonate solution (50 mL) and extracted with dichloromethane (100 mL). The combined organic layers were washed with water (50 mL), brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude residue was purified by flash column chromatography (silica gel, eluting with dichloromethane / methanol: 40 / 1 to 20 / 1) to give N-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl)methyl)tetrahydro-2H-pyran-4-amine as a yellow solid.

[1216] To a solution of 5-((tert-butoxycarbonyl)(prop-2-yn-1-yl)amino)picolinic acid (1 eq) and pyrrolidine (1.2 eq) in dichloromethane (5 mL) was added triethylamine (2 eq) and HATU (1.20 eq). The reaction was stirred at 20 °C for 1 h and water (20 mL) was added. The mixture was extracted with dichloromethane (3 x 20 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluting with petroleum ether / ethyl acetate: 4 / 1 to 2 / 1) to give tert-butyl prop-2-yn-1-yl(6-(pyrrolidine-1-carbonyl)pyridin-3-yl)carbamate as a light yellow oil. This crude product was used in the next step without further purification.

[1217] A solution of tert-butyl prop-2-yn-1 -yl(6-(pyrrolidin-1 - carbonyl)pyridin-3-yl)carbamate (1 equiv) in HCl / ethyl acetate (5 mL) was stirred at room temperature for 2 h. The reaction mixture was then concentrated in vacuo to give crude (5-(prop-2-yn-1 -ylamino)pyridin-2-yl)(pyrrolidin-1 -yl)methanone as a yellow solid. This crude product was used in the next step without further purification.

[1218] In a similar manner to that described in Example 205, N-{[2-iodo-1 -(2,2,2- trifluoroethyl)-1 H-indol-5-yl]methyl}oxan-4-amine was coupled with (5-(prop-2-yn-1 - ylamino)pyridin-2-yl)(pyrrolidin-1 -yl)methanone to give N-[3-(5-{[(oxan-4-yl)amino]methyl}- 1 -(2,2,2-trifluoroethyl)-1 H-indol-2-yl)prop-2-yn-1 -yl]-6-(pyrrolidin-1 - carbonyl)pyridin-3-amine.

[1219] LC-MS (ES + ,m / z): 540.3 [(M+H) + ]

[1220] The following compounds were prepared starting from 2-iodo-1 -(2,2,2- trifluoroethyl)-1 H-indole-5-carbaldehyde and using the procedure described in Example 266:

[1221]

[1222]

[1223]

[1224] Example 288 Preparation of 2-methyl-2-{5-[(3-{5-[(oxan-4-yl)amino]-1 -(2,2,2- trifluoroethyl)-1 H-indol-2-yl}prop-2-yn-1 -yl)amino]pyridin-2-yl}propanenitrile

[1225]

[1226] 2-methyl-2-{5-[(3-{5-[(oxan-4-yl)amino]-1 -(2,2,2-trifluoroethyl)-1 H-indol-2- yl}prop-2-yn-1 -yl)amino]pyridin-2-yl}propanenitrile was prepared starting from 5-bromo- 1 H-indole and using a similar procedure to that described in Example 1.

[1227] LC-MS (ES+ m / z): 496.3 [(M+H) + ]

[1228] Example 289 Preparation of 2-{5-[(3-{4-[(1 -acetylpiperidin-4-yl)amino]-1 -(2,2,2- trifluoroethyl)-1 H-indol-2-yl}prop-2-yn-1 -yl)amino]pyridin-2-yl}-2-methylpropanenitrile

[1229]

[1230] Synthesis scheme

[1231]

[1232] To a solution of 4-nitro-indole (1 equiv) in dimethylformamide was added sodium hydride (1.50 equiv) and benzenesulfonyl chloride (1.0 equiv). The mixture was stirred at 0-30 °C for 2 h and quenched with 0 °C aqueous ammonium chloride solution. The mixture was filtered, the solid washed with water and petroleum ether, and the resulting solution concentrated and dried in vacuo to give 4-nitro-1-(phenylsulfonyl)-1 H-indole.

