TW201427970A - Kinase inhibitors and method of treating cancer with same - Google Patents

Abstract

The invention is a compound represented by Structural Formula (I): or a pharmaceutically acceptable salt thereof. Values for the variables are provided herein. Also included is a phannaceutical composition comprising the compound represented by Structural Fonnula (I) and a pharmaceutically acceptable carrier or diluent and methods of treating a subject with cancer with the compound of Structural Formula (I).

Description

Kinase inhibitors and methods of using same for treating cancer Related application

This application claims the benefit of U.S. Provisional Application No. 61/813,011, filed on Apr. 17, 2013. This application also claims the benefit of International Application No. PCT/CA2012/000955, filed on October 12, 2012. All teachings of both of these applications are incorporated herein by reference.

Protein kinases have been the subject of extensive research to search for new therapeutic agents for various diseases such as cancer. Protein kinases are known to mediate intracellular signal transduction by effecting the transfer of phosphonium groups from nucleoside triphosphates to protein receptors involved in signaling pathways. There are a variety of kinases and pathways through which extracellular and other stimuli cause various cellular responses to occur within the cell.

Human TTK protein kinase (TTK), also known as tyrosine sulphate kinase, bispecific protein kinase TTK, monopolar spindle 1 (Mps1), and phosphotyrosine acid collection of sulphate kinase (PYT), A conserved multispecific kinase capable of phosphorylating serine, threonine and tyrosine residues, which is expressed in E. coli (Mills et al , J, Biol Chem. 22(5): 16000-16006 (1992)). TTK mRNA is not expressed in most physiologically normal tissues of humans ( Id ). TTK mRNA is expressed in some rapidly proliferating tissues (such as testis and thymus) and in some tumors (for example, TTK mRNA is not expressed in renal cell carcinoma, expressed in 50% breast cancer samples, and expressed in testicular tumors and ovarian cancer samples) ( Id ). TTK is expressed in some cancer cell lines and tumors relative to normal counterparts ( Id.; see also WO 02/068444 A1).

Thus, agents that inhibit protein kinases, specifically TTK, have the potential to treat cancer. There is a need for other agents that can act as protein kinase inhibitors, specifically TTK inhibitors.

In addition, cancer recurrence, drug resistance or cancer metastasis is one of the main challenges in cancer therapy. Cancer patients who respond well to initial anticancer therapies often develop resistance and secondary tumors that cause disease recurrence. Current research evidence suggests that the ability of tumor growth and proliferation depends on the smaller subtype of cells within the tumor. These cells are referred to as tumor initiating cells (TIC) or cancer stem cells. TIC is considered to be responsible for drug resistance, cancer recurrence, and cancer metastasis. Compounds that inhibit the growth and survival of such tumor-initiating cells can be used to treat cancer, metastasize, or prevent cancer recurrence. Therefore, there is a need for new compounds that inhibit the growth and survival of tumor-initiating cells.

The Applicants have now discovered that certain carbazole compounds are potent kinase inhibitors such as TTK protein kinase, polo-like kinase 4 (PLK4) and Aurora kinase (see Examples B, E and F). The present inventors have now also found that these oxazole compounds have potent anticancer activity against breast cancer cells, colon cancer cells and ovarian cancer cells in cell culture studies (see Examples C-D). Based on these findings, the carbazole compounds and their pharmaceutical groups are disclosed herein. Adults and methods of treating cancer using such carbazole compounds.

A specific example of the invention is a compound represented by structural formula (I): Or a pharmaceutically acceptable salt thereof.

-NR ¹ R ^{2 is taken} together to form a bridged bicyclic group which optionally contains an epoxy atom and optionally an alkyl, haloalkyl, hydroxy, alkoxy, halo, hydroxyalkyl or alkoxyalkyl group. Substituent; or -NR ¹ R ² bonded together to form an oxetane group, a tetrahydrofuranyl group, a tetrahydropyranyl group, a hydroxyethyl group or an alkoxyethyl group N, and optionally substituted by an alkyl group C Piper And R ³ is i) (C ₂ -C ₆ )alkyl optionally substituted by hydroxy, alkoxy or halo; or ii) optionally alkyl, haloalkyl, hydroxy, alkoxy or a halo-substituted cycloalkyl group; and R ⁴ is a phenyl group or a monocyclic heteroaryl group, each of which is optionally substituted with one to three groups represented by R ⁵ ; each R ^{5 is} independently selected from halo, alkyl And haloalkyl, hydroxy, alkoxy, -CN, -NO ₂ , -haloalkoxy and -NR ^a R ^a ; and each R ^{a is} independently hydrogen or alkyl.

Another embodiment of the invention is a compound represented by structural formula (Ia):

Or a pharmaceutically acceptable salt thereof, wherein: R ³ is i) (C ₂ -C ₆ )alkyl optionally substituted by hydroxy, alkoxy or halo; or ii) optionally alkyl, halo alkyl, hydroxy, alkoxy or halo substituted with the cycloalkyl; and R ⁴ is phenyl or monocyclic heteroaryl, each optionally substituted by R ⁵ represents a group of by one to three; each R ⁵ Independently selected from halo, alkyl, haloalkyl, hydroxy, alkoxy, -CN, -NO ₂ , -haloalkoxy, and -NR ^a R ^a ; and each R ^{a is} independently hydrogen or alkyl .

Another embodiment of the present invention is a pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent and a compound represented by the above formula (I) or (Ia) or a pharmaceutically thereof thereof Acceptable salt.

Another embodiment of the invention is a method of treating an individual having cancer comprising administering to the individual an effective amount of a compound of formula (I) or (Ia) or a pharmaceutically acceptable salt thereof.

Another embodiment of the invention is a method of inhibiting TTK activity in an individual in need of inhibition of TTK activity, comprising administering to the individual an effective amount of a compound represented by Structural Formula (I) or (Ia) or a pharmaceutically acceptable thereof Acceptable salt.

Another embodiment of the invention is the use of structural formula (I) or (Ia) for therapy A compound represented by the use thereof or a pharmaceutically acceptable salt thereof. In some embodiments, the therapy is for treating an individual having cancer. Alternatively, the therapy is used to inhibit TTK activity in an individual in need of inhibition of TTK activity.

Another embodiment of the present invention is the use of a compound represented by Structural Formula (I) or (Ia) or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating an individual having cancer.

Another embodiment of the present invention is the use of a compound represented by Structural Formula (I) or (Ia) or a pharmaceutically acceptable salt thereof for the production of TTK activity for inhibiting an individual in need of inhibition of TTK activity. Pharmacy.

[Details of the Invention]

In one embodiment, the teachings of the present invention relate to a compound represented by Structural Formula (I), (Ia), or a pharmaceutically acceptable salt thereof. Alternatively, the invention relates to a compound represented by structural formula (II), (III), (IV) or (V):

The pharmaceutically acceptable salts of the compounds represented by the structural formulae (II), (III), (IV) and (V) are also included in the present invention. The values of the variables in Structural Formulas (II) and (III) are the same as described for Structural Formula (I). The values of the variables in Structural Formulas (IV) and (V) are the same as described for Structural Formula (Ia).

In a first specific embodiment, the invention relates to a compound represented by the formula (I), (II) or (III), wherein the bridged bicyclic group formed by -NR ¹ R ² is [3.3.1] or [ 3.2.1] Bridged bicyclic groups, and the values of the remaining variables are the same as described above for Structural Formula (I). Typically, a bridged bicyclic group contains one ring nitrogen atom, zero or one epoxy atom and the remaining ring atoms of a total of eight or nine ring atoms are carbon atoms. Alternatively, -NR ¹ R ^{2 is taken} together to form a phenyl group substituted with oxetanyl, tetrahydrofuranyl, tetrahydropentanyl, hydroxyethyl or alkoxyethyl N and optionally substituted by alkyl C Base, and the values of the remaining variables are the same as described above for Structural Formula (I).

In a second specific embodiment, the invention relates to a compound represented by structural formula (I), (Ia), (II), (III), (IV) or (V) wherein R ⁴ is phenyl, pyridyl a pyrimidinyl or thienyl group, each of which is optionally substituted with one to three groups represented by R ⁵ , and the values of the remaining variables are as above for the structural formula (I) or (Ia) or the first specific example (for The same as described in the compound represented by the formula (I), (II) or (III).

In a third specific embodiment, the invention relates to a compound represented by structural formula (I), (II) or (III), wherein -NR ¹ R ² consists of Representation, and the value of the remaining variables is the same as described above for Structural Formula (I) or in the first specific example or in the second specific example.

In a fourth specific embodiment, the invention relates to a compound represented by structural formula (I), (Ia), (II), (III), (IV) or (V) wherein R ³ is cyclopropyl, different a propyl group, a cyclopentyl group, an isobutyl group, a cyclopropylmethyl group or a 2,2-dimethylpropan-1-yl group, and the values of the remaining variables are as above for the structural formula (I) or the first specific example ( For a compound represented by Structural Formula (I), (II) or (III)) or a second specific example or a third specific example (for a compound represented by Structural Formula (I), (II) or (III) The same as described in ). Alternatively, R ³ is cyclopropyl, cyclopentyl or isobutyl.

In a fifth specific embodiment, the invention relates to a compound represented by structural formula (I), (Ia), (II), (III), (IV) or (V) wherein R ⁴ is pyridin-2-yl Or 2-chlorophenyl, and the value of the remaining variables is the same as above for the structural formula (I), the first specific example (for the compound represented by the structural formula (I), (II) or (III)) The preferred embodiment or the third embodiment (for the compound represented by the formula (I), (II) or (III)) or the same as described in the fourth embodiment.

In a sixth specific embodiment, the invention relates to a compound represented by structural formula (I), (II) or (III), wherein -NR ¹ R ² is And the value of the remaining variable is the same as described above for the structural formula (I) or the first specific example, or the second specific example or the third specific example or the fourth specific example or the fifth specific example.

In a seventh specific embodiment, the invention relates to a compound represented by structural formula (I), (Ia), (II), (III), (IV) or (V), wherein R ⁵ is chloro or fluoro, and The value of the remaining variable is in the above or against the structural formula (I) or the first specific example (for the compound represented by the structural formula (I), (II) or (III)) or the second specific example or the third specific example (for compounds represented by Structural Formula (I), (II) or (III)) or in the fourth specific example or in the fifth specific example or the sixth specific example (for the structural formula (I), (II) The same as described in the compound represented by (III).

The invention also includes compounds that are depicted by structure and/or by name in the examples. The present invention includes the neutral form of such compounds as well as pharmaceutically acceptable salts thereof. The treatment with such compounds and/or the use of such compounds includes the neutral form of such compounds as well as the pharmaceutically acceptable salts thereof.

The term "alkyl" used alone or as part of a larger portion (such as "alkoxy" or "haloalkyl" and the like) means saturated aliphatic straight chain or Branched chain monovalent hydrocarbon group. Unless otherwise stated, an alkyl group typically has from 1 to 6 carbon atoms, i.e., (C ₁ -C ₆ )alkyl. As used herein, "(C ₁ -C ₆₎ alkyl _{((C 1 -C 6) alkyl} ) " means a group having from 1 to 6 carbon atoms in a straight chain or branched chain configuration.

"Alkoxy" means an alkyl group bonded through an oxygen-bonded atom represented by -O-alkyl. For example, "(C ₁ -C ₄ )alkoxy ((C ₁ -C ₄ )alkoxy)" includes methoxy, ethoxy, propoxy and butoxy.

The terms "haloalkyl" and "haloalkoxy" mean an alkyl or alkoxy group, optionally substituted by one or more halogen atoms. The term "halogen" means F, Cl, Br or I. The halogen in the haloalkyl or haloalkoxy group is preferably F.

"Hydroxyalkyl" is an alkyl group substituted with a hydroxyl group.

"Alkoxyalkyl" is an alkyl group substituted with an alkoxy group.

"Cycloalkyl" means a saturated aliphatic cyclic hydrocarbon group which typically contains from 3 to 8 ring carbon atoms, that is, a (C ₃ -C ₈ )cycloalkyl group. (C ₃ -C ₈ )cycloalkyl includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.

The terms "heteroaryl", "heteroaromatic", "heteroaryl ring", "heteroaryl group", "heteroaromatic ring" and "Heteroaromatic groups" are used interchangeably herein. A "monocyclic heteroaryl" is a heteroaryl group having one ring. "Monocyclic heteroaryl" means a ring having five or six heteroatoms selected from carbon and at least one (typically from 1 to 4, more typically 1 or 2) heteroatoms (eg, oxygen, nitrogen or sulfur). An aromatic ring group of an atom.

Examples of the monocyclic 5- to 6-membered heteroaryl group include a furyl group (e.g., 2-furyl, 3-furyl), an imidazolyl group (e.g., N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl). ),different Azolyl (eg 3-iso) Azolyl, 4-iso Azolyl, 5-iso Azolyl), Diazolyl (eg 2- Diazolyl, 5- Diazolyl), Azolyl (eg 2- Azolyl, 4- Azolyl, 5- Azolyl), pyrazolyl (eg 3-pyrazolyl, 4-pyrazolyl), pyrrolyl (eg 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl), pyridyl (eg 2-pyridyl) , 3-pyridyl, 4-pyridyl), pyrimidinyl (eg 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl), anthracene Base (eg 3-嗒 a thiazolyl group (eg, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl), isothiazolyl, triazolyl (eg, 2-triazolyl, 5-triazolyl), tetrazolyl (eg, Tetrazolyl) and thienyl (eg 2-thienyl, 3-thienyl).

As used herein, the term "bridged bicyclic group" refers to a ring system comprising two rings sharing at least three adjacent ring atoms. Typically, the bridged bicyclic group is a ring atom (typically 8-9) selected from carbon, nitrogen and oxygen, typically a nitrogen atom, 0-1 oxygen atoms and the remaining 7 carbon atoms 10 member group. The bridged bicyclic group has a bridgehead atom and a bridge bond. The bridgehead atom is bonded to three other ring atoms and is typically carbon. The bridge is the ring atom between the bridgeheads. The nomenclature of bridged bicyclic groups provides parentheses, which are numbers separated by periods in parentheses. These numbers are in descending order as the number of carbon atoms between each bridgehead. For example, in bicyclo [2.2.1] heptane, the molecule has three bridge bonds having two, two and one carbon atoms, indicated by the prefix bicyclo [2.2.1]. The bridged bicyclic group of the present invention is preferably [3.3.1] or [3.2.1]. An exemplary bridged bicyclic group is

The term "substituted C (C -substituted)" means a substituent on a carbon atom by. In the case of a ring group such as a bridged bicyclic group, " C substituted" means substituted on a ring carbon atom. The term "N-substituted (N -substituted)" means a substituent on the nitrogen atom via. In the case of a ring group such as a bridged bicyclic group, " N substitution" means substitution over a ring nitrogen atom.

The teachings of the present invention also include various isomers and mixtures thereof. "Isomer" means a compound having the same composition and molecular weight but differing in physical and/or chemical properties. Structural differences can be in the ability to construct (geometric isomers) or to rotate polarized planes (stereoisomers).

Certain compounds described herein may exist in various stereoisomeric or tautomeric forms. Stereoisomers are compounds that differ only in their spatial configuration. The present teachings encompass all such forms, including compounds in substantially pure enantiomers, racemic mixtures, and tautomeric forms (including structurally undepicted forms). When a disclosed compound is named or depicted by a structure without specifying stereochemistry, it is understood that the name or structure encompasses all possible stereoisomers, tautomers, geometric isomers, or combinations thereof.

When a geometric isomer is depicted by a name or structure, it is understood that the geometric isomer purity of the named or depicted geometric isomer is at least 60%, 70%, 80% by weight, 90% by weight, 99% by weight or 99.9% by weight purity. Geometric isomer purity is determined by dividing the weight of the named or depicted geometric isomer in the mixture by the total weight of all geometric isomers in the mixture.

