Polo-like kinase 1 inhibitors

Abstract

The present invention relates to inhibitors of polo-like kinase 1("PLK1"). In particular, the present invention relates to compounds that selectively inhibit the activity of PLK1, pharmaceutical compositions comprising a therapeutically effective amount of the compounds, and methods of use therefor, such as methods for treating PLK1-mediated diseases and disorders, such as cancer.

Description

POLO-LIKE KINASE 1 INHIBITORSFIELD OF THE INVENTIONThe present invention pertains to inhibitors of polo-like kinase 1 (" PLK1"). Specifically, the present invention concerns compounds that selectively inhibit the activity of PLK1, pharmaceutical compositions comprising a therapeutically effective amount of said compounds, and methods of application thereof, such as methodologies for treating PLKl-mediated diseases and disorders, including cancer.BACKGROUND OF THE INVENTION

[0001] TP53 is the most frequently mutated and inactivated gene in cancer, resulting in decreased levels of functional p53. Since existing approaches designed to target p53 directly have shown limited clinical success, there remains a high unmet medical need for new therapeutics to address cancers driven by changes in p53 biology

[0002] The TP53 gene, which encodes the p53 protein, is a critical tumor suppressor that plays a vital role in maintaining genomic stability and preventing cancer development. Its significance in cancer biology has been demonstrated through several studies.

[0003] Cell cycle regulation: p53 functions as a "guardian of the genome" by inhibiting cell division when DNA damage is detected. For instance, in response to UV radiation-induced DNA damage, p53 activates the expression of p21. This protein inhibits cyclin-dependent kinases and arrests the cell cycle, allowing time for DNA repair.

[0004] Apoptosis induction: When DNA damage is irreparable, p53 initiates programmed cell death (apoptosis) to eliminate potentially cancerous cells. In severe DNA damage caused by chemotherapeutic agents such as cisplatin, p53 upregulates pro-apoptotic genes, including BAX and PUMA, resulting in cell death and preventing the propagation of damaged cells.

[0005] Senescence promotion: p53 induces cellular senescence, a permanent cell cycle arrest state, in response to oncogenic stress. For example, when the RAS oncogene is activated, p53 initiates senescence to prevent uncontrolled cell proliferation as a tumor formation barrier.

[0006] Metabolic regulation: p53 influences cellular metabolism, often altered in cancer cells. It can inhibit glycolysis and promote oxidative phosphorylation, counteracting the Warburg effectobserved in many cancers. This metabolic control contributes to the maintenance of normal cellular functions and energy balance.

[0007] Angiogenesis inhibition: p53 suppresses tumor angiogenesis by downregulating pro- angiogenic factors such as VEGF and upregulating anti-angiogenic factors, including thrombospondin- 1. This function limits the blood supply to tumors, restricting their growth and potential for metastasis.

[0008] DNA repair coordination p53 facilitates DNA repair by regulating genes involved in various repair pathways. For instance, it upregulates XPC and DDB2, which are crucial for nucleotide excision repair and contribute to the maintenance of genomic integrity.

[0009] Stem cell regulation by p53 plays a role in maintaining stem cell populations and preventing their malignant transformation. In hematopoietic stein cells, p53 regulates self-renewal and differentiation, preventing the accumulation of mutations that could lead to leukemia.

[0010] The high frequency of TP53 mutations in cancer underscores its importance as a tumor suppressor. For example, in ovarian cancer, TP53 mutations are present in over 90% of cases, contributing to chemoresistance and poor prognosis.

[0011] In Li-Fraumeni syndrome, inherited TP53 mutations significantly increase the risk of developing multiple types of cancer throughout life, highlighting the crucial role of this gene in cancer prevention.

[0012] In lung cancer, TP53 mutations are associated with increased genomic instability and metastatic potential, demonstrating their importance in maintaining cellular homeostasis.

[0013] Given the central role of p53 in cancer biology, developing effective strategies to restore or mimic its function in tumors with small-molecule therapeutic intervention makes targeting p53 activity viaPLKl a valuable endeavor.SUMMARY OF THE INVENTION

[0014] The compounds of Formula (I) are as follows:Formula (I)wherein R1, R2, R3, R6, and R7are as defined herein, and pharmaceutically acceptable salts thereof. Also provided are pharmaceutical compositions comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient or elixir.

[0015] Provided are methods for treating PLK1 -mediated diseases or disorders in a subject in need thereof, comprising administering a therapeutically effective amount of a compound of formula (I), a pharmaceutical salt thereof, or a pharmaceutical composition thereof.

[0016] Also provided herein is a use of a compound of Formula (I), or a pharmaceutically acceptable salt or thereof, as defined herein in the manufacture of a medication for the inhibition ofPLKl activity.