[1233] To a -78 °C stirred solution of 4-nitro-1-(phenylsulfonyl)-1 H-indole (1.0 equiv) in anhydrous tetrahydrofuran was added a solution of lithium diisopropylamide (2 M, 3.0 equiv). The resulting mixture was stirred at -78 °C for 60 min and a solution of iodine in tetrahydrofuran (1.5 equiv) was added dropwise. After stirring at -78 °C for 30 min, the mixture was quenched with saturated ammonium chloride and extracted with ethyl acetate. The combined organic layers were washed with aqueous Na2S203 and brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give crude 2-iodo-4-nitro-1-(phenylsulfonyl)-1 H-indole which was used without further purification.

[1234] Following a similar procedure to that described in Example 205, 2-iodo-4-nitro-1-(phenylsulfonyl)-1 H-indole was treated with potassium carbonate in methanol to give 2-iodo-4-nitro-1 H-indole.

[1235] To a solution of 2-iodo-4-nitro-lH-indole (1.0 equiv) in tetrahydrofuran at 0 °C was added sodium hydride (5.0 equiv). The mixture was stirred at 0-25 °C for 30 min, and to the reaction mixture was added CF3CH2OTf (4.0 equiv). After the reaction was stirred at room temperature for 2 h, the reaction was quenched by the addition of water at 0 °C and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by recrystallization from a mixture of ethyl acetate and petroleum ether to give 2-iodo-4-nitro-l-(2,2,2-trifluoroethyl)-lH-indole as a yellow solid.

[1236] To a solution of 2-iodo-4-nitro-l-(2,2,2-trifluoroethyl)-lH-indole (1.0 equiv) in acetic acid was added Fe (6.0 equiv). The mixture was stirred at 50 °C for 2 h, then poured into water at 0 °C (Fe powder was removed using a magnet) and filtered to give the crude product. The crude product was taken up in ethyl acetate and washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The concentrated solution was added dropwise to petroleum ether to precipitate the product. By filtration, 2-iodo-l-(2,2,2-trifluoroethyl)-lH-indol-4-amine was obtained as a yellow solid.

[1237] To a solution of 2-methyl-2-(5-(prop-2-yn-l-ylamino)pyridin-2-yl)propanenitrile (693 mg, 2.94 mmol, prepared from 2-(5-amino pyridin-2-yl)-2-methylpropanenitrile and propargyl bromide) in dimethyl sulfoxide (5 mL) was added N-isopropylpropan-2-amine (892 mg, 8.82 mmol) and CuI (84 mg, 441 μmol). Then 2-iodo-l-(2,2,2-trifluoroethyl)indol-4-amine (500 mg, 1.47 mmol) and tetrakis(triphenylphosphine)palladium(0) (170 mg, 147 μmol) were added. The resulting reaction mixture was stirred at room temperature under a nitrogen atmosphere for 2 h, then quenched with EDTA solution (5 mL). The reaction mixture was extracted with ethyl acetate (2 x 5 mL), and the combined organic layers were concentrated in vacuo. The residue was purified by preparative thin layer chromatography (silica gel, eluted with a 1 / 1 mixture of ethyl acetate / petroleum ether) to give 2-(5-((3-(4-amino-l-(2,2,2-trifluoroethyl)-lH-indol-2-yl)prop-2-yn-l-yl)amino)pyridin-2-yl)-2-methylpropanenitrile as a black-brown solid (1.0 g, 73.25% yield, 88.6% purity).

[1238] To a solution of 2-(5-((3-(4-amino-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1- yl)amino)pyridin-2-yl)-2-methylpropanenitrile (150 mg, 364.59 pmol) in ethanol (3 mL) was added 1-acetylpiperidin-4-one (5 eq) and tetraethoxy titanium (83 mg, 364.59 pmol). The mixture was stirred at 50 °C for 2 h and sodium cyanoborohydride (115 mg, 1.82 mmol) was added. After stirring the reaction at 50 °C for 1 h, the reaction mixture was poured into a saturated solution of sodium bicarbonate (10 mL). The resulting mixture was then filtered and the filtrate was extracted with ethyl acetate (2 x 20 mL). The combined organic layers were concentrated in vacuo and the crude residue was purified by preparative HPLC to give 2-{5-[(3-{4-[(1-acetylpiperidin-4-yl)amino]-1-(2,2,2- trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}-2-methylpropanenitrile as a yellow solid.