A racemic mixture means that 50% is one enantiomer and 50% is the corresponding enantiomer. The present invention teaches all enantiomerically-pure, enantiomerically-enriched, diastereomeric, and diastereomeric properties of the compounds described herein. Concentrated and racemic mixtures and diastereomeric mixtures.

Enantiomeric and diastereomeric mixtures can be crystallized in the form of a chiral salt complex by means of well-known methods such as chiral phase gas chromatography, chiral phase high performance liquid chromatography or Crystallization in a chiral solvent resolves to its component enantiomer or stereoisomer. Enantiomers and diastereomers can also be obtained from diastereomeric or enantiomerically pure intermediates, reagents and catalysts by well-known asymmetric synthetic methods.

When a compound is designated by the name or structure indicating a single enantiomer, the compound is at least 60%, 70%, 80%, 90%, 99% or 99.9% optically pure (also known as "unspecified" unless otherwise indicated. Enantiomerically pure). Optical purity is the weight of the enantiomer designated or depicted in the mixture divided by the total weight of the two enantiomers in the mixture.

When the stereochemistry of a disclosed compound is named or depicted by a structure, and the named or depicted structure encompasses more than one stereoisomer (eg, in the form of a diastereomeric pair), the inclusion of the stereoisomerism Either one of the bodies or any mixture of the stereoisomers contained therein is included. It is further understood that the stereoisomers of the named or depicted stereoisomers are at least 60%, 70%, 80%, 90%, 99%, or 99.9. weight%. The stereoisomeric purity in this case is determined by dividing the total weight of the mixture of stereoisomers encompassed by the name or structure by the total weight of the mixture of all stereoisomers.

Pharmacologically acceptable salts of the compounds disclosed herein are included in the teachings of the present invention. The disclosed compounds have a basic amine group and are therefore capable of forming a pharmaceutically acceptable salt with a pharmaceutically acceptable acid. Suitable pharmaceutically acceptable acid addition salts of the compounds described herein include inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, metaphosphoric acid, nitric acid and sulfuric acid, and organic acids such as acetic acid, benzenesulfonic acid, benzene. Formic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glycolic acid, isethionic acid, lactic acid, lactobionic acid, maleic acid, malic acid, methanesulfonic acid, succinic acid, p-toluene a salt of sulfonic acid and tartaric acid). A compound of the present invention having an acid group such as a carboxylic acid can form a pharmaceutically acceptable salt with a pharmaceutically acceptable base. Suitable pharmaceutically acceptable basic salts include ammonium salts, alkali metal salts such as sodium and potassium salts, and alkaline earth metal salts such as magnesium salts and calcium salts. Compounds having a quaternary ammonium group also contain counter anions such as chloride, bromide, iodide, acetate, perchlorate and the like. Other examples of such salts include hydrochloride, hydrobromide, sulfate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, tartrate [For example (+)-tartrate, (-)-tartrate or mixtures thereof (including racemic mixtures)], succinates, benzoates and salts of amino acids such as glutamic acid.

The compounds described herein inhibit a variety of kinases including TTK, PLK (such as PLK4), Aurora A, Aurora B, CHK (such as CHK2), ALK, cKit (V560G), JNK3, MELK, and MUSK. Thus, the compounds described herein are generally suitable for the treatment of diseases or conditions associated with such kinases. In some embodiments, the compounds described herein inhibit TTK, PLK (such as PLK4), and/or Aurora B kinase. In some embodiments, the compounds described herein are Inhibition of ALK, Aurora B, cKit (V560G), JNK3, MELK and/or MUSK kinase.

In one embodiment, the compounds described herein are TTK, PLK, Aurora A, Aurora B, and/or CHK inhibitors and are useful in the treatment of diseases associated with such kinases, such as cancer. Alternatively, the compounds described herein are TTK inhibitors and are useful in the treatment of diseases associated with TTK, such as cancer. In another alternative embodiment, the compounds described herein are Aurora A and/or B inhibitors and are useful for inhibiting Aurora A and/or B activity for the treatment of various conditions such as cancer. In another specific embodiment, the compounds described herein are PLK inhibitors and are useful for inhibiting PLK activity for the treatment of various conditions such as cancer. Typically, PLK is PLK4, PLK2 and/or PLK1. In one embodiment, the PLK is PLK1 and/or PLK4. In another embodiment, the PLK is PLK4. In another alternative embodiment, the compounds described herein are CHK inhibitors and are useful for inhibiting CHK activity for the treatment of various conditions such as cancer. In another alternative embodiment, the compounds described herein are ALK inhibitors and are useful for inhibiting ALK activity for the treatment of various conditions such as cancer. In another alternative embodiment, the compounds described herein are cKit (V560G) inhibitors and are useful for inhibiting cKit (V560G) activity for the treatment of various conditions such as cancer. In another alternative embodiment, the compounds described herein are JNK3 inhibitors and are useful for inhibiting JNK3 activity for the treatment of various conditions such as cancer. In another alternative embodiment, the compounds described herein are MELK inhibitors and are useful for inhibiting MELK activity for the treatment of various conditions such as cancer. In another alternative embodiment, the compounds described herein are MUSK inhibitors and are useful for inhibiting MUSK activity for the treatment of various conditions such as cancer.

Another aspect of the teachings of the present invention relates to a method of treating an individual having cancer comprising administering to the individual an effective amount of a compound described herein. In a specific example, The compounds described herein inhibit tumor growth. For example, the compounds described herein inhibit the growth of tumors that overexpress at least one of TTK, PLK, Aurora A, Aurora B, CHK, ALK, cKit (V560G), JNK3, MELK, and MUSK.

In one embodiment, the compounds described herein inhibit the growth of tumors that overexpress TTK.

Cancers that can be treated by the methods of the present invention, including reducing the likelihood of recurrence, include lung cancer, breast cancer, colon cancer, brain cancer, neuroblastoma, prostate cancer, melanoma, glioblastoma multiforme, ovary Cancer, lymphoma, leukemia, melanoma, sarcoma, paraneoplasia, osteosarcoma, blastoma, glioma and mesothelioma. In one embodiment, the cancer is selected from the group consisting of leukemia, acute myeloid leukemia, chronic myelogenous leukemia, breast cancer, brain cancer, colon cancer, colorectal cancer, head and neck cancer, hepatocellular carcinoma, lung adenocarcinoma, metastatic melanoma, pancreas Dirty cancer, prostate cancer, ovarian cancer and kidney cancer. In one embodiment, the cancer is lung cancer, colon cancer, brain cancer, neuroblastoma, prostate cancer, melanoma, pleomorphic glioblastoma or ovarian cancer. In another embodiment, the cancer is lung cancer, breast cancer, colon cancer, brain cancer, neuroblastoma, prostate cancer, melanoma, pleomorphic glioblastoma or ovarian cancer. In another embodiment, the cancer is breast cancer, colon cancer, and lung cancer. In another embodiment, the cancer is breast cancer. In another embodiment, the cancer is a basal subtype breast cancer or a luminal B subtype breast cancer. In another embodiment, the cancer is a basal subtype of breast cancer that overexpresses TTK. In another embodiment, the basal subtype breast cancer is ER (estrogen receptor), HER2, and PR (progesterone receptor) negative breast cancer. In another embodiment, the cancer is a soft tissue cancer. "Soft tissue cancer" is a technically accepted term encompassing tumors of any soft tissue derived from the body. Such soft tissues connect, support or surround various structures and organs of the body including, but not limited to, smooth muscle, bone Skeletal muscle, tendon, fibrous tissue, adipose tissue, blood vessels and lymphatic vessels, perivascular tissue, nerves, mesenchymal cells, and synovial tissue. Therefore, soft tissue cancer can be cancer of adipose tissue, muscle tissue, nerve tissue, joint tissue, blood vessels, lymphatic vessels, and fibrous tissue. Soft tissue cancer can be benign or malignant. Generally, malignant soft tissue cancer is called sarcoma or soft tissue sarcoma. There are many types of soft tissue tumors, including lipoma, lipoblastoma, sputum lipoma, liposarcoma, leiomyomas, leiomyosarcoma, rhabdomyomas, rhabdomyosarcoma, neurofibromas, schwannoma/neurilemoma, nerves Tumor, malignant schwannomas, neurofibrosarcoma, neurosarcoma, nodular tenosynovitis, synovial sarcoma, hemangioma, glomus tumor, angioendothelioma, hemangioendothelioma, angiosarcoma, Kaposi sarcoma, Lymphangioma, fibroids, fibroids, superficial fibromatosis, fibroblastoma, fibrosarcoma, fibromatosis, cutaneous fibrosarcoma (DFSP), malignant fibrous histiocytoma (MFH), myxomas, Granulosa cell tumor, malignant stromal tumor, soft alveolar sarcoma, epithelial sarcoma, clear cell sarcoma and fibroproliferative small cell tumor. In a specific embodiment, the soft tissue cancer is a sarcoma selected from the group consisting of fibrosarcoma, gastrointestinal sarcoma, leiomyosarcoma, dedifferentiated liposarcoma, pleomorphic liposarcoma, malignant fibrous histiocytoma, round cell sarcoma and Synovial sarcoma.

In another embodiment, the cancer treated by the disclosed method is breast cancer, colon cancer or ovarian cancer.

In some embodiments, the present teachings provide methods for inhibiting the growth of tumor-initiating cells or reducing the likelihood of cancer recurrence in an individual undergoing anti-cancer therapy. The method comprises the steps of: a) evaluating the individual to determine if the cancer is in remission; and b) if the cancer is in remission; then administering an effective amount of TTK to the individual An inhibitor (for example, a compound represented by the formula (I) or (Ia)). If the cancer is not in remission, the method further comprises, optionally, continuing the anti-cancer therapy until the cancer enters the remission phase and subsequently administering an effective amount of a TTK inhibitor (eg, a compound represented by Structural Formula (I) or (Ia) ) step b).

As used herein, the term "tumor-initiating cell" or "TIC" refers to a cell that has self-renewal and proliferative capacity present in some tumors. These cells are sometimes referred to as cancer stem cells (CSCs) and are observed to share certain characteristics with normal stem cells, including stem cell-like phenotypes and functions. In some embodiments, TIC is characterized by its ability to form a tumor after xenografting in immunodeficient mice.

In some embodiments, the present teachings provide methods of inhibiting the growth of a tumor-initiating cell or reducing the likelihood of cancer recurrence in an individual in which the cancer is in remission, comprising administering to the individual an effective amount of a TTK inhibitor (eg, a compound represented by the formula (I) or (Ia)).

In some specific examples, for example, where an individual is treated to reduce the likelihood of cancer recurrence, the individual has been treated with an anti-cancer therapy. Alternatively, the individual has been treated with an anti-cancer therapy and the individual is in remission.

In some embodiments, the present teachings provide a method of treating an individual having cancer comprising administering to the individual an effective amount of a compound represented by Structural Formula (I) or (Ia) in combination with an effective anti-cancer therapy. In one embodiment, the cancer is a metastatic cancer. "Metastatic cancer" is a cancer that has spread from its primary site to other parts of the body.

In another embodiment, the present teachings are directed to methods of treating an individual having a drug resistant cancer. "drug-resistant cancer" is for one, two, Three, four, five or more cancers that are typically used to treat cancer drugs that are not reactive. In one embodiment, the drug resistant cancer is mediated by the growth of tumor initiating cells.

Suitable methods known in the art can be used to assess an individual to determine if the cancer is in remission. For example, the size of a tumor and/or tumor marker (usually a tumor-associated protein) can be monitored to determine the status of the cancer. The size of the tumor can be monitored using imaging devices such as X-ray, MRI, CAT scan, ultrasound, mammography, PET and the like or via biopsy.

For the methods described herein, such as co-administration methods, the anti-cancer therapy is selected from the group consisting of surgery, radiation therapy, immunotherapy, endocrine therapy, gene therapy, and administration of an anticancer agent. Alternatively, the anti-cancer therapy is radiation therapy. In another alternative, the anti-cancer therapy is immunotherapy. In another alternative, the anti-cancer therapy is the administration of an anti-cancer agent. In another alternative, the anti-cancer therapy is surgery.

Radiation therapy kills, destroys or treats cancer using radiation. Exemplary radiation therapy includes, but is not limited to, gamma radiation, neutron beam radiation therapy, electron beam radiation therapy, proton therapy, brachytherapy, and radioisotope therapy (ie, systemic radioisotope therapy).

Endocrine therapy is a therapy that adds, blocks, or removes hormones. For example, chemotherapeutic agents that block the production or activity of estrogen have been used to treat breast cancer. In addition, hormone stimulation of the immune system has been used to treat specific cancers, such as renal cell carcinoma and melanoma. In one embodiment, the endocrine therapy comprises administering a natural hormone, a synthetic hormone, or other synthetic molecule that blocks or enhances the body's natural hormone production. In another embodiment, endocrine therapy includes removing glands that produce certain hormones.

As used herein, gene therapy is the insertion of a gene into an individual's cells and organisms. Tissues to treat diseases such as cancer. Exemplary gene therapies include, but are not limited to, germline gene therapy and somatic gene therapy.

Immunotherapy (also known as biologic therapy/biotherapy, immunotherapy, or biological therapy) is the treatment of disease using a portion of the immune system. Immunotherapy can help the immune system recognize cancer cells or increase the response to cancer cells. Immunotherapy includes active and passive immunotherapy. Active immunotherapy stimulates the body's own immune system, while passive immunotherapy typically uses components of the immune system that are produced outside the body.

Examples of active immunotherapy include, but are not limited to, vaccines including cancer vaccines, tumor cell vaccines (autologous or allogeneic), dendritic cell vaccines, antigen vaccines, anti-idiotypic vaccines, DNA vaccines, viral vaccines or Tumor-infiltrating lymphocyte (TIL) vaccine with interleukin-2 (IL-2), or lymphocyte activating killer (LAK) cell therapy.

Examples of passive immunotherapy include, but are not limited to, monoclonal antibodies containing toxins and targeted therapies. Individual antibodies include naked antibodies and conjugated monoclonal antibodies (also known as tag antibodies, labeled antibodies or loaded antibodies). The naked monoclonal antibody does not have an attached drug or radioactive material and the conjugated monoclonal antibody is conjugated to, for example, a chemotherapeutic drug (chemical label), a radioactive particle (radioactive label), or a toxin (immunotoxin). Examples of such naked monoclonal antibody drugs include, but are not limited to, rituximab (Rituximab; Rituxan), one for the treatment of, for example, B-cell non-Hodgkin lymphoma Antibody to CD20 antigen; trastuzumab (Herceptin), an antibody against HER2 protein for the treatment of, for example, advanced breast cancer; alemtuzumab; Campath, An antibody against CD52 antigen for treating, for example, B cell chronic lymphocytic leukemia (B-CLL); cetuximab (cetuximab; Erbitux), for example with irinotecan Used in combination to treat antibodies against EGFR proteins such as advanced colorectal cancer and head and neck cancer; and bevacizumab (Avastin), which is an anti-angiogenic therapy that acts against VEGF proteins and, for example, Chemotherapy is used in combination to treat, for example, metastatic colorectal cancer. Examples of conjugated monoclonal antibodies include, but are not limited to, the radiolabeled antibody temimumab (Ibritumomab tiuxetan; Zevalin), which delivers radioactivity directly to cancerous B lymphocytes and is used to treat, for example, B cells. Non-Hodgkin's lymphoma; radiolabeled antibody tositumomab (Toxitumomab; Bexxar) for the treatment of, for example, certain types of non-Hodgkin's lymphoma; and immunotoxin gemtuzumab Gemtuzumab ozogamicin; Mylotarg, which contains calichemycin and is used to treat, for example, acute myeloid leukemia (AML). BL22 is a conjugated monoclonal antibody for the treatment of, for example, hairy cell leukemia, for the treatment of immunotoxins such as leukemia, lymphoma and brain tumors, and radiolabeled antibodies such as, for example, OncoScint for colorectal cancer and ovarian cancer, and for example ProstaScint for prostate cancer.

