Methods and compounds for restoring mutant p53 function

Abstract

Mutations of oncogenes and tumor suppressor genes promote the development and progression of cancer. The present disclosure describes compounds and methods for restoring wild type function of p53 mutants. The compounds of the present invention can bind to mutant p53 and restore the ability of the p53 mutant to bind to 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 application of the Chinese patent application with the application date of February 17, 2017, application number 201780013450.6, and invention name “Methods and compounds for restoring the function of mutant p53” (the application date of the corresponding PCT application is February 17, 2017, and application number PCT / US2017 / 018511).

[0002] Cross-references

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

[0004] Sequence Listing

[0005] This application contains a sequence listing, which has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy was created on February 17, 2017, is named 44727-702_601_SL.txt, and is 2,540 bytes in size. Background Art

[0006] Cancer is a multifactorial disease characterized by uncontrolled cell proliferation, tumor formation, growth, and, in some cases, metastasis. Cells harboring activated oncogenes, damaged genomes, or other cancer-promoting alterations are prevented from replicating by an elaborate tumor suppressor network. A central component of this tumor suppressor network is p53, one of the most potent tumor suppressors in cells. Both wild-type and mutant forms of p53 are implicated in the progression of cancer.

[0007] Incorporation by reference

[0008] Each patent, publication, and non-patent document cited in this application is hereby incorporated by reference in its entirety, as if each were individually incorporated by reference. Summary of the Invention

[0009] In some embodiments, the present invention provides a compound of the following formula, or a pharmaceutically acceptable salt thereof:

[0010]

[0011] in:

[0012] - Each are independently a single bond or a double bond;

[0013] -X 1 CR 5 , CR 5R 6 、N、NR 5 , O, S, C=O, C=S or with Q 1 the attached carbon atom;

[0014] -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;

[0015] -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;

[0016] -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;

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

[0018] where X 1 、X 2 、X 3 and X 4 At least one of them is 1 the attached carbon atom;

[0019] -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;

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

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

[0022] -R 1 -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 is hydrogen or halogen;

[0028] -Each R 19 and R 20 -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 is 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 is 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 is hydrogen.

[0033] In some embodiments, the present invention provides a method for 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.

[0034] In some embodiments, the present invention 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 compound increases the ability of the p53 mutant to bind DNA, wherein the cell expresses the p53 mutant.

[0035] In some embodiments, the present invention 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 as compared to the ability of the p53 mutant to bind DNA in the absence of the compound.

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

[0037] 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;

[0038] b) contacting the labeled mutant p53 portion 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 of the compound based on the fluorescence resonance energy transfer 150 value;

[0042] 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

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

[0044] In some embodiments, the present invention provides a method for 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 by at least about 50% in an assay, wherein the assay comprises:

[0045] 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;

[0046] b) contacting the labeled mutant p53 portion 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 of the compound based on the fluorescence resonance energy transfer 150 value;

[0050] 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

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

[0052] In some embodiments, the present invention provides a method of inducing apoptosis in a cell, 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 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;

[0054] b) contacting the labeled mutant p53 portion 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 of the compound based on the fluorescence resonance energy transfer 150 value;

[0058] f) Comparison of the SC of the compounds 150 The SC values ​​of the control samples were 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] Figure 1 Shown is a protein DNA binding assay of mutant p53 in the presence of compounds of the present invention.

[0069] Figure 2 Protein DNA binding assay of mutant p53 in the presence of the compounds of the present invention is shown to demonstrate the specificity of the compounds of the present invention. DETAILED DESCRIPTION

[0070] The present invention provides compounds and methods for restoring wild-type function to mutant p53. The compounds of the present invention can bind to mutant p53 and restore the mutant p53's ability to bind DNA. Restoration of mutant p53 activity can allow activation of p53's downstream effectors, leading to inhibition of cancer progression. The present invention also provides methods for 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 major categories, including, for example: carcinomas, which can arise from cells that line the inside and outside of the body, such as the lungs, breast, and colon; sarcomas, which can arise from cells located in bone, cartilage, fat, connective tissue, muscle, and other supporting tissues; lymphomas, which can arise in the lymph nodes and immune system tissue; 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 nearly any tissue of the body and contain unique characteristics, the basic processes that lead to cancer are likely similar across all forms of the disease. Cancer begins when cells break free from their normal constraints on cell division and begin to grow and divide uncontrollably. Genetic mutations in cells can eliminate the cell's ability to repair damaged DNA or initiate apoptosis, leading 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 attrition. Programmed cell death, or apoptosis, represents the primary mechanism of cell attrition. Cancer cells can evade apoptosis through various strategies, such as inhibiting p53 function and 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 activating or inhibiting their activity abnormally, further promoting uncontrolled cell division. Oncogenes contribute to cell growth, while tumor suppressor genes slow cell division by repairing damaged DNA and activating apoptosis. Cellular oncogenes that may mutate in cancer include, for example, Cdk1, Cdk2, Cdk3, Cdk4, Cdk6, EGFR, PDGFR, VEGF, HER2, Raf kinase, K-Ras, and myc. Tumor suppressor genes that may mutate in cancer include, for example, BRCA1, BRCA2, cyclin-dependent kinase inhibitor 1C, retinoblastoma protein (pRb), PTEN, p16, 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 stresses including, for example, UV radiation, hypoxia, oncogene activation, and DNA damage. p53 has multiple mechanisms that inhibit cancer progression, including, for example, initiation of apoptosis, maintenance of genomic stability, cell cycle arrest, induction of senescence, and inhibition of angiogenesis. Due to its critical role in tumor suppression, p53 is inactivated in almost all cancers, either through direct mutation or through interference with related signaling pathways involved in tumor suppression. Homozygous deletion of the p53 gene occurs in almost all types of cancer, including breast cancer, colon cancer, and lung cancer. The presence of certain p53 mutations in several types of human cancer may be associated with a less favorable patient prognosis.

[0077] In the absence of stress signals, p53 levels are maintained low by its interaction with the E3 ubiquitin ligase Mdm2. In unstressed cells, Mdm2 targets p53 for degradation by the proteasome. Under stressful conditions, the interaction between Mdm2 and p53 is disrupted, leading to p53 accumulation. A key event leading to p53 activation is phosphorylation of the N-terminal domain of p53 by protein kinases, which transduce upstream stress signals. Phosphorylation of p53 induces conformational changes that promote p53 DNA binding and allow transcription of downstream effectors. Activation of p53 can induce, for example, intrinsic and extrinsic apoptotic pathways, cell cycle arrest, senescence, and DNA repair. p53 can activate proteins involved in the above-mentioned 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 inhibit the transcription of multiple genes including, for example, c-MYC, cyclin B, VEGF, RAD51, and hTERT.

[0078] Each chain of the p53 tetramer is composed of several functional domains, including the transactivation domain (amino acids 1-100), the DNA binding domain (amino acids 101-306), and the tetramerization domain (amino acids 307-355), which are highly mobile and largely unstructured. Most p53 cancer mutations are located in the protein's DNA-binding core domain, which contains a central β-sandwich structure of antiparallel β-sheets that serves as the basic scaffold for the DNA binding surface. The DNA binding surface consists of two β-turn loops, L2 and L3, stabilized by zinc ions, such as at Arg175 and Arg248, and a loop-sheet-helix motif. Together, these structural elements form an extended DNA-binding surface that is rich in positively charged amino acids and specifically contacts various p53 response elements.

[0079] Since p53 mutations are prevalent in almost all types of cancer, the reactivation of wild-type p53 function in cancer cells may be an effective therapy. P53 mutations located in the DNA binding domain of the protein or the periphery of the DNA binding surface lead to abnormal protein folding required for DNA recognition and binding. Mutations in p53 can occur at, for example, amino acids Val143, His168, Arg175, Tyr220, Gly245, Arg248, Arg249, Phe270, Arg273, and Arg282. P53 mutations that can eliminate 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 thermodynamically destabilize the folded protein at body temperature. Wild-type function of p53 mutants can be restored by binding them to compounds that shift 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); glutamate (E, Glu); glutamine (Q, Gln); 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 present invention

[0082] The compounds of the present invention can selectively bind to p53 mutants and 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 present invention selectively bind to the p53 Y220C mutant. The Y220C mutant is a temperature-sensitive mutant that binds to DNA at lower temperatures and denatures at body temperature. The compounds of the present 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 present invention may 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 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 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 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 up to about 86%, at least or up to about 87%, at least or up to about 88%, at least or up to about 89%, at least or up to about 90%, at least or up to about 91%, at least or up to about 92%, at least or up to about 93%, at least or up to about 94%, at least or up to about 95%, at least or up to about 96%, at least or up to about 97%, at least or up to about 98%, at least or up to about 99%, at least or up to about 100%, at least or up to about 125%, at least or up to about 150%, at least or up to about 175%, at least or up to about 200%, at least or up to about 225%, or at least or up to about 250%.

[0097] The compounds described herein can increase the activity of a p53 mutant, i.e., for example, by at least or up to about 2-fold, at least or up to about 3-fold, at least or up to about 4-fold, at least or up to about 5-fold, at least or up to about 6-fold, at least or up to about 7-fold, at least or up to about 8-fold, at least or up to about 9-fold, at least or up to about 10-fold, at least or up to about 11-fold, at least or up to about 12-fold, at least or up to about 13-fold, at least or up to about 14-fold, at least or up to about 15-fold, at least or up to about 16-fold, at least or up to about 17-fold, at least or up to about 18-fold, at least or up to about 19-fold, at least or up to about 20-fold, at least or up to about 21-fold, at least or up to about 22-fold, at least or up to about 23-fold, at least or up to about 24-fold, at least or up to about 25-fold, at least or up to about 26-fold, at least or up to about 27-fold, at least or up to about 28-fold, at least or up to about 29-fold, at least or up to about 30-fold, at least or up to about 31-fold, at least or up to about 32-fold, at least or up to about 33-fold, at least or up to about 34-fold, at least or up to about 35-fold, at least or up to about 36-fold, at least or up to about 37-fold, at least or up to about 38-fold, at least or up to about 39-fold, at least or up to about 40-fold, at least or up to about 41-fold, at least or up to about 42-fold, at least or At least or up to about 18 times, at least or up to about 19 times, at least or up to about 20 times, at least or up to about 25 times, at least or up to about 30 times, at least or up to about 35 times, at least or up to about 40 times, at least or up to about 45 times, at least or up to about 50 times, at least or up to about 55 times, at least or up to about 60 times, at least or up to about 65 times, at least or up to about 70 times, at least or up to about 75 times, at least or up to about 80 times, at least or up to about 85 times, at least or up to about 90 times, at least or up to about 95 times, at least or up to about 100 times, at least or up to about 110 times, at least or up to about 120 times, at least or up to about 130 times, at least or up to about 140 times, at least or up to about 150 times, at least or up to about 160 times, at least or up to about 170 times, at least or up to about 180 times, at least or up to about 190 times, at least or up to about 200 times, at least or up to about 250 times, at least or up to about 300 times, at least or up to about 350 times, at least or up to about 400 times, at least or up to about 450 times, at least or up to about 500 times, at least or up to about 550 times, at least or up to about 600 times, at least or up to about 650 times, at least or up to about 700 times, at least or up to about 750 times, at least or up to about 800 times, at least or up to about 800 times, In some embodiments, the amount of the pharmaceutical composition can be increased by at least or up to about 100 times, at least or up to about 750 times, at least or up to about 800 times, at least or up to about 850 times, at least or up to about 900 times, at least or up to about 950 times, at least or up to about 1,000 times, at least or up to about 1,500 times, at least or up to about 2,000 times, at least or up to about 3,000 times, at least or up to about 4,000 times, at least or up to about 5,000 times, at least or up to about 6,000 times, at least or up to about 7,000 times, at least or up to about 8,000 times, at least or up to about 9,000 times, or at least or up to about 10,000 times.

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

[0099] Compounds of the present invention

[0100] Non-limiting examples of compounds of the present invention include any of the following formulae:

[0101]

[0102] In some embodiments, the compound of the present invention is a compound of the formula:

[0103]

[0104] Each of these are independently single bonds or double bonds; X 1 CR 5 , CR 5 R 6 、N、NR 5 , O, S, C=O, C=S or with Q 1 Attached carbon atom; X 2 CR 7 , CR 7 R 8 、N、NR 7 , O, S, C=O, C=S or with Q 1 Attached carbon atom; X 3 CR 9 , CR 9 R 10 、N、NR 9 , O, S, C=O, C=S or with Q 1 Attached carbon atom; X 4 CR 11 , CR 11 R 12 、N、NR 11 , O, S, C=O, C=S or with Q 1 Attached carbon atom; X 5 CR 13 , N or NR 13 ; where X 1 、X 2 、X 3 and X 4 At least one of them is 1 Attached carbon atom; 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 is a bond; m is 1, 2, 3 or 4; Y is N, O or absent; R 1 -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 is 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 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; 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 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 20C(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; and each R 23 and R 24 is independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or is hydrogen.

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

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

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

[0108]

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

[0110]

[0111] where R 1 -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.

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

[0113]

[0114] where R 1 -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 is hydrogen.

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

[0116] In some embodiments, 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. 1 In some embodiments, Q 1 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 an aryl group, and R 17 In some embodiments, Q 1 is C1-alkylene, R 16 is an aryl group, and R 17 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 heterocyclic. 17 is phenyl, indolyl, piperidinyl, imidazolyl, thiazolyl, morpholinyl, pyrrolyl or pyridinyl.

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

[0124]

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

[0126]

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

[0128]

[0129] Each Z 1 and Z 2 Independently for CR x or N; each R x 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 25 and R 26 are 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 is hydrogen or halogen.

[0130] In some embodiments, Z 1 is N. In some embodiments, Z 1 and Z 2 is N. In some embodiments, each R 25 and R 26 In some embodiments, R 25 for In some embodiments, R 25 SO2R 21 In some embodiments, R 25 SO2R 21 , where R 21 In some embodiments, R 25 SO2R 21 , where R 21 It is a methyl group.

[0131] Non-limiting examples of compounds of the present invention 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 invention 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 invention 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 、R2b 、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′ are 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 2 is independently alkylene, alkenylene, alkynylene, heteroalkylene, cycloalkylene, heterocycloalkylene, cycloarylene, or heterocycloalkylene.

[0153] In some embodiments, L 1 is an alkylene group and L 2 For esters.

[0154] The compounds herein may 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, azide, sulfoxide, sulfone, sulfonamide, carboxyl, carboxaldehyde, imine, alkyl, halo-alkyl, alkenyl, halo-alkenyl, alkynyl, halo-alkynyl, alkoxy, aryl, aryloxy, aralkyl, arylalkoxy, heterocyclic, acyl, acyloxy, carbamate, amide, urea, epoxy, and ester groups.

[0156] Non-limiting examples of alkyl and alkylene groups include straight chain, branched chain, and cyclic alkyl and alkylene groups. Alkyl or alkylene groups 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] Branched chain alkyl groups include any straight chain alkyl group substituted with any number of alkyl groups. Non-limiting examples of branched chain alkyl groups include isopropyl, isobutyl, sec-butyl, and tert-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 bicyclics and higher fused, bridged and spiro systems. Cyclic alkyl groups can be substituted by any number of straight chain, branched or cyclic alkyl groups. Non-limiting examples of cyclic alkyl groups include cyclopropyl, 2-methyl-cyclopropyl-1-yl, cyclopropyl-2-ene-1-yl, cyclobutyl, 2,3-dihydroxycyclobut-1-yl, cyclobut-2-ene-1-yl, cyclopentyl, cyclopent-2-ene-1-yl, cyclopent-2,4-diene-1-yl, cyclohexyl, cyclohex-2-ene-1-yl, cycloheptyl, cyclooctyl, 2,5-dimethylcyclopent-1-yl, 3,5-dichlorocyclohex-1-yl, 4-hydroxycyclopent-2,4-diene-1-yl. cyclohexan-1-yl, 3,3,5-trimethylcyclohexan-1-yl, octahydropentenyl, octahydro-1H-indenyl, 3a,4,5,6,7,7a-hexahydro-3H-inden-4-yl, decahydroazulyl, 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 chain and cyclic alkenyl groups. The one or more alkenyl groups of the alkenyl group can be, for example, E, Z, cis, trans, terminal or exo-methylene groups. 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 、C 17 、C 18 、C 19 、C 20 、C21 、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 、C 28 、C 29 、C 30 、C 31 、C32 、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 substituted with any number of halogen atoms, such as fluorine, chlorine, bromine, and iodine atoms. Halo-alkenyl can be any alkenyl substituted with any number of halogen atoms. Halo-alkynyl can be any alkynyl substituted with any number of halogen atoms.

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

[0165] Aryl can be heterocyclic or non-heterocyclic. Aryl can be monocyclic or polycyclic. Aryl can be substituted by any number of substituents described herein such as hydrocarbon groups, alkyl groups, alkoxy groups and halogen atoms. Non-limiting examples of aryl include phenyl, toluoyl, naphthyl, pyrrolyl, pyridyl, imidazolyl, thienyl and furyl. Non-limiting examples of substituted aryl 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 such as N, O, S, P, Si, B or any other heteroatoms that are not carbon.Heterocycle can be substituted by 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, pyrans and tetrahydropyran.

[0168] Non-limiting examples of heterocycles include heterocyclic units having a monocyclic 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-1H-azepine and ii) a heterocyclic unit having two or more rings one of which is a heterocycle, non-limiting examples of which include hexahydro-1H-pyrrolidinyl, 3a,4,5,6,7,7a-hexahydro-1H-benzo[d]imidazolyl, 3a,4,5,6,7,7a-hexahydro-1H-indolyl, 1,2,3,4-tetrahydroquinolinyl, and decahydro-1H-cycloocta[b]pyrrolyl.