[0017] Also provided herein is the use of a compound of Formula (I), or a pharmaceutically acceptable salt or solvate thereof, as defined herein, in the manufacture of a medication for the treatment of a PLKl-mediated disease or disorder.DETAILED DESCRIPTION OF THE INVENTION

[0018] The present invention relates to inhibitors of polo-like kinase 1 (‘TLK1”). These novel PLK1 inhibitors offer a promising approach for targeted cancer therapy by selectively disruptingcell cycle progression in malignant cells. Preclinical studies have demonstrated their potent antiproliferative effects across various cancer cell lines, with an exceptionally high efficacy observed in aggressive tumor types. Further research is ongoing to elucidate the full spectrum of their therapeutic potential and optimize their pharmacokinetic properties for clinical development. In particular, the present invention relates to compounds that covalently inhibit the activity of PLK1, pharmaceutical compositions comprising a therapeutically effective amount of the compounds, and methods of use, such as methods for treating plkl-mediated diseases and disorders, such as cancer.PLK1

[0019] Polo-like Kinase 1 (PLK1), a member of the polo subfamily of serine / threonine protein kinases, is vital for various cellular functions, including regulation of cell cycle progression and DNA damage response (DDR). PLK1 overexpression, which undermines cellular checkpoints, thereby resulting in heightened cell proliferation, has been observed across a large spectrum of human cancers, solid tumors, and hematologic cancers and is associated with poor clinical prognosis and resistance to chemotherapy. In contrast, PLK1 depletion or inhibition leads to mitotic arrest and DNA damage, often accompanied by cell death, and enhances the sensitivity of cancer cell viability to irradiation. The critical role of PLK1 in cancer is widely accepted to be unassailable.

[0020] As a component of the DDR, PLK1 plays a role in the oscillatory behavior in the activity of p53, a regulator tumor suppressor protein essential for DNA stability. It is known as the guardian of the genome. The kinase activity of PLK1 promotes proteasomal degradation of p53 through the E3 ligases TOPORS and MDM2, with PLK1 depletion or inhibition resulting in elevated cellular levels of p53. In addition, active PLK1 can physically bind to and inhibit p53 activity to transactivate the transcription of tumor suppressor genes. Polo-like kinase 3 (PLK3), another member of the polo subfamily of kinases, plays a role opposite to that of PLK1 in p53 activity; PLK3-mediated phosphorylation of p53 increases its transactivation function. The implications of this research are significant for understanding the complex regulatory mechanisms of p53, a crucial tumor suppressor protein. The opposing roles of PLK1 and PLK3 in modulating p53 activity suggest a delicate balance between the cellular responses to DNA damage. This knowledge could potentially lead to new therapeutic strategies for cancer treatment, mainlytargeting the PLKl-p53 axis. Further investigation into the interplay between these kinases and p53 may reveal novel approaches for enhancing p53 function in cancer cells or manipulating the DNA damage response pathway for therapeutic benefits.

[0021] While pan-PLK inhibitors can have off-setting effects on p53 function, PLKl-specific inhibitors, the PLKl-specific inhibitors of the present disclosure, can selectively increase p53 levels and activity. TP53 is mutated in -50% of human cancers, more than any other gene, and mutations in TP53 are among the most common emergent alterations in refractory cancers ( Hsiehchen et al., (2022) Nat Comm. 13: 7477). Since no effective p53-based therapeutic has been successfully translated into clinical treatment, the PLKl-specific inhibitors of the present disclosure may lead to therapeutic benefits by raising p53 levels in cancer cells, including but not limited to p53-defective or p53-altered.DEFINITIONS

[0022] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one skilled in the art to which this invention belongs. All patents, patent applications, and publications referred to herein are incorporated by reference to the extent that they are consistent with the present disclosure. Terms and ranges have generally defined definitions unless expressly defined otherwise.

[0023] For simplicity, chemical moieties are defined and referred to as univalent chemical moieties (e.g., alkyl, aryl, etc.). Nevertheless, such terms may also convey the corresponding multivalent moieties under the appropriate structural circumstances apparent to those skilled in the art. All atoms are understood to have their standard number of valences for bond formation (i e., 4 for carbon, 3 for N, 2 for O, and 2, 4, or 6 for S, depending on the oxidation state of the S).

[0024] As used herein, a “PLK1” refers to the mammalian enzyme polo-like kinase 1.

[0025] As used herein, a “PLK1 inhibitor” refers to compounds of the present invention that are represented by Formula (I) as described herein. These compounds can negatively modulate or inhibit all or a portion of the enzymatic activity of PLK1 by forming a covalent adduct between the compound of Formula (I) and PLK1.