Alternatively, the anti-cancer therapies described herein include administration of an anti-cancer agent. An "anti-cancer agent" is a compound which, when administered in an effective amount to an individual having cancer, may partially or substantially achieve one or more of the following: arresting and reducing cancer growth. The degree of cancer (eg, reducing tumor size), inhibiting the growth rate of cancer, and improving or improving clinical symptoms or indicators associated with cancer, such as tissue or serum components, or prolonging the lifespan of an individual.

Anticancer agents suitable for use in the methods described herein include any anticancer agent that has been approved for the treatment of cancer. In one embodiment, the anticancer agent includes, but is not limited to, a targeting antibody, an angiogenesis inhibitor, an alkylating agent, an antimetabolite, a vinca alkaloid, a taxane, a ghost Podophyllotoxin, topoisomerase inhibitor, hormone antibiotic Tumor agents and other anti-tumor agents.

In one embodiment, anticancer agents useful in the methods described herein include, but are not limited to, paclitaxel, docetaxel, 5-fluorouracil, trastuzumab, lapatinib (lapatinib), bevacizumab, letrozole, goserelin, tamoxifen, cetuximab, panitumumab, gemcitabine , capecitabine, irinotecan, oxaliplatin, carboplatin, cisplatin, doxorubicin, epirubicin, cyclophosphine Indamine, methotrexate, vinblastine, vincristine, melphalan, cytarabine, etoposide, daunorubicin, Bole Bleomycin, mitomycin, and adriamycin, and combinations thereof.

In a specific example, an anticancer agent and a compound represented by the formula (I) or (Ia) are administered simultaneously. When administered simultaneously, the anticancer agent and compound can be administered in the same formulation or in different formulations. Alternatively, the compound and the additional anticancer agent are administered separately at different times.

In one embodiment, an individual in the methods described herein has not previously been treated with a TTK inhibitor (eg, a compound represented by Structural Formula (I) or (Ia)).

The term "effective amount" means that a beneficial or desired result is produced when administered to an individual, including clinical outcomes, such as inhibiting, inhibiting, or ameliorating cancer in an individual (eg, by clinical symptoms or cancer cells). The amount determined.

The term "inhibiting the growth of tumor-initiating cell" refers to preventing or reducing the proliferation rate and/or survival rate of tumor-initiating cells.

As used herein, the term "reducing the likelihood of cancer recurrence" The likelihood of recurrence of a cancer) means inhibiting or delaying the recurrence of cancer at or near the primary site and/or at the secondary site after the remission period. The term also means that in the case of the treatments described herein, cancer is less likely to recur than in the absence of such treatment.

As used herein, the term "remission" refers to the state of a cancer that has disappeared or is undetectable, typically associated with a clinical condition or indicator associated with cancer, after the individual has successfully treated with an anti-cancer therapy.

As used herein, "treating a subject with a cancer" includes partially or substantially achieving one or more of the following: arresting cancer growth, reducing the extent of cancer (eg, reducing tumor size). Inhibiting the growth rate of cancer, improving or improving clinical symptoms or indicators associated with cancer (such as tissue or serum components) or prolonging the lifespan of an individual; and reducing the likelihood of cancer recurrence.

The effective amount of a compound taught herein will generally vary depending on various factors such as a given drug or compound, pharmaceutical formulation, route of administration, type of disease or condition, characteristics of the individual or subject being treated, and the like, However, it can still be determined in a conventional manner by those skilled in the art. An effective amount of a compound of the present teachings can be readily determined by one of ordinary skill in the art using conventional methods known in the art.

In one embodiment, an effective amount of a compound taught herein is in the range of from about 0.1 mg to about 1000 mg per kilogram of body weight, or from about 1 mg to about 500 mg per kilogram of body weight, and in another alternative, per kilogram of body weight From about 20 mg to about 300 mg. In another embodiment, an effective amount of a compound taught herein is in the range of from about 0.5 mg/m ² to about 5000 mg/m ² , or from about 5 mg/m ² to about 2500 mg/m ² , and in another alternative In the scheme, it is from about 50 mg/m ² to about 1000 mg/m ² . Skilled artisans should be aware that certain factors may affect the dosage required to effectively treat an individual suffering from cancer or reduce the likelihood of cancer recurrence. Such factors include, but are not limited to, the severity of the disease or condition, prior treatment, general health and/or age of the individual, and other diseases present.

Furthermore, for the methods described herein, including treating an individual having cancer or reducing the likelihood of cancer recurrence, a "treatment" or dosing regimen of an effective amount of an individual using a compound of the present teachings can be a single The composition is administered or contains a series of administrations. For example, a compound of the present teachings can be administered at least once a week. However, in another embodiment, the compound can be administered to the individual from about once a week to once daily for a given treatment. The length of the treatment period depends on a number of factors, such as the severity of the disease, the age of the patient, the concentration and activity of the compound taught by the present invention, or a combination thereof. It is also understood that the effective dosage of the compound for treatment may be increased or decreased during the course of a particular treatment regimen. Dose changes can be caused by standard diagnostic assays known in the art and become apparent. In some cases, long-term investment may be required.

"Subject" is a mammal, preferably a human, but may also be an animal that requires veterinary treatment, such as a companion animal (such as a dog, a cat, and the like), a farm animal (such as a cow, a sheep, a pig, or a horse). And similar animals) and laboratory animals (eg, rats, mice, guinea pigs, and the like).

As will be understood by those skilled in the art, the compounds taught herein can be administered to a patient in a variety of forms depending on the route of administration chosen. The compounds of the present teachings can be administered, for example, by oral, parenteral, buccal, sublingual, nasal, rectal, patch, pumping or transdermal administration and correspondingly formulated pharmaceutical compositions. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, intrapulmonary, intrathecal, transrectal and topical modes. Parenteral administration can be carried out by continuous infusion over a selected period of time.

The compounds taught herein can be suitably formulated for administration to an individual. Pharmaceutical composition. The pharmaceutical compositions taught by the present invention include, as appropriate, one or more pharmaceutically acceptable carriers and/or diluents such as lactose, starch, cellulose and dextrose. Other excipients such as flavoring agents; sweeteners; and preservatives such as methylparaben, ethylparaben, propylparaben and butylparaben may also be included. A more complete list of suitable excipients can be found in the Handbook of Pharmaceutical Excipients (5th Ed., Pharmaceutical Press (2005)). Those skilled in the art will know how to prepare formulations suitable for the various routes of administration. The conventional procedures and ingredients for selecting and preparing suitable formulations are described, for example, in Remington's Pharmaceutical Sciences (2003-20th Edition) and in The United States Pharmacopeia: The National Formulary (USP 24 NF19), published in 1999. The carrier, diluent and/or excipient are "acceptable" in the sense of being compatible with the other ingredients of the pharmaceutical composition and not deleterious to the recipient.

Typically, for oral therapeutic administration, the compounds taught by the present invention may be combined with excipients and ingestible lozenges, buccal tablets, dragees, capsules, elixirs, suspensions, syrups, powders and the like. The form of the object is used.

Typically, for parenteral administration, solutions of the compounds of the present teachings can be prepared generally in water suitably mixed with a surfactant such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycol, DMSO, and mixtures thereof, and in oils in the presence or absence of alcohol. Under ordinary conditions of storage and use, such preparations contain a preservative to prevent the growth of microorganisms.

Sterile aqueous solutions or dispersions and sterile powders of the compounds described herein for the temporary preparation of sterile injectable solutions or dispersions are usually suitable for injectable use.

For nasal administration, the compounds taught by the present invention can be formulated as aerosols, drops Agents, gels and powders. Aerosol formulations typically comprise a solution or fine suspension of the active substance in a physiologically acceptable aqueous or nonaqueous solvent and are usually presented in a sterile form in a single or multiple doses in a sealed container which may take the form of a filter cartridge Or refill with an atomizing device for use. Alternatively, the sealed container can be a single dispensing device, such as a single dose nasal inhaler or aerosol dispenser, equipped with a metering valve intended to be disposed of after use. When the dosage form comprises an aerosol dispenser, it will contain a propellant, which may be a compressed gas such as compressed air or an organic propellant such as a chlorofluorocarbon. Aerosol dosage forms can also be in the form of a pump-spray.

For buccal or sublingual administration, the compounds of the present teachings can be formulated into lozenges, buccal tablets or tablets with carriers such as sugar, acacia, tragacanth or gelatin and glycerin.

For rectal administration, the compounds described herein may be formulated in the form of a suppository containing a conventional suppository base such as cocoa butter.

The compounds of the present invention can be prepared by methods known to those skilled in the art as illustrated by the following general schemes and procedures and subsequent preparation examples. All starting materials are either commercially available or prepared by methods known to those skilled in the art and as described below.

A general synthetic method for 1H-carbazole cores has been reviewed in the literature (Schmidt A. et al. Eur . J. Org . Chem. 2008, 4073-4095).

In a general synthetic procedure, the compound III described herein can be prepared in a sequence from 3-halo-1H-indazole-5-carboxylic acid II according to the following Reaction Scheme 1 , which sequence comprises the formation of guanamine, followed by A cross-coupling reaction with a suitable cross-coupling partner R ² Met (eg, ArB(OR') ₂ , ArSnR ₃ ) in the presence of a metal catalyst (eg, PdCl ₂ (dppf) or Pd(PPh ₃ ) ₄ ).

The composition of the intermediate boronic ester VI described herein may be prepared via a suitable aryl halide of V borylation of action based on aniline, aniline based on the aryl halide of V in turn via the appropriately substituted dihalobenzene IV of Intercatalytic amination preparation ( Scheme 2 ).

Although nortropinol and norpseudotorpine are commercially available, nortropinone and its analogs may be via p-halanilide VII , 3-oxoglutaric acid and the appropriate two. The multicomponent condensation of an aldehyde or its precursor ( VIII ) is prepared as shown in Scheme 3 . Subsequent reduction of the ketone ( IX ) thus obtained gives the corresponding bridged bicyclol X.

The enantiomerically pure (typically >98% ee) 3-iodo-1H-indazole-5-carboxamide described herein can be prepared by chiral preparative chromatography as outlined in Scheme 4 . The racemic compound ( XI ) is isolated to prepare.

The racemic amine ( XIII ) described herein can be synthesized in three steps as outlined in Scheme 5A . The nucleophilic addition of an aldehyde to an aldehyde using an organometallic reagent such as Grignard, organolithium or an organozinc reagent produces a secondary alcohol XII . Subsequent oxidation to the corresponding ketone followed by a reductive amination step produces the desired racemic amine ( XIII ). Alternatively, the desired racemic amine described herein can be obtained by condensation of an organometallic reagent with an organic nitrile followed by one-pot synthesis using the reduction of an imine intermediate as shown in Scheme 5B .

Alternatively, the enantiomerically pure 3-iodo-1H-indazole-5-carbamimidamine XI can be via 3-halo-1H-indazole-5-carboxylic acid and the enantiomerically pure amine XIII. The indoleamine is coupled to prepare ( Scheme 6 ). The enantiomerically pure amine can be obtained by separation of the racemic amine by chiral preparative chromatography or by recrystallizing the salt with a chiral acid such as, for example, tartaric acid, mandelic acid and benzhydryl tartaric acid. obtain.

In addition, the enantiomerically pure amines described herein can be synthesized using asymmetric nucleophilic addition of a carbon anion to the chiral imines XIVa , b ( Scheme 7 ). In this method, the desired chiral amine can be synthesized in two ways by the role of the fragment which acts as a nucleophile in the addition step and acts as a fragment of the electrophile. The chiral auxiliary acts as a chiral guiding group to generally give the addition product XV with higher diastereoselectivity, or can be further enriched using standard purification methods. The chiral auxiliary is removed to give the desired optically active amine XIII . The enantiomeric excess of the amines described herein can be further improved by recrystallization.

A variety of chiral auxiliaries ( Scheme 8A ) can be employed in the synthesis of chiral imines XIVa , b . The process developed by Ellman involves the condensation of a third butyl sulfinamide with an aldehyde to give the intermediate XVI ( Scheme 8B ; Reference: Chem . Rev. 2010 , 110, 3600). The Grignard reagent was added stereoselectively and the auxiliaries were removed under mildly acidic conditions. Further examples of chiral auxiliaries commonly employed in this process are 1-amino-2-methoxymethylpyrrolidine (Reference: Tetrahedron: Asymmetry 1997 , 8, 1895) and phenylglycolamine (Reference: J .Org.Chem . 1991 , 56 1340)

The compounds described herein can be prepared in a manner similar to the general procedures described above or the detailed procedures described in the Examples herein.

The invention is illustrated by the following examples, which are not intended to be limited in any way.

Example A: Synthesis General method

Commercially available starting materials, reagents and solvents are used as they are. In general, the anhydrous reaction is carried out under an inert atmosphere such as nitrogen or argon. PoraPak ^® Rxn CX refers to a commercially available cation exchange resin available from Waters.

The microwave reaction was carried out using a Biotage Initiator microwave reactor. The progress of the reaction is usually monitored by TLC using a Merck silica gel plate and observed by UV at 254 nm by analytical HPLC or by LCMS (Bruker Exquire 4000). Rapid column chromatography purification of the intermediate or final product using 230-400 mesh silicone 60 from EMD chemicals or Silicycle, or using a KP-SIL or HP-SIL cerium dioxide filter cartridge, or KP-NH alkaline modification The ruthenium dioxide and the Biot Isolera corresponding to the samplet were purified. Reverse phase RPHPLC purification uses about 5%-30% MeCN or MeOH/0.05% TFA-H ₂ O to 70%-90% MeCN or MeOH/0.05% over a period of 20-40 minutes at a flow rate of 30-50 mL/min. TFA-H ₂ O was carried out on a Varian PrepStar Model SD-1 HPLC system with a Varian Monochrom 10u C-18 reverse phase column. Also the use of reverse-phase purified equipped with a KP-C18-H column of Biotage Isolera, MeOH / 0.1% TFA to use between 10% to 95% / H ₂ O for. Proton NMR was recorded on a Bruker 400 MHz spectrometer and mass spectra were obtained using a Bruker Esquire 4000 spectrometer. The optical rotation was measured at a given sample concentration (c, unit g/100 mL) using an AA-55 polarimeter from Optical Activity with a 2.5 x 100 mm uncased stainless steel tube on a sodium D line (589.44 nM).

Compound names were generated using software constructed in CambridgeSoft-PerkinElmer's ChemBioDraw Ultra version 11.0 or 12.0.

abbreviation:

Preparation of starting materials

General Method A (Hydrazine Coupling)

Treatment of 3-iodo-1H-indazole-5-carboxylic acid (1.0 eq.), DIPEA (3.0-5.0 eq.) and RR'NH with TBTU or BOP-Cl (1.05 eq.) with one-time addition at 0 ° C or rt. (1.00-1.05 equivalents) of DMF solution. The reaction was stirred and allowed to slowly warm to room temperature. After stirring for several hours or overnight, the crude reaction was then diluted with H ₂ O. In most embodiments, the (H ₂ O) precipitate is filtered and washed to obtain the desired material in the desired purity, or the material is purified directly by preparative HPLC or/and flash chromatography.

General Method B (Suzuki-Miyaura cross coupling)

3-iodo-1H-carbazole (1.0 eq.), aryl boronic acid or borate ester (1.2 eq.), base and palladium catalyst (0.05 equivalents, for example, and PdCl ₂ dppf.DCM, Pd(PPh ₃ ) ₄ ) in a solvent The mixture was degassed with Ar and heat sealed in a Biotage microwave reactor. The crude material was filtered through a pad of celite using Celite. In most of the examples, purification by preparative HPLC gave the target material.