[0169] Non-limiting examples of heteroaryl groups include: i) heteroaryl rings 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, pyridinyl, 3-methylpyridinyl, and 4-dimethylaminopyridinyl; and ii) heteroaryl rings containing Non-limiting examples of heteroaryl rings wherein one ring is a fused heteroaryl ring 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 of the compounds herein may be purified. A compound herein may 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 % 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 invention can be used to treat cancer in a subject. For example, the compounds of the present invention can slow the proliferation of cancer cell lines or kill cancer cells. Non-limiting examples of cancers that can be treated by the compounds of the present invention include: acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, AIDS-related cancers, AIDS-related lymphomas, anal cancer, appendix cancer, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain tumors such as cerebellar astrocytoma, cerebral astrocytoma / malignant glioma, ependymoma, medulloblastoma, supratentorial primitive neuroectodermal tumor, visual pathway and hypothalamic glioma, breast cancer, bronchial adenoma, Burkitt's lymphoma, cancer of unknown primary site, central nervous system lymphoma, cerebellar astrocytoma, Cervical cancer, childhood cancers, chronic lymphocytic leukemia, chronic myeloid 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, stomach cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, gliomas, 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, squamous neck cancer with occult metastasis of the primary site, oral cancer, multiple endocrine neoplasia syndrome, myelodysplastic syndrome, 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, islet cell pancreatic cancer, paranasal sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, Pineal astrocytoma, pineal germ cell tumor, pituitary adenoma, pleuropulmonary blastoma, plasmacytoma, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell carcinoma, transitional cell carcinoma of the renal pelvis and ureter, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, skin cancer, Merkel cell skin cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, gastric cancer, T-cell lymphoma, laryngeal cancer, thymoma, thymic carcinoma, thyroid cancer, trophoblastoma (pregnancy), cancer of unknown primary site, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom's macroglobulinemia, and Wilms' tumor.

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

[0173] Pharmaceutically acceptable salts

[0174] The present invention provides for the use of pharmaceutically acceptable salts 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 acid or an inorganic acid. The base added to the compound to form the base addition salt can be an organic base or an 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 the compounds of the invention. The inorganic base consists of a metal cation paired with a basic counterion, such as hydroxide, carbonate, bicarbonate, or phosphate. The metal can be an alkali metal, alkaline earth metal, transition metal, or 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, sodium salt, potassium salt, cesium salt, cerium salt, magnesium salt, manganese salt, iron salt, calcium salt, strontium salt, cobalt salt, titanium salt, aluminum salt, copper salt, cadmium salt, or zinc salt.

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

[0178] In some embodiments, the ammonium salt is a triethylamine salt, a diisopropylamine 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 pyridinium salt, a pipyrrazole salt, a piperazole salt, an imidazole salt, a pyrazine salt, or a pipyrazine salt.

[0179] Acid addition salts can be produced by adding acid to the compounds of this invention. 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, glucuronic acid (glucaronic acid), sugar 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 hydrochloride, hydrobromide, hydroiodide, nitrate, nitrite, sulfate, sulfite, phosphate, isonicotinate, lactate, salicylate, tartrate, ascorbate, gentisate, gluconate, glucuronate, saccarate, formate, benzoate, glutamate, pantothenate, acetate, propionate, butyrate, fumarate, succinate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, citrate, oxalate, or maleate.

[0181] Pharmaceutical composition of the present invention

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

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

[0184] The pharmaceutical compositions of the present invention can be any of the pharmaceutical compounds described herein in combination with other chemical components such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and / or excipients. The pharmaceutical compositions facilitate administration of the compounds to an organism. The pharmaceutical compositions can be administered as pharmaceutical compositions in therapeutically effective amounts in various forms and routes, including, for example, intravenous, subcutaneous, intramuscular, oral, parenteral, ocular, subcutaneous, transdermal, nasal, vaginal, and topical administration.

[0185] The pharmaceutical composition can be used in a local manner, for example, via direct injection of the compound into an organ, optionally in the form of a reservoir or sustained release formulation or implant. The pharmaceutical composition can be provided in the form of a quick-release formulation, in the form of an extended-release formulation, or in the form of an intermediate-release formulation. The quick-release form can provide immediate release. Extended-release formulations can provide controlled-release or sustained delayed release.

[0186] For oral administration, pharmaceutical composition can be prepared by combining active compound with pharmaceutically acceptable carrier or excipient.Such carrier can be used to prepare liquid, gel, syrup, elixir, slurry or suspension for oral ingestion by subject.Non-limiting examples of the solvent used in oral soluble preparations can include water, ethanol, isopropyl alcohol, saline, normal saline, DMSO, dimethylformamide, potassium phosphate buffer, phosphate buffered saline (PBS), sodium phosphate buffer, 4-2-hydroxyethyl-1-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 the cosolvent used in oral soluble preparations can include sucrose, urea, cremaphor, DMSO and potassium phosphate buffer.

[0187] Pharmaceutical preparations can be formulated for intravenous administration. The pharmaceutical composition can be in the form of a sterile suspension, solution or emulsion in an oily or aqueous vehicle suitable for parenteral injection, and can contain preparatons such as suspending agents, stabilizers and / or dispersants. Pharmaceutical preparations for parenteral administration include aqueous solutions of the active compound in water-soluble form. The suspension of the active compound can be prepared into an oily injection suspension. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. The suspension can also contain suitable stabilizers or agents that increase the solubility of the compound to allow the preparation of highly concentrated solutions. Alternatively, the active ingredient can be in powder form for reconstitution with a suitable vehicle such as sterile pyrogen-free water before use.

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

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

[0190] The compounds can also be formulated in 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, a low melting wax such as a mixture of fatty acid glycerides optionally in combination with cocoa butter may be melted.

[0191] In practicing the treatment or use methods provided herein, a therapeutically effective amount of a compound described herein is administered in the form of a pharmaceutical composition to a subject suffering from the disease or condition to be treated. 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 efficacy 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 a mixture.

[0192] Pharmaceutical compositions can be formulated using one or more physiologically acceptable carriers comprising excipients and adjuvants that facilitate processing of the active compound into pharmaceutically acceptable preparations. The formulation can vary depending on the route of administration chosen. Pharmaceutical compositions containing the compounds described herein can be prepared, for example, by mixing, dissolving, emulsifying, encapsulating, embedding, or compression processes.

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

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

[0195] Non-limiting examples of dosage forms suitable for use with the present invention 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 invention include binders, disintegrants, anti-adherents, antistatic agents, surfactants, antioxidants, coating agents, colorants, plasticizers, preservatives, suspending agents, emulsifiers, antimicrobial agents and spheronizers, and any combination thereof.

[0197] The compositions of the present invention can be, for example, quick-release or controlled-release preparations. Quick-release preparations can be formulated to allow compounds to work quickly. Non-limiting examples of quick-release preparations include easily soluble preparations. Controlled-release preparations can be such pharmaceutical preparations, which have been adjusted so that the release rate and release profile of the active agent can be matched with physiological and chronotherapy requirements, or have been formulated to realize that the active agent is released at a programmed rate. Non-limiting examples of controlled-release preparations include particles, delayed-release particles, hydrogels (for example, synthetic or natural origin), other gelling agents (for example, the dietary fiber forming a gel), preparations based on matrix (for example, preparations comprising a polymeric material with at least one active ingredient dispersed therein), particles in the matrix, polymeric mixtures and particulate matter.

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

[0199] Controlled release formulations can be sustained release forms. Sustained release forms can be formulated to maintain, for example, the effects of a compound over an extended period of time. Sustained release forms can be formulated to provide an effective dose of any compound described herein (e.g., to provide a physiologically effective blood profile) within about 4, about 8, about 12, about 16, or about 24 hours.

[0200] Non-limiting examples of pharmaceutically acceptable excipients can be found in, for example, 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, HA and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th ed. (Lippincott Williams & Wilkins 1999), each of which is herein incorporated by reference in its entirety.

[0201] The multiple therapeutic agents may be administered in any order or simultaneously. In some embodiments, the compounds of the invention are administered in combination with another therapeutic agent, before, or after the treatment. If administered simultaneously, the multiple therapeutic agents may be provided in a single, unified form or in multiple forms, such as multiple individual pills. The agents may be packaged together in a single package or individually in multiple packages. One or all of the therapeutic agents may be administered in multiple doses. If not administered simultaneously, the time between multiple doses may vary up to about one month.

[0202] The therapeutic agent as described herein can be used before, during or after the disease or the patient's condition occurs, and the time selection of applying the composition containing the therapeutic agent can be changed. For example, the composition can be used as a prophylactic agent and can be continuously applied to a subject susceptible to the condition or disease to reduce the possibility of the disease or the patient's condition occurring. The composition can be applied to the subject as quickly as possible during or after the onset of symptoms. The administration of the therapeutic agent can be started within the first 48 hours of the onset of symptoms, within the first 24 hours of the onset of symptoms, within the first 6 hours of the onset of symptoms, or within 3 hours of the onset of symptoms. Any preparation as described herein can be used, via any practical approach, such as by any approach as described herein for initial administration.

[0203] After a disease or condition is detected or suspected, the compound can be administered as soon as it is available and for the length of time necessary to treat the disease, for example, from about 1 month to about 3 months. In some embodiments, the compound can be administered for a period of 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 The length of treatment can vary for each subject.

[0204] The pharmaceutical compositions described herein can be in the form of unit dosage forms suitable for single administration of precise doses. In a unit dosage form, the formulation is divided into unit doses containing an appropriate amount of one or more compounds. The unit dose can be in the form of a package containing discrete amounts of the formulation. Non-limiting examples are packaged injections, vials, or ampoules. Aqueous suspension compositions can be packaged in non-resealable single-dose containers. Resealable multi-dose containers can be used, for example, with or without preservatives. Formulations for injection can be presented in unit dosage form, for example, in ampoules or multi-dose containers with preservatives.

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

[0206] Depending on the intended mode of administration, the pharmaceutical composition may be in the form of a solid, semisolid or liquid dosage form, e.g., tablet, suppository, pill, capsule, powder, liquid, suspension, lotion, cream or gel, e.g., in the form of a unit dosage form suitable for single administration of a precise dose.

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

[0208] Non-limiting examples of pharmaceutically active agents suitable for combination with the compositions of the present invention include anti-infective agents, i.e., aminoglycosides, antiviral agents, antimicrobial agents, anticholinergics / antispasmodics, antidiabetics, antihypertensives, antineoplastic agents, cardiovascular agents, central nervous system agents, coagulation regulators, hormones, immunosuppressants, immunosuppressants, and ophthalmic agents.

[0209] Compounds can be delivered via liposome technology. Using liposomes as drug carriers can increase the therapeutic index of a compound. Liposomes are composed of natural phospholipids and may contain mixed lipid chains with surfactant properties (e.g., egg phosphatidylethanolamine). Liposome design can employ surface ligands to attach to unhealthy tissues. Non-limiting examples of liposomes include multilamellar vesicles (MLVs), small unilamellar vesicles (SUVs), and large unilamellar vesicles (LUVs). The physicochemical properties of liposomes 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. The optimal liposome properties depend on the route of administration: large-sized liposomes show good retention upon local injection, while small-sized liposomes are more suitable for passive targeting. PEGylation reduces the uptake of liposomes by the liver and spleen and prolongs circulation time, thereby increasing localization to inflamed sites due to the enhanced permeability and retention (EPR) effect. In addition, the liposome surface can be modified to achieve selective delivery of encapsulated drugs to specific target cells. Non-limiting examples of targeting ligands include monoclonal antibodies, vitamins, peptides, and polysaccharides that are specific for receptors concentrated on the surface of cells associated with the disease.

[0210] Non-limiting examples of dosage forms suitable for use in the present invention 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 invention include granulating agents, binders, lubricants, disintegrants, sweeteners, glidants, anti-adherents, antistatic agents, surfactants, antioxidants, gums, coating agents, colorants, flavorings, coating agents, plasticizers, preservatives, suspending agents, emulsifiers, plant cellulosic materials, and spheronizers, and any combination thereof.

[0211] The composition of the present invention can be packaged into a test kit. In some embodiments, the test kit includes written instructions for the administration / use of the composition. The written material can be, for example, a label. The written material can suggest the conditions and methods of administration. The instructions provide the subject and the attending physician with the best guidance for obtaining the best clinical results from the 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 agencies.

[0212] Drug administration

[0213] The pharmaceutical compositions described herein can be in the form of unit dosage forms suitable for single administration of precise doses. In a unit dosage form, the formulation is divided into unit doses containing an appropriate amount of one or more compounds. The unit dose can be in the form of a package containing discrete amounts of the formulation. Non-limiting examples are liquids in vials or ampoules. Aqueous suspension compositions can be packaged in non-reclosable single-dose containers. Reclosable multi-dose containers can be used, for example, in combination with preservatives. Formulations for parenteral injection can be presented in unit dosage form, for example, in ampoules or multi-dose containers with preservatives.

[0214] The compounds described herein can be administered in an amount 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. In some embodiments, the present invention provides an iodine-containing pharmaceutical composition comprising: a ...

[0215] The compounds described herein can be administered in 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 6 In some embodiments, the present invention relates to an amount of at least one pharmaceutical composition comprising: about 100 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 of pharmaceutical composition.

[0216] In some embodiments, the dosage can be expressed as the amount of drug divided by the mass of the subject, for example, 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 invention provides embodiments including but not limited to the following:

[0218] 1. A compound of the following formula, or a pharmaceutically acceptable salt thereof:

[0219]

[0220] in:

[0221] - Each are independently a single bond or a double bond;

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

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

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

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

[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 carbon atom to which it is attached;

[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 、-SR 21 、-

[0240] 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;

[0241] -Each R 19 and R 20 C(O)R 23 、-C(O)OR 23 、-C(O)NR 23 R 24 、-

[0242] 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;

[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 is 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 is hydrogen.

[0246] 2. The compound according to embodiment 1, wherein X 3 For Q 1 The carbon atom to which it is attached, 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 according to embodiment 1, wherein the compound is a compound of the following 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 For alkyl.

[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 CR 13 , N or NR 13 ;

[0297] where X 1 、X 2 、X 3 and X 4 At least one of them is 1 the carbon atom to which it is attached;

[0298] -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;

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

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

[0301] -R 1 -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 heterocyclic group,

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

[0305] -Each R 3 and R 4 are 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 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

[0307] R 3 does not exist;

[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 Independently for-

[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 is hydrogen or halogen;

[0311] -Each R 19 and R 20 -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 is hydrogen or halogen;

[0313] -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;

[0314] and

[0315] -Each R 23 and R 24 is independently alkyl, alkenyl, alkynyl, aryl, heteroaryl, or heterocyclyl, each of which is independently substituted or unsubstituted, or is 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 to the p53 mutant compared to wild-type p53.

[0320] 45. The method of embodiment 40, wherein the therapeutically effective amount is from 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 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:

[0322] 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;

[0323] b) contacting the labeled mutant p53 portion 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 the fluorescence resonance energy transfer;

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

[0327] 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

[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 the cell.

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

[0335] 53. The method of embodiment 46, wherein the therapeutically effective amount is from 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.

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

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

[0344] 62. The method of embodiment 46, wherein the subject is 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 tagged p53 is His-tagged.

[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. A method as described in 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)-1H-indole

[0356] To a solution of 5-bromo-indole (60 g, 309 mmol) in tetrahydrofuran (600 mL) was added NaH (18.5 g, 464 mmol, 60% in mineral oil) at 0°C. The mixture was stirred at 0°C for 1 h. PhSO Cl (65 g, 370 mmol) in tetrahydrofuran (600 mL) was then 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 a 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, eluted 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)-1H-indole

[0358] To a stirred solution of 5-bromo-1-(phenylsulfonyl)-1H-indole (28 g, 83.28 mmol) in anhydrous tetrahydrofuran (500 mL) at -70°C was added a solution of LDA (2 M, 62.46 mL) dropwise. After the mixture was stirred at 0°C for 2 h, the progress of anion formation was checked by adding D2O to an equal portion. The solution was then cooled to -70°C and a solution of I2 (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 ammonium chloride solution (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-1H-indole

[0360] The mixture of 5-bromo-2-iodo-1-(phenylsulfonyl)-1H-indole (30.00g, 61.67mmol) and potassium carbonate (2M, 100mL) in methyl alcohol (300mL) was stirred at 90 ℃ for 2h. After confirming that the reaction was complete by thin layer chromatography (petroleum ether / ethyl acetate 5 / 1), 80% solvent was removed, and reaction mixture was filtered to obtain thick 5-bromo-2-iodo-1H-indole (13g, yellow solid). This crude product was used for next reaction without the need for further purification.

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

[0362] To a mixture of 5-bromo-2-iodo-1H-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) at 0°C and 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 that the reaction was complete. The reaction mixture was poured into an 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 phases were washed with brine (2 x 200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give crude 5-bromo-1-ethyl-2-iodo-1H-indole (10 g, yellow solid). The crude product was used directly without further purification.

[0363] 1 H NMR (400MHz MeOD): δ7.60-7.65(dd,J=1.92Hz,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-1H-indol-2-yl)-2-propynyl]aniline

[0365] The flask was charged with copper (I) iodide (544.2 mg, 2.86 mmol) and triethylamine (3.62 g, 35.73 mmol), and crude 5-bromo-1-ethyl-2-iodo-1H-indole (5.00 g,

[0366] To the mixture was added 4-[ ... 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]aniline (3.10 g, 49.13% yield) as a yellow solid.

[0367] 1 H NMR (400MHz, MeOD) δ7.68 (s, 1H), 7.24-7.30 (m, 4H), 7.11-7.13 (d, J = 4.0Hz, 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-1H-indole-5-carbaldehyde

[0369] To a solution of [3-(5-bromo-1-ethyl-1H-indol-2-yl)-2-propynyl] aniline (500mg, 1.42mmol) in tetrahydrofuran (10.00mL) cooled to -78°C under nitrogen, n-butyl lithium (2.5M, 3.41mL) was added once. The mixture was stirred at -78°C for 30min, then 4-morpholinecarboxaldehyde (1.63g, 14.20mmol) was added once at -78°C. The mixture was stirred for 1.5h, and thin layer chromatography (petroleum ether / ethyl acetate, 5 / 1) showed that the reaction was complete. Residue was poured into aqueous ammonium chloride ice-water (20mL, w / w=1 / 1) and stirred for 3min. The aqueous phase was extracted with ethyl acetate (2x 10mL). 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-anilino-1-propynyl)-1-ethyl-1H-indole-5-carbaldehyde (600 mg) as a yellow oil. The product was used directly in the next step without further purification.