[0026] As used herein, the term “hydroxyl” refers to -OH

[0027] As used herein, the term “hydroxyalkyl” refers to -alkylene-OH.

[0028] As used herein, the term “alkoxy” refers to -OC1-C3 alkyl.

[0029] The term "halogen" or "halo" as employed herein refers to chlorine, bromine, fluorine, or iodine.

[0030] The term "alkyl," as employed herein, refers to straight and branched chain aliphatic groups having from 1 to 12 carbon atoms. As such, “alkyl” encompasses C1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11and C12groups. Examples of alkyl groups include, without limitation, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl.

[0031] The term "alkylene" group is an alkyl group, as defined hereinabove, that is positioned between and serves to connect two other chemical groups. Examples of alkylene groups include, without limitation, methylene, ethylene, propylene, and butylene.

[0032] The term "alkynyl," as employed herein, refers to straight and branched chain aliphatic groups comprising at least one carbon-carbon triple bond with 2 to 6 carbon atoms. As such, “alkynyl” encompasses C2, C3, C4, C5, and C6groups. Examples of alkynyl groups include, without limitation, ethinyl, propynyl, butynyl, pentynyl and hexynyl.

[0033] The term “haloalkyl” refers to an alkyl chain in which one or more hydrogens have been replaced by a halogen. Exemplary haloalkyls are trifluoromethyl, difluoromethyl,fl urochloromethyl, and fluoromethyl.

[0034] The term "cycloalkyl," employed herein, is a saturated and partially unsaturated cyclic hydrocarbon group with 3 to 12 carbons. As such, “cycloalkyl” includes C3, C4, C5, C6, C7, C8, C9, C10, C11and C12cyclic hydrocarbon groups. Examples of cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.

[0035] An "aryl" group is a C6-C14 aromatic moiety comprising one to three aromatic rings. As such, “aryl” includes C6, CIO, C13, and C14 cyclic hydrocarbon groups. An exemplary arylgroup is a C6-C10 aryl group. Particular aryl groups include, without limitation, phenyl, naphthyl, anthracenyl, and fluorenyl. An “aryl” group also includes fused multicyclic (e g., bicyclic) ring systems in which one or more of the fused rings is non-aromatic, provided that at least one ring is aromatic, such as indenyl.

[0036] A "heterocyclyl" or "heterocyclic" group is a mono- or bicyclic (fused or spiro) ring structure having from 3 to 12 atoms (3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 atoms), for example 4 to 8 atoms, wherein one or more ring atoms are independently -C(O)-, N, NRA, O, or S, and the remainder of the ring atoms are quaternary or carbonyl carbons. Examples of heterocyclic groups include, without limitation, epoxy, oxiranyl, oxetanyl, azetidinyl, aziridinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, piperazinyl, imidazolidinyl, thiazolidinyl, thiatanyl, dithianyl, trithianyl, azathianyl, oxathianyl, dioxolanyl, oxazolidinyl, oxazolidinonyl, decahydroquinolinyl, piperidonyl, 4-piperidonyl, 1,4-diazapanyl, 1 -methyl- 1,4-diazapanyl, l,4-dimethyl-l,4-di(ll-oxidaneyl)-114,414-piperazinyl, thiomorpholinyl, dimethyl-morpholinyl, and morpholinyl. Specifically excluded from the scope of this term are compounds having adjacent ring O and / or S atoms.

[0037] As used herein, “L-heterocyclyl” refers to a heterocyclyl group covalently linked to another group via an alkylene linker.

[0038] In certain instances, formulas of the inventive compounds may include certain notations. These notations generally refer to various isomeric forms of the depicted compounds and intend to mean the following:“&1” -racemic chiral center 1;“&2” -racemic chiral center 2;“&3” - racemic chiral center 3;“Abs” - absolute chiral center;“Or 1” -chiral center 1 is either R or S but not racemic;“Or 2” -chiral center 2 is either R or S but not racemic;“Or 3” -chiral center 3 is either R or S but not racemic.

[0039] As used herein, “an effective amount” of a compound is an amount that is sufficient to modulate or inhibit the activity of PLK1 negatively.

[0040] As used herein, a “therapeutically effective amount” of a compound is an amount that is sufficient to ameliorate or, in some manner, reduce a symptom or stop or reverse the progression of a condition, e.g., cancer, or negatively modulate or inhibit the activity of PLK1. Such amount may be administered as a single dosage or may be administered according to a regimen, whereby it is effective.

[0041] As used herein, “treatment” means any manner in which the symptoms or pathology of a condition, disorder, or disease in a patient are ameliorated or otherwise beneficially altered.