General Method B2 (Suzuki-Miyaura Cross Coupling with PdCl ₂ dppf.DCM-Na ₂ CO ₃ )

Aqueous Na ₂ CO ₃ (2M, 3-4 mmol) was added to 5-substituted-3-iodo-1H-carbazole (1.0 mmol), aryl boronic acid or boronic ester (1.0-1.4 mmol) and PdCl ₂ dppf. DCM (0.1 mmol) in a mixture of PhMe: EtOH (1:1, 20 mL), heated under Ar in a Biotage microwave reactor, oil bath or reaction zone at a temperature between 100 ° C and 130 ° C . The crude material using MeOH (or acetone / MeOH or EtOAc) flushing pad filtration or partitioned between EtOAc and H ₂ O through diatomaceous earth, followed by drying (Na ₂ SO ₄ or MgSO ₄₎ and evaporated and purified by chromatography .

General Method C (boronation of aryl halides): use B ₂ pin ₂ /Pd

A mixture of aryl iodide or aryl bromide (1 equivalent), bis(pinacolyl)diboron (1.2 to 1.5 equivalents), KOAc (3 equivalents) and DMF or DMSO was purged with Ar for 10 minutes. [1,1'-PdCl ₂ dppf*CH ₂ Cl ₂ (3-5 mol%) was added, and the vial was sealed and heated at 85 ° C - 100 ° C for 2-3 hours. The product was partitioned between EtOAc and saturated aqueous NaHCO _3, washed with brine, dried over MgSO or ₂ SO _{4 4} Na, filtered and concentrated to dryness. The crude product was purified by flash chromatography toiel

General Method D (boration of aryl halides): use HBpin/Pd

Under Ar, aryl iodide or aryl bromide (1.0 mmol) in NEt ₃ (3.0 mmol) and HB pin (1.5 mmol), S-Phos (0.040 mmol) and Cl ₂ Pd(CH ₃ CN) ₂ (0.010 mmol) were added to the solution in (1.0 mL), and the reaction was heated to 110 ° C for 3 hours. The mixture was then transferred to a EtOAc (10mL) of a separatory funnel, and washed with NaHCO ₃ (saturated) (2 × 10mL), H 2 O (10mL) and brine (10mL). The organic layer was dried over MgSO _4, filtered, and the solvent removed to give the pinacol boronic ester, which was directly used in the next step.

General Method E (reductive amination)

Add NaBH ₃ CN (4 mmol) to a solution of the arylalkyl ketone (1 mmol) and NH ₄ OAc (12 mmol) in MeOH (4-5 mL), and heat the reaction mixture at 60 ° C. hour. Was added aqueous NaOH (2M, 15mL), and the product was extracted in to _{Et 2 O (3 × 40mL)} . The combined Et ₂ O layer was washed with H ₂ O (10mL) and brine (10 mL), dried (Na ₂ SO _4), filtered, and concentrated to dryness, and purified by chromatography or used in the form of a crude product.

General Method F (Copper Catalytic Amination of Aryl Halides)

1,4-diiodo benzene, or 1-bromo-4-iodobenzene (1.0 eq), CuI (20mol%), BINOL (20mol%) and K ₃ PO ₄ (2 equiv.) Is fed into a microwave vial. The vial was capped and then evacuated and backfilled with Ar. Dialkylamine (1.2 equivalents) and DMF were then added. The resulting mixture was stirred at room temperature for 2 to 4 days. The mixture was diluted with EtOAc, filtered over EtOAc EtOAc. The crude product was purified by flash chromatography toiel

General Method G (One-pot synthesis of cyclopropylmethylamine using aryl nitrile)

Bromocyclopropane (2 equivalents) was added to a microwave vial fed with Mg powder (2 equivalents) and THF. The resulting mixture was stirred at room temperature for 30 minutes, then a solution of aryl nitrile (1 eq.) in THF was added. It was subjected to microwave treatment at 100 ° C for 10 minutes, cooled to room temperature and added dropwise at 0 ° C to a solution of cold NaBH ₄ (2 eq.) in MeO. The resulting mixture was stirred at room temperature for 15 minutes, treated with H ₂ O, extracted with DCM and by Biotage SiO ₂ column: (gradient MeOH / DCM 0% -30%) , to give the desired product.

General method H (synthesis of tert-butyl sulfinylimine)

Add aryl or alkyl aldehyde (1.2 eq.) to stirred (S)-t-butylsulfinyl decylamine (1.0 eq.) and flame dried CuSO ₄ (2.2 eq.) in anhydrous CH ₂ Cl ₂ In the suspension. The resulting mixture was stirred at room temperature for 69 hours. The reaction mixture was filtered through a pad of celite and extracted with CH ₂ Cl ₂ . The combined organic extracts were concentrated under reduced pressure to give a crude material. Using cyclohexane as eluant EtOAc- by repeated flash chromatography (SiO ₂₎ to obtain the desired product.

General Method I (Removal of the sulfinamide protecting group)

The HCl (2.0M in Et ₂ O in 2.0 equiv.) Was carefully added to a stirred solution of 0 ℃ sulfinyl amine (1.0 eq.) In MeOH the solution. After the addition was completed, the cooling bath was removed, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure and Et ₂ O was added and a white precipitate formed. The precipitate was filtered off and washed with ₂ O Et dried under reduced pressure and used to obtain a crude product.

General Method J (Hydrate Formation)

It was added to final compound in MeOH 0 ℃ of the solution containing the 1M HCl Et ₂ O (1-2.1 equivalents, depending on the number of basic centers may be). After stirring at 0 ° C for 1 minute, it was concentrated to remove the solvent. MeOH was added and the solvent was removed. This was repeated twice to give the desired hydrochloride salt or the hydrochloride salt as a yellow solid.

Intermediate:

Synthesis of (1R,3R,5S)-8-(4-iodophenyl)-8-azabicyclo[3.2.1]oct-3-ol

Oxetan-3-one (10.8 g, 150 mmol) was added to a solution of 1-(4-bromophenyl)piperazine (24.1 g, 100 mmol) in DCM (400 mL) and a precipitate formed immediately. NaBH(OAc) ₃ (31.8 g, 150 mmol) and HOAc (1 mL) were added sequentially. The resulting mixture was stirred overnight at room temperature. Followed by extraction (200mL × 2) with saturated NaHCO, H ₂ O (100mL) wherein the stopper and stirred for 5 minutes ₃ (100mL), with DCM. The combined extracts were washed with the H ₂ O (300mL) and using dried over Na ₂ SO _4. The solvent was removed to give 1-(4-bromophenyl)-4-(oxetan-3-yl)piperazine (28.53 g, 96%). ^{_{1 H NMR (400MHz, CDCl 3}} ) δ ppm 7.36 (d, J = 9.2Hz, 2H), 6.80 (d, J = 8.8Hz, 2H), 4.71 (t, J = 6.4Hz, 2H), 4.66 (t , J = 5.8 Hz, 2H), 3.56 (five peaks, J = 6.4 Hz, 1H), 3.21 (t, J = 5.0 Hz, 4H), 2.50 (t, J = 5.0 Hz, 4H). MS ESI 296.1 [M+H] ⁺ , [C ₁₃ H ₁₇ BrN ₂ O + H] ⁺

Synthesis of (1R,3R,5S)-8-(4-iodophenyl)-8-azabicyclo[3.2.1]oct-3-ol

The title compound was obtained by using nortropinol (2.0 g, 15.7 mmol), 1,4-diiodobenzene (7.78 g, 23.5 mmol), K ₃ PO ₄ (10 g, 47 mmol), BINOL (0.9 g) at 45 °C. , 3.14 mmol), CuI (0.6 g, 3.14 mmol) and anhydrous DMF (48 mL) were synthesized for 18 hours according to General Procedure. After completion of the reaction, the solid sludge was filtered through celite and washed with EtOAc. Purification by flash chromatography (0%-45%EtOAc /EtOAc) The title compound was isolated as a pale pink solid (1.14 g, first crop). The filtrate was concentrated and then purified by flash chromatography (5% -90% MeOH / H 2 O), and then to give 1.12 g of (Batch 2) (of the combined Yield: 2.26g, 43.6%). ^{_{1 H NMR (400MHz, CDCl 3}} ) δ ppm 7.45-7.48 (m, 2 H), 6.52-6.55 (m, 2H), 4.14 (br.s, 2H), 4.01 (br.s, 1H), 2.29- 2.34 (m, 2H), 2.17-2.23 (m, 2H), 2.05-2.07 (m, 2H), 1.56-1.60 (m, 2H), 1.42 (d, J = 2.4 Hz, 1H); MS ESI 330.0 [ M+H] ⁺ , [C ₁₃ H ₁₆ INO+H] ⁺ calculated 330.2

Synthesis of (1R,5S)-3-(4-iodophenyl)-8-oxa-3-azabicyclo[3.2.1]octane

The title compound was obtained by using 8-oxa-3-azabicyclo[3.2.1]octane hydrochloride (100 mg, 0.66 mmol), 1,4-diiodobenzene (330 mg, 1.0 mmol), K at 24 °. ₃ PO ₄ (567 mg, 2.67 mmol), BINOL (38 mg, 0.13 mmol), CuI (26 mg, 0.13 mmol) and anhydrous DMF (6 mL) The title compound was isolated as a white solid (95 mg, 45%) after EtOAc. ^{_{1 H NMR (400MHz, CDCl 3}} ) δ ppm 7.51 (d, J = 8.3Hz, 2 H), 6.58 (d, J = 8.3Hz, 2 H), 4.49 (br.s, 2 H), 3.28 (d , J =11.3 Hz, 2 H), 3.00 (d, J = 11.3 Hz, 2 H), 1.84-2.05 (m, 4 H); MS ESI 316.2 [M+H] ⁺ , [C ₁₂ H ₁₄ INO+ H] ⁺ calculated value 316.0

The following intermediates were synthesized according to the general method F:

Synthesis of (1R,5S)-8-(4-bromophenyl)-8-azabicyclo[3.2.1]oct-3-one

(, 345mmol 16.8g) in the in H ₂ O (200mL) was added concentrated HCl solution of 2,5-dimethoxytetrahydrofuran (43.5g, 330mmol) and 3-oxo-glutaric acid (24mL). The resulting solution was stirred at room temperature for 30 minutes and then cooled to 0 °C. A solution of 4-bromoaniline (51.6 g, 300 mmol) in MeOH (250 mL) was added over 20 min. After the addition, the resulting mixture was stirred overnight at room temperature. Concentrated HCl (6 mL) was added and the reaction was heated at <RTI ID=0.0></RTI><RTIgt; After stopping with K ₂ CO ₃ /H ₂ O (27.6 g / 200 mL) to a pH of about 7, it was stirred at room temperature for 45 minutes. The precipitate was collected by suction filtration, washed with H ₂ O, MeOH rinsed and dried, to give a brown solid of the title compound (73.98g, 88%). ^{_{1 H NMR (400MHz, CDCl 3}} ) δ ppm 7.40 (d, J = 8.8Hz, 2H), 6.77 (d, J = 8.8Hz, 2H), 4.48-4.44 (m, 2H), 2.65 (dd, J = 15.6, 4.0 Hz, 2H), 2.33 (d, J = 15.6 Hz, 2H), 2.14 - 2.07 (m, 2H), 1.85-1.78 (m, 2H). MS ESI 279.9 [M+H] ⁺ , [C ₁₃ H ₁₄ BrNO+H] ⁺ calc.

Synthesis of (1R,3R,5S)-8-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaboron) (dioxaborolan)-2-yl)phenyl)-8-azabicyclo[3.2.1]oct-3-ol

To (1R,3r,5S)-8-(4-iodophenyl)-8-azabicyclo[3.2.1]oct-3-ol (13.16g, 40mmol), dicyclohexyl (2' under Ar ,6'-dimethoxy[1,1'-biphenyl]-2-yl)phosphine (987 g, 2.4 mmol, 6 mol%), Pd(CH ₃ CN) ₂ Cl ₂ (156 mg, 0.6 mmol, 1.5 mol %) in anhydrous two Et ₃ N (16.8 mL, 120 mmol, 3 equivalents) and 4,4,5,5-tetramethyl-1,3,2-diboron were added sequentially to the suspension in (120 mL). (17.4 mL, 3 equivalents). After the addition was completed, the resulting mixture was heated in an oil bath at 110 ° C for 5 hours. After cooling to room temperature, the reaction mixture was diluted with DCM (100mL) and washed with saturated aqueous NaHCO ₃ was slowly suspension to a pH of about 8. The reaction mixture was then taken through celite, washed with DCM (200 mL) and evaporated. The aqueous layer was extracted with DCM (100mL), and the combined organic layers were dried (Na ₂ SO ₄₎ and concentrated. The title compound (11.619 g) was obtained as a white solid. The wet mill was purified by a Biotage cartridge (EtOAc / hexanes 5% to 50%) to yield 1.88 g of pale yellow off white solid. The total amount of the title compound C was 13.50 g (quantitative yield, some pinacol was present). ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.69 (d, J = 8.4 Hz, 2H), 6.74 (d, J = 8.8 Hz, 2H), 4.28 - 4.23 (m, 2H), 4.03 - 3.97 (m, 1H), 2.36-2.28 (m, 2H), 2.27-2.19 (m, 2H), 2.11-2.05 (m, 2H), 1.60 (d, J = 14.8 Hz, 2H), 1.33 (s, 12H). MS ESI 330.1 [M+H] ⁺ , [C ₁₉ H ₂₈ BNO ₃ + H] ⁺ calc. 330.2

The following intermediates were synthesized according to the synthesis of (1R,5S)-8-(4-bromophenyl)-8-azabicyclo[3.2.1]octan-3-one:

The following intermediates are synthesized according to the general method C (or D, when indicated):

Synthesis of (1R,3R,5S)-9-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaboron) -2-yl)phenyl)-9-azabicyclo[3.3.1]non-3-ol

To (1R,5S)-9-(4-bromophenyl)-9-azabicyclo[3.3.1]indol-3-one (6.2 g, crude) in DCM (100 mL) NaBH ₄ (5.7 g, 15 mmol) was added to a solution (60 mL). The resulting mixture was stirred at 0 ° C for 10 minutes and then at room temperature for 30 minutes. After aqueous work-up, the residue was purified by flash chromatography (EtOAc /EtOAc elut Heterobicyclo[3.3.1]non-3-ol (1.30 g). MS ESI 295.9 [M+H] ⁺ , [C ₁₄ H ₁₈ BrNO+H] ⁺ calc. 296.1

The title compound (727 mg, beige solid) was obtained using (1R,5S)-9-(4-bromophenyl)-9-azabicyclo[3.3.1]indole-3-ol as a brown solid. 1.30 g, 4.4 mmol) and HBpin (1.9 mL) were prepared. ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.67 (d, J = 8.8 Hz, 2H), 6.82 (d, J = 8.8 Hz, 2H), 4.36 - 4.27 (m, 2H), 3.85 - 3.75 (m, 1H), 2.48-2.38 (m, 2H), 1.83-1.73 (m, 2H), 1.63-1.43 (m, 6H), 1.33 (s, 12H). MS ESI 344.1 [M+H] ⁺ , [C ₂₀ H ₃₀ BNO ₃ + H] ⁺ calc. 344.2

Synthesis of (1R,5S,7S)-9-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaboron) -2-yl)phenyl)-3-oxa-9-azabicyclo[3.3.1]non-7-ol

(1R,5S)-9-(4-Bromophenyl)-3-oxa-9-azabicyclo[3.3.1]non-7-one (0.98 g, 3.34 mmol) in THF at 0 °C in the (20mL) and MeOH (0.5mL) was added NaBH ₄ (380mL, 10mmol) to. The resulting mixture was heated at 50 ° C for 30 minutes. After cooling to room temperature, diluted with H ₂ O and extracted with DCM, to give greenish yellow solid of (1R, 5S, 7S) -9- (4- bromophenyl) -3-oxa-9 Azabicyclo[3.3.1]non-7-ol. MS ESI 298.0 [M+H] ⁺ , [C ₁₃ H ₁₆ BrNO ₂ +H] ⁺

The title compound (336 mg, off-white solid) was obtained using the above solid (1R,5S)-9-(4-bromophenyl)-3-oxa-9-azabicyclo[3.3.1]壬-7- Prepared with alcohol and HBpin (1.45 mL, 10 mmol). ¹ H NMR (400 MHz, CDCl ₃ ) δ ppm 7.72 (d, J = 8.4 Hz, 2H), 6.79 (d, J = 8.8 Hz, 2H), 6.65 (d, J = 12.4 Hz, 1H), 4.02-3.92 (m, 6H), 2.32 - 2.24 (m, 2H), 1.78 (d, J = 14.8 Hz, 2H), 1.34 (s, 12H). MS ESI 346.1 [M+H] ⁺ , [C ₁₉ H ₂₈ BNO ₄ + H] ⁺ calc. 346.2

Synthesis of (S)-N-(1-(2-fluorophenyl)-3-methylbutyl)-3-iodo-1H-indazole-5-carboxamide

A.(R,E)-N-(2-fluorobenzylidene)-2-methylpropane-2-sulfinamide

The title compound was obtained by using 2-fluorobenzaldehyde (10.0 g, 80.5 mmol), (S)-t-butylsulfinyl decylamine (12.2 g, 100 mmol), flame dried CuSO ₄ (16 g, 100 mmol) and MgSO ₄ (29g, 240mmol) of DCM (150mL) was synthesized. The resulting mixture was stirred at room temperature for 72 hours. The reaction mixture was filtered through a pad of diatomaceous earth and the pad rinsed with _{CH 2 Cl 2 (5 × 100mL} ). The combined org. By flash chromatography (Biotage Isolera, 100g HP-SIL , 0% -15% EtOAc / hexanes) to afford the clean product as a pale yellow oil ^{(11.3g, 61%) 1 H} NMR (300MHz, CDCl ₃ ) δ ppm 8.91 (s, 1H), 8.01 (dt, J = 7.6, 1.6 Hz, 1 H), 7.48-7.54 (m, 1 H), 7.23-7.27 (m, 1 H), 7.14-7.19 ( m, 1 H), 1.28 (s, 9 H).