[1239] LC-MS (ES + ,m / z): 537.1 [(M+H) + ]

[1240] The following compounds were prepared starting from 2-iodo-1-(2,2,2-trifluoroethyl)-1H- indol-4-amine and using a similar procedure to that described in Example 289:

[1241]

[1242]

[1243]

[1244]

[1245]

[1246]

[1247]

[1248] Example 342 Preparation of N-(1-methylpiperidin-4-yl)-2-{3-[(6-methylpyridin-3-yl)amino]prop-1-yn-1-yl}- 1-(2,2,2-trifluoroethyl)-1H-indol-4-amine

[1249]

[1250] To a 25°C mixture of 2-iodo-l-(2,2,2-trifluoroethyl)-lH-indol-4-amine (8 g, 23.52 mmol) in ethanol (70 mL) was added l-methylpiperidin-4-one (13.31 g, 117.60 mmol, 13.7 mL) and tetraethoxytitanium (26.83 g, 117.60 mmol, 24.39 mL). The reaction mixture was heated to 50°C and stirred at 50°C for 12 h and to the reaction at 25°C was added sodium cyanoborohydride (7.39 g, 117.60 mmol). After stirring at 25°C for 2 h, the reaction was diluted with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluting with dichloromethane / methanol: 40 / 1 to 10 / 1) to give 2-iodo-N-(l-methylpiperidin-4-yl)-l-(2,2,2-trifluoroethyl)-lH-indol-4-amine (2.17 g, 19.64% yield, 93.1% purity) as a yellow solid.

[1251] Reacting 2-iodo-N-(l-methylpiperidin-4-yl)-l-(2,2,2-trifluoroethyl)-lH-indol-4-amine with 6-methyl-N-(prop-2-yn-l-yl)pyridin-3-amine (prepared from 6-methylpyridin-3-amine) under Sonogashira coupling conditions gave N-(l-methylpiperidin-4-yl)-2-{3-[(6-methylpyridin-3-yl)amino]prop-l-yn-l-yl}-l-(2,2,2-trifluoroethyl)-lH-indol-4-amine.

[1252] LC-MS (ES + ,m / z): 456.2 [(M+H) + ]

[1253] The following compounds were prepared starting from 2-iodo-l-(2,2,2-trifluoroethyl)-lH-indol-4-amine and using similar methods to those described in Example 342:

[1254]

[1255]

[1256]

[1257]

[1258]

[1259]

[1260]

[1261] Example 395 : 5-[(3-{4-[(1,1-dioxo-1 lambda 6 Preparation of 5-[(3-{4-[(1,1-dioxo-1 lambda

[1262]

[1263] To a solution of 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine (500 mg, 1.06 mmol) in ethanol (15 mL) was added 1,1-dioxothiolan-4-one (471 mg, 3.18 mmol) and tetraethoxy titanium (1.21 g, 5.29 mmol, 1.1 mL). The mixture was stirred at 50 °C for 12 h and to the reaction was added sodium cyanoborohydride (665 mg, 10.59 mmol). After the reaction was stirred at 80 °C for 2 h, the reaction mixture was diluted with ethyl acetate (15 mL) and poured into saturated sodium bicarbonate solution (40 mL). The mixture was extracted with ethyl acetate (3 x 40 mL). The combined organic layers were washed with brine (3 x 120 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography to give 4-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)tetrahydro-2H-thiopyran 1,1-dioxide (450 mg, 69% yield, 77% purity) as a light red solid.

[1264] Refluxing 4-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)tetrahydro-2H- thiopyran 1,1-dioxide with 5-(prop-2-yn-1-ylamino)pyridinecarbonitrile (prepared from 5-amino-pyridinecarbonitrile and propargyl bromide) under Sonogashira coupling conditions gave 5-[(3-{4-[(1,1-dioxo-1 lambda 6 - thiacyclohexan-4-yl)amino]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridine-2-carbonitrile.