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

[0371] To a mixture of 2-(3-anilino-1-propynyl)-1-ethyl-1H-indole-5-carbaldehyde (200 mg, 529.15 μmol, 1 eq) and 1-methylpiperidin-4-amine (53.5 mg, 529.15 μmol, 1 eq) in dichloromethane (10 mL) was added anhydrous magnesium sulfate (318.5 mg, 2.65 mmol, 5 eq) at 25°C under nitrogen. The mixture was stirred at 25°C for 60 min, then NaBH(OAc) (336.5 mg, 1.59 mmol, 3 eq) was added and the reaction mixture was stirred for 5 h. LCMS showed that 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 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 (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% HCl) to afford 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 2 : Preparation of 1-anilino-3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}-2-propyne

[0374]

[0375] In a manner analogous to the method 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 3: Preparation of 1-anilino-3-{1-ethyl-5-[(tetrahydro-2H-pyran-4-ylamino)methyl]-1H-indol-2-yl}-2-propyne

[0378]

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

[0380] Example 4 : Preparation of 1-anilino-3-[5-(benzylaminomethyl)-1-ethyl-1H-indol-2-yl]-2-propyne

[0381]

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

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

[0384] Example 5 : Preparation of 3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}-1-(p-fluorophenylamino)-2-propyne

[0385]

[0386] Preparation of [3-(5-bromo-1-ethyl-1H-indol-2-yl)-2-propynyl](p-fluorophenyl)amine

[0387] To a solution of 5-bromo-1-ethyl-2-iodo-indole (800 mg, 2.29 mmol, 1 equiv) in tetrahydrofuran (5 mL) was added 4-fluoro-N-prop-2-ynyl-aniline (341 mg, 2.29 mmol, 1 equiv), triethylamine (694 mg, 6.86 mmol, 951 μL, 3 equiv), copper (I) iodide (44 mg, 228.58 μmol, 0.1 equiv) and tetrakis (triphenylphosphine) palladium (0) (264 mg, 228.58 μmol, 0.1 equiv) at 25°C. The mixture was stirred for 2 h, then poured into EDTA solution (2 M, 20 mL) and stirred for an additional 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) and dried over anhydrous sodium sulfate. The solid was filtered off and the solvent was removed in vacuo. 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-1-ethyl-indol-2-yl)prop-2-ynyl]-4-fluoro-aniline (750 mg, 88.2% yield) as a yellow solid.

[0388] Preparation of 1-ethyl-2-[3-(4-fluoroanilino)prop-1-ynyl]indole-5-carbaldehyde

[0389] To a solution of N-[3-(5-bromo-1-ethyl-indol-2-yl)prop-2-ynyl]-4-fluoro-aniline (700 mg, 1.89 mmol, 1 equiv) in tetrahydrofuran (5.00 mL) cooled to -78 ° C was added n-butyllithium (2.5 M, 3.02 mL, 4 equiv). After the mixture was stirred at -78 ° C for 0.5 h, morpholine-4-carboxaldehyde (1.09 g, 9.43 mmol, 944 μL, 5 equiv) was added. 2 h after the addition, thin layer chromatography showed that the reaction was complete. The reaction mixture was poured into an aqueous ammonium chloride solution (5 mL) and extracted with ethyl acetate (5 mL). The combined organic phase was washed with water (5 mL) and brine (5 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, eluting with petroleum ether / ethyl acetate 1 / 1) to give 1-ethyl-2-[3-(4-fluoroanilino)prop-1-ynyl]indole-5-carbaldehyde (300 mg, 49.6% yield) as a yellow solid.

[0390] In a manner analogous to the method described in Example 1, 1-ethyl-2-[3-(4-fluoroanilino)prop-1-ynyl]indole-5-carbaldehyde was reacted with methylamine to give 3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}-1-(p-fluorophenylamino)-2-propyne.

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

[0392] Example 6 : Preparation of 3-{1-ethyl-5-[(tetrahydro-2H-pyran-4-ylamino)methyl]-1H-indol-2-yl}-1-(p-fluorophenylamino)-2-propyne

[0393]

[0394] Following the procedure described in Example 1, 1-ethyl-2-[3-(4-fluoroanilino)prop-1-ynyl]indole-5-carbaldehyde was reacted with 4-aminotetrahydropyran to give 3-{1-ethyl-5-[(tetrahydro-2H-pyran-4-ylamino)methyl]-1H-indol-2-yl}-1-(p-fluorophenylamino)-2-propyne.

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

[0396] Example 7 : Preparation of 1-(p-chlorophenylamino)-3-{1-ethyl-5-[(1-methyl-4-piperidinylamino)methyl]-1H-indol-2-yl}-2-propyne

[0397]

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

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

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

[0401] Example 8 : Preparation of 3-{1-ethyl-5-[(tetrahydro-2H-pyran-4-ylamino)methyl]-1H-indol-2-yl}-1-(6-methyl-3-pyridylamino)-2-propyne

[0402]

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

[0404] Following the procedure described in Example 1, 1-ethyl-2-{3-[(6-methylpyridin-3-yl)amino]prop-1-yn-1-yl}-1H-indole-5-carbaldehyde was reacted with 4-aminotetrahydropyran to give 3-{1-ethyl-5-[(tetrahydro-2H-pyran-4-ylamino)methyl]-1H-indol-2-yl}-1-(6-methyl-3-pyridinylamino)-2-propyne.

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

[0406] Example 9 : Preparation of 3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}-1-(6-methyl-3-pyridylamino)-2-propyne

[0407]

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

[0409] In a manner analogous to the method described in Example 1, 1-ethyl-2-{3-[(6-methylpyridin-3-yl)amino]prop-1-yn-1-yl}-1H-indole-5-carbaldehyde was reacted with methylamine to give 3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}-1-(6-methyl-3-pyridinylamino)-2-propyne.

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

[0411] Example 10: Preparation of 3-{1-ethyl-5-[(1-methyl-4-piperidinylamino)methyl]-1H-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}-1H-indole-5-carbaldehyde was prepared in a manner similar to that described for the preparation of 1-ethyl-2-[3-(4-fluoroanilino)prop-1-ynyl]indole-5-carbaldehyde in Example 6.

[0414] Following the procedure described in Example 1, 1-ethyl-2-{3-[(2-methylpyridin-4-yl)amino]prop-1-yn-1-yl}-1H-indole-5-carbaldehyde was reacted with 1-methylpiperidin-4-amine to give 3-{1-ethyl-5-[(1-methyl-4-piperidinylamino)methyl]-1H-indol-2-yl}-1-(2-methyl-4-pyridinylamino)-2-propyne.

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

[0416] Example 11 : Preparation of 3-[5-(benzylaminomethyl)-1-ethyl-1H-indol-2-yl]-1-(2-methyl-4-pyridylamino)-2-propyne

[0417]

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

[0419] Following the procedure described in Example 1, 1-ethyl-2-{3-[(2-methylpyridin-4-yl)amino]prop-1-yn-1-yl}-1H-indole-5-carbaldehyde was reacted with benzylamine to give 3-[5-(benzylaminomethyl)-1-ethyl-1H-indol-2-yl]-1-(2-methyl-4-pyridinylamino)-2-propyne.

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

[0421] Example 12 :Preparation of N-(3-{5-[(diethylamino)methyl]-1-ethyl-1H-indol-2-yl}prop-2-yn-1-yl)aniline

[0422]

[0423] In a manner analogous to the method described in Example 1, 2-(3-anilino-1-propynyl)-1-ethyl-1H-indole-5-carbaldehyde was reacted with diethylamine to give N-(3-{5-[(diethylamino)methyl]-1-ethyl-1H-indol-2-yl}prop-2-yn-1-yl)aniline.

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

[0425] Example 13 : Preparation of 4-chloro-N-(3-{5-[(diethylamino)methyl]-1-ethyl-1H-indol-2-yl}prop-2-yn-1-yl)aniline

[0426]

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

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

[0429] Example 14 :Preparation of N-({1-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H-indol-5-yl}methyl)oxetan-3-amine

[0430]

[0431] In a manner similar to the method described in Example 1, 2-(3-anilino-1-propynyl)-1-ethyl-1H-indole-5-carbaldehyde was reacted with oxetan-3-amine to give N-({1-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H-indol-5-yl}methyl)oxetan-3-amine. LC-MS (ES + ,m / z):360.2[(M+H)+ ]

[0432] Example 15 :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 manner similar to the method described in Example 1, 2-(3-anilino-1-propynyl)-1-ethyl-1H-indole-5-carbaldehyde was reacted with 2-methylpropylamine to give N-[3-(1-ethyl-5-{[(2-methylpropyl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]aniline. LC-MS (ES + ,m / z):360.3[(M+H) + ]

[0435] Example 16 :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 manner similar to the method described in Example 1, 2-(3-anilino-1-propynyl)-1-ethyl-1H-indole-5-carbaldehyde 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 17 : Preparation of N-({1-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H-indol-5-yl}methyl)-1-methanesulfonylpiperidin-4-amine

[0439]

[0440] Following the procedure described in Example 1, 2-(3-anilino-1-propynyl)-1-ethyl-1H-indole-5-carbaldehyde was reacted with 1-methanesulfonylpiperidin-4-amine to give N-({1-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H-indol-5-yl}methyl)-1-methanesulfonyl-piperidin-4-amine.

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

[0442] Example 18 :Preparation of N-(3-{1-ethyl-5-[(ethylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)aniline

[0443]

[0444] Following the procedure described in Example 1, 2-(3-anilino-1-propynyl)-1-ethyl-1H-indole-5-carbaldehyde was reacted with ethylamine to give N-(3-{1-ethyl-5-[(ethylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)aniline.

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

[0446] Example 19 : Preparation of N-{3-[5-({[2-(dimethylamino)ethyl]amino}methyl)-1-ethyl-1H-indol-2-yl]prop-2-yn-1-yl}aniline

[0447]

[0448] In a manner analogous to the method described in Example 1, 2-(3-anilino-1-propynyl)-1-ethyl-1H-indole-5-carbaldehyde was reacted with 2-dimethylaminoethylamine to give N-{3-[5-({[2-(dimethylamino)ethyl]amino}methyl)-1-ethyl-1H-indol-2-yl]prop-2-yn-1-yl}aniline.

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

[0450] Example 20 : 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]pyridin-3-amine

[0451]

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

[0453] Following the procedure described in Example 1, 2-{3-[(6-tert-butylpyridin-3-yl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indole-5-carbaldehyde was reacted with 1-methylpiperidin-4-amine to give 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 21 :Preparation of N-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5-yl)methyl]oxane-4-amine

[0456]

[0457] Following the procedure described in Example 1, 1-ethyl-2-[3-(4-chloroanilino)prop-1-ynyl]indole-5-carbaldehyde was reacted with 4-aminotetrahydropyran to provide N-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5-yl)methyl]oxan-4-amine.

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

[0459] Example 22 : 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-carbaldehyde was prepared in a manner similar to that described for the preparation of 1-ethyl-2-[3-(4-fluoroanilino)prop-1-ynyl]indole-5-carbaldehyde in Example 6.

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

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

[0463] Example 23 : Preparation of 4-[(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)amino]benzonitrile

[0464]

[0465] At 25 ° C, to a solution of N-[(1-ethyl-2-iodo-1H-indol-5-yl)methyl]amino 2,2-dimethylpropanoic acid methyl ester (1 equivalent) in dimethyl sulfoxide (5.0 mL), p-(2-propynylamino)benzonitrile (1 equivalent), copper (I) iodide (0.47 equivalent), diisopropylamine (3 equivalents) and tetrakis (triphenylphosphine) palladium (0) (0.1 equivalent) were added, and the mixture was stirred for 2 hours under a nitrogen atmosphere. After LCMS showed that the reaction was complete, the reaction mixture was poured into EDTA solution (20 mL) and stirred for 2 hours. 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 solid was filtered off and the filtrate was concentrated in vacuo to obtain 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 afford methyl N-({2-[3-(p-cyanophenylamino)-1-propynyl]-1-ethyl-1H-indol-5-yl}methyl)amino 2,2-dimethylpropanoate as a yellow oil.

[0466] To a solution of methyl N-({2-[3-(p-cyanophenylamino)-1-propynyl]-1-ethyl-1H-indol-5-yl}methyl)amino 2,2-dimethylpropanoate (1 equiv.) in acetonitrile (2 mL) was added BiCl3 (4 equiv.) at 50 ° C., and the mixture was stirred under a nitrogen atmosphere for 1 h. LCMS showed that the reaction was complete. The reaction mixture was poured into EDTA solution (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 solid was 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 afford p-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}-2-propynylamino)benzonitrile as a white solid.

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

[0468] Example 24 : Preparation of 4-tert-butyl-N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)benzamide

[0469]

[0470] 4-tert-Butyl-N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)benzamide was prepared in a manner analogous to the method described in Example 23 using (2-propynylamino)[p-(tert-butyl)phenyl]carbaldehyde.

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

[0472] Example 25 : Preparation of 4-chloro-N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)-3-fluorobenzamide

[0473]

[0474] 4-Chloro-N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)-3-fluorobenzamide was prepared in a manner analogous to the method described in Example 23 using (4-chloro-3-fluorophenyl)(2-propynylamino)carbaldehyde.

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

[0476] Example 26 : Preparation of 4-cyano-N-({1-ethyl-2-[3-(phenylformamido)prop-1-yn-1-yl]-1H-indol-5-yl}methyl)-N-methylbenzamide

[0477]

[0478] (3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}-2-propynylamino)phenylcarbaldehyde was prepared in a manner analogous to the method described in Example 23 using phenyl(2-propynylamino)carbaldehyde.

[0479] To a solution of (3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}-2-propynylamino)phenylcarboxaldehyde (1 equiv.) and triethylamine (3 equiv.) in dichloromethane (5 mL) was added 4-cyanobenzoyl chloride (1 equiv.) at 25 ° C., and the mixture was stirred under a nitrogen atmosphere for 5 h. LCMS showed that the reaction was 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), and the organic phase was washed with water (50 mL) and brine (50 mL) and dried over anhydrous sodium sulfate. The solid was 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-(phenylformamido)prop-1-yn-1-yl]-1H-indol-5-yl}methyl)-N-methylbenzamide as a white solid.

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

[0481] Example 27 : Preparation of 3-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)-1-[4-(trifluoromethyl)phenyl]urea

[0482]

[0483] 3-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)-1-[4-(trifluoromethyl)phenyl]urea was prepared in a manner analogous to the method 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 28 :Preparation of N-{[1-(2-chloroethyl)-2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1H-indol-5-yl]methyl}oxane-4-amine

[0486]

[0487] To a solution of 1H-indole-5-carboxaldehyde (85 g, 585.56 mmol, 1 equivalent) in dichloromethane (100 mL) was added potassium hydroxide (65.38 g, 1.17 mol, 2 equivalents) and n-Bu4NSO4 (198.82 g, 585.56 mmol, 1 equivalent). The mixture was stirred at 25 ° C for 30 min, then PhSO2Cl (155.13 g, 878.34 mmol, 112.41 mL, 1.5 equivalents) was added. The mixture was stirred at 25 ° C for 1 h, and thin layer chromatography showed that the reaction was complete. The reaction mixture was poured into an 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 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 / dichloromethane: 15 / 1) to give 1-(phenylsulfonyl)indole-5-carbaldehyde (124 g, 71% yield) as a white solid.

[0488] To a solution of 1-(phenylsulfonyl)indole-5-carbaldehyde (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) (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, and thin layer chromatography showed the reaction was complete. The mixture was poured into aqueous sodium bicarbonate (100 mL) and extracted with dichloromethane (3×50 mL). The combined organic layers were washed with aqueous sodium bicarbonate (3×50 mL) and brine (50 mL) and dried over anhydrous sodium sulfate. The solid was filtered off and the filtrate was concentrated in vacuo to give N-[[1-(phenylsulfonyl)indol-5-yl]methyl]-tetrahydropyran-4-amine (4.6 g) as an off-white gum. The 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 equivalent) in dioxane (60 mL) was added (Boc)2O (4.61 g, 21.11 mmol, 4.85 mL, 1.70 equivalents), and the reaction mixture was stirred at 90 ° C for 1 h. Thin layer chromatography showed that the reaction was complete. The reaction mixture was concentrated in vacuo and the crude product 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 equivalent) in tetrahydrofuran (6 mL) was added LDA (2 M, 1.31 mL, 2.5 equivalents) in several small portions, and the reaction mixture was stirred at -78 ° C for 1 h. A solution of iodine (293.2 mg, 1.16 mmol, 1.1 equivalents) in 1 mL of tetrahydrofuran was added, and the reaction mixture was stirred at -78 ° C for another 1 h. LCMS showed a conversion of approximately 20%. The reaction mixture was poured into an aqueous ammonium chloride solution (100 mL) and extracted with dichloromethane (3 x 30 mL). The combined organic layer was 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. The crude product was used directly without further purification.