B.(S)-N-((S)-1-(2-Fluorophenyl)-3-methylbutyl)-2-methylpropane-2-sulfinamide

Isobutylmagnesium bromide (2.0 M in Et ₂ O, 8.25 mL, 16.5 mmol) was carefully added to stirred dimethylzinc (1.2 M in toluene, 15.6 mL, 18.7 mmol) in. Anhydrous THF (30 mL) was added, and the mixture was stirred at room temperature for 30 minutes, then slowly added dropwise to -78 ° C (R,E)-N-(2-fluorobenzamide) over 30 minutes. A solution of 2-methylpropane-2-sulfinamide (2.5 g, 11 mmol) in dry THF (30 mL). Once the addition was complete, the mixture was stirred at this temperature for 3 hours. The reaction was quenched by the addition of a saturated aqueous solution of NH ₄ Cl (50 mL). H ₂ O (200 mL) was added and the mixture was extracted with Et ₂ O (2×75 mL). The combined the organic extracts were washed with brine (50mL), dried (Na ₂ SO ₄₎ and concentrated under reduced pressure to give the crude product as a clear yellow oil of (3.05g, 97%, and the product of co-additive Me 4:1 mixture of the product). ^{_{1 H NMR (400MHz, CDCl 3}} ) δ 7.36-7.40 (m, 0.25 H), 7.29-7.33 (m, 1 H), 7.22-7.29 (m, 1.25 H), 7.11-7.17 (m, 1.25 H), 7.02-7.06 (m, 1.25 H), 4.79-4.82 (m, 0.25 H), 4.56-4.61 (m, 1 H), 3.53 (m, 1.25 H), 1.63-1.68 (m, 0.91 H), 1.55- 1.60 (m, 1.50 H), 1.44-1.54 (m, 1.30 H), 1.21 (s, 9 H), 0.90-0.95 (m, 6 H), [C ₁₅ H ₂₄ FNOS + H] ⁺

C. (S)-1-(2-fluorophenyl)-3-methylbutan-1-amine

Removal of the chiral auxiliary protecting group by using HCl (2.0 M in Et ₂ O, 25 mL) and (S)-N-((S)-1-(2-fluorophenyl)-3-methylbutyl A solution of 2-methylpropane-2-sulfinamide (3.05 g, 10.6 mmol) in MeOH (30 mL). After the addition was completed, the cooling bath was removed and the mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure and crystallite crystal crystal By flash chromatography (Biotage isolera 120g C18,5% -80% MeOH / H 2 O) to give a white solid hydrochloride salt of the title compound (1.52g, 64%, 97% ee (S)). ¹ H NMR (400 MHz, CD ₃ OD ) δ ppm 7.46-7.52 (m, 2H), 7.32 (t, J = 7.6 Hz, 1H), 7.22-7.26 (m, 1H), 4.62-4.66 (m, 1H) , 1.93-1.98 (m, 1H), 1.79-1.86 (m, 1H), 1.37-1.47 (m, 1H), 0.93-0.98 (m, 6H). The ee of the compound was determined by the following: chiral HPLC, Daicel Chiralpak OD-H, 3:97 v/v 0.5% DEA-IPA: hexane, 1.0 mL/min, λ = 254 nm, R _t = 6.08 min (R) , R _t = 6.89 min (S).

Synthesis of (S)-3-methyl-1-(pyridin-2-yl)butan-1-amine

Addition of racemic 3-methyl-1-(pyridin-2-yl)butene-1- to a solution of hot (L)-DBTA (7.2 g, 20 mmol) in MeOH (75 mL) A solution of the amine (3.3 g, 20 mmol) in MeOH (30 mL). After the addition, the resulting suspension was stirred under reflux for 5 minutes and cooled in air for about 5 minutes. The resulting precipitate was collected by EtOAc (EtOAc) (EtOAc) (L)-DBTA salt of butan-1-amine (1.95 g, 95.6% ee). The ee of the compound was acetonitrile aliquoted with acetonitrile and chiral HPLC: Daicel Chiralpak AD-H, 90:10 v/v hexane-IPA (+0.5% Et ₃ N), 1.0 ml min ^-1 , λ = 254 nm, R _t = 5.8 min (R), R _t = 7.5 min (S) The product was analyzed for determination.

To a suspension of the above salt (1.9 g) in MeOH (5 mL) A clear solution is formed. After dilution with H ₂ O (50mL), the aqueous layer with DCM (30mL × 2) and extracted, and the combined organic layers were dried (Na ₂ SO _4), and the solvent removed to afford the desired amine as a colorless oil (705 mg, 21%).

Synthesis of (S)-1-(2-chlorophenyl)-3-methylbutan-1-amine

The fed with Me ₂ Zn in Ar (1.2M in PhMe in, 12.5mmol, 1.5 equiv) was carefully added the RBF i -BuMgBr (2.0M in Et ² O in, 7.5mL 1.5 equivalents). After diluting with THF (10 mL), the resulting mixture was stirred at room temperature for 30 min. The mixture was added to the stirred (S,E)-N-(2-chlorobenzylidene)-2-methylpropane-2-sulfinamide (2.44 g, at -78 °C over 10 min. 10 mmol, 1 eq.) in THF (50 mL). , Which was stirred for 3 hours at -78 deg.] C after addition, then quenched with saturated NH ₄ Cl and warmed to room temperature. Extraction with Et ₂ O gave crude 1-((S)-1-((R)-t-butylsulfinyl)-4-methylpentan-2-yl) as a colorless oil. 2-Chlorobenzene (3.06 g). ¹ H NMR indicated about 21% methylation by-product.

The mixture was dissolved in MeOH (30mL), cooled to 0 ℃ and treated with aqueous 1M HCl Et containing the ₂ O (20mL, 20mmol) process. After stirring for 30 minutes, it was concentrated to dryness and purified by reverse phase Biotage (MeOH / H ₂ O 5% to 90%) to give a white solid of the title compound (1.41g, 60%, HCl salt) . ^{_{1 H NMR (400MHz, CDCl 3}} ) δ ppm 7.61 (dd, J = 7.6,1.6Hz, 1H), 7.55 (dd, J = 7.8,1.4Hz, 1H), 7.49 (dt, J = 7.5,1.5Hz, 1H), 7.44 (dt, J = 7.6, 2.0 Hz, 1H), 4.93-4.35 (m, 1H, partially buried in H ₂ O), 2.00-1.91 (m, 1H), 1.90 to 1.82 (m, 1H) ), 1.55-1.43 (m, 1H), 1.00 (d, J = 6.4 Hz, 3H), 0.97 (d, J = 6.8 Hz, 3H). MS ESI 181.0 [M+H] ⁺ , [C ₁₁ H ₁₆ ClN+H-NH ₃ ] ⁺

The following intermediates were synthesized via reductive amination using General Procedure E:

(R,E)-N-(cyclopentylmethylene)-2-methylpropane-2-sulfinamide

The title compound utilizes cyclopentanecarboxaldehyde (15.0 g, 152.8 mmol, 1.0 eq.), (R)-t-butylsulfinyl decylamine (24.1 g, 198.7 mmol, 1.3 eq.) and flame dried CuSO ₄ (73.2) g, 458.5 mmol, 3.0 eq.) was synthesized according to general procedure H. The resulting mixture was stirred at room temperature for 71 hours. The reaction mixture was filtered through a pad of diatomaceous earth and the pad rinsed with _{CH 2 Cl 2 (5 × 100mL} ). The combined organic extracts were concentrated to dryness crystals crystals As eluent (SiO ₂₎ was purified by flash chromatography to afford the clean product as a pale yellow oil (23.8g, 78% isolated yield) 9 EtOAc- cyclohexane: Use 1. ^{_{1 H NMR (300MHz, CDCl 3}} ) δ ppm 7.99 (d, J = 5.5Hz, 1H), 3.02-2.87 (m, 1H), 1.97-1.78 (m, 2H), 1.78-1.55 (m, 6H), 1.18 (s, 9H).

The following sulfinamides are synthesized according to the synthesis of ( R,E )-N-(cyclopentylmethylene)-2-methylpropane-2-sulfinamide using the general method H:

Large-scale asymmetric synthesis of (S)-1-(cyclopentyl)-1-(2-pyridyl)methylamine hydrochloride

A.(R _S )-N-((S)-cyclopentyl ((pyridin-2-yl)methyl)-2-methylpropane-2-sulfinamide

A solution of 2-bromopyridine (11.5 g, 73 mmol in dry THF (50 mL)) was carefully added to i- PrMgCl. LiCl (1.3 M in THF, 56 mL, 73 mmol). The resulting solution was stirred at room temperature for 2 hours, after which it was added dropwise to -42 ° C ( R,E )-N-(cyclopentylmethylene)-2-methylpropane-2 over 30 minutes. - sulfinyl amine (11.3g, 56mmol) in dry the CH ₂ Cl ₂ (100mL) solution. The resulting mixture was slowly warmed to room temperature and stirred at room temperature overnight. The reaction was quenched by the addition of a saturated aqueous solution of NH ₄ Cl and mixture was extracted with CH ₂ Cl ₂ . After the solvent was removed, the residue was crystalljjjjjjjjjj The impure eluted fractions were re-purified with EtOAc (EtOAc EtOAc) Total: 3.347g (21% ^{yield) (; 1 H NMR (400MHz} , CDCl 3) δ ppm 8.56 (d, J = 4.5Hz, 1H), 7.63 (dt, J = 1.0,7.5Hz, 1H), 7.22 (d, J = 7.5 Hz, 1H), 7.16 (dd, J = 4.5, 7.5 Hz, 1H), 4.26 (dd, J = 5.0, 8.5 Hz, 1H), 3.95 (d, J = 5.0 Hz, 1H) , 2.44-2.31 (m, 1H), 1.94-1.83 (m, 1H), 1.68-1.44 (m, 5H), 1.44-1.32 (m, 1H), 1.30-1.17 (m, 1H), 1.13 (s, 9H).

B. (S)-1-(Cyclopentyl)-1-(2-pyridyl)methylamine hydrochloride

The title compound utilizes HCl (2.0M in Et ₂ O, 31.4 mL, 62.8 mmol) and ( R _S )-N-(( S )-cyclopentyl (pyridin-2-yl)methyl)-2-methyl A solution of propane-2-sulfinamide (8.8 g, 31.4 mmol) in MeOH (100 mL). After the addition was completed, the cooling bath was removed and the mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure and the residue was suspended in (125 mL of) in Et ₂ O. The precipitate was filtered off and washed with and the Et ₂ O ₍₂ × 125mL) and dried under reduced pressure to afford the crude product as a white solid (7.7g, 95.0% ee (S )). The crude product was recrystallized from t-BuOMe (150 mL),EtOAc (EtOAc) The crystals formed after cooling of the solution (3.3 g, 99.0% ee( s )) were collected by filtration, and the filtrate was concentrated under reduced pressure and recrystallized from t-BuOMe (100 mL) and MeOH (150 mL). A second crop of crystals (1.3 g, 98.0% ee( S )) was collected by filtration to afford 4.6 g, yield of 69% isolated yield. ¹ H NMR (400 MHz, D ₂ O + NaOH) δ ppm 8.81 (d, J = 5.5 Hz, 1H), 8.55 (t, J = 8.0 Hz, 1H), 8.06 (d, J = 8.0 Hz, 1H), 7.99 (t, J = 6.5 Hz, 1H), 4.53 (d, J = 10.5 Hz, 1H), 2.63-2.50 (m, 1H), 2.11-2.01 (m, 1H), 1.84-1.40 (m, 6H) , 1.24-1.12 (m, 1H). The ee of the compound was determined by aliquoting an aliquot with AcCl (see below for the synthesis example) and chiral HPLC: Daicel Chiralpak AD-H, 80:20 v/v heptane-EtOH ( +0.2% Et ₃ N), 1.0 mL/min, λ = 230 nm, R _t = 9.5 min (R), R _t = 25.4 min (S) Analysis product (S)- and (R)-N- (cyclopentane) (pyridin-2-yl)methyl)acetamide.

C. (racemic)-N-(cyclopentyl(pyridin-2-yl)methyl)acetamide

Add AcCl (0.10g, 1.25mmol) to a stirred solution of _{Et 3 N (0.35mL, 0.25g,} 2.5mmol, 2.2 eq) and (rac) -1-cyclopentyl-1- (2-pyridyl ) methylamine hydrochloride (0.25g, 1.14mmol) in CH of the ₂ Cl ₂ (5mL) suspension. The resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was washed with _{H 2 O (10%, 3} × 3mL) , and dried over Na ₂ SO ₄ dried and concentrated under reduced pressure to give the crude product as of a clear yellow oil (0.20 g), the crude product was rapid crystallization . ¹ H NMR (300MHz, CDCl ₃ ) δ ppm 8.53 (d, J = 5.0 Hz, 1H), 7.62 (dt, J = 1.5, 7.5 Hz, 1H), 7.28 (t, J = 7.5 Hz, 1H), 7.16 (d, J = 5.0 Hz, 1H), 6.72 (br d, J = 7.0 Hz, 1H), 4.93 (t, J = 9.0 Hz, 1H), 2.37-2.20 (m, 1H), 2.00 (s, 3H) ), 1.80-1.10 (m, 8H); HPLC: Daicel Chiralpak AD-H, 80:20 v/v heptane-EtOH (+0.2% Et ₂ NH), 1.0 mL min ^-1 , 210 nm, R _t = 9.5 Min, R _t = 19.2 min.