[1265] LC-MS (ES + , m / z): 502.2 [(M+H) + ]

[1266] The following compounds were prepared starting from 2-iodo-1-(2,2,2- trifluoroethyl)-1H-indol-4-amine and using similar methods as described in Example 396:

[1267]

[1268]

[1269]

[1270]

[1271]

[1272]

[1273]

[1274]

[1275] Example 455 Preparation of 2-{4-[(2-{3-[(4-methane sulfonylphenyl)amino]prop-1-yn-1-yl}-1- (2,2,2-trifluoroethyl)-1H-indol-4-yl)amino]piperidin-1-yl}-N,N-dimethylacetamide

[1276]

[1277] Synthesis scheme:

[1278]

[1279] To a solution of 4-(methylsulfonyl)aniline (15 g, 87.61 mmol) in dioxane (100 mL) was added di-tert-butyl dicarboxylate (57.36 g, 262.82 mmol). The mixture was stirred at 110 °C for 10 h, then concentrated in vacuo to give crude tert-butyl N-(4-methylsulfonylphenyl)carbamate (20 g, 73.71 mmol, 84% yield). This crude product was used without further purification.

[1280] To a solution of tert-butyl N-(4-methylsulfonylphenyl)carbamate (17 g, 62.65 mmol) in dimethylformamide (260 mL) cooled to 0 °C was added sodium hydride (7.52 g, 187.96 mmol, 60% in mineral oil). The mixture was stirred at 0 °C for 30 min, then propargyl bromide (22.36 g, 187.96 mmol, 16.2 mL) was added to the reaction. After stirring at 0 °C for a further 40 min, the reaction mixture was poured into water (300 mL) and extracted with ethyl acetate (2 x 100 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluting with petroleum ether / ethyl acetate: 10 / 1 to 1 / 1) to give tert-butyl (4-(methylsulfonyl)phenyl)(prop-2-yn-1-yl)carbamate (16.8 g, 87% yield) as a white solid.

[1281] To a 25 °C mixture of 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine (1 eq) in ethanol was added tert-butyl 4-oxopiperidine-1-carboxylate (5 eq) and tetraethoxytitanium (5 eq). The mixture was stirred at 50 °C for 12 h, and sodium cyanoborohydride (5 eq) was added to the reaction. After stirring at 25 °C for 2 h, the mixture was poured into saturated sodium bicarbonate solution and stirred for 30 min. The reaction mixture was then filtered through Celite and washed with ethyl acetate. The aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluting with petroleum ether / ethyl acetate: 100 / 0 to 25 / 1) to give tert-butyl 4-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)piperidine-1-carboxylate.

[1282] To a solution of tert-butyl (4-(methylsulfonyl)phenyl)(prop-2-yn-1-yl)carbamate (2 eq) in dimethyl sulfoxide was added N-isopropylpropan-2-amine (30 eq), CuI (2 eq), tert-butyl 4-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)piperidine-1-carboxylate (1 eq) and Pd(Ph3)4 (0.25 eq). After stirring the reaction under a nitrogen atmosphere at 20 °C for 1 h, the mixture was poured into a saturated EDTA solution. The biphasic mixture was stirred at room temperature for 20 min, then extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography to give tert-butyl 4-((2-(3-((tert-butoxycarbonyl)(4-(methylsulfonyl)phenyl)amino)prop-1-yn-1-yl)-1-(2,2,2- trifluoroethyl)-1H-indol-4-yl)amino)piperidine-1-carboxylate as a light yellow solid.

[1283] To a solution of tert-butyl 4-((2-(3-((tert-butoxycarbonyl)(4-(methylsulfonyl)phenyl)amino)prop-1-yn-1-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)piperidine-1-carboxylate (1 eq) in ethyl acetate was added HC1 / ethyl acetate and the resulting mixture was stirred at 20 °C for 1 h. The mixture was then concentrated in vacuo to give 2-(3-((4-(methylsulfonyl)phenyl)amino)prop-1-yn-1-yl)-N-(piperidin-4-yl)-1-(2,2,2- trifluoroethyl)-1H-indol-4-amine as a hydrochloride salt (light yellow solid).