[0491] To a solution of tert-butyl N-[[1-(phenylsulfonyl)-2-iodo-indol-5-yl]methyl]-N-tetrahydropyran-4-yl-carbamate (2.5 g, 4.19 mmol, 1 equivalent) in methanol (50 mL) was added potassium carbonate (2.9 g, 20.95 mmol, 5 equivalents) and the reaction mixture was stirred at 80° C. for 5 h. Thin layer chromatography showed that the reaction was complete. The reaction mixture was poured into 100 mL of ice water and acidified to pH=4 with 37% HCl. The mixture was extracted with dichloromethane (3 x 30 mL), and 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-1H-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 tert-butyl N-[(2-iodo-1H-indol-5-yl)methyl]-N-tetrahydropyran-4-yl-carbamate (300 mg, 90% purity, 591.69 μmol, 1 eq) in dichloroethane (58.6 mg, 591.69 μmol, 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 μmol, 1 eq). The reaction mixture was stirred at 25 ° C for 6 h. Thin layer chromatography showed that 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-[[1-(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-aniline (122 mg, 736.68 μmol, 1.5 eq) in dimethyl sulfoxide (5 mL) was added N-isopropylpropan-2-amine (149.09 mg, 1.47 mmol, 207.07 μL, 3.00 eq) and copper (I) iodide (28.06 mg, 147.34 μmol, 0.3 eq) followed by N-[[1-(2-chloroethyl)-2-iodo-indol-5-yl]methyl]-N-tetrahydropyran-4-yl-carbamic acid tert-butyl ester (260 mg, 491.12 μmol, 1 eq) and tetrakis(triphenylphosphine)palladium(0) (56.8 mg, 49.11 μmol, 0.1 eq). The reaction mixture was stirred at 25° C. for 1 h after which thin layer chromatography indicated the reaction was complete. In 4- hydroxy-2- pyridine-2-ol (200mg, 71% yield) 4- hydroxy-2- pyridine-3 ...

[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 equivalent) in acetonitrile (2 mL) was added trichlorobismuthane (204 mg, 646.88 μmol, 42.94 μL, 4 equivalents), and the reaction mixture was stirred at 50 ° C for 2 h. LCMS showed that the reaction was complete. An EDTA aqueous solution (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 layer was washed with water (3 x 10 mL) and brine (10 mL), and dried over anhydrous sodium sulfate. The solid was 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 29 : 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-methylpropionitrile

[0497]

[0498] To a solution of 2-methyl-2-[p-(2-propynylamino)phenyl]propionitrile (2 equivalents) and [(1-ethyl-2-iodo-1H-indol-5-yl)methyl](1-methyl-4-piperidinyl)amino 2,2-dimethylpropanoate (100 mg, 1 equivalent) in dimethyl sulfoxide (2 mL) was added N-isopropylpropan-2-amine (3 equivalents) and copper (I) iodide (0.2 equivalents), followed by tetrakis(triphenylphosphine)palladium (0) (0.1 equivalents). After the reaction mixture was stirred at 25 ° C for 1 h, the mixture was poured into an EDTA aqueous 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 solid was 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, eluting 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 two-phase 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 solid was filtered off and the filtrate was concentrated in vacuo. The crude residue was purified by reverse phase HPLC (C18, eluting 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-methylpropionitrile.

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

[0501] Example 30 : Preparation of 4-cyano-N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)benzamide

[0502]

[0503] 4-Cyano-N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)benzamide was prepared in a manner analogous to the method described in Example 23 using p-[(2-propynylamino)carbonyl]benzonitrile.

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

[0505] Example 31 : Preparation of N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)-6-methylpyridine-3-carboxamide

[0506]

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

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

[0509] Example 32 : Preparation of 3-[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]-1-phenylurea

[0510]

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

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

[0513] Example 33 : Preparation of N-[(2-{3-[(4-chloro-3-fluorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-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 manner similar to the method 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 34 : 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-methylpropionitrile

[0518]

[0519] To a solution of 2-methyl-2-[5-(2-propynylamino)-2-pyridyl]propionitrile (2.5 equiv) in dimethyl sulfoxide (2 mL) was added N-isopropylpropan-2-amine (3 equiv) and copper (I) iodide (0.3 equiv), followed by 1-ethyl-2-iodo-1H-indole-5-carbaldehyde (100 mg, 1 equiv) and tetrakis(triphenylphosphine)palladium (0) (0.1 equiv). After the reaction mixture was stirred at 25 ° C. under a nitrogen atmosphere for 1 h, the mixture was poured into an EDTA aqueous 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 sulfate. The solid was 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-pyridinyl}-2-methylpropionitrile.

[0520] To a solution of 2-{5-[3-(1-ethyl-5-formyl-1H-indol-2-yl)-2-propynylamino]-2-pyridyl}-2-methylpropionitrile (1 equiv.) in dichloromethane / methanol (4 mL, 1 / 1 mixture) were added 1-methyl-4-piperidinylamine (4 equiv.) and anhydrous magnesium sulfate (15 equiv.) respectively. The mixture was stirred at 25°C for 12 h, and then sodium bicarbonate (1 equiv.) and NaBHCN (4 equiv.) were added at 0°C. The reaction mixture was stirred at 0°C for 2 h, and then quenched with a saturated solution of sodium bicarbonate (10 mL). The mixture was extracted with ethyl acetate (2 x 30 mL), and the combined organic phases were washed with water (30 mL) and brine (30 mL), and dried over anhydrous sodium sulfate. The solid was 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 2-(4-{[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}phenyl)-2-methylpropionitrile.

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

[0522] Example 35 : 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 manner similar to the method described in Example 28 using 1-methylpiperidin-4-amine.

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

[0526] Example 36 : 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] 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 was prepared in a manner analogous to the method described in Example 34 using N-2-propynyl[6-(tert-butyl)-3-pyridinyl]amine.

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

[0530] Example 37 : 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-methylpropionic acid

[0531]

[0532] Ethyl 2-(4-{[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}phenyl)-2-methylpropanoate was prepared using ethyl 2-methyl-2-[p-(2-propynylamino)phenyl]propanoate in a manner similar to that described in Example 34. The 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}-1H-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 38 : Preparation of 3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)-N-methylprop-2-ynamide

[0535]

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

[0537] To a solution of 1-(methylamino)-2-propyn-1-one (3 equivalents) in dimethyl sulfoxide (4 mL) was added N-isopropylpropan-2-amine (3 equivalents) and copper (I) iodide (0.2 equivalents), followed by 1-ethyl-2-iodo-1H-indole-5-carboxaldehyde (100 mg, 1 equivalent) and tetrakis(triphenylphosphine)palladium (0) (0.1 equivalents). After the reaction mixture was stirred at 20 ° C for 12 h under a nitrogen atmosphere, the mixture was poured into an EDTA aqueous solution (10 mL). The biphasic mixture was stirred at 25 ° C for 2 h and continued to stand for 4 h, then extracted 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 solid was 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, eluted with a gradient of 10 / 1 to 5 / 1 petroleum ether / ethyl acetate) to give a yellow oil. The 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 equiv.) in dichloromethane (4 mL) were added 1-methyl-4-piperidinylamine (7 equiv.) and anhydrous magnesium sulfate (15 equiv.) respectively. The mixture was stirred at 25 ° C for 12 h, then NaBH CN (2 equiv.) was added at 0 ° C. 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 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 solid was 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-methylprop-2-ynamide.

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

[0540] Example 39 : Preparation of ethyl 2-(4-{[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}phenyl)-2-methylpropanoate

[0541]

[0542] Ethyl 2-(4-{[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}phenyl)-2-methylpropanoate was prepared in a manner similar to the method described in Example 34 using ethyl 2-methyl-2-[p-(2-propynylamino)phenyl]propanoate.

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

[0544] Example 40 : Preparation of 2-(5-{[3-(1-ethyl-5-{[(oxacyclohexane-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile

[0545]

[0546] To a solution of 2-methyl-2-[5-(2-propynylamino)-2-pyridyl]propionitrile (1 equiv.) in dimethyl sulfoxide (2 mL) was added N-isopropylpropan-2-amine (3 equiv.) and copper (I) iodide (0.47 equiv.), followed by 1-ethyl-2-iodo-1H-indole-5-carbaldehyde (100 mg, 1 equiv.) and tetrakis(triphenylphosphine)palladium(0) (0.1 equiv.). After the reaction mixture was stirred at 40 ° C. under a nitrogen atmosphere for 2 h, the mixture was poured into an EDTA aqueous solution (20 mL). The two-phase mixture was stirred at 25 ° C. for 2 h and 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 solid was 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-1H-indol-2-yl)-2-propynylamino]-2-pyridinyl}-2-methylpropionitrile.

[0547] To a solution of 2-{5-[3-(1-ethyl-5-formyl-1H-indol-2-yl)-2-propynylamino]-2-pyridyl}-2-methylpropionitrile (1 equivalent) in dichloromethane (2 mL) was added tetrahydro-2H-pyran-4-ylamine (8 equivalents) and anhydrous magnesium sulfate (10 equivalents) respectively. The mixture was stirred at 25 ° C for 2 h, and then NaBHCN (10 equivalents) was added. The reaction mixture was stirred at 25 ° C for 1 h, and 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 solid was 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, eluting 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 41 : 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 manner similar to the method described in Example 34 using N-2-propynyl(p-tolyl)amine.

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

[0553] Example 42 : 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] 4-{[3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}benzonitrile was prepared in a manner analogous to the method described in Example 34 using p-(2-propynylamino)benzonitrile.

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

[0557] Example 43 : Preparation of 3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)-N-phenylprop-2-ynamide

[0558]

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

[0560] Example 44 :Preparation of N-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5-yl)methyl]-1-methanesulfonylpiperidin-4-amine

[0561]

[0562] A flask was charged with copper (I) iodide (0.3 equivalents) and triethylamine (2.5 equivalents), and a solution of N-2-propynyl (p-chlorophenyl) amine (2 equivalents) and 5-bromo-1-ethyl-2-iodo-1H-indole (1 equivalent) in tetrahydrofuran (10 mL) was added under nitrogen, followed by the addition of tetrakis (triphenylphosphine) palladium (0) (0.05 equivalents). The reaction mixture was stirred at 25° C. for 1 h. Thin layer chromatography (petroleum ether / ethyl acetate: 5 / 1) showed that the reaction was complete. The reaction was diluted with 50 mL of ethyl acetate and 100 mL of 2M EDTA, and the two-phase 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 solid was filtered off and the filtrate was concentrated under reduced pressure. The crude residue was purified by flash column chromatography (silica gel, eluting with a 30 / 1 to 20 / 1 petroleum ether / ethyl acetate gradient) 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 equivalent) in tetrahydrofuran (10 mL) was added n-butyllithium (2.5 M, 6 equivalents) in one portion under nitrogen and -78°C. The mixture was stirred at -78°C for 30 min, and then 4-morpholinecarboxaldehyde (10 equivalents) 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 that the reaction was 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 (2x10 mL). The combined organic phases were washed with brine (2x10 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-carbaldehyde as a yellow oil.

[0564] To a mixture of 2-[3-(p-chlorophenylamino)-1-propynyl]-1-ethyl-1H-indole-5-carboxaldehyde (1 equivalent) and 1-(methylsulfonyl)-4-piperidinylamine (2 equivalents) in dichloromethane (10 mL) was added anhydrous magnesium sulfate (5 equivalents) at 25°C under nitrogen. The mixture was stirred at 25°C for 60 min, then NaBH(OAc) (3 equivalents) was added. The reaction mixture was stirred for 5 h, and LCMS showed that the reaction was complete. The reaction mixture was poured into ice-water (10 mL, w / w=1 / 1), and the two-phase mixture was stirred for 3 min, then separated. The aqueous phase was extracted with dichloromethane (3x 5 mL). The combined organic phases were washed with brine (3x5 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 preparative HPLC (C18 silica gel, eluting 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-methanesulfonylpiperidin-4-amine.

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

[0566] Example 45 : 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] Following the procedure analogously to that described in Example 44, 2-[3-(p-chlorophenylamino)-1-propynyl]-1-ethyl-1H-indole-5-carbaldehyde 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 46 : 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] 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 was prepared in a manner similar to the method described in Example 29 using (2-propynylamino)[6-(tert-butyl)-3-pyridinyl]carbaldehyde.

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

[0574] Example 47 :Preparation of N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)-4-(trifluoromethyl)aniline

[0575]

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

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

[0578] Example 48 :Preparation of N-[(1-ethyl-2-{3-[(4-methylphenyl)amino]prop-1-yn-1-yl}-1H-indol-5-yl)methyl]oxane-4-amine

[0579]

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

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

[0582] Example 49 :Preparation of N-(3-{1-ethyl-4-[(methylamino)methyl]-1H-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-1H-indole (~100 mg, 1.0 eq, prepared from 4-bromo-1H-indole in a manner similar to that described in Example 1) in tetrahydrofuran (10 mL) was added under nitrogen, followed by the addition of 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 that the reaction was complete. The reaction was diluted with ethyl acetate (50 mL) and EDTA (2 M, 100 mL), and the two-phase mixture was stirred at 25 ° C for 3 h. The reaction mixture was extracted with ethyl acetate (3 x 40 mL). The organic extract was washed with saturated brine (150 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 30 / 1 to 20 / 1 gradient) to give N-[3-(4-bromo-1-ethyl-1H-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-1H-indol-2-yl)prop-2-yn-1-yl]aniline (1 equivalent) in tetrahydrofuran (10 mL) was added n-butyllithium (2.5 M, 6 equivalents) at -78 ° C under nitrogen. The mixture was stirred at -78 ° C for 30 min, and 4-morpholinecarboxaldehyde (10 equivalents) was added at -78 ° C. The reaction mixture was stirred at -78 ° C for 1.5 h. TLC (petroleum ether: ethyl acetate = 5: 1) showed that the reaction was 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 afford crude 1-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H-indole-4-carbaldehyde as a yellow oil. The product was used directly in the next step without further purification. LC-MS (ESI) + ,m / z):275.1[(M-CHO) + ]

[0587] To a mixture of 1-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H-indole-4-carbaldehyde (1 equivalent) and methylamine (1 equivalent) in dichloromethane (10 mL) was added anhydrous magnesium sulfate (5 equivalents) in one portion at 25°C under nitrogen. The mixture was stirred at 25°C for 60 min, then NaBH(OAc)3 (3 equivalents) 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]-1H-indol-2-yl}prop-2-yn-1-yl)aniline.

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

[0589] Example 50 :Preparation of N-[3-(1-ethyl-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]aniline

[0590]

[0591] 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 manner similar to that described in Example 49 using 4-(pyrrolidin-1-yl)piperidine.

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

[0593] Example 51 : Preparation of N-({1-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H-indol-4-yl}methyl)-1-methylpiperidin-4-amine

[0594]

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

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

[0597] Example 52 : 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 manner analogous to the method 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 53 : 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 manner analogous to the method 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 54 :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 55 : 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 56 : 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 57: Preparation 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-methylpropionitrile

[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-methylpropionitrile was prepared in a manner similar to the method described in Example 49 using 1,4'-bipiperidin-4-amine and 2-methyl-2-[5-(prop-2-yn-1-ylamino)pyridin-2-yl]propionitrile.

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

[0621] Example 58 :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] Following the procedure described in Example 1, 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.

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

[0625] Example 59 :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] Following the procedure described in Example 1, 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.

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

[0629] Example 60 : Preparation of 4-chloro-N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)-3-fluoroaniline

[0630]

[0631] Following the procedure described in Example 1, 4-chloro-N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)-3-fluoroaniline was prepared.

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

[0633] Example 61 : Preparation of 6-tert-butyl-N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)pyridine-3-carboxamide

[0634]

[0635] Following the procedure described in Example 1, 6-tert-butyl-N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)pyridine-3-carboxamide was prepared.

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

[0637] Example 62 : Preparation of N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)benzamide

[0638] Following the procedure described in Example 1, N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)benzamide was prepared.

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

[0640] Example 63 : Preparation of 3-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)-1-(4-methylphenyl)urea

[0641]

[0642] Following the procedure described in Example 1, 3-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)-1-(4-methylphenyl)urea was prepared.

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

[0644] Example 64 : Preparation of 4-chloro-N-(3-{1-ethyl-5-[(methylamino)methyl]-1H-indol-2-yl}prop-2-yn-1-yl)aniline

[0645]

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

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

[0648] Example 65 : Preparation of 4-{[3-(1-ethyl-5-{[(oxacyclohexane-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}benzonitrile

[0649]

[0650] Following the procedure described in Example 28, 4-{[3-(1-ethyl-5-{[(oxan-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]amino}benzonitrile was prepared.

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

[0652] Example 66 :Preparation of N-[(2-{3-[(4-chloro-3-fluorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5-yl)methyl]oxane-4-amine

[0653]

[0654] Following the procedure described in Example 28, N-[(2-{3-[(4-chloro-3-fluorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5-yl)methyl]oxan-4-amine was prepared.

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

[0656] Example 67 : Preparation of 3-[3-(1-ethyl-5-{[(oxacyclohexane-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]-1-phenylurea

[0657]

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

[0659] Example 68 : Preparation of 6-tert-butyl-N-[3-(1-ethyl-5-{[(oxacyclohexane-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]pyridin-3-amine

[0660]

[0661] Following the procedure described in Example 28, 6-tert-butyl-N-[3-(1-ethyl-5-{[(oxan-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-yl]pyridin-3-amine was prepared.

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

[0663] Example 69 :4-{[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5-yl)methyl]amino}-1λ 6 -Preparation of 1,1-thiol-1,2-dione

[0664]

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

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

[0667] Example 70 : Preparation of N-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5-yl)methyl]-1-(2-methanesulfonylethyl)piperidin-4-amine

[0668]

[0669] Following the procedure described in Example 28, N-[(2-{3-[(4-chlorophenyl)amino]prop-1-yn-1-yl}-1-ethyl-1H-indol-5-yl)methyl]-1-(2-methanesulfonylethyl)piperidin-4-amine was prepared.

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

[0671] Example 71 : 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] Following the procedure 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 72 :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] Following the procedure described in Example 28, 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.

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

[0679] Example 73 : 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] Following the procedure described in Example 28, 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.

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

[0683] Example 74 : 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)propionitrile

[0684]

[0685] Following the procedure described in Example 28, 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 was prepared.

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

[0687] Example 75 : Preparation of 2-[5-({3-[1-(2-fluoroethyl)-5-{[(oxacyclohexane-4-yl)amino]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile

[0688]

[0689] Following the procedure described in Example 28, 2-[5-({3-[1-(2-fluoroethyl)-5-{[(oxan-4-yl)amino]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile was prepared.