Large scale asymmetric synthesis of (S)-1-(2-chlorophenyl)-1-isopropylmethylamine hydrochloride

A.(S _S )-N-((S)-1-(2-chlorophenyl)-2-methylpropyl)-2-methylpropane-2-sulfinamide

The i -PrMgCl (2.0M in THF, 46.2mL, 92.3mmol) was carefully added to a stirred solution at room temperature of Me ₂ Zn (in PhMe in 82mL, 98.4mmol 1.2M,) of. The resulting solution was stirred at room temperature for 30 minutes, and by subsequently added dropwise over 30 minutes to a stirred solution of -78 ℃ (S, E) - N - (2- chlorobenzylidene) -2-methylpropane A solution of -2-sulfinamide (15.0 g, 61.5 mmol) in dry THF (350 mL). After complete addition, the reaction mixture was stirred for 3 hours at -78 deg.] C, followed by careful addition of aqueous saturated NH ₄ Cl (200mL) suspension. The mixture was extracted with Et ₂ O (3×100 mL). With brine (100 mL) washed with the organic extracts were combined and dried (Na ₂ SO _4). The organic layer was concentrated under reduced pressure afforded crude material (17.9 g, quantitative yield, 16:1d.r. (S _S ,S)-(S _S ,R) as a white solid without any further Purification is used. ¹ H NMR (300MHz, CDC l ₃ ) δ ppm 7.38-7.15 (m, 4H), 4.46 (t, J = 8.0 Hz, 1H), 3.75 (br d, J = 8.0 Hz, 1H), 2.28-2.15 (m, 1H), 1.22 (s, 9H), 1.01 (d, J = 6.5 Hz, 3H), 0.85 (d, J = 6.5 Hz, 3H).

B. (S)-1-(2-Chlorophenyl)-1-isopropylmethylamine hydrochloride

The title compound was obtained using HCl (2.0M in Et ₂ O, 61.0 mL, 122.0 mmol) and (S _S )-N-((S)-1-(2-chlorophenyl)-2-methylpropyl)- A solution of 2-methylpropane-2-sulfinamide (17.8 g, 61.0 mmol) in MeOH (175 mL) After the addition was completed, the cooling bath was removed and the mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure and Et ₂ O (250mL) and a white precipitate formed. The precipitate was filtered off and washed with and the Et ₂ O ₍₂ × 200mL) and dried under reduced pressure to give a white solid of the crude product (11.8g, 88.7% ee (S )). The crude product was recrystallized from t-BuOMe (300 mL) and MeOH (48 mL). After cooling overnight, only a small amount of crystals were formed, which was removed by filtration. The filtrate was concentrated under reduced pressure and after about half of the volume had been removed, a second crop of solid appeared, which was also removed by filtration. The two batches of crystals were found to be racemic by chiral HPLC. The filtrate was concentrated to dryness and recrystallised from t-BuOMe (300mL) and MeOH (33mL) again. Also, when the solution was cooled, only a small amount of crystals were formed, like the second batch of solids formed when the solution was concentrated under reduced pressure, and the crystals were removed by filtration. The remaining filtrate was concentrated to dryness and suspended in t-BuOMe (200 mL) and filtered. The resulting white solid was dried under reduced pressure and washed with _{Et 2 O (3 × 150mL)} , to give a white solid of the purified product (9.0g, 67% isolated yield, 97% ee (S)) . ¹ H NMR (400 MHz, D ₂ O + NaOH) δ ppm 7.59-7.41 (m, 4H), 4.60 (d, J = 9.5 Hz, 1H), 2.44-2.30 (m, 1H), 1.18 (d, J = 6.5 Hz, 3H), 0.85 (d, J = 6.5 Hz, 3H; HPLC: Daicel Chiralpak AD-H, 97:3 v/v heptane-EtOH (+0.1% Et ₃ N), 1.0 mL/min, λ = 280 nm, R _t =6.0 min (S), R _t =7.3 min (R).

Synthesis of (R)-N-((S)-cyclopropyl)(thiophen-3-yl)methyl)-2-methylpropane-2-sulfinamide

Add cyclopropyl(thiophen-3-yl)methanone (0.29 mL, 2.4 mmol) to (R)-2-methylpropane-2-sulfinamide (0.242 g, 2.0 mmol) and Ti (Oi-) Pr) 4 (0.83 mL), 2.8 mmol. The reaction was heated at 70 ° C under Ar overnight. The reaction was then cooled to -48 &lt;0&gt;C and &lt;RTI ID=0.0&gt;&gt; The reaction was allowed to warm to rt and stirred for 4 h. The reaction was quenched by the addition of MeOH, diluted with brine and filtered over EtOAc. The filtrate was concentrated and purified by flash chromatography on SiO ₂ followed by two respective, first by using: DCM: MeOH (97: 3 ) and then with Et20: hexane (4: 1), to give a white solid of (R -N-((S)-cyclopropyl(thiophen-3-yl)methyl)-2-methylpropane-2-sulfinamide (0.175 g, 24%). ¹ H NMR (400 MHz, CDCl ₃ ) d ppm 7.27-7.32 (m, 2 H), 7.17 (dd, J = 4.77, 1.51 Hz, 1 H), 3.81 (dd, J = 8.66, 4.14 Hz, 1 H) , 3.48 (br.s., 1 H), 1.24 (s, 9 H), 1.13-1.26 (m., 1 H), 0.67-0.78 (m, 1 H), 0.46-0.64 (m, 2 H) , 0.30 (s, 1H);

MS ESI 258.0 [M + H] +, [C 12 H 19 INOS 2 + H] + 258.1 calcd.

Synthesis of (S)-cyclopropyl(thiophen-3-yl)methylamine

Add HCl (1.0 M in Et 2 O, 0.98 mL, 0.98 mmol) to (R)-N-((S)-cyclopropyl(thiophen-3-yl)methyl)-2-methyl A solution of propane-2-sulfinamide (0.125 g, 0.49 mmol) in MeOH (1 mL). The reaction was stirred at this temperature for 2 hours, concentrated under reduced pressure and dissolved in _{hexane: Et 2 O (1: 1} v / v) in. The precipitate formed was filtered off and washed with _{hexane: Et 2 O (1: 1} v / v) rinsing, to give hydrochloride salt of (S) - cyclopropyl (thiophen-3-yl) methanamine (white solid , 0.069g, 74%). ¹ H NMR (400 MHz, CD ₃ OD) d ppm 7.48-7.58 (m, 2 H), 7.25 (dd, J = 4.77, 1.51 Hz, 1 H), 3.73 (d, J = 10.04 Hz, 1 H), 1.29-1.43 (m, 1 H), 0.66-0.86 (m, 2 H), 0.51-0.61 (m, 1 H), 0.37-0.50 (m, 1 H); MS ESI 136.9 [M-NH ₂ ] ⁺ , [C ₈ H ₁₁ NS-NH ₂ ] ⁺ calculated value 137.0

Synthesis of (S)-N-(cyclopropyl(phenyl)methyl)-3-iodo-1H-indazole-5-carboxamide

The title compound was obtained using 3-iodo-1H-indazole-5-carboxylic acid (79 mg, 0.79 mmol), (S)- cyclopropylbenzylamine hydrochloride (50 mg, 0.27 mmol), TBTU (87 mg, 0.27 mmol), DIPEA (0.14 mL, <RTI ID=0.0></RTI></RTI><RTIID=0.0> ¹ H NMR (400 MHz, CD ₃ OD ) δ ppm 8.09 (s, 1 H), 7.95 (dd, J = 8.9, 1.6 Hz, 1 H), 7.56 (d, J = 8.8 Hz, 1 H), 7.43 7.50 (m, 2 H), 7.33 (t, J = 7.6 Hz, 2 H), 7.24 (t, J = 7.3 Hz, 1 H), 4.46 (d, J = 9.5 Hz, 1 H), 1.34-1.46 (m, 1 H), 0.66 (d, J = 8.0 Hz, 2 H), 0.48 (m, 2 H); MS ESI 418.1 [M+H] ⁺ , [C ₁₈ H ₁₆ IN ₃ O+H] ⁺ Calculated value 418.0.

Synthesis of N-(1-(2-chlorophenyl)-2-methylpropyl)-3-iodo-1H-indazole-5-carboxamide

A. 1-(2-Chlorophenyl)-2-methylpropan-1-ol

A solution of 2-chlorobenzaldehyde (2.75 g) in Et ₂ O (30 mL) was slowly added to a solution of isopropylmagnesium bromide at 0 ° C (from 0.98 g of magnesium and 4.85 g of 2-bromopropane in 70 mL of anhydrous) Obtained in Et ₂ O). The reaction mixture was stirred at 0 ℃ 1 h, and then with 25% NH ₄ Cl solution (100 mL) suspension. The organic layer was separated and aqueous brine evaporated with EtOAc The combined organic layers were washed with H ₂ O and brine, dried (Na ₂ SO ₄₎ and concentrated in vacuo. By flash chromatography _{(SiO 2, 0% -25%} EtOAc / hexanes) to give the title compound (clear colorless oil, 1.5g, 41%). ^{1 H NMR (400MHz, DMSO-} d 6) δ 7.52 (d, J = 7.2Hz, 1H) .7.37-7.31 (m, 2H), 7.25-7.21 (m, 1H), 5.27 (d, J = 4.4Hz , 1H), 4.68 (dd, J = 5.2 Hz, 1H), 1.88-1.80 (m, 1H), 0.86 (dd, J = 17.2 Hz, J = 6.8 Hz, 6H).

B. 1-(2-Chlorophenyl)-2-methyl-propan-1-one

A solution of 1-(2-chlorophenyl)-2-methylpropan-1-ol (1.5 g in 15 mL DCM) was added to a suspension of PCC (2.62 g in 30 mL DCM) at 25 °C. The reaction was monitored by TLC. The reaction was completed in 2 hours. Et ₂ O (120 mL) was added and the reaction mixture was stirred 15 min. The supernatant was decanted, dried (Na ₂ SO ₄₎ and concentrated in vacuo. By flash chromatography _{(SiO 2, 0% -10%} EtOAc / hexanes) to give the title compound (clear colorless oil, 1.24g, 82%). ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.41-7.27 (m, 4H), 3.37-3.30 (m, 1H), 1.19 (d, J = 6.8 Hz, 6H).

C. 1-(2-Chlorophenyl)-2-methylpropan-1-amine

The title compound was prepared from 1-(2-chlorophenyl)-2-methyl-propan-1-one (1.5 g, 8.2 mmol). MeOH was evaporated and the aqueous solution was added 3M NaOH (100mL), was extracted using EtOAc (2 × 100mL), and purified by flash chromatography _{(SiO 2, 0-25% DCM /} MeOH), to give the title compound (colorless oil, 348mg ,twenty three%). ^{1 H NMR (400MHz, DMSO-} d 6) δ ppm 7.38-7.21 (m, 4H), 4.87 (br.s, 2H), 4.16 (d, J = 8.0Hz, 1H), 2.12-2.04 (m, 1H ), 1.02 (d, JJ = 6.4 Hz, 3H), 0.82 (d, J = 6.8 Hz, 3H); MS ESI 184.08. [M+H] ⁺ , [C ₁₀ H ₁₄ ClN+H] ⁺ Calculated value 184.08 .

D. N-(1-(2-Chlorophenyl)-2-methylpropyl)-3-iodo-1H-indazole-5-carboxamide

The title compound was obtained by using 1-(2-chlorophenyl)-2-methylpropan-1-amine (0.55 g, 2.99 mmol), DMF (11 mL), 3-iodo-1H-indazole-5-carboxylic acid ( 863 mg, 2.99 mmol), DIPEA (2.09 mL, 11.98 mmol) and TBTU (960 mg, 2.99 mmol) were synthesized according to General Method A. The resulting stirred reaction mass at 25 deg.] C 12 hours and then it was suspended in H ₂ O (440mL). The solid was collected by filtration and washed with H ₂ O, to give the title compound (cream solid, 1.29g, 95%). ^{1 H NMR (400MHz, DMSO-} d 6) δ 13.76 (s, 1H), 8.91 (d, J = 8.8Hz, 1H), 7.91 (d, J = 9.2Hz, 1H), 7.74-7.66 (m, 1H ), 7.59 (d, J = 8.8 Hz, 1H), 7.46-7.22 (m, 4H), 5.27 (t, J = 9.2 Hz, 1H), 2.20-2.15 (m, 1H), 1.08 (d, J = 6.0Hz, 3H), 0.79 (d , J = 6.8Hz, 3H); MS ESI 454 [M + H] +, [C 18 H 17 ClIN 3 O + H] + calcd 454.

Synthesis of (R)-2-cyclopropyl-N-(3-iodo-1H-indazol-5-yl)-2-(pyridin-2-yl)acetamide

The title compound was obtained using EtOAc (EtOAc) ¹ H NMR (400 MHz, CD ₃ OD ) δ ppm 8.51-8.55 (m, 1 H), 8.13-8.16 (m, 1 H), 7.95-8.00 (m, 1 H), 7.81-7.86 (m, 1 H) ), 7.53-7.61 (m, 2 H), 7.30-7.36 (m, 1 H), 4.49-4.53 (m, 1 H), 1.38-1.48 (m, 1 H), 0.68-0.75 (m, 1 H) ), 0.51-0.64 (m, 3 H ); MS ESI [m + H] + 419.0, [C 17 H 15 INO 4 + H] + calcd 419.04.

Synthesis of (S)-2-cyclopropyl-N-(3-iodo-1H-indazol-5-yl)-2-(pyridin-2-yl)acetamide

The title compound was obtained using EtOAc (EtOAc) ¹ H NMR (400 MHz, CD ₃ OD ) δ ppm 8.51-8.55 (m, 1 H), 8.13-8.16 (m, 1 H), 7.95-8.00 (m, 1 H), 7.81-7.86 (m, 1 H) ), 7.53-7.61 (m, 2 H), 7.30-7.36 (m, 1 H), 4.49-4.53 (m, 1 H), 1.38-1.48 (m, 1 H), 0.68-0.75 (m, 1 H) ), 0.51-0.64 (m, 3 H ); MS ESI [m + H] + 419.0, [C 17 H 15 INO 4 + H] + calcd 419.04.

Synthesis of (S)-N-(cyclopropyl(2-fluorophenyl)methyl)-3-iodo-1H-indazole-5-carboxamide

The title compound was used at 0 ° C using 3-iodo-1H-indazole-5-carboxylic acid (358 mg, 1.23 mmol), (S)-cyclopropyl(2-fluorophenyl)methylamine hydrochloride (250 mg, 1.23 mmol) ), BOP-Cl (576 mg, 1.3 mmol), DIPEA (1.08 mL, 6.19 mmol) and DMF (5 mL) were synthesized according to General Method A. The reaction was stirred and allowed to warm slowly to rt and stirred at 24 ° C for 3 h. The reaction was concentrated and purified by flash chromatography (Biotage isolera 60g C18-HS, 5% -90% MeOH / 0.1% TFA.H 2 O), to give an off-white solid of the title compound (405mg, 75%). ¹ H NMR (400 MHz, CD ₃ OD ) δ ppm 8.08 (s, 1 H), 7.95 (dd, J = 8.8, 1.6 Hz, 1 H), 7.56-7.58 (m, 2 H), 7.26-7.31 (m) , 1 H), 7.17 (t, J = 7.6 Hz, 1 H), 7.09 (t, J = 10 Hz, 1 H), 4.76 (d, J = 9.2 Hz, 1 H), 1.41-1.50 (m, 1 H), 0.66-0.72 (m, 1 H), 0.58-0.62 (m, 1 H), 0.50-0.56 (m, 2 H); MS ESI 436.2 [M+H] ⁺ , [C ₁₈ H ₁₅ FIN ₃ O+H] ⁺ calculated value 436.