[1284] A mixture of 2-(3-((4-(methylsulfonyl)phenyl)amino)prop-1-yn-1-yl)-N-(piperidin-4-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine (1 eq, hydrochloride salt), 2-chloro-N,N-dimethyl-acetamide (3 eq) and potassium carbonate (3 eq) in acetonitrile was stirred at 50 °C for 0.5 h. The solvent was removed and the residue was diluted with water and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC to give 2-{4-[(2-{3-[(4-methane sulfonylphenyl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino]piperidin-1-yl}-N,N-dimethylacetamide as a light yellow solid.

[1285] LC-MS (ES + ,m / z): 590.4 [(M+H) + ]

[1286] The following compounds were prepared starting from 2-iodo-1 -(2,2,2- trifluoroethyl)-1 H-indol-4-amine and using similar methods to those described in Example 455:

[1287]

[1288]

[1289]

[1290]

[1291]

[1292]

[1293] Example 498 Preparation of 2-[5-({3-[1 -(cyanomethyl)-4-[(1 -methylpiperidin-4- yl)amino]-1 H-indol-2-yl]prop-2-yn-1 -yl}amino)pyridin-2-yl]-2-methylpropanenitrile

[1294]

[1295] Synthesis scheme:

[1296]

[1297] A flask was charged with Cul (89 mg, 465.6 pmol) and diisopropylamine (222 mg, 2.2 mmol, 309 pL) and a solution of tert-butyl (2-iodo-lH-indol-4-yl)(l- methylpiperidin-4-yl)carbamate (200 mg, 439.2 pmol, prepared from 2-iodo-4-nitro-lH- indole) and tert-butyl (6-(2-cyanopropan-2-yl)pyridin-3-yl)(prop-2-yn-l-yl)carbamate (197 mg, 658.9 pmol) in dimethyl sulfoxide (3 mL) was added under nitrogen followed by tetrakis(triphenylphosphine)palladium(0) (61 mg, 52.7 pmol). After the reaction was stirred at 45 °C for 1 h, the reaction was diluted with 20 mL ethyl acetate and 50 mL 1 M EDTA and stirred at 25 °C for 1 h. The reaction was extracted with ethyl acetate (3 x 10 mL) and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate and concentrated in vacuo. The crude residue was purified by preparative thin layer chromatography (silica gel eluting with petroleum ether / ethyl acetate: 1 / 1) to give tert-butyl (2-(3-((tert-butoxycarbonyl)(6-(2-cyanopropan-2-yl)pyridin-3-yl)amino)prop- 1-yn-1-yl)-lH-indol-4-yl)(l-methylpiperidin-4-yl)carbamate (200 mg, 319.1 pmol, 73% yield) as a yellow solid.

[1298] To a mixture of tert-butyl (2-(3-((tert-butoxycarbonyl)(6-(2-cyanopropan-2-yl)pyridin-3- yl)amino)prop-1-yn-1-yl)-lH-indol-4-yl)(l-methylpiperidin-4-yl)carbamate (200 mg, 319 pmol) in dimethylformamide (3 mL) cooled to 0 °C was added sodium hydride (38 mg, 957 pmol, 60% in mineral oil) and 2-bromoacetonitrile (147 mg, 957 pmol). The reaction was stirred at 0 °C for 2 h and poured into ice water (50 mL, 1 / 1 w / w). The reaction mixture was then extracted with ethyl acetate (3 x 10 mL). The combined organic layers were washed with brine (3 x 10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give tert-butyl (2-(3-((tert-butoxycarbonyl)(6-(2-cyanopropan-2-yl)pyridin-3-yl)amino)prop- 1-yn-1-yl)-l-(cyanomethyl)-lH-indol-4-yl)(l-methylpiperidin-4-yl)carbamate (200 mg, crude) as a yellow oil.