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

[0691] Example 76 : Preparation of 3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-ol

[0692]

[0693] Following the procedure described in Example 28, 3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-ol was prepared.

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

[0695] Example 77 : Preparation of 2-[5-({3-[1-(2-chloroethyl)-5-{[(oxacyclohexane-4-yl)amino]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile

[0696]

[0697] Following the procedure described in Example 28, 2-[5-({3-[1-(2-chloroethyl)-5-{[(oxan-4-yl)amino]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile was prepared.

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

[0699] Example 78 : Preparation of 2-[5-({3-[1-(2,2-difluoroethyl)-5-{[(oxacyclohexane-4-yl)amino]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile

[0700]

[0701] Following the procedure 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 was prepared.

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

[0703] Example 79 : 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] Following the procedure 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 was prepared.

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

[0707] Example 80: 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] Synthesis scheme:

[0710]

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

[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, after which sodium borohydride was added in small portions. The mixture was stirred at 0 ° C for 10 min, then allowed to warm to 25 ° C under additional 10 min stirring. 1N sodium hydroxide solution (20 mL) was slowly added under stirring for 30 min, and the mixture was then 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 layer was 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 (2.15 g, 96% crude yield) as a yellow gum. The crude product was used directly without further purification.

[0713] Under nitrogen and 0 ℃, be charged with dichloromethane (60mL) and aluminum chloride (2.27g, 17.01mmol) in a dry flask (250mL). Add { [1- (phenylsulfonyl) -1H- indol-5-yl] methyl} -tetrahydro -2H- pyran -4- ylamine (2.0g, 4.86mmol) in dichloromethane (15mL) dropwise, and after addition, stir the mixture for 10min. Then add acetyl chloride (0.36mL, 0.39g, 5.10mmol) in dichloromethane (15mL) dropwise, and stir the resulting mixture for 10min at 0 ℃. Then make the mixture warm up to 25 ℃, and continue to stir for 3h. The reaction mixture is decanted, and the residue is treated with ice / water (30mL), and then 1N sodium hydroxide solution (30mL) is added. The mixture is stirred for 30min and extracted with ethyl acetate (3x 50mL). 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 afford 1-{1-(phenylsulfonyl)-5-[(tetrahydro-2H-pyran-4-ylamino)methyl]-1H-indol-3-yl}-1-ethanone (1.92 g, 87% crude yield) as a white foaming solid.

[0714] Under nitrogen and 0 ℃, trifluoroacetic acid (30mL) is charged into dry flask (250mL), and sodium borohydride (2.60g, 69.01mmol) is added portionwise under vigorous stirring. 1-{1-(phenylsulfonyl)-5-[(tetrahydro-2H-pyrans-4-ylamino)methyl]-1H-indol-3-yl}-1-ethanone (1.90g, 4.60mmol) in dichloromethane (30mL) is added dropwise at 0 ℃, and the resulting mixture is warmed to 25 ℃. Stirring is continued for 5h. The reaction mixture is quenched with ice / water (20mL), and alkalized with 1N sodium hydroxide (450mL) in 30min while stirring. The mixture was then extracted with dichloromethane (3 x 30 mL), and the organic layer was washed with water (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to afford {[3-ethyl-1-(phenylsulfonyl)-1H-indol-5-yl]methyl}-tetrahydro-2H-pyran-4-ylamine (1.60 g, 86% crude yield) as a light brown foamy solid.

[0715] To {[3-ethyl-1-(phenylsulfonyl)-1H-indol-5-yl]methyl}-tetrahydro-2H-pyran-4-ylamine (0.96 g, 4.02 mmol) in dichloromethane (50 mL) was added Boc-anhydride (0.96 g, 4.42 mmol) dropwise, 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, elution with hexanes / ethyl acetate 4 / 1) to give tert-butyl (3-ethyl-1-(phenylsulfonyl)-1H-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-1-(phenylsulfonyl)-1H-indol-6-yl)methyl(tetrahydro-2H-pyran-4-yl)carbamate (196 mg, 0.40 mmol) in tetrahydrofuran (10 mL) at -78 ° C and nitrogen was added lithium diisopropylamide (1 M, 1.18 mL, 3 equivalents) dropwise while stirring at -78 ° C for 1 h. A solution of iodine (105 mg, 0.413 mmol, 1.05 equivalents) in 1 mL of tetrahydrofuran was added dropwise, and the reaction mixture was stirred at -78 ° C for an additional 5 min after the addition. The reaction mixture was poured into an aqueous ammonium chloride solution (5 mL) and extracted with ethyl acetate (3x30 mL). The combined organic layers were washed with water (3x30 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-1-(phenylsulfonyl)-1H-indol-6-yl)methyl(tetrahydro-2H-pyran-4-yl)carbamate (270 mg) as a yellow oil. The crude product was used directly without further purification.

[0717] To a solution of (3-ethyl-2-iodo-1-(phenylsulfonyl)-1H-indol-6-yl)methyl(tetrahydro-2H-pyran-4-yl)carbamic acid tert-butyl ester (248 mg, 0.4 mmol, 1 equivalent) in methanol (6.8 mL) was added 2M 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 (3x 30 mL). The combined organic layer was washed with water (2x 30 mL) and brine (2x 30 mL) and dried over anhydrous magnesium sulfate. The solid was filtered off and the filtrate was concentrated in vacuo to give crude (3-ethyl-2-iodo-1H-indol-6-yl)methyl(tetrahydro-2H-pyran-4-yl)carbamic acid tert-butyl ester (200 mg) as an orange oil. The 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 iodide (I) (5 mg, 25 μmol) at room temperature and under nitrogen, while stirring for 5 min. The mixture was then stirred for 1 h at 25 ° C using tert-butyl (3-ethyl-2-iodo-1H-indol-6-yl) methyl (tetrahydrochysene-2H-pyrans-4-yl) carbamate (60 mg, 125 μmol, 1 equivalent). The reaction mixture was stirred for 1 h at 25 ° C. The reaction was quenched with ice / water (10 mL). Ethyl acetate (30 mL) and an EDTA aqueous solution (20 mL) were added, and the mixture was stirred for 2 h. The mixture is extracted with ethyl acetate (3x 30mL), and the combined organic layer is washed with water (3x 10mL) and salt solution (10mL), and dried over anhydrous magnesium sulfate.Solid is filtered off and the filtrate is concentrated in vacuo.The crude residue is purified by preparative HPLC to obtain tert-butyl N-({3-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H-indol-6-yl}methyl)-N-(oxacyclohexane-4-yl)carbamate (40mg) as a pale yellow solid.

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

[0720] Example 81 : 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] Following the procedure 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 82 : Preparation of 3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-yn-1-ylbenzoate

[0725]

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

[0727] Example 83 : 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-methylpropionitrile

[0728]

[0729] Following the procedure 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-methylpropionitrile was prepared.

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

[0731] Example 84 : 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] Following a procedure analogous to that described in Example 28, N-(6-chloropyridin-3-yl)-3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-ynamide was prepared.

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

[0735] Example 85 : Preparation of N-[6-(1-cyano-1-methylethyl)pyridin-3-yl]-3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-ynamide

[0736]

[0737] Following the procedure described in Example 28, N-[6-(1-cyano-1-methylethyl)pyridin-3-yl]-3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-ynamide was prepared.

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

[0739] Example 86 :Preparation of N-({3-ethyl-2-[3-(phenylamino)prop-1-yn-1-yl]-1H-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 μmol, prepared as described in Example 80) in acetonitrile (1 mL) was added trichlorobismuthane (50 mg, 158.7 μmol, 42.94 μL), and the reaction mixture was stirred at 50 ° C for 25 min. EDTA aqueous solution (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 (3x10 mL), and the combined organic layers were washed with water (3x 10 mL) and brine (10 mL), and dried over anhydrous magnesium sulfate. The solid was filtered off and the filtrate was concentrated in vacuo. The crude residue was immediately purified by preparative HPLC (C18 silica gel, eluting 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 (5 mg, 57% yield) as a yellow solid.

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

[0743] Example 87 : 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-methylpropionitrile

[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-methylpropionitrile was prepared in a manner similar to the method 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 88 : 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-methylpropionitrile

[0748]

[0749] Following the procedure described in Example 28, 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-methylpropionitrile was prepared.

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

[0751] Example 89 : Preparation of 2-[5-({3-[5-({[1-(2-methanesulfonylethyl)piperidin-4-yl]amino}methyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile

[0752]

[0753] Following the procedure described in Example 28, 2-[5-({3-[5-({[1-(2-methanesulfonylethyl)piperidin-4-yl]amino}methyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile was prepared.

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

[0755] Example 90 : Preparation of 2-(5-{[3-(5-{[(1-acetylpiperidin-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile

[0756]

[0757] Following the procedure described in Example 28, 2-(5-{[3-(5-{[(1-acetylpiperidin-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile was prepared.

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

[0759] Example 91 : Preparation of 2-{5-[(3-{5-[({1-[2-(dimethylamino)acetyl]piperidin-4-yl}amino)methyl]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}-2-methylpropionitrile

[0760]

[0761] Following the procedure described in Example 28, 2-{5-[(3-{5-[({1-[2-(dimethylamino)acetyl]piperidin-4-yl}amino)methyl]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}-2-methylpropionitrile was prepared.

[0762] LC-MS(ES + ,m / z):594.4[(M+H) + ]

[0763] Example 92 : Preparation of 2-methyl-2-(5-{[3-(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)propionitrile

[0764]

[0765] 2-Methyl-2-(5-{[3-(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)propanenitrile was prepared in a manner similar to that described in Example 28.

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

[0767] Example 93 : Preparation of 2-methyl-2-{5-[(3-{5-[(methylamino)methyl]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile

[0768]

[0769] Following the procedure described in Example 28, 2-methyl-2-{5-[(3-{5-[(methylamino)methyl]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}propanenitrile was prepared.

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

[0771] Example 94 : Preparation of 6-chloro-N-[3-(5-{[(1-methylpiperidin-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]pyridin-3-amine

[0772]

[0773] Following the procedure described in Example 28, 6-chloro-N-[3-(5-{[(1-methylpiperidin-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-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 95 : Preparation of 6-chloro-N-[3-(5-{[(oxacyclohexane-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]pyridin-3-amine

[0776]

[0777] Following the procedure described in Example 28, 6-chloro-N-[3-(5-{[(oxan-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]pyridin-3-amine was prepared.

[0778] LC-MS(ES + ,m / z):477.1[(M+H) + ]

[0779] Example 96: Preparation of 2-[5-({3-[1-(cyclopropylmethyl)-5-{[(oxacyclohexane-4-yl)amino]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile

[0780]

[0781] Following the procedure described in Example 28, 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.

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

[0783] Example 97 : 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-methylpropionitrile

[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-methylpropionitrile was prepared in a manner similar to the method described in Example 1 using 4-bromo-indole.

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

[0787] Example 98 : 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}propionitrile

[0788]

[0789] 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}propionitrile was prepared in a manner similar to the method described in Example 1 using 4-bromo-indole.

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

[0791] Example 99 : 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-methylpropionitrile

[0792]

[0793] 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-methylpropionitrile was prepared in a manner similar to the method described in Example 1 using 4-bromo-7-fluoro-indole.

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

[0795] Example 100 : Preparation of 2-methyl-2-(5-{[3-(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)propionitrile

[0796]

[0797] 2-Methyl-2-(5-{[3-(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)propanenitrile was prepared in a manner similar to the method described in Example 1 using 4-bromo-indole.

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

[0799] Example 101 :Preparation of 2-(5-{[3-(4-{[4-(4-hydroxypiperidin-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-methylpropionitrile

[0800]

[0801] 2-(5-{[3-(4-{[4-(4-hydroxypiperidin-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-methylpropionitrile was prepared in a manner similar to the method described in Example 1 using 4-bromo-indole.

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

[0803] Example 102 : Preparation of N-(6-cyanopyridin-3-yl)-3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-ynamide

[0804]

[0805] Following a procedure analogous to that described in Example 28, N-(6-cyanopyridin-3-yl)-3-(1-ethyl-5-{[(1-methylpiperidin-4-yl)amino]methyl}-1H-indol-2-yl)prop-2-ynamide was prepared.

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

[0807] Example 103 : Preparation of 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-ynamide

[0808]

[0809] Following the procedure described in Example 28, 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-ynamide was prepared.

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

[0811] Example 104 :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-ynamide

[0812]

[0813] Following the procedure described in Example 28, 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-ynamide was prepared.

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

[0815] Example 105 : Preparation of 2-methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(oxiran-2-ylmethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propionitrile

[0816]

[0817] Following the procedure described in Example 28, 2-methyl-2-(5-{[3-(5-{[(oxan-4-yl)amino]methyl}-1-(oxiran-2-ylmethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propanenitrile was prepared.

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

[0819] Example 106: 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 107 : 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 108 : 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] 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 in a manner similar to the method described in Example 1 using 4-bromo-indole.

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

[0832] Example 109 : 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)propionitrile

[0833]

[0834] 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 in a manner similar to the method described in Example 1 using 4-bromo-indole.

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

[0836] Example 110 : 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-methylpropionitrile

[0837]

[0838] Following the procedure 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-methylpropionitrile was prepared.

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

[0840] Example 111: Preparation 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-methylpropionitrile

[0841]

[0842] 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-methylpropionitrile was prepared using 4-bromo-7-fluoro-indole in a manner similar to that described in Example 1. LC-MS (ES + ,m / z):581.4[(M+H) + ]

[0843] Example 112 : 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]propionitrile

[0844]

[0845] Following the procedure 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 113 : 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-methylpropionitrile

[0848]

[0849] Following the procedure described in Example 28, 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-methylpropionitrile was prepared.

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

[0851] Example 114 : Preparation of 2-[5-({3-[5-({[1-(2-methoxyethyl)piperidin-4-yl]amino}methyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile

[0852]

[0853] Following the procedure described in Example 28, 2-[5-({3-[5-({[1-(2-methoxyethyl)piperidin-4-yl]amino}methyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile was prepared.

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

[0855] Example 115 : Preparation of 2-[5-({3-[5-({[4-(dimethylamino)cyclohexyl]amino}methyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile

[0856]

[0857] Following the procedure described in Example 28, 2-[5-({3-[5-({[4-(dimethylamino)cyclohexyl]amino}methyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile was prepared.

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

[0859] Example 116 : Preparation of 2-methyl-2-{5-[(3-{5-[({1-[2-(morpholin-4-yl)acetyl]piperidin-4-yl}amino)methyl]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile

[0860]

[0861] Following the procedure described in Example 28, 2-methyl-2-{5-[(3-{5-[({1-[2-(morpholin-4-yl)acetyl]piperidin-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.

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

[0863] Example 117 : Preparation of 2-(5-{[3-(4-{[(2-methoxyethyl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-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)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile was prepared in a manner similar to the method described in Example 1 using 4-bromo-indole.

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

[0867] Example 118 : Preparation of 2-methyl-2-{5-[(3-{4-[(methylamino)methyl]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile

[0868]

[0869] 2-Methyl-2-{5-[(3-{4-[(methylamino)methyl]-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.

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

[0871] Example 119 : Preparation of 2-{5-[(3-{4-[(4-acetylpiperazin-1-yl)methyl]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}-2-methylpropionitrile

[0872]

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

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

[0875] Example 120 : Preparation of 2-methyl-2-[5-({3-[4-(morpholin-4-ylmethyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propionitrile

[0876]

[0877] 2-Methyl-2-[5-({3-[4-(morpholin-4-ylmethyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propanenitrile was prepared in a manner analogous to the method described in Example 1 using 4-bromo-indole.

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

[0879] Example 121: Preparation 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-methylpropionitrile

[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-methylpropionitrile was prepared in a manner similar to the method described in Example 1 using 4-bromo-indole.

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

[0883] Example 122 : 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-methylpropionitrile

[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-methylpropionitrile was prepared in a manner similar to the method described in Example 1 using 4-bromo-indole.

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

[0887] Example 123 : 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]propionitrile

[0888]

[0889] 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 was prepared using 4-bromo-indole in a manner similar to that described in Example 1. LC-MS (ES + ,m / z):622.4[(M+H) + ]

[0890] Example 124 : 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] Following the procedures described in Example 81 and Example 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 125 : 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-carboxylic acid methyl ester

[0895]

[0896] Following the procedure 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}picolinate was prepared.

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

[0898] Example 126: Preparation 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] Following the procedure described in Example 28, 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.

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

[0902] Example 127 :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] Following the procedure described in Example 28, 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.

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

[0906] Example 128 :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] Following the procedure described in Example 28, 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 was prepared.

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

[0910] Example 129 : Preparation of 2-[(5-{[3-(5-{[(oxacyclohexane-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)formamido]acetic acid

[0911]

[0912] 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)formamido]acetic acid was prepared in a manner similar to that described in Example 28.

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

[0914] Example 130 : 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-carboxylic acid

[0915]

[0916] 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}picolinic acid was prepared.

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

[0918] Example 131:Preparation of N-(2-methanesulfonylethyl)-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

[0919]

[0920] Following the procedure described in Example 28, N-(2-methanesulfonylethyl)-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.

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

[0922] Example 132 : Preparation of 2-[5-({3-[1-(cyanomethyl)-5-{[(oxacyclohexane-4-yl)amino]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile

[0923]

[0924] Following the procedure described in Example 28, 2-[5-({3-[1-(cyanomethyl)-5-{[(oxan-4-yl)amino]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropanenitrile was prepared.

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

[0926] Example 133 : Preparation of 2-methyl-2-[5-({3-[1-(2-methylpropyl)-5-{[(oxacyclohexane-4-yl)amino]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propionitrile

[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] Example 134 : 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 136:Preparation 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] Following the procedure described in Example 28, 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.

[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] Following the procedure described in Example 28, 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.

[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] Following the procedure described in Example 28, 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 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)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridine-2-carboxamide

[0951]

[0952] Following the procedure described in Example 28, N-(1-methylpiperidin-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.