Synthesis of (S)-N-(1-(2-fluorophenyl)-3-methylbutyl)-3-iodo-1H-indazole-5-carboxamide

The title compound utilizes (S)-1-(2-fluorophenyl)-3-methylbutan-1-amine hydrochloride (1.0 g, 0.46 mmol), 3-iodo-1H-indazole-5-carboxylic acid ( 1.32 mg, 0.46 mmol), TBTU (1.55 g, 0.48 mmol), DIPEA (3.01 mL, 1.84 mmol) and DMF (15 mL) were synthesized according to General Method A. The crude reaction was then diluted After stirring for 4 hours with H ₂ O, filtered and washed with H ₂ O, obtained as a cream solid of the title compound (1.8g, 87%). ¹ H NMR (400 MHz, CD ₃ O D ) δ ppm 8.95 (d, J = 8.4 Hz, 1H), 8.08 (s, 1 H), 7.93 (d, J = 8.8 Hz, 1H), (m, 1 H ), 7.60-7.50 (m, 1 H), 7.47-7.56 (m, 1 H), 7.23-7.31 (m, 1 H), 7.13-7.21 (m, 2 H), 5.41-5.53 (m, 1 H) ), 1.84-1.92 (m, 1 H), 1.60-1.72 (m, 1 H), 1.47-1.58 (m, 1 H), 0.89-0.99 (m, 6 H); MS ESI 452.2 [M+H] ⁺ ,[C ₁₉ H ₁₉ FIN ₃ O+H] ⁺ calculated value 452.06

The following intermediates were synthesized according to General Method A:

Synthesis of N-(1-(2-chlorophenyl)-3-methylbutyl)-3-iodo-1H-indazole-5-carboxamide

A. 1-(2-Chlorophenyl)-3-methylbutan-1-amine

Using general procedure G: The batch was fed with Et added to the Mg-containing powder (7.20g, 300mmol) of the ₂ O (100mL) of the RBF i -BuBr (32.6mL, 300mmol). After the addition, it was stirred at room temperature for 1 hour, and then PhMe (100 mL) containing 2-chlorobenzonitrile (20.63 g, 150 mmol) was slowly added. The resulting mixture was refluxed (oil temperature 70 ° C) for 3 hours and cooled to room temperature. At -78 deg.] C was slowly added to a cold mixture of NaBH ₄ in MeOH in the solution. After the addition, the cold bath was removed and the mixture was stirred for 45 minutes, then quenched with 2 M aqueous HCl at 0 ° C and adjusted to pH 2 and diluted with H ₂ O. After extraction with EtOAc, EtOAc (EtOAc m. g). MS ESI 181.0 [M+H] ⁺ , [C ₁₁ H ₁₆ ClN+H-NH ₃ ] ⁺

B. N-(1-(2-Chlorophenyl)-3-methylbutyl)-3-iodo-1H-indazole-5-carboxamide

The title compound (2.78 g, pale-yellow solid) using 1-(2-chlorophenyl)-3-methylbutan-1-amine (1.98 g, 10 mmol), 3-iodo-1H-carbazole- 5-carboxylic acid (2.88 g, 10 mmol) was prepared. ^{1 H NMR (400MHz, DMSO-} d 6) δ ppm 13.70 (s, 1H), 9.01 (d, J = 8.4Hz, 1H), 8.12-8.09 (m, 1H), 7.96 (dd, J = 8.8,1.6 Hz, 1H), 7.62-7.57 (m, 2H), 7.41 (dd, J = 8.0, 1.2 Hz, 1H), 7.34 (dt, J = 8.0, 1.2 Hz, 1H), 7.24 (dt, J = 7.6, 1.6 Hz, 1H), 5.60-5.52 (m, 1H), 1.88-1.72 (m, 2H), 1.49-1.41 (m, 1H), 0.97 (d, J = 6.4 Hz, 3H), 0.95 (d, J) =6.4 Hz, 3H). MS ESI 468.4 [M+H] ⁺ , [C ₁₉ H ₁₉ ClIN ₃ O+H] ⁺ Calculated 468.0

The following enantiomerically pure intermediates were prepared by preparative chiral supercritical fluid chromatography (SFC) by separation of racemic compounds:

Example A1: N-((R)-cyclopropyl(pyridin-2-yl)methyl)-3-(4-((1R,3R,5S)-3-hydroxy-8-azabicyclo[3.2. 1]oct-8-yl)phenyl)-1H-indazole-5-carboxamide

Crude material (before hydrolysis of decylamine) ( R )-N-(cyclopropyl(pyridin-2-yl)methyl)-3-iodo-1H-indazole-5-carboxamide (4.954 g, 11.85 mmol And (1R,5S)-8-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaboron) A mixture of 2-yl)phenyl)-8-azabicyclo[3.2.1]oct-3-ol (4.29 g, 13.04 mmol, 1.1 eq.) was used sequentially with EtOH/PhMe (100 mL / 50 mL), 1M Na ₂ CO ₃ aqueous solution (23.7 mL, 2.7 mmol, 2 eq.) and Pd(PPh ₃ ) ₄ (685 mg, 0.593 mmol, 5 mol%). The resulting mixture was refluxed under Ar (an oil bath temperature at 110 ° C) for 1.5 days; LCMS showed that the reaction was incomplete. A 1 M aqueous Na ₂ CO ₃ solution (14.2 mL, 1.2 eq.) and Pd(PPh ₃ ) ₄ ( 685 mg, 0.593 mol) were then added and the mixture was refluxed (oil bath temperature at 110 ° C) for 1 day. LCMS indicated the reaction was complete, H ₂ O (100mL) and brine (50 mL), followed by addition of EtOAc (300mL). When separated, the aqueous layer (60mL × 2) and extracted with EtOAc, and the combined organic layers were dried over Na ₂ SO ₄ and concentrated. The resulting dark brown oil was purified by flash chromatography using two Biotage 100 g SiO ₂ column (gradient: EtOAc/hexanes 10% to 100% then MeOH/DCM 0% to 10%). The impure wash fractions were combined and subjected to a second purification by flash chromatography. All the cleansing fractions were combined and dried to give crystalljjjjjjjjjj ¹ H NMR (400 MHz, MeOH-d4) δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ Hz, 2H), 7.80 (dt, J = 8.0, 1.6 Hz, 1H), 7.57 (d, J = 8.8 Hz, 1H), 7.53 (d, J = 8.0 Hz, 1H), 7.30 (dd, J = 5.2) , 2.0 Hz, 1H), 6.94 (d, J = 8.8 Hz, 2H), 4.50 (d, J = 9.2 Hz, 1H), 4.25 (br, s, 2H), 3.93 (t, J = 4.8 Hz, 1H) ), 2.39-2.36 (m, 2H), 2.26-2.18 (m, 2H), 2.07-2.02 (m, 2H), 1.63 (d, J = 14.8 Hz, 2H), 1.46-1.36 (m, 1H), 0.71-0.66 (m, 1H), 0.61-0.50 (m, 3H); MS ESI [m + H] + 494.2, [C 30 H 32 N 2 O 5 + H] + 494.2 calcd. This was converted to the 2-hydrochloride salt (3.018 g, quantitative yield) using General Method J. ¹ H NMR (400 MHz, MeOH-d4) δ ppm 8.23 (s, 1H), 7.96 (s, 1H), 7.72 (dd, J = 8.4, 1.6 Hz, 1H), 7.67 (d, J = 8.0 Hz, 1H) ), 7.48 (t, J = 7.8 Hz, 2H), 7.32 - 7.27 (m, 3H), 6.49 (br, s, 1H, NH), 6.40 (t, J = 6.2 Hz, 1H, NH), 4.08- 4.02 (m, 2H), 3.44 (t, J = 11.8 Hz, 2H), 3.33 (t, J = 6.4 Hz, 2H), 2.97 - 2.89 (m, 1H), 2.10 - 1.97 (m, 1H), 1.81 -1.75 (m, 2H), 1.53-1.42 (m, 2H), 0.92 - 0.86 (m, 2H), 0.67 - 0.62 (m, 2H). HPLC (2 hydrochloride, >99% ee eluted in 7.9 min): Daicel Chiralpak IA, 60:40 v/v hexane-iPrOH (+0.5% Et ₂ NH), 1.0 mL min ^-1 , 254 nm, R _t (R) = 7.9 min, R _t (S) = 10.6 min. (2 hydrochloride) = -75.3 (c = 0.46, MeOH).

Example A2: 3-(4-((1R,5S)-3-oxa-8-azabicyclo[3.2.1]oct-8-yl)phenyl)-N-((S)-cyclopentyl (pyridin-2-yl)methyl)-1H-indazole-5-carboxamide

Crude material (before hydrolysis of guanamine) (S)-N-(cyclopentyl(pyridin-2-yl)methyl)-3-iodo-1H-indazole-5-carboxamide (200 mg, 0.44 mmol) And (1R,5S)-8-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaboron a mixture of -2-yl)phenyl))-3-oxa-8-azabicyclo[3.2.1]octane (138 mg, 0.44 mmol, 1 eq.) was taken in EtOH/PhMe (2 mL / 2 mL) Na ₂ CO ₃ saturated aqueous solution (2 mL) and Pd(PPh ₃ ) ₄ (26 mg, 0.022 mmol, 5 mol%) were treated. The resulting mixture was heated in a Biotage microwave reactor at 125 °C for 3 hours under Ar; LCMS indicated the reaction was completed. H ₂ O (10 mL) and brine (5 mL) and EtOAc (20 mL). When separated, the aqueous layer (10mL × 3) and extracted with EtOAc, and the combined organic layers were dried over Na ₂ SO ₄ and concentrated. The resulting yellow oil was purified using Biotage 50g SiO ₂ column (gradient: EtOAc / Hexane 0 to 100%) is purified by flash chromatography. The purest fractions were combined and repurified by reverse phase chromatography using a Biotage 50g C18 column (gradient: ACN / water 10% to 90%). All pure extracts were combined and dried to give a pale yellow solid. The title compound was converted to the <RTI ID=0.0></RTI></RTI><RTIID=0.0> ¹ H NMR (400 MHz, CD ₃ OD ) δ ppm 8.78 (d, J = 6.0 Hz, 1 H), 8.61 (s, 1 H), 8.55 (t, J = 7.9 Hz, 1 H), 8.09 (d, J = 8.0 Hz, 1 H), 7.81-7.98 (m, 4 H), 7.60 (d, J = 8.3 Hz, 1 H), 7.08 (br.s., 2 H), 5.01 (d, J = 10.8) Hz, 1 H), 4.25 (br.s., 2 H), 3.94 (d, J = 11.0 Hz, 2 H), 3.58 (br.s., 2 H), 2.55-2.68 (m, 1 H) , 2.15-2.26 (m, 1 H), 2.00-2.14 (m, 4 H), 1.67-1.84 (m, 3 H), 1.53-1.67 (m, 2 H), 1.39-1.49 (m, 1 H) , 1.18-1.29 (m, 1 H) ; MS ESI 330.1 [m + H] + 508.4, [C 33 H 33 N 5 O 2 + H] + 508.3 calcd. HPLC (2 hydrochloride, >99% ee, eluted at 16.6 min): Daicel Chiralpak IA, 60:40 v/v hexane-iPrOH (+0.5% Et ₂ NH), 1.0 mL min ^-1 , 254 nm, R _t (R) = 9.7 min, R _t (S) = 16.6 min. (2 hydrochloride) = +90.9 (c = 0.48, MeOH).

Example A3: N-((S)-cyclopropyl(pyridin-2-yl)methyl)-3-(4-((1R,3R,5S)-3-hydroxy-8-azabicyclo[3.2. 1]oct-8-yl)phenyl)-1H-indazole-5-carboxamide

( R )-N-(cyclopropyl(pyridin-2-yl)methyl)-3-iodo-1H-indazole-5-carboxamide (600 mg, 1.44 mmol) and (1R,3S,5S)- 8-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaboron) A mixture of 2-yl)phenyl)-8-azabicyclo[3.2.1]oct-3-ol (520 mg, 1.58 mmol, 1.1 eq.) was used in EtOH/PhMe (10 mL/5 mL), 1M Na ₂ An aqueous solution of CO ₃ (2.87 mL, 2.87 mmol, 2 eq.) and Pd(PPh ₃ ) ₄ (83 mg, 0.072 mmol, 5 mol%). The resulting mixture was heated in a microwave at 125 °C for 4 hours; this reaction was repeated 4 times on the same scale (1.44 mmol) and repeated on a 0.48 mmol scale.

All six parts of the combined reactants through diatomaceous earth, diluted with EtOAc and washed with H ₂ O and brine. After drying and evaporation of the solvent, it was concentrated and purified by flash chromatography using a Biotage 100 g SiO ₂ column (gradient: EtOAc/hexane 0% to 100% then MeOH/DCM 0% to 20%). The title compound (free base, 1.97 g, 53%) eluted ¹ H NMR (400 MHz, MeOH- d ₄ ) δ ppm 8.66 (s, 1H), 8.52 (d, J = 5.0 Hz, 1H), 7.95 (dd, J = 8.8, 1.0 Hz, 1 H), 7.91-7.77 (m, 3H), 7.56 (dd, J = 13.2, 8.4 Hz, 2H), 7.32 (dd, J = 7.2, 5.4 Hz, 1H), 7.00 (d, J = 8.8 Hz, 2H), 4.52 (d, J = 9.3 Hz, 1H), 4.37 (br, s, 2H), 4.21-4.10 (m, 1H), 2.14 - 2.02 (m, 2H), 1.89-1.69 (m, 6H), 1.47-1.34 (m, 1H), 0.70 (d, J = 2.3 Hz, 1H), 0.63-0.47 (m, 3H).

The containing 1M HCl _{Et 2 O (8.2mL, 8.2mmol,} 2.05 eq) was added dropwise to the above-mentioned compound (free base, 1.97g, 4.00mmol) in MeOH (15mL) in the solution at the -10 ℃. After stirring at -10 °C for 2 minutes, it was concentrated to remove the solvent. MeOH was added and the solvent was removed. This operation was repeated twice to give the desired 2-hydrochloride salt (2.30 g, quantitative yield) as a yellow solid. ¹ H NMR (400 MHz, MeOH- d ₄ ) δ δ 8.81 (br, s, 2H), 8.67 (t, J = 7.5 Hz, 1H), 8.33 (d, J = 8.0 Hz, 1H), 8.26 (d, J = 7.8 Hz, 2H), 8.04-8.02 (m, 2H), 7.78-7.64 (m, 3H), 4.73 (br, s, 2H), 4.52 (d, J = 10.0 Hz, 1H), 4.34 - 4.19 (m, 1H), 2.37-2.01 (m, 8H), 1.61 (br, s, 1H), 0.94 (br.s., 1H), 0.84-0.61 (m, 3H); MS ESI [M+H] ⁺ 494.3, [C ₃₀ H ₃₂ N ₂ O ₅ + H] ⁺ calc. HPLC (2 hydrochloride, >99% ee, eluted at 16.1 min): Daicel Chiralpak IA, 60:40 v/v hexane-iPrOH (+0.5% Et ₂ NH), 1.0 mL min ^-1 , 254 nm, R _t (R) = 10.7 min, R _t (S) = 16.1 min. (2 hydrochloride) = +82° (c = 0.30, MeOH).

The following compound examples were prepared according to the synthesis described for Examples A1, A2 and A3 and using General Method B or B2:

Example B: TTK inhibition analysis

Active TTK was purchased from Invitrogen as an amine-based GST fusion of full length human TTK. Amino-terminal 6-histamine sumo-labeled human TTK (residues 1-275) was expressed in E. coli and purified to >95% homogeneity by Ni ²⁺ agarose, gel filtration and ion exchange chromatography. .