[1299] To a mixture of tert-butyl (2-(3-((tert-butoxycarbonyl)(6-(2-cyanopropan-2-yl)pyridin-3- yl)amino)prop-1-yn-1-yl)-1-(cyanomethyl)-1H-indol-4-yl)(1-methylpiperidin-4-yl)carbamate (100 mg, 1 equiv) was added, in one portion, bismuth trichloride (15 equiv) in acetonitrile (5 mL) at 50 °C under nitrogen atmosphere. The mixture was stirred at 50 °C for 1 h, poured into a sodium bicarbonate ice-water mixture (30 mL, 1 / 1 w / w) and to the resulting mixture was added EDTA solution (2 M, 50 mL). The biphasic mixture was stirred at room temperature for 2 h, then separated into the aqueous and organic phases. The aqueous phase was extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with brine (3 x 20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC to give 2-[5-({3-[1-(cyanomethyl)-4-[(1-methylpiperidin-4-yl)amino]-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile as a yellow solid. LC-MS (ES+) m / z): 466.3 [(M+H)]. + ,m / z):466.3 [(M+H) + ]

[1300] The following compounds were prepared starting from 2-iodo-4-nitro-1H-indole and using similar methods to those described in Example 498:

[1301]

[1302]

[1303] Example 513 Preparation of 1-[6-(1-cyano-1-methylethyl)pyridin-3-yl]-3-(3-{4-[(1-methylpiperidin-4- yl)amino]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)urea

[1304]

[1305] To a mixture of propargyl bromide (2.1 g, 38.1 mmol, 2.44 mL) in acetonitrile (80 mL) was added pyridine (7.54 g, 95.3 mmol, 7.69 mL) at 0 °C. The mixture was stirred at 0 °C for 1 h and to the reaction was added phenyl chloroformate (6.0 g, 38.3 mmol). After the reaction was stirred at 0 °C for 1 h, the reaction mixture was poured into water (100 mL) and the aqueous phase was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (3 x 50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluting with petroleum ether / ethyl acetate: 20 / 1 to 5 / 1) to give phenyl prop-2-yn-1-ylcarbamate (5 g, 75% yield) as a white solid.

[1306] To a mixture of phenyl prop-2-yn-1-ylcarbamate (100 mg, 570.8 μmol) in acetonitrile (3 mL) was added triethylamine (144 mg, 1.4 mmol, 198 μL) and 2-(5- aminopyridin-2-yl)-2-methylpropanenitrile (92 mg, 570.8 μmol) at 20 °C. The mixture was stirred at 50 °C for 2 h and at 100 °C for 12 h. The reaction mixture was poured into water (10 mL) and the aqueous phase was extracted with ethyl acetate (3 x 10 mL). The combined organic layers were washed with brine (3 x 10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative thin layer chromatography (silica gel, eluting with petroleum ether / ethyl acetate: 1 / 1) to give 1-(6-(2-cyanopropan-2-yl)pyridin-3-yl)-3-(prop-2-yn-1-yl)urea (90 mg, 46% yield) as a white solid.

[1307] To a 25 °C mixture of l-(6-(2-cyanopropan-2-yl)pyridin-3-yl)-3-(prop-2-yn-l- yl)urea (90 mg) in dimethyl sulfoxide (2 mL) was added diisopropylamine (160.5 mg, 1.59 mmol), Cul (60.4 mg, 317.3 μmol), 2-iodo-N-(l-methylpiperidin-4-yl)-l-(2,2,2- trifluoroethyl)-lH-indol-4-amine (80 mg, 158.6 μmol) and tetrakis(triphenylphosphine)palladium(0) (37 mg, 31.7 μmol). The mixture was stirred at 25 °C for 1 h and diluted with ethyl acetate (10 mL) before being poured into a 2 N EDTA solution (30 mL) and stirred for 2 h. The aqueous phase was extracted with ethyl acetate (2 x 30 mL). The combined organic layers were washed with brine (3 x 30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC to give l-[6-(l-cyano-l-methylethyl)pyridin-3-yl]-3-(3-{4-[(l- methylpiperidin-4-yl)amino]-l-(2,2,2-trifluoroethyl)-lH-indol-2-yl}prop-2-yn-l- yl)urea (19.6 mg, 22% yield) as a white solid.