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

[0954] Example 140 : Preparation of N-[6-(1-cyano-1-methylethyl)pyridin-3-yl]-3-[1-(2-fluoroethyl)-5-{[(oxacyclohexane-4-yl)amino]methyl}-1H-indol-2-yl]prop-2-ynamide

[0955]

[0956] Following the procedure described in Example 28, N-[6-(1-cyano-1-methylethyl)pyridin-3-yl]-3-[1-(2-fluoroethyl)-5-{[(oxan-4-yl)amino]methyl}-1H-indol-2-yl]prop-2-ynamide was prepared.

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

[0958] Example 141: 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-methylpropionitrile

[0959]

[0960] 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-methylpropionitrile was prepared in a manner similar 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-(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-methylpropionitrile

[0962]

[0963] Synthesis scheme:

[0964]

[0965] To a solution of 4-bromo-6-fluoro-indole (2g, 9.38mmol) in anhydrous tetrahydrofuran (30mL) cooled in an ice bath, sodium hydride (0.452g, 11.3mmol, 60% in mineral oil) was added. The reaction mixture was stirred under nitrogen for 10min, followed by addition of benzenesulfonyl chloride (1.44mL, 11.3mmol). The black solution was warmed to room temperature over 4h. TLC and LCMS indicated that the reaction was complete. Saturated aqueous ammonium chloride solution was slowly added, 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 ground with ethanol. The resulting beige solid was collected by filtration to obtain 4-bromo-6-fluoro-1-(phenylsulfonyl)-1H-indole (2.6g, 79% yield).

[0966] To a solution of 4-bromo-6-fluoro-1-(phenylsulfonyl)-1H-indole (0.6 g, 1.7 mmol) in anhydrous tetrahydrofuran (10 mL) cooled at -78 ° C under a nitrogen atmosphere was added a 1M lithium diisopropylamide solution (2.7 mL, 2.7 mmol) in tetrahydrofuran / hexane. The reaction mixture was kept at -78 ° C for 90 min. The solution of iodine (0.427 mg, 1.7 mmol) in tetrahydrofuran (10 mL) was then slowly added (over 2 min). The reaction mixture was warmed to room temperature over 14 h. LCMS indicated that the reaction was complete. A saturated aqueous ammonium chloride solution was slowly added, 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-1-(phenylsulfonyl)-1H-indole. The crude solid was used for the next step without further purification.

[0967] 2M K is added to a suspension of crude 4-bromo-7-fluoro-2-iodo-1-(phenylsulfonyl)-1H-indole (1.7mmol) in methanol (30mL) CO aqueous solution (10mL, 20mmol). The reaction mixture is heated at 60°C for 18h. Methanol is removed in vacuo. The remaining aqueous phase is diluted with saturated aqueous ammonium chloride solution and extracted with ethyl acetate (2x). The combined organic phases are washed with salt water, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification of the crude residue occurs by flash column chromatography (ISCO 24g, gradient elution with ethyl acetate and hexane). Product is eluted with 20% ethyl acetate in hexane. Fractions are combined to obtain 4-bromo-6-fluoro-2-iodo-1H-indole (310mg, 54% yield) as an oil.

[0968] To a solution of 4-bromo-6-fluoro-2-iodo-1H-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, 2,2,2-trifluoroethyl trifluoromethanesulfonate (0.170 mL, 1.2 mmol) was added. The reaction mixture was stirred at room temperature for 14 h. A saturated aqueous ammonium chloride solution was slowly added, 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 vacuum dried to give 4-bromo-7-fluoro-2-iodo-1-(2,2,2-trifluoroethyl)-1H-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-1-(2,2,2-trifluoroethyl)-1H-indole (0.59 mmol) was added 2-methyl-2-[5-(2-propynylamino)-2-pyridyl]propionitrile (207 mg, 0.89 mmol), CuI (22 mg, 0.12 mmol), Pd(PPh 3 ) 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 (2×) with ethyl acetate. 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, gradient elution with ethyl acetate and hexane). The desired fraction was eluted with 50% ethyl acetate in hexane. Fractions were combined to give 2-(5-{3-[4-bromo-7-fluoro-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]-2-propynylamino}-2-pyridyl)-2-methylpropionitrile (120 mg, 41% yield).

[0970] To a solution of 2-(5-{3-[4-bromo-7-fluoro-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]-2-propynylamino}-2-pyridinyl)-2-methylpropionitrile (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. under a nitrogen atmosphere for 10 h. 0.5 M EDTA (4 mL) and ethyl acetate (10 mL) were added to the reaction mixture, which was stirred at room temperature for 1 h. The organic phase was extracted (2×) with ethyl acetate. 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 12g, eluting with a gradient of ethyl acetate and hexanes) to afford recovered starting material (60 mg) and the desired 2-(5-{3-[7-fluoro-4-formyl-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]-2-propynylamino}-2-pyridinyl)-2-methylpropionitrile (40 mg, 75% yield based on starting material recovery) 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-methylpropionitrile (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 then concentrated. The crude material was purified by preparative HPLC (Combiflash™ EZ-prep system, gradient elution with water (formic acid 0.1%) and acetonitrile (formic acid 0.1%) 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-methylpropionitrile (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-methylpropionitrile

[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-methylpropionitrile was prepared in a manner similar to the method 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)propionitrile

[0977]

[0978] 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 was prepared in a manner similar to the method described in Example 28 using 1,6-diaza-1H-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-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-methylpropionitrile

[0981]

[0982] 2-(5-{[3-(5-{[4-(Dimethylamino)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-methylpropionitrile was prepared in a manner similar to the method described in Example 28 using 1,6-diaza-1H-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-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-methylpropionitrile

[0985]

[0986] Synthesis scheme:

[0987]

[0988] 5- bromo-2- chloro-3-nitro-pyridine (20.2 g, 85.07 mmol, 1 equivalent) was dissolved in anhydrous tetrahydrofuran (100 mL) under a nitrogen atmosphere, and the solution was cooled to -78 ° C. To the solution was added bromo (vinyl) magnesium (1 M, 340 mL, 4 equivalents), and the reaction mixture was stirred at -78 ° C for 4 h. LCMS analysis showed that the reaction was complete. The reaction was quenched with saturated ammonium chloride (150 mL). The aqueous phase was extracted with ethyl acetate (3 x 150 mL), and the combined organic layer was dried over magnesium sulfate, filtered and concentrated. The crude product was purified by flash column chromatography (silica gel, eluted with a 1 / 1 mixture of ethyl acetate and petroleum ether) to obtain 4- bromo-7- chloro-1H- pyrrolo- [2,3-c] pyridine (4.12 g, 21% yield) as a yellow solid.

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

[0990] The resulting reaction mixture was 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-(benzenesulfonyl)-4-bromo-7-chloro-pyrrolo[2,3-c]pyridine (2.03 g, 59% yield) as a light yellow solid.

[0991] To a solution (20 mL) of 1-(benzenesulfonyl)-4-bromo-7-chloro-pyrrolo[2,3-c]pyridine (400 mg, 1.08 mmol) in tetrahydrofuran cooled to -78 ° C was added lithium diisopropylamide (2 M, 1.62 mL) dropwise. After addition, the mixture was stirred at -78 ° C for 1 h, and 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, then distributed 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 layer was 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 dichloromethane / petroleum ether: 1 / 1) to afford 1-(benzenesulfonyl)-4-bromo-7-chloro-2-iodo-pyrrolo[2,3-c]pyridine (240 mg, 45% yield) as a light yellow solid.

[0992] Under a nitrogen atmosphere, a mixture of 1-(benzenesulfonyl)-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. 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] 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-methylpropionitrile was prepared using 4-bromo-7-chloro-2-iodo-1H-pyrrolo[2,3-c]pyridine in the manner described in Examples 1 and 49.

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

[0995] Following a similar procedure to that described in Example 142, the following compounds were prepared:

[0996]

[0997]

[0998] Starting from 2-iodo-1H-indole-4-carbaldehyde (prepared from 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-(oxiran-2-ylmethyl)-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propionitrile

[1002] 2-Iodo-1H-indole-4-carbaldehyde was reacted with 2-(bromomethyl)oxirane followed by coupling with 2-methyl-2-{5-[(prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile in a manner analogous to the methods described in Examples 1 and 49. Reductive amination of the resulting intermediate using 4-(pyrrolidin-1-yl)piperidine afforded 2-methyl-2-[5-({3-[1-(oxiran-2-ylmethyl)-4-{[4-(pyrrolidin-1-yl)piperidin-1-yl]methyl}-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]propionitrile.

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

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

[1005]

[1006] Following the procedures 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-methylpropionitrile 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] Following the procedures described in Examples 80 and 86, 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 was prepared.

[1019] LC-MS(ES + ,m / z):469.3[(M+H) + ]

[1020] Example 171 : Preparation of 2-methyl-2-(5-{[3-(6-{[(1-methylpiperidin-4-yl)amino]methyl}-3-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propionitrile

[1021]

[1022] 2-Methyl-2-(5-{[3-(6-{[(1-methylpiperidin-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 in a manner similar to that described in Examples 80 and 86.

[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)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}-2-methylpropionitrile

[1025]

[1026] Synthesis scheme:

[1027]

[1028] Sodium hydride (1.39 g, 34.70 mmol, in mineral oil 60%) is added to a 0°C solution of 4-bromo-6-chloro-indole (4 g, 17.35 mmol) in dimethylformamide (60 mL). The reaction is stirred at 0°C for 1 hour, and 2-(trimethylsilyl)ethoxymethyl chloride (SEMCl, 4.34 g, 26.03 mmol) is added to the reaction mixture. The reaction is stirred at 20°C for 1 hour, and a saturated solution of ammonium chloride (50.0 mL) is added. The reaction mixture is extracted with dichloromethane (3 x 100 mL). The combined organic layers are 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, eluted 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 for 12 h in a sealed tube. The reaction mixture was treated with water (10 mL) and extracted with ethyl acetate (3x50 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, elution with dichloromethane / methanol: 50 / 1) to give 6-chloro-N-(1-methylpiperidin-4-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-indol-4-amine (1.6 g, 33% yield, 90% purity) as a yellow oil.

[1030] To a solution of 6-chloro-N-(1-methylpiperidin-4-yl)-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-indole-4-amine (1.55 g, 3.55 mmol, 90% purity) in t-BuOH (21 mL) was added di-tert-butyl diformate (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, eluted with dichloromethane / methanol: 100 / 1) to give tert-butyl N-(6-chloro-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-indole-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 (1 M 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 N-(6-chloro-1H-indol-4-yl)-N-(1-methylpiperidin-4-yl)carbamic acid tert-butyl ester (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. Benzenesulfonyl chloride (345 mg, 1.95 mmol) was added to the reaction mixture. The ice bath was removed, the reaction mixture was warmed 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 (2x20 mL). The combined organic layer was 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-(phenylsulfonyl)-6-chloro-1H-indol-4-yl]-N-(1-methylpiperidin-4-yl)-carbamate (800 mg, 88% yield, 72% purity) as a yellow oil.

[1033] To N-[1-(benzenesulfonyl)-6-chloro-1H-indol-4-yl]-N-(1-methylpiperidin-4-yl)-t-butyl carbamate (260 mg, 371.40 μmol, 72% purity) in tetrahydrofuran (6 mL) -65 ° C solution of lithium diisopropylamide (2M in tetrahydrofuran, 520 μ L). The reaction was stirred at -65 ° C for 1.5 h, and iodine solution (144 mg, 567.35 μmol in 1.5 mL tetrahydrofuran) was added dropwise to the reaction. The reaction was stirred at -65 ° C for 0.5 h, and a saturated ammonium chloride solution (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-(benzenesulfonyl)-6-chloro-2-iodo-1H-indol-4-yl]-N-(1-methylpiperidin-4-yl)-carbamic acid tert-butyl ester (700 mg) as a yellow solid. The crude product was used directly in the next step without further purification.

[1034] To a solution of N-[1-(benzenesulfonyl)-6-chloro-2-iodo-1H-indol-4-yl]-N-(1-methylpiperidin-4-yl)-t-butyl carbamate (900 mg) in methanol (15 mL) is added aqueous potassium carbonate solution (0.5 mL, 2 M). The reaction is then stirred at 80 ° C for 30 min, cooled to room temperature, and concentrated. The residue is extracted with ethyl acetate (3x 10 mL). The combined organic layer is washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product is purified by preparative thin-layer chromatography (silica gel, eluted with dichloromethane / methanol containing 0.1% triethylamine: 10 / 1) to obtain N-(6-chloro-2-iodo-1H-indol-4-yl)-N-(1-methylpiperidin-4-yl)-t-butyl carbamate (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-methylpropionitrile was prepared in a manner similar to the method described in Example 28 using tert-butyl N-(6-chloro-2-iodo-1H-indol-4-yl)-N-(1-methylpiperidin-4-yl)carbamate.

[1036] LC-MS(ES + ,m / z):543.3[(M+H) + ]

[1037] Example 173: Preparation 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-methylpropionitrile (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 several 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 a 1:1 solution of 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 μmol, 80% purity) in dichloromethane (3 mL) was added diisopropylethylamine (56 mg, 436.29 μmol) and HATU (50 mg, 130.89 μmol). Subsequently, 1-methylpiperidin-4-amine (20 mg, 174.52 μmol) was added to the reaction mixture. 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-methylpropionitrile

[1043]

[1044] Following the procedure described in Example 173, 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-methylpropionitrile was prepared.

[1045] LC-MS(ES + ,m / z):541.2[(M+H) + ]

[1046] Example 175: Preparation 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)-1H-indole-4-amine

[1047]

[1048] 6-Fluoro-2-{3-[(6-methanesulfonylpyridin-3-yl)amino]prop-1-yn-1-yl}-N-(1-methylpiperidin-4-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine was prepared in a manner analogous to that described in Example 172 using 4-bromo-6-fluoro-indole.

[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)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}-2-methylpropionitrile

[1051]

[1052] 2-{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]pyridin-2-yl}-2-methylpropionitrile was prepared in a manner similar to that described in Example 172 using 4-bromo-6-fluoro-indole.

[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)-1H-indol-2-yl}prop-2-yn-1-yl)amino]-N-(pyridin-3-yl)pyridine-2-carboxamide

[1055]

[1056] 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 manner analogous to that described in Example 172 using 4-bromo-6-fluoro-indole.

[1057] LC-MS(ES + ,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)propionitrile

[1059]

[1060] Following the procedure described in Example 142, 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.

[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}propionitrile

[1063]

[1064] Synthesis scheme:

[1065]

[1066] To a 0 ° C mixture of sodium hydride (4.41g, 110.34mmol, 60% in mineral oil) in tetrahydrofuran (20mL) was added a solution of 4-nitro-1H-pyrrolo[2,3-b]pyridine (6g, 36.78mmol) in tetrahydrofuran (5mL). The mixture was stirred at 0 ° C for 1h, and then benzenesulfonyl chloride (9.74g, 55.17mmol, 7.06mL) was added. The mixture was stirred at 0 ° C for 1h, and then quenched by adding saturated aqueous ammonium chloride solution. The reaction mixture was extracted with ethyl acetate (3x 20mL). The combined organic layer was washed with brine (5mL), 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 1-(phenylsulfonyl)-4-nitro-1H-pyrrolo[2,3-b]pyridine (4.90 g, 44% yield) as a yellow solid.

[1067] A solution of 1-(benzenesulfonyl)-4-nitro-pyrrolo[2,3-b]pyridine (1 g, 3.30 mmol) in tetrahydrofuran (10 mL) was added to a solution of 2,2,6,6-tetramethylpiperidinium lithium (1.70 g, 11.55 mmol) in tetrahydrofuran (27 mL) at -78 ° C. The mixture was stirred at -78 ° C for 1 h, and 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 adding a 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 1-(phenylsulfonyl)-2-iodo-4-nitro-1H-pyrrolo[2,3-b]pyridine (600 mg) which was used without further purification.

[1068] To a solution of 1-(benzenesulfonyl)-2-iodo-4-nitro-1H-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 solid was filtered off and the resulting filtrate was concentrated. The crude product was purified by flash column chromatography (silica gel, eluted with petroleum ether / ethyl acetate: 1 / 1) to give the product 2-iodo-4-nitro-1H-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 trifluoromethanesulfonic acid 2,2,2-trifluoroethyl ester (1.81 g, 7.80 mmol). The resulting reaction mixture was stirred at 0 ° C for 1 h, and 2-iodo-4-nitro-1H-pyrrolo[2,3-b]pyridine (451 mg, 1.56 mmol) was added to the reaction mixture. The mixture was stirred at 0 ° C for 1 h and quenched by adding saturated aqueous ammonium chloride solution. The resulting mixture was extracted with ethyl acetate (3x20 mL). The combined organic layer was 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, eluted 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 μmol) 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 layer was 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, eluted with petroleum ether / ethyl acetate: 30 / 1 to 5 / 1) to obtain 2-iodo-1-(2,2,2-trifluoroethyl)-1H-pyrrolo[2,3-b]pyridine-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}propionitrile (180 mg, 762.30 μmol) in dimethyl sulfoxide (2 mL) was added N-isopropylpropan-2-amine (536 μL, 3.81 mmol). 2-iodo-1-(2,2,2-trifluoroethyl)pyrrolo[2,3-b]pyridin-4-amine (130 mg, 381.15 μmol), CuI (7 mg, 38.11 μmol), and tetrakis(triphenylphosphine)palladium(0) (44 mg, 38.11 μmol) were then added to the reaction mixture. The reaction was stirred at 25° C. under a 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 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, eluted with petroleum ether / ethyl acetate: 1 / 2) to give 2- [5- [3- [4-amino -1- (2,2,2- trifluoroethyl) pyrrolo [2,3-b] pyridin-2-yl] prop-2-ynylamino] -2-pyridyl] -2-methyl-propionitrile (130 mg, 83% yield) as a yellow solid.