TTK activity was measured using an indirect ELISA detection system. GST-TTK (0.68 nM) in a 96-well microtiter plate pre-coated with amino-terminal 6-histamine-sodium-tagged TTK (amino acid residue 1-275) in 16 μM ATP (Sigma catalog number A7699), Incubation was carried out in the presence of 50 mM Hepes pH 7.2, 1 mM EGTA, 10 mM MgCl ₂ and 0.1% Pluronic. The reaction was allowed to occur for 30 minutes, then the plate was washed 5 times with wash buffer (phosphate buffered saline supplemented with 0.2% Tween 20) and incubated with a 1:3000 dilution of primary antibody (Cell Signaling Cat. No. 9381). minute. The plate was washed 5 times with wash buffer, incubated for 30 minutes in the presence of a secondary antibody (BioRad Cat. No. 1721019, 1:3000 concentration) conjugated to horseradish peroxidase, washed 5 more times with wash buffer and at TMB Incubation in the presence of the substrate (Sigma catalog number T0440). The colorimetric reaction was allowed to continue for 5 minutes, followed by the addition of a stop solution (0.5 N sulfuric acid) and detection at 450 nm by using a monochromator or filter based disk reader (corresponding to Molecular Devices M5 or Beckman DTX880) To quantify.

Compound inhibition was determined at a fixed concentration (10 [mu]M) or a variable inhibitor concentration (typically 0.5 [mu]M to 0.001 [mu]M in a 10 point dose reaction titration). Compounds were pre-incubated for 5 minutes in the presence of enzyme, followed by the addition of ATP and the remaining activity was quantified using the activity assay described above. The % inhibition of the compound was determined using the following formula: % inhibition = 100 x (1 - (experimental value - background value) / (high activity control - background value)). IC ₅₀ values were determined using the formula (A + (B + (1 + ((x / C) ^ D))))) using a nonlinear 4-point log-curve fit (XLfit4, IDBS), where A = background value, B = range , C = inflection point, D = curve fitting parameter.

In the following Table 1, IC ₅₀ of the compounds exemplified is given range. For values less than or equal to 0.1 μM; values greater than 0.1 μM and less than or equal to 0.5 μM; and values greater than 0.5 μM, the IC ₅₀ ranges are indicated as "A", "B", and "C", respectively.

Example C: Cancer cell line data of exemplary compounds of the invention

Breast cancer cells (MDA-MB-231), colon cancer cells (HCT116) and ovarian cancer cells (PA-1) were inoculated 24 hours before compound coverage (depending on cell growth rate, 1000 to 4000 in 80 μl per well) ) to a 96-well plate. Compounds were prepared as 10 mM stock solutions in 100% DMSO with DMEM (Dulbecco's Modified Eagle's Medium) cell growth medium (Invitrogen, Burlington) containing 10% FBS (fetal calf serum) , ON, Canada) was diluted to a concentration ranging from 50 nM to 250 μM. Aliquots (20 μl) of each concentration were overlaid onto 80 μl of pre-seeded cells in 96-well plates to form a final concentration of 10 nM to 50 μM. The cells were cultured for 5 days, followed by Sulfurhodamine B assay (SRB) to determine the cytostatic activity of the compounds.

Sulfadone rhodamine B (purchased from Sigma, Oakville, ON, Canada) is a water soluble dye that binds to the basic amino acid of cellular proteins. Thus, the colorimetric measurement of the combined dye provides an estimate of the total protein quality associated with the number of cells. The cells were fixed in situ by gently aspirating the medium and adding 50 μl of ice-cold 10% trichloroacetic acid (TCA) per well and incubating at 4 ° C for 30-60 minutes. Allowed to air dry and was washed with H ₂ O five such discs 5 minutes. A staining reaction was completed by adding 1 μm (v/v) acetic acid containing 50 μl of a 0.4% (w/v) SRB solution to each well and incubating at room temperature for 30 minutes. After staining, the discs were washed four times with 1% acetic acid to remove unbound dye and then allowed to air dry for 5 minutes. The dye was dissolved in each well with 100 μl of 10 mM Tris pH 10.5. The absorbance was read at 570 nm.

The percentage (%) of relative growth inhibition was calculated by comparison with cells treated with DMSO only (100%). GI ₅₀ for measuring cytotoxic activity of the compounds having. GI _{50 was} calculated using GraphPad PRISM software (GraphPad Software, Inc., San Diego, CA, USA). GI ₅₀ (growth inhibition) is the concentration of a compound that causes 50% inhibition of cell growth.

In Table 1 below, the range of GI ₅₀ values for examples of compounds for breast cancer cell lines (MDA-MB-231), colon cancer cell lines (HCT116), and ovarian cancer cell lines (PA-1) are given. The example compounds demonstrate different growth inhibition/cell killing activities against cells of breast, colon and ovarian cancer. For values less than or equal to 0.1 μM; values greater than 0.1 μM and less than or equal to 0.5 μM; and values greater than 0.5 μM, the GI ₅₀ ranges are indicated as "A", "B", and "C", respectively.

Example D: Colony and ovarian cancer tumor initiation cell data for exemplary compounds

Materials and Methods: Non-tissue or tissue culture treated T-75 flasks and 96-well plates were purchased from VWR. Vitamin B-27 supplement, MEM NEAA (minimum essential medium non-essential amino acid), sodium pyruvate, L-glutamic acid, N2 supplement, penicillin-streptomycin and amphotericin B (fungizone/amphotericin B ) was obtained from Invitrogen. Lipid mixture, heparin and EGF Was purchased from Sigma; bFGF was from BD Biosciences. Tumor-derived tumor-derived cells (TIC) were treated with non-tissue culture-treated T-75 flasks containing 0.2×B-27 supplement, 4 μg/ml heparin, 1×MEM NEAA, 1× sodium pyruvate, 1 mM branamide Acid, 10 pg/μl bFGF, 20 pg/μl EGF, 1×N2 supplement, lipid mixture, penicillin-streptomycin and amphotericin B in DMEM:F12 medium were routinely maintained. Ovarian TIC was routinely maintained in tissue culture treated T-75 flasks in DMEM:F12 medium containing 1×B-27 supplement, 4 μg/ml heparin, 20 pg/μl bFGF, 20 pg/μl EGF, and penicillin-streptomycin. .

Analysis Protocol: The compound of the dissolved and diluted further described herein for measuring GI in ₅₀ cell culture medium in DMSO. Colon TIC was trypsinized and seeded at 4,000 cells/well into non-tissue culture treated 96 well plates. After 24 hours, the compounds were added to the cell culture at various concentrations and the final concentration of DMSO was adjusted to 0.1%. The cells were then cultured for 9 days at 37 °C. Ovarian TICs were trypsinized and seeded at 1,000 cells/well into tissue culture treated 96 well plates. After 24 hours, the compounds were added to the cell culture at various concentrations and the final concentration of DMSO was adjusted to 0.1%. The cells were then cultured for 6 days at 37 °C. Cell viability was assessed by Alamar Blue assay: 10 [mu]l Alamar Blue was added to each well. After incubation for 4 hours at 37 ° C, fluorescence was recorded at excitation 544 and emission 590. GI ₅₀ (growth inhibition) was calculated using GraphPad Prism 4.0 software. The list of cell growth inhibition data for the compounds described herein is as follows.

In the following Table 1, compounds given in Examples ₅₀ value ranges for the TIC (12 colon and ovarian 2393A) of GI. For values less than or equal to 0.1 μM; values greater than 0.1 μM and less than or equal to 0.5 μM; and values greater than 0.5 μM, the GI ₅₀ ranges are indicated as "A", "B", and "C", respectively.

NA: not available

Example E: Selectivity for 270 human kinase groups

The compounds of the invention (A32, A33, A1 and A11) selected by Millipore were evaluated for their inhibitory activity against 270 different human kinase groups. The % inhibition was determined by the formula: inhibition % = 100 × (1 - (experimental value - background value) / (high activity control - background value)). As shown in the table below, only seven kinases were inhibited >80% at 1 [mu]M.

From the inhibition data in Table 2, it is apparent that certain kinases such as Aurora-B, cKit (V560G), JNK3, MAPK2, MELK, Met (Y1248C) and MUSK are strongly inhibited by the compounds of the present invention at a concentration of 1 μM, i.e., high. Its measured TTK IC ₅₀ > 2 orders of magnitude. Other kinases, including AKT (also known as PKB), PKA, PDK1, p70S6K, ROCK-I, ROCK-II, and JNK2, inhibited < or = 50% at this concentration.

Example F: PLK4 inhibition assay

The active PLK4 was purified from the E. coli expression system in the form of an amino-terminal GST fusion of residues 1-391 of human PLK4. After overnight incubation at 15 °C, proteins were purified from clarified cell extracts using glutathione sepharose, gel permeation chromatography and ion exchange (Resource Q). The resulting protein was dephosphorylated with lambda phosphatase (NEB Cat. No. P0753) and resolved from phosphatase using glutathione agarose. The dephosphorylated GST-PLK4 was stored in aliquots at -80 °C until use.

PLK4 activity was measured using an indirect ELISA detection system. Dephosphorylated GST-PLK4 (4 nM) in a 96-well microtiter plate pre-coated with MBP (Millipore Cat. No. 30-011) in 15 μM ATP (Sigma Catalog No. A7699), 50 mM HEPES-Na ²⁺ pH 7.4, 10 mM Incubation in the presence of MgCl ₂ , 0.01% Brij 35 (Sigma Cat. No. 03-3170). The reaction was allowed to occur for 30 minutes, then the plate was washed 5 times with wash buffer (50 mM TRIS-Cl pH 7.4 and 0.2% Tween 20) and incubated with a 1:3000 dilution of primary antibody (Cell Signaling Cat. No. 9381). minute. The plate was washed 5 times with wash buffer, incubated for 30 minutes in the presence of a secondary antibody (BioRad Cat. No. 1721019, 1:3000 concentration) conjugated to horseradish peroxidase, washed 5 more times with wash buffer and at TMB Incubation in the presence of the substrate (Sigma catalog number T0440). The colorimetric reaction was allowed to continue for 5 minutes, followed by the addition of a stop solution (0.5 N sulfuric acid) and detection at 450 nm by using a monochromator or filter based disk reader (corresponding to Molecular Devices M5 or Beckman DTX880) To quantify.

Compound inhibition was determined at a fixed concentration (10 [mu]M) or a variable inhibitor concentration (typically 50 [mu]M to 0.1 [mu]M in a 10 point dose reaction titration). Compounds were pre-incubated for 15 minutes in the presence of enzyme, followed by the addition of ATP and the remaining activity was quantified using the activity assay described above. The % inhibition of the compound was determined using the following formula: % inhibition = 100 x (1 - (experimental value - background value) / (high activity control - background value)). IC ₅₀ values were determined using the formula (A + (B + (1 + ((x / C) ^ D))))) using a nonlinear 4-point log-curve fit (XLfit4, IDBS), where A = background value, B = range , C = inflection point, D = curve fitting parameter.

Certain compounds of the present invention exhibit IC ₅₀ values of less than 0.05μM PLK4 inhibiting thereto and which include: A9, A10, A11, A16 , A19, A29, A46, A47, A48, A49, A55 and A56.

Claims (28)

A compound represented by the following structural formula: Or a pharmaceutically acceptable salt thereof, wherein: -NR ¹ R ^{2 is taken} together to form a bridged bicyclic group, which optionally contains an epoxy atom and optionally an alkyl group, a haloalkyl group, a hydroxyl group, an alkoxy group. , halo, hydroxyalkyl or alkoxyalkyl substituted; or -NR ¹ R ² bonded together to form oxetane, tetrahydrofuranyl, tetrahydropyranyl, hydroxyethyl or alkoxy Substituted by a base N and optionally substituted with an alkyl C And R ³ is i) (C ₂ -C ₆ )alkyl optionally substituted by hydroxy, alkoxy or halo; or ii) optionally alkyl, haloalkyl, hydroxy, alkoxy or a halo-substituted cycloalkyl group; and R ⁴ is a phenyl group or a monocyclic heteroaryl group, each of which is optionally substituted with one to three groups represented by R ⁵ ; each R ^{5 is} independently selected from halo, alkyl And haloalkyl, hydroxy, alkoxy, -CN, -NO ₂ , -haloalkoxy and -NR ^a R ^a ; and each R ^{a is} independently hydrogen or alkyl. The compound of claim 1, wherein the compound is represented by the following structural formula: Or a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein the compound is represented by the following structural formula: Or a pharmaceutically acceptable salt thereof. The compound of any one of clauses 1 to 3, wherein the bridged bicyclic group formed by -NR ¹ R ² is a [3.3.1] or [3.2.1] bridged bicyclic group. The compound of any one of claims 1 to 3, wherein -NR ¹ R ² is oxetane, tetrahydrofuranyl, tetrahydropyranyl, hydroxyethyl or alkoxy Substituted by a base N and optionally substituted with an alkyl C base. The compound of any one of claims 1 to 3, wherein R ⁴ is phenyl, pyridyl, pyrimidinyl or thienyl, each of which is optionally substituted by one to three groups represented by R ⁵ . The compound of any one of claims 1 to 4 or 6, wherein -NR ¹ R ² is: The compound of any one of clauses 1 to 7 wherein R ³ is cyclopropyl, isopropyl, cyclopentyl, isobutyl, cyclopropylmethyl or 2,2-dimethyl Propyl-1-yl. A compound according to claim 8 wherein R ³ is cyclopropyl, cyclopentyl or isobutyl. The compound of any one of clauses 1 to 9, wherein R ⁴ is pyridin-2-yl or 2-chlorophenyl. The compound of any one of claims 1 to 3 or 6 to 10, wherein -NR ¹ R ² is The compound of any one of clauses 1 to 10 wherein R ⁵ is chlorine or fluorine. A compound represented by the following structural formula: Or a pharmaceutically acceptable salt thereof, wherein: R ³ is i) (C ₂ -C ₆ )alkyl optionally substituted by hydroxy, alkoxy or halo; or ii) optionally alkyl, halo alkyl, hydroxy, alkoxy or halo substituted with the cycloalkyl; and R ⁴ is phenyl or monocyclic heteroaryl, each optionally substituted by R ⁵ represents a group of by one to three; each R ⁵ Independently selected from halo, alkyl, haloalkyl, hydroxy, alkoxy, -CN, -NO ₂ , -haloalkoxy, and -NR ^a R ^a ; and each R ^{a is} independently hydrogen or alkyl . A compound according to claim 13 wherein the compound is represented by the following structural formula: Or a pharmaceutically acceptable salt thereof. A compound according to claim 13 wherein the compound is represented by the following structural formula: Or a pharmaceutically acceptable salt thereof. The compound of any one of clauses 13 to 15, wherein R ⁴ is phenyl, pyridyl, pyrimidinyl or thienyl, each of which is optionally substituted by one to three groups represented by R ⁵ . The compound of any one of claims 13 to 16, wherein R ³ is cyclopropyl, isopropyl, cyclopentyl, isobutyl, cyclopropylmethyl or 2,2-dimethyl Propyl-1-yl. A compound according to claim 17, wherein R ³ is cyclopropyl, cyclopentyl or isobutyl. The compound of any one of claims 13 to 18, wherein R ⁴ is pyridin-2-yl or 2-chlorophenyl. A compound represented by the following structural formula: Or a pharmaceutically acceptable salt thereof. A compound represented by the following structural formula: Or a pharmaceutically acceptable salt thereof. A compound represented by the following structural formula: Or a pharmaceutically acceptable salt thereof. A compound represented by the following structural formula: Or a pharmaceutically acceptable salt thereof. A compound represented by the following structural formula: Or a pharmaceutically acceptable salt thereof. A compound represented by the following structural formula: Or a pharmaceutically acceptable salt thereof. The compound of any one of claims 1 to 3, wherein -NR ¹ R ^{2 is} bonded together to form a bridged bicyclic group, optionally containing an epoxy atom and optionally an alkyl group, a halogen Substituent, hydroxy, alkoxy, halo, hydroxyalkyl or alkoxyalkyl. A pharmaceutical composition comprising a compound according to any one of claims 1 to 26 and a pharmaceutically acceptable carrier or diluent. A method of treating an individual having cancer comprising: administering to the individual an effective amount of a compound according to any one of claims 1 to 26.