[1308] LC-MS (ES + ,m / z): 552.4 [(M+H) + ]

[1309] The following compounds were prepared starting from 2-iodo-4-nitro-lH-indole and using similar methods to those described in Example 513:

[1310]

[1311]

[1312] Example 522 : 4-[(2-{3-[(6-methanesulfonylpyridin-3-yl)amino]prop-l-yn-l-yl}-l-(2,2,2- trifluoroethyl)-lH-indol-4-yl)amino]-lλ 6 Preparation of 4-[(2-{3-[(6-methanesulfonylpyridin-3-yl)amino]prop-l-yn-l-yl}-l- (2,2,2-trifluoroethyl)-lH-indol-4-yl)amino]-lλ

[1313]

[1314] Synthesis scheme:

[1315]

[1316] A mixture of 6-(methylsulfonyl)pyridin-3-amine (5 g, 29.04 mmol) and di-tert- butyl dicarbonate (95.07 g, 435.60 mmol) in dioxane (80 mL) was stirred at 110 °C for 72 h. The solvent was removed and the crude product was purified by flash column chromatography (silica gel, eluting with petroleum ether / ethyl acetate: 5 / 1 to 1 / 1) to afford tert-butyl (6-(methylsulfonyl)pyridin-3- yl)carbamate (6 g, 75.86% yield) as a yellow solid.

[1317] To a solution of tert-butyl (6-(methylsulfonyl)pyridin-3-yl)carbamate (1.7 g, 6.24 mmol) in dimethylformamide (20 mL) was added sodium hydride (624 mg, 15.60 mmol, 60% in mineral oil). After the reaction was stirred at 0 °C for 30 min, propargyl bromide (1.86 g, 12.48 mmol, 1.34 mL) was added and the mixture was stirred at 0 °C for 1 h under nitrogen atmosphere. The mixture was quenched by the addition of saturated ammonium chloride solution (20 mL) and the mixture was extracted with dichloromethane (2 x 20 mL). The combined organic layers were washed with water (20 mL) and brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluting with petroleum ether / ethyl acetate: 1 / 1) to afford tert-butyl (6-(methylsulfonyl)pyridin-3-yl)(prop-2-yn-1-yl)carbamate (1.8 g) as a yellow solid.

[1318] A solution of tert-butyl (6-(methylsulfonyl)pyridin-3-yl)(prop-2-yn-1-yl)carbamate (6 g, 19.33 mmol) in HCl / ethyl acetate (60 mL) was stirred at 25 °C for 12 h. The mixture was filtered and concentrated in vacuo to afford 6-(methylsulfonyl)-N-(prop-2-yn-1-yl)pyridin-3-amine hydrochloride (3 g, crude) as a yellow solid which was used without further purification.

[1319] To a solution of 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine (1.50 g, 6.08 mmol) in dimethyl sulfoxide (20 mL) was added diisopropylamine (2.56 g, 25.33 mmol, 3.56 mL), CuI (1.02 g, 5.37 mmol), 6-(methylsulfonyl)-N-(prop-2-yn-1-yl)pyridine-3-amine hydrochloride (1.72 g, 5.07 mmol) and tetrakis(triphenylphosphine)palladium(0) (703 mg, 608 μmol). The reaction mixture was stirred at 25 ° C. for 1 h under a nitrogen atmosphere and then poured into EDTA solution (20 mL) and ethyl acetate (20 mL). The two-phase mixture was stirred at room temperature for 1 h and then separated. The aqueous layer was extracted with ethyl acetate (2x20 mL). The combined organic layer was washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluted with petroleum ether / ethyl acetate: 1 / 1) to give 2-(3-((6-(methylsulfonyl)pyridin-3-yl)amino)prop-1-yn-1-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine (2 g, 93% yield) as a dark brown oil.