[1072] To a solution of 1-methylpiperidin-4-one (11 mg, 97 μmol, 11 μL) in ethanol (2 mL) and acetic acid (7.3 mg, 121.25 μmol, 7 μL) was added 2-[5-[3-[4-amino-1-(2,2,2-trifluoroethyl)pyrrolo[2,3-b]pyridin-2-yl]prop-2-ynylamino]-2-pyridinyl]-2-methyl-propionitrile (10 mg, 24.25 μmol) and Molecular sieves (30 mg). The mixture was stirred at 50 ° C for 12 h, and sodium cyanoborohydride (15 mg, 242.50 μmol) 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 (3x 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 give 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}propionitrile (3 mg, 22% yield). LC-MS (ES+, m / z): 510.3[(M+H)+]

[1073] Example 180: Preparation 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-methylpropionitrile

[1074]

[1075] Following the procedure described in Example 141, 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-methylpropionitrile was prepared.

[1076] LC-MS (ES+, m / z): 543.4 [(M+H)+]

[1077] Example 181 : Preparation of 2-(5-{[3-(7-chloro-5-{[(oxacyclohexane-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile

[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-methylpropionitrile was prepared starting from 5-bromo-7-chloro-1H-indole using a method similar 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-methylpropionitrile

[1082]

[1083] Starting from 5-bromo-7-chloro-1H-indole, 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-methylpropionitrile was prepared using a method similar to that 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-methylpropionitrile

[1086]

[1087] Starting from 4-bromo-7-fluoro-1H-indole and following a procedure analogous to that described in Example 172, 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-methylpropionitrile was prepared.

[1088] LC-MS (ES+, m / z): 527.3 [(M+H)+]

[1089] Example 184 : Preparation of 2-(5-{[3-(7-fluoro-5-{[(oxacyclohexane-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)-2-methylpropionitrile

[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-methylpropionitrile was prepared starting from 5-bromo-7-fluoro-1H-indole and using a method similar to that described in Example 1.

[1092] LC-MS(ES + ,m / z):528.3[(M+H) + ]

[1093] Example 185: Preparation of 2-(5-{[3-(7-fluoro-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-methylpropionitrile

[1094]

[1095] Starting from 5-bromo-7-fluoro-1H-indole and using a method similar to that described in Example 1, 2-(5-{[3-(7-Fluoro-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-methylpropionitrile was prepared.

[1096] LC-MS(ES + ,m / z):541.3[(M+H) + ]

[1097] Example 186 : Preparation of 2-methyl-2-(5-{[3-(5-{[(oxacyclohexane-4-yl)amino]methyl}-1-(2,2,2-trifluoroethyl)-1H-1,3-benzimidazol-2-yl)prop-2-yn-1-yl]amino}pyridin-2-yl)propionitrile

[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). 2,2,2-trifluoroethylamine (3.87 g, 39.03 mmol, 3.07 mL) was then added to the reaction mixture. The reaction mixture was stirred at 100 ° C for 1 h, cooled to room temperature, and concentrated. Water (500 mL) was added to the mixture, 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. NaBH4 (1.02 g, 27.09 mmol) was added to the solution. 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 layer was 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)oxan-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)oxan-4-amine (3 g, 8.46 mmol) in dichloromethane (70 mL) were added triethylamine (3.52 mL, 25.38 mmol) and di-tert-butyl diformate (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 dichloromethane (3 To the mixture of 4-nitro-2-[(2,2,2-trifluoroethyl)amino]phenyl]methyl)-N-(oxane-4-yl)carbamic acid tert-butyl ester (3.90 g, 96% yield) was obtained by flash column chromatography (silica gel, eluted with dichloromethane / methanol: 200 / 1).

[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 for 30 min under a hydrogen atmosphere (15 psi). 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). Carbon disulfide (6.30 g, 82.74 mmol, 5 mL) was then added to the reaction, 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 12M HCl. The reaction mixture was extracted with dichloromethane (3x 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, eluting 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 N-(oxan-4-yl)-N-{[2-mercapto-1-(2,2,2-trifluoroethyl)-1H-1,3-benzimidazol-5-yl]methyl}carbamic acid tert-butyl ester (150 mg, 329.97 μmol) in acetic acid (1.70 mL) and HBr (69 μL, 442.16 μmol, 35% solution in acetic acid) was added dropwise bromine (1.32 mmol, 68 μL) in acetic acid (0.5 mL). HBr (154 μL, 35% in acetic acid) in acetic acid (1 mL) was then added to the reaction, 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, elution 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-1-(2,2,2-trifluoroethyl)-1H-1,3-benzimidazol-5-yl]methyl}oxane-4-amine (30 mg, 68.84 μmol) and di-tert-butyl dicarboxylate (35 mg, 158.33 μmol) 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-1-(2,2,2-trifluoroethyl)-1H-1,3-benzimidazol-5-yl]methyl}-N-(oxane-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]propionitrile (50 mg, 212.41 μmol), diisopropylamine (1.42 mmol, 200 μL) and CuI (1.2 mg, 6.40 μmol) in dimethyl sulfoxide (2.5 mL) was added tert-butyl N-{[2-bromo-1-(2,2,2-trifluoroethyl)-1H-1,3-benzimidazol-5-yl]methyl}-N-(oxan-4-yl)carbamate (35 mg, 63.98 μmol) and tetrakis(triphenylphosphine)palladium(0) (7 mg, 6.40 μmol, 0.10 equiv). The reaction mixture was stirred at 80° C. under a nitrogen atmosphere for 1 h and then treated with saturated EDTA solution (5 mL). The two-phase mixture was stirred at 20° C. for 1 h and then extracted with dichloromethane (3×10 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) to give 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 (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 saturated EDTA solution (10 mL). The two-phase mixture was stirred at 20 ° C for 1 h and extracted with ethyl acetate (3 x 20 mL). The combined organic layer was 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}oxane-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 μmol) in dimethylformamide (5 mL) was added 3-phenylprop-2-ynal (174 mg, 1.34 mmol, 163 μL). Potassium hydrogen sulfate (Oxone) (274 mg, 446.16 μmol) was then 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 Na2SO3 solution (5 mL), and the mixture was stirred at 20° C. for 1 h. The mixture was then extracted with ethyl acetate (3×10 mL). The combined organic layers were washed with water (3×10 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, elution 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] Using the method described in Example 186, 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 deprotected with bismuth trichloride to afford N-{[2-(2-phenylethynyl)-1-(2,2,2-trifluoroethyl)-1H-1,3-benzimidazol-5-yl]methyl}oxan-4-amine (35 mg, light yellow 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)propionitrile

[1117]

[1118] To a 0 ° C solution of 1H-pyrrolo [3,2-b] pyridine-5-carboxylic acid methyl ester (3.01g, 17.05mmol) in dimethylformamide (30mL) was added sodium hydride (890mg, 22.17mmol, 60% in mineral oil). The mixture was stirred at 0 ° C for 30min, and PhSO2Cl (2.2mL, 17.05mmol) was added. The ice bath was then removed, and the mixture was warmed to room temperature and stirred at room temperature overnight. The reaction mixture was poured into ice / water (300mL) and stirred for 1h. The solid was filtered, washed with water (100mL) and hexane (50mL), and the resulting solution was dried overnight to obtain 1- (benzenesulfonyl) -1H- pyrrolo [3,2-b] pyridine -5-carboxylic acid methyl ester (3.50g, 66% yield) as an off-white solid.

[1119] 1-(Benzenesulfonyl)-1H-pyrrolo[3,2-b]pyridine-5-carboxylic acid methyl ester (500mg, 1.58mmol) was dissolved in anhydrous tetrahydrofuran (50mL) and cooled to -78°C under a nitrogen atmosphere. DIBAL solution (1.8M, 5.3mL, 9.48mmol) was added dropwise, and the reaction mixture was stirred at -78°C for 20min. The mixture was then poured into a mixture of water (20mL), 1N sodium hydroxide (20mL) and brine (20mL) and stirred for 10min. The mixture was then extracted with ethyl acetate (3x 30mL). The combined organic layer was washed with water, dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, eluting 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)propionitrile was prepared using 1-(benzenesulfonyl)-1H-pyrrolo[3,2-b]pyridine-5-carbaldehyde and 2-methyl-2-{5-[(prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile in a manner analogous to the methods described in Examples 1 and 49.

[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] 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 was prepared in a manner analogous to the method described in Example 179 using 5-methyl-4-nitro-1H-indole and 2-methyl-2-{5-[(prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile.

[1133] LC-MS(ES + ,m / z):523.0[(M+H) + ]

[1134] Example 192 :Preparation of N-[1-(2-methanesulfonylethyl)piperidin-4-yl]-2-{3-[(4-methanesulfonylphenyl)amino]prop-1-yn-1-yl}-5-methyl-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine

[1135]

[1136] N-[1-(2-Methanesulfonylethyl)piperidin-4-yl]-2-{3-[(4-methanesulfonylphenyl)amino]prop-1-yn-1-yl}-5-methyl-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.

[1137] LC-MS(ES + ,m / z):625.3[(M+H) + ]

[1138] Example 193 : Preparation of 4-[(3-{5-methyl-4-[(piperidin-4-yl)amino]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]benzene-1-sulfonamide

[1139]

[1140] 4-[(3-{5-methyl-4-[(piperidin-4-yl)amino]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]benzene-1-sulfonamide was prepared in a manner analogous to the method described in Example 179 using 5-methyl-4-nitro-1H-indole and 4-[(prop-2-yn-1-yl)amino]benzene-1-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 196: Preparation of 2-[5-({3-[4-(methoxymethyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile

[1151]

[1152] 2-[5-({3-[4-(methoxymethyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile was prepared in a manner analogous to the methods described in Examples 1 and 49 using 2-iodo-1H-indole-4-carbaldehyde (prepared from methyl 1H-indole-4-carboxylate) and 2-methyl-2-{5-[(prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile.

[1153] LC-MS(ES + ,m / z):441.2[(M+H) + ]

[1154] Example 197 : Preparation of 2-[5-({3-[4-(cyanomethyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile

[1155]

[1156] 2-[5-({3-[4-(Cyanomethyl)-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl]prop-2-yn-1-yl}amino)pyridin-2-yl]-2-methylpropionitrile was prepared in a manner analogous to the methods described in Examples 1 and 49 using 2-iodo-1H-indole-4-carbaldehyde (prepared from methyl 1H-indole-4-carboxylate) and 2-methyl-2-{5-[(prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile.

[1157] LC-MS(ES + ,m / z):436.2[(M+H) + ]

[1158] Example 198 : Preparation of 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]propionitrile

[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)indole-5-carboxylate (300 mg, 594.14 μmol) in methanol (4.5 mL) was added sodium hydroxide (5 M, 810 μL), 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 1N HCl. The reaction mixture was then extracted with ethyl acetate (3×10 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)indole-5-carboxylic acid as a 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)indole-5-carboxylic acid (1 equiv) was coupled with morpholine (2 equiv) using HATU (2 equiv) and diisopropylethylamine (3 equiv) 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]propionitrile 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]propionitrile

[1164]

[1165] Following the procedure described in Example 198, 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.

[1166] LC-MS(ES + ,m / z):523.3[(M+H)+ ]

[1167] Example 200 : Preparation of 2-{5-[(3-{5-[4-(dimethylamino)piperidine-1-carbonyl]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}-2-methylpropionitrile

[1168]

[1169] Following the procedure described in Example 198, 2-{5-[(3-{5-[4-(dimethylamino)piperidine-1-carbonyl]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}-2-methylpropionitrile was prepared.

[1170] LC-MS(ES + ,m / z):551.4[(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] Following the procedure described in Example 198, 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.

[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-(oxacyclohexane-4-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-5-carboxamide

[1176]

[1177] Following the procedure 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)propionitrile

[1180]

[1181] 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)propionitrile was prepared using 1-(1H-indol-5-yl)ethan-1-one and 2-methyl-2-{5-[(prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile in a manner similar to that described in Examples 1 and 49.

[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}propionitrile

[1184]

[1185] To a solution of 2-(5-aminopyridin-2-yl)-2-methylpropionitrile (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 propargyl bromide (7.5 mL, 87.44 mmol) was added to the reaction mixture. The resulting reaction mixture was stirred at 25°C for another 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, eluted 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 colorless 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 and then quenched with aqueous sodium bicarbonate. The resulting mixture was concentrated and the residue was washed with petroleum ether (2 x 5 mL) to give 2-methyl-2-{5-[(prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile (1.50 g, 94% yield, hydrochloride) 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}propionitrile was prepared using 4-chloro-1H-pyrrolo[3,2-c]pyridine and 2-methyl-2-{5-[(prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile.

[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]propionitrile

[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 adding 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 methyl 1H-indole-5-carboxylate (300 g, crude product) 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 adding saturated ammonium chloride (1000 mL) at 0 ° C, and extracted with ethyl acetate (3x1000 mL). The combined organic layer was 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-(benzenesulfonyl)-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 for 5 h under a nitrogen atmosphere. The solid was 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] By (2- iodo-1H- indol-5-yl) methanol (15.83g, 34.79mmol) and MnO2 (24.20g, 278.32mmol) in dichloromethane (150mL) is stirred at 30 DEG C under nitrogen atmosphere for 5h.The reaction mixture is filtered using diatomaceous earth and washed with dichloromethane (2x 150mL).The filtrate is washed with brine, 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: 10 / 1) to obtain 2- iodo-1H- indole-5-carboxaldehyde (8g, 66.17% yield, 78% purity) as a yellow solid. The crude product is used without further purification.

[1201] To a solution of 2-iodo-1H-indole-5-carboxaldehyde (3 g, 8.63 mmol) in tetrahydrofuran (40 mL) cooled to 0 ° C., sodium hydride (1.04 g, 25.90 mmol, 60% in mineral oil) was added. The mixture was stirred at 0 ° C. for 30 min under a nitrogen atmosphere, and 2,2,2-trifluoroethyl trifluoromethanesulfonate (5.01 g, 21.58 mmol) was added dropwise to the reaction mixture. The ice bath was then removed, and the reaction mixture was stirred for 1 h under a nitrogen atmosphere, 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]propionitrile (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 CuI (291 mg, 1.53 mmol). 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-5-carbaldehyde (1.80 g, 5.10 mmol) and tetrakis(triphenylphosphine)palladium(0) (589 mg, 510 μmol) were then added to the reaction mixture. The mixture was stirred at room temperature for 2 h under a nitrogen atmosphere. 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, elution 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-methylpropionitrile (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-methylpropionitrile (1 equiv) and morpholine (1 equiv) in dichloromethane (2 mL) and methanol (2 mL) was added magnesium sulfate (15 equiv). After the mixture was stirred at room temperature for 12 h, sodium bicarbonate (1 equiv) and sodium cyanoborohydride (5 equiv) were added to the reaction mixture, the reaction was stirred at room temperature for 1 h, and 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] In a similar manner to that described in Example 205, the following compounds were prepared:

[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 equivalent) in dichloromethane (20 mL) was added tetrahydro-2H-pyran-4-amine (8 equivalents) and magnesium sulfate (10 equivalents). The reaction mixture was then stirred at room temperature under a nitrogen atmosphere for 2 h. Sodium cyanoborohydride (4 equivalents) was then added to the reaction, and the mixture was stirred under a nitrogen atmosphere for 1 h. The reaction mixture was poured into an 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 equiv) and pyrrolidine (1.2 equiv) in dichloromethane (5 mL) was added triethylamine (2 equiv) and HATU (1.20 equiv). 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, eluted 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. The crude product was used in the next step without further purification.

[1217] A solution of propan-2-yn-1-yl (6- (pyrrolidine -1- carbonyl) pyridin-3-yl) tert-butyl carbamate (1 equivalent) in HCl / ethyl acetate (5 mL) was stirred at room temperature for 2 h. The reaction mixture was then concentrated in vacuo to give a crude (5- (propan-2-yn-1-ylamino) pyridin-2-yl) (pyrrolidin-1-yl) ketone as a yellow solid. The crude product was used in the next step without further purification.

[1218]

[0266] Following the procedure analogous to that described in Example 205, N-{[2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-5-yl]methyl}oxan-4-amine was coupled with (5-(prop-2-yn-1-ylamino)pyridin-2-yl)(pyrrolidin-1-yl)methanone to provide 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.

[1219] LC-MS(ES + ,m / z):540.3[(M+H) + ]

[1220] Starting from 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-5-carbaldehyde and using the procedure described in Example 266, the following compound was prepared:

[1221]

[1222]

[1223]

[1224] Example 288 : Preparation of 2-methyl-2-{5-[(3-{5-[(oxacyclohexane-4-yl)amino]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}propionitrile

[1225]

[1226] Starting from 5-bromo-1H-indole and using a method similar to that described in Example 1, 2-methyl-2-{5-[(3-{5-[(oxan-4-yl)amino]-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}propanenitrile was prepared.

[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)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridin-2-yl}-2-methylpropionitrile

[1229]

[1230] Synthesis scheme

[1231]

[1232] To a solution of 4-nitro-indole (1 equivalent) in dimethylformamide was added sodium hydride (1.50 equivalents) and benzenesulfonyl chloride (1.0 equivalents). The mixture was stirred at 0-30°C for 2 hours and quenched with 0°C aqueous ammonium chloride. The mixture was filtered, the solid was washed with water and petroleum ether, and the resulting solution was concentrated and dried under vacuum to give 4-nitro-1-(phenylsulfonyl)-1H-indole.

[1233] To a stirred solution of 4-nitro-1-(phenylsulfonyl)-1H-indole (1.0 equiv) in anhydrous tetrahydrofuran at -78°C was added a solution of lithium diisopropylamide (2M, 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 Na2S2O3 and brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to afford crude 2-iodo-4-nitro-1-(phenylsulfonyl)-1H-indole, which was used without further purification.

[1234] Following a procedure analogous to that described in Example 205, 2-iodo-4-nitro-1-(phenylsulfonyl)-1H-indole was treated with potassium carbonate in methanol to provide 2-iodo-4-nitro-1H-indole.

[1235] To a solution of 2-iodo-4-nitro-1H-indole (1.0 equiv.) in tetrahydrofuran was added sodium hydride (5.0 equiv.) at 0°C. The mixture was stirred at 0-25°C for 30 min, and CF3CH2OTf (4.0 equiv.) was added to the reaction mixture. After the reaction was stirred at room temperature for 2 h, the reaction was quenched by adding 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-1-(2,2,2-trifluoroethyl)-1H-indole as a yellow solid.

[1236] To a solution of 2-iodo-4-nitro-1-(2,2,2-trifluoroethyl)-1H-indole (1.0 equivalent) in acetic acid, Fe (6.0 equivalent) is added. The mixture is stirred at 50 ° C for 2 h, then poured into 0 ° C water (using a magnet to remove Fe powder) and filtered to obtain a crude product. The crude product is absorbed in ethyl acetate and washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated. The concentrated solution is added dropwise to petroleum ether to precipitate the product. 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine as a yellow solid is obtained by filtration.

[1237] To a solution of 2-methyl-2-(5-(prop-2-yn-1-ylamino)pyridin-2-yl)propionitrile (693 mg, 2.94 mmol, prepared from 2-(5-aminopyridin-2-yl)-2-methylpropionitrile 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). 2-iodo-1-(2,2,2-trifluoroethyl)indol-4-amine (500 mg, 1.47 mmol) and tetrakis(triphenylphosphine)palladium(0) (170 mg, 147 μmol) were then added. The resulting reaction mixture was stirred at room temperature for 2 h under a nitrogen atmosphere and then quenched with EDTA solution (5 mL). The reaction mixture was extracted with ethyl acetate (2x5 mL), and the combined organic layers were concentrated in vacuo. The residue was purified by preparative thin layer chromatography (silica gel, eluting with a 1 / 1 mixture of ethyl acetate / petroleum ether) to give 2-(5-((3-(4-amino-1-(2,2,2-trifluoroethyl)-1H-indol-2-yl)prop-2-yn-1-yl)amino)pyridin-2-yl)-2-methylpropionitrile (1.0 g, 73.25% yield, 88.6% purity) as a dark brown solid.

[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-methylpropionitrile (150 mg, 364.59 μmol) in ethanol (3 mL) was added 1-acetylpiperidin-4-one (5 equivalents) and tetraethoxytitanium (83 mg, 364.59 μmol). The mixture was stirred at 50 ° C for 2 h, and sodium cyanoborohydride (115 mg, 1.82 mmol) was added. After the reaction was stirred 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 (2x20 mL). The combined organic layers were concentrated in vacuo, and the crude residue was purified by preparative HPLC to afford 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-methylpropionitrile as a yellow solid.

[1239] LC-MS(ES + ,m / z):537.1[(M+H) + ]

[1240] Starting from 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine and using a procedure similar to that described in Example 289, the following compound was prepared:

[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-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine (8 g, 23.52 mmol) in ethanol (70 mL) was added 1-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 sodium cyanoborohydride (7.39 g, 117.60 mmol) was added to the reaction at 25°C. After stirring for 2 h at 25°C, 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-(1-methylpiperidin-4-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine (2.17 g, 19.64% yield, 93.1% purity) as a yellow solid.

[1251] 2-Iodo-N-(1-methylpiperidin-4-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine is reacted with 6-methyl-N-(prop-2-yn-1-yl)pyridin-3-amine (prepared from 6-methylpyridin-3-amine) under Sonogashira coupling conditions to give 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.

[1252] LC-MS(ES + ,m / z):456.2[(M+H) + ]

[1253] Starting from 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine and using a method similar to that described in Example 342, the following compound was prepared:

[1254]

[1255]

[1256]

[1257]

[1258]

[1259]

[1260]

[1261] Example 395 :5-[(3-{4-[(1,1-dioxo-1λ 6 Preparation of [4-(2,2,2-trifluoroethyl)-1H-indol-2-yl}prop-2-yn-1-yl)amino]pyridine-2-carbonitrile

[1262]

[1263] To a solution of 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indole-4-amine (500 mg, 1.06 mmol) in ethanol (15 mL) was added 1,1-dioxothianilide-4-one (471 mg, 3.18 mmol) and tetraethoxytitanium (1.21 g, 5.29 mmol, 1.1 mL). The mixture was stirred at 50 ° C for 12 h, and sodium cyanoborohydride (665 mg, 10.59 mmol) was added to the reaction. After the reaction was stirred at 80 ° C for 2 h, the reaction mixture was diluted with ethyl acetate (15 mL) and poured into a saturated sodium bicarbonate solution (40 mL). The mixture was extracted with ethyl acetate (3 x 40 mL). The combined organic layer was 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] 4-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)tetrahydro-2H-thiopyran 1,1-dioxide was reacted with 5-(prop-2-yn-1-ylamino)picolinonitrile (prepared from 5-amino-picolinonitrile and propargyl bromide) under Sonogashira coupling conditions to give 5-[(3-{4-[(1,1-dioxo-1λ 6 -thiazole-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] Starting from 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine and using a method similar to that described in Example 396, the following compound was prepared:

[1267]

[1268]

[1269]

[1270]

[1271]

[1272]

[1273]

[1274]

[1275] Example 455 :Preparation of 2-{4-[(2-{3-[(4-methanesulfonylphenyl)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 diformate (57.36 g, 262.82 mmol). The mixture was stirred at 110° C. for 10 h and then concentrated in vacuo to give the crude product, tert-butyl N-(4-methylsulfonylphenyl)carbamate (20 g, 73.71 mmol, 84% yield). The 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, and then propargyl bromide (22.36 g, 187.96 mmol, 16.2 mL) was added to the reaction. After stirring at 0° C. for an additional 40 min, the reaction mixture was poured into water (300 mL) and extracted with ethyl acetate (2×100 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, eluted 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 equivalent) in ethanol was added tert-butyl 4-oxopiperidine-1-carboxylate (5 equivalents) and tetraethoxytitanium (5 equivalents). The mixture was stirred at 50°C for 12 hours, and sodium cyanoborohydride (5 equivalents) was added to the reaction. After stirring at 25°C for 2 hours, the mixture was poured into a saturated sodium bicarbonate solution and stirred for 30 minutes. 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 equiv) in dimethyl sulfoxide was added N-isopropylpropan-2-amine (30 equiv), CuI (2 equiv), tert-butyl 4-((2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino)piperidine-1-carboxylate (1 equiv), and Pd(Ph3)4 (0.25 equiv). After the reaction was stirred at 20°C under a nitrogen atmosphere for 1 h, the mixture was poured into a saturated EDTA solution. The biphasic mixture was stirred at room temperature for 20 min and 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 afford 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 equiv) in ethyl acetate was added HCl / ethyl acetate, and the resulting mixture was stirred at 20° C. for 1 h. The mixture was then concentrated in vacuo to afford 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 the hydrochloride salt (pale 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 equivalent, hydrochloride), 2-chloro-N,N-dimethyl-acetamide (3 equivalents) and potassium carbonate (3 equivalents) in acetonitrile was stirred at 50 ° C for 0.5h. 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-methanesulfonylphenyl)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] Starting from 2-iodo-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine and using a method similar to that described in Example 455, the following compound was prepared:

[1287]

[1288]

[1289]

[1290]

[1291]

[1292]

[1293] Example 498 : Preparation of 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-methylpropionitrile

[1294]

[1295] Synthesis scheme:

[1296]

[1297] The flask was charged with CuI (89 mg, 465.6 μmol) and diisopropylamine (222 mg, 2.2 mmol, 309 μL), and under nitrogen, tert-butyl (2-iodo-1H-indol-4-yl) (1-methylpiperidin-4-yl) carbamate (200 mg, 439.2 μmol, prepared from 2-iodo-4-nitro-1H-indole) and (6-(2-cyanoprop-2-yl)pyridin-3-yl) (prop-2-yn-1-yl) carbamate (197 mg, 658.9 μmol) in dimethyl sulfoxide (3 mL) was added, followed by the addition of tetrakis(triphenylphosphine)palladium(0) (61 mg, 52.7 μmol). After the reaction was stirred at 45° C. for 1 h, the reaction was diluted with 20 mL of ethyl acetate and 50 mL of 1 M EDTA and stirred at 25° C. for 1 h. The reaction mixture was stirred at 4 ℃ for 2 hours, and the mixture was stirred for 3 hours at 4 ℃ for 1 hour.Then the mixture was stirred for 2 hours at 4 ℃ for 3 hours.The reaction mixture was extracted with ethyl acetate (3x 10mL) and washed with saturated brine (50mL), dried over anhydrous sodium sulfate and concentrated in vacuo.The crude residue was purified by preparative thin-layer chromatography (silica gel, eluted with petroleum ether / ethyl acetate: 1 / 1) to obtain (2-(3-((tert-butoxycarbonyl)(6-(2-cyanoprop-2-yl)pyridin-3-yl)amino)prop-1-yn-1-yl)-1H-indol-4-yl) (1-methylpiperidin-4-yl) tert-butyl carbamate (200mg, 319.1 μmol, 73% yield) as a yellow solid.

[1298] To a mixture of tert-butyl (2-(3-((tert-butoxycarbonyl)(6-(2-cyanoprop-2-yl)pyridin-3-yl)amino)prop-1-yn-1-yl)-1H-indol-4-yl)(1-methylpiperidin-4-yl)carbamate (200 mg, 319 μmol) in dimethylformamide (3 mL) cooled to 0° C. was added sodium hydride (38 mg, 957 μmol, 60% in mineral oil) and 2-bromoacetonitrile (147 mg, 957 μmol). 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 (3×10 mL). The combined organic layers were washed with brine (3 x 10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to afford tert-butyl (2-(3-((tert-butoxycarbonyl)(6-(2-cyanoprop-2-yl)pyridin-3-yl)amino)prop-1-yn-1-yl)-1-(cyanomethyl)-1H-indol-4-yl)(1-methylpiperidin-4-yl)carbamate (200 mg, crude) as a yellow oil.

[1299] To a mixture of tert-butyl (2-(3-((tert-butoxycarbonyl)(6-(2-cyanoprop-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 eq) was added bismuth trichloride (15 eq) in acetonitrile (5 mL) at 50 ° C. under a nitrogen atmosphere. The mixture was stirred at 50 ° C. for 1 h, poured into a mixture of sodium bicarbonate and ice water (30 mL, 1 / 1 w / w), and EDTA solution (2M, 50 mL) was added to the resulting mixture. The two-phase mixture was stirred at room temperature for 2 h and then separated into an aqueous phase and an organic phase. The aqueous phase was extracted with ethyl acetate (3x20 mL). The combined organic layers were washed with brine (3x 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-methylpropionitrile as a yellow solid. LC-MS (ES + ,m / z):466.3[(M+H) + ]

[1300] Starting from 2-iodo-4-nitro-1H-indole and using a method similar to that described in Example 498, the following compound was prepared:

[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] Pyridine (7.54g, 95.3mmol, 7.69mL) was added to a mixture of propargyl bromide (2.1g, 38.1mmol, 2.44mL) in acetonitrile (80mL) at 0°C. The mixture was stirred at 0°C for 1h, and phenyl chloroformate (6.0g, 38.3mmol) was added to the reaction. After the reaction was stirred at 0°C for 1h, the reaction mixture was poured into water (100mL), and the aqueous phase was extracted with ethyl acetate (3x 50mL). The combined organic layer was washed with brine (3x 50mL), 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: 20 / 1 to 5 / 1) to obtain phenyl propan-2-yn-1-ylcarbamate (5g, 75% yield) as a white solid.

[1306] To a mixture of phenyl propane-2-alkynes-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-methylpropionitrile (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 layer was 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, elution with petroleum ether / ethyl acetate: 1 / 1) to give 1-(6-(2-cyanoprop-2-yl)pyridin-3-yl)-3-(prop-2-yn-1-yl)urea (90 mg, 46% yield) as a white solid.

[1307] To a mixture of 1-(6-(2-cyanoprop-2-yl)pyridin-3-yl)-3-(prop-2-yn-1-yl)urea (90 mg) in dimethyl sulfoxide (2 mL) at 25° C., diisopropylamine (160.5 mg, 1.59 mmol), CuI (60.4 mg, 317.3 μmol), 2-iodo-N-(1-methylpiperidin-4-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine (80 mg, 158.6 μmol) and tetrakis(triphenylphosphine)palladium(0) (37 mg, 31.7 μmol) were added. The mixture was stirred at 25° C. for 1 h and diluted with ethyl acetate (10 mL), then poured into a 2N 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 afford 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 (19.6 mg, 22% yield) as a white solid.

[1308] LC-MS(ES + ,m / z):552.4[(M+H) + ]

[1309] Starting from 2-iodo-4-nitro-1H-indole and using a method similar to that described in Example 513, the following compound was prepared:

[1310]

[1311]

[1312] Example 522 :4-[(2-{3-[(6-methanesulfonylpyridin-3-yl)amino]prop-1-yn-1-yl}-1-(2,2,2-trifluoroethyl)-1H-indol-4-yl)amino]-1λ 6 -Preparation of 1,1-thiol-1,2-dione

[1313]

[1314] Synthesis scheme:

[1315]

[1316] A mixture of 6-(methylsulfonyl)pyridin-3-amine (5 g, 29.04 mmol) and di-tert-butyl diformate (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, eluted with petroleum ether / ethyl acetate: 5 / 1 to 1 / 1) to give 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 a nitrogen atmosphere. The mixture was quenched by adding saturated ammonium chloride solution (20 mL) and 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 give 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 give 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] Starting from 2-(3-((6-(methylsulfonyl)pyridin-3-yl)amino)prop-1-yn-1-yl)-1-(2,2,2-trifluoroethyl)-1H-indol-4-amine and using a method similar to that described in Example 522, the following compound was prepared:

[1323]

[1324] Example 530 : Preparation of 2-{3-[(4-methanesulfonylphenyl)amino]prop-1-yn-1-yl}-N-(oxacyclohexane-4-yl)-1-(2,2,2-trifluoroethyl)-1H-indole-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)...

Claims

1. A compound of the following formula, or a pharmaceutically acceptable salt thereof: in: - Each are independently a single bond or a double bond; -X 1 CR 5 , CR 5 R 6 , N, NR 5 , O, S, C=O, C=S or with Q 1 The carbon atom to which it is attached; -X 2 CR 7 , CR 7 R 8 , N, NR 7 , O, S, C=O, C=S or with Q 1 The carbon atom to which it is attached; -X 3 CR 9 , CR 9 R 10 , N, NR 9 , O, S, C=O, C=S or with Q 1 The carbon atom to which it is attached; -X 4 CR 11 , CR 11 R 12 , N, NR 11 , O, S, C=O, C=S or with Q 1 The carbon atom to which it is attached; -X 5 CR 13 , N or NR 13 ; Where X 1 , X 2 , X 3 and X 4 At least one of them is related to Q 1 The carbon atom to which it is attached; -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 is 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 、-OR 16 、-SR 16 、- NR 16 R 17 、-NR 16 C(O)R 16 、-OC(O)R 16 、C=O、C=S、-CN、- S i 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl or heterocyclic group, each of which is independently substituted or unsubstituted, or is 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, 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 R 3 does not exist; - 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 - 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 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; and - Each R 23 and R 24 is independently alkyl, alkenyl, alkynyl, aryl, heteroaryl or heterocyclyl, each of which is independently substituted or unsubstituted, or is hydrogen.

2. The compound of claim 1, wherein X 3 For Q 1 The carbon atom to which it is attached, and Y is N or O.

3. The compound of claim 1, wherein m is 1, and Y is N.

4. The compound according to claim 1, wherein the compound is a compound of the following formula:

5. The compound as claimed in claim 1, wherein R 1Alkyl, alkenyl, -C(O)R 16 、-C(O)OR 16 or -C(O)NR 16 R 17 .

6. The compound according to claim 1, wherein the compound is a compound of the following formula:

7. The compound according to claim 1, wherein the compound is a compound of the following formula:

8. The compound according to claim 1, wherein the compound is a compound of the following formula:

9. 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 to DNA, wherein the cell expresses the p53 mutant, wherein the compound is a compound of the formula: in: - Each are independently a single bond or a double bond; -X 1 CR 5 , CR 5 R 6 , N, NR 5 , O, S, C=O, C=S or with Q 1 The carbon atom to which it is attached; -X 2 CR 7 , CR 7 R 8 , N, NR 7 , O, S, C=O, C=S or with Q 1 The carbon atom to which it is attached; -X 3 CR 9 , CR 9 R 10 , N, NR 9 , O, S, C=O, C=S or with Q 1 The carbon atom to which it is attached; -X 4 CR 11 , CR 11 R 12 , N, NR 11 , O, S, C=O, C=S or with Q 1 The carbon atom to which it is attached; -X 5 CR 13 , N or NR 13 ; Where X 1 , X 2 , X 3 and X 4 At least one of them is related to Q 1 The carbon atom to which it is attached; -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 is 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 、-OR 16 、-SR 16 、- NR 16 R 17 、-NR 16 C(O)R 16 、-OC(O)R 16 、C=O、C=S、-CN、- S i 16 R 17 R 18 , alkyl, alkenyl, alkynyl, aryl, heteroaryl or heterocyclic group, each of which is independently substituted or unsubstituted, or is 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, 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 R 3 does not exist; - 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 - 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 -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 are independently alkyl, alkenyl, alkynyl, aryl, heteroaryl or heterocyclyl, each of which is independently substituted or unsubstituted, or is 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 is hydrogen.

10. 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 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, wherein the assay comprises: 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; b) contacting the labeled mutant p53 portion with biotin-labeled DNA and streptavidin conjugated with a fluorescent energy donor in the test chamber; c) illuminating the test chamber with light that promotes fluorescence resonance energy transfer; d) detecting the fluorescence resonance energy transfer; e) determining the SC of the compound based on the fluorescence resonance energy transfer 150 value; f) Comparison of the SC of the compounds 150 The SC values ​​of the control samples 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; as well as g) determining the level of activation of protein-DNA binding in the presence of said compound based on said comparison.

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