[1320] To a solution of 2-(3-((6-(methylsulfonyl)pyridin-3-yl)amino)prop-1-yn-1-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine (200 mg, 473.46 μmol) in ethanol (2 mL) was added Ti(EtO)4 (301 mg, 2.37 mmol, 311 μL) and tetrahydro-4H-thiopyran-4-one 1,1-dioxide (351 mg, 2.37 mmol). The reaction was stirred at 50 ° C for 12 h. Sodium cyanoborohydride (89 mg, 1.42 mmol) was then added to the reaction mixture, and the reaction was stirred at 50 ° C for 1 h under a nitrogen atmosphere. The mixture was poured into a cold saturated sodium bicarbonate solution (5 mL). The mixture was filtered through Celite, and the filtrate was extracted with ethyl acetate (2 x 10 mL). The combined organic layers were washed with water (10 mL) and brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC to give 4-[(2-{3-[(6-methanesulfonylpyridin-3-yl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino]-1λ as a yellow oil. 6 -thiazole-1,1-dione.

[1321] LC-MS(ES + ,m / z):555.2[(M+H) + ]

[1322] The following compounds were prepared starting from 2-(3-((6-(methylsulfonyl)pyridin-3- yl)amino)prop-1 -yn-1 -yl)-1 -(2,2,2-trifluoroethyl)-1 H-indol-4-amine and using similar procedures to those described in Example 522:

[1323]

[1324] Example 530 Preparation of 2-{3-[(4-methanesulfonylphenyl)amino]prop-1 -yn-1 -yl}-N-(oxan-4-yl)-1 -(2,2,2- trifluoroethyl)-1 H-indol-4-amine

[1325]

[1326] To a solution of 4-(methylsulfonyl)-N-(prop-2-yn-1 -yl)aniline (369.2 mg, prepared from 4-(methylsulfonyl)aniline and propargyl bromide) in dimethyl sulfoxide (1 mL) was added N-isopropylpropan-2-amine (268 mg), CuI (50 mg), 2-iodo-1 -(2,2,2-trifluoroethyl)-1 H-indol-4- amine (300 mg) and tetrakis(triphenylphosphine)palladium(0) (102 mg). The mixture was stirred at 35 °C for 4 h, poured into saturated sodium bicarbonate solution (30 mL) and filtered. The filtrate was extracted with ethyl acetate (2 x 10 mL), the combined organic layers were poured into saturated EDTA solution (20 mL) and the resulting biphasic mixture was stirred for 1 h. Once the biphasic mixture had separated into an organic layer and an aqueous layer, the organic layer was concentrated under reduced pressure and the residue was purified by preparative thin layer chromatography to give 2-(3-((4-(methylsulfonyl)phenyl)amino)prop-1 -yn-1 -yl)-1 -(2,2,2- trifluoroethyl)-1 H-indol-4-amine as a yellow solid (250 mg, 68% yield).

[1327] Using similar procedures to those described in Example 455, 2-(3-((4-(methylsulfonyl)phenyl)amino)prop-1 -yn-1 -yl)-1 -(2,2,2-trifluoroethyl)-1 H-indol-4-amine was coupled with tetrahydro-4H-pyran-4-one.

[1328] LC-MS (ES + ,m / z): 506.2 [(M+H) + ]

[1329] The following compounds were prepared starting from 2-iodo-1 -(2,2,2-trifluoroethyl)-1 H-indol-4- amine and using similar procedures to those described in Example 522:

[1330]

[1331]

[1332]

[1333]

[1334]

[1335]

[1336]

[1337]

[1338]

[1339]

[1340]

[1341]

[1342]

[1343]

[1344]

[1345]

[1346] ...

Claims

1. A compound of the formula: in: -Q 1 is the key; -R 2 is an ethyl group which is unsubstituted or substituted by halogen; -R 3 、R 13 and R 16 Each is hydrogen; -R 4 is piperidinyl or tetrahydropyranyl, each of which is unsubstituted or substituted by halogen and alkane One or more substitutions in the alkyl group, wherein the alkyl group is selected from the group consisting of methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl and decyl; -R 17 is phenyl which is unsubstituted or substituted with one or more of halogen, cyano, methoxy, ethoxy, sulfonamide, amide, sulfone, methyl, ethyl, isopropyl and tert-butyl; or a pharmaceutically acceptable salt thereof.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 4 is piperidinyl which is unsubstituted or substituted with one or more of halogen and alkyl, wherein the alkyl is selected from methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl and decyl.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 2 It is an ethyl group substituted by fluorine.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is 。

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