[0042] As used herein, amelioration of the symptoms of a particular disorder by administration of a particular compound or pharmaceutical composition refers to any lessening, whether permanent or temporary', lasting or transient, that can be attributed to or associated with the administration of the composition.COMPOUNDS

[0043] In certain aspects of the present disclosure, provided is a compound of Formula (I):Formula (I)wherein:R1is hydrogen, halogen, hydroxy, aniline, C1-C6 alkyl, cycloalkyl, and haloalkyl;R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more alkoxy groups;R3is hydrogen, heterocyclyl optionally substituted with one or more R4, -NH-L1- N(RARB), -NH-L2-cycloalkyl or -NH-L2-heterocyclyl, wherein the cycloalkyl or the heterocyclyl is optionally substituted with one or more R5;R6is OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3;R7is NH2, NHCH3, or CH3;each R4is independently C1-C4 alkyl, cyclopropyl, hydroxyalkyl,?-N(RARB), halogen, O, O-CH3, NH2, -C(O)O-t-butyl; cycloalkyl optionally substituted with NH2, -L3-aryl; or -L4-heterocyclyl;each R5is independently C1-C4 alkyl, N(CH3)(CH3), L2-aryl, or -L2-heterocyclyl, wherein L2-heterocyclyl is optionally substituted with one or more C1-C4 alkyl, C1-C4 alkylcycloalkyl, C(0)CH3 or aryl;each L1is C1-C4 alkylene;each L2is a bond or C1-C4 alkylene;each L3is -CH2-O-CH2-;each L4is a bond or methylene;each RAis hydrogen or C1-C3 alkyl;each RBis hydrogen or C1-C3 alkyl;and pharmaceutically acceptable salts thereof.

[0044] In certain aspects for compounds of Formula (I), R1is halogen. In one embodiment, the halogen is chlorine, fluorine, or bromine.

[0045] In certain aspects for compounds of Formula (I), R1is hydroxy.

[0046] In certain aspects for compounds of Formula (1), R1is C1-C6 alkyl. In one embodiment, the C1-C6 is alkyl methyl, ethyl, or isopropyl.

[0047] In certain aspects for compounds of Formula (I), R1is cycloalkyl. In one embodiment, the cycloalkyl is cyclopropyl.

[0048] In certain aspects for compounds of Formula (I), R1is haloalkyl. In one embodiment, the haloalkyl is fluoromethyl, difluoromethyl, or trifluoromethyl.

[0049] In certain aspects for compounds of Formula (I), R1is aniline. In one embodiment, the aniline is NH2.

[0050] In certain aspects for compounds of Formula (I), R2is alkoxy. In one embodiment, the alkoxy is methoxy or propoxy.

[0051] In certain aspects for compounds of Formula (I), R2is -O-haloalkyl. In one embodiment, the haloalkyl is difluoromethyl.

[0052] In certain aspects for compounds of Formula (I), R3is a heterocyclyl optionally substituted with one or more R4.

[0053] In one embodiment, the heterocyclyl is 1,4-diazapanyl, 1-m ethyl- 1,4-diazapanyl or 1,4-dimethyl- 1,4-di(l 1 -oxidaneyl)-114,414-piperazinyl.

[0054] In another embodiment, the heterocyclyl is pyrrolidinyl, piperidinyl, or piperazinyl, each optionally substituted with one or more R4, wherein R4is C1-C4 alkyl, hydroxyalkyl, -L1-N(RARB), -C(O)O-t-butyl, -L3-aryl, or -L4-heterocyclyl.

[0055] In one embodiment, the heterocyclyl is pyrrolidinyl substituted with one R4, and R4is L4-heterocyclyl, wherein L4is methylene, and the heterocyclyl is pyrrolidinyl.

[0056] In another embodiment, the heterocyclyl is piperidinyl substituted with one R4, and R4is L4-heterocyclyl, wherein L4is a bond, and the heterocyclyl is pyrrolidinyl. In one embodiment, the heterocyclyl is piperazinyl and R4is C1-C4 alkyl, hydroxyalkyl, -L'-NfR^R15), -C(O)O-t-butyl; or -L3-aryl.

[0057] In one embodiment, the piperazinyl is substituted with one R4, and R4is C1-C4 alkyl, wherein the C1-C4 alkyl is methyl or ethyl.

[0058] In one embodiment, the piperazinyl is substituted with one R4, and R4is -L1-N(RARB).

[0059] In one embodiment, the piperazinyl is substituted with one R4, and R4is -C(O)O-t-butyl.

[0060] In another embodiment, the piperazinyl is substituted with two R4groups, wherein one R4group is C1-C4 alkyl, and the second R4group is hydroxyalkyl or -L3-aryl.

[0061] In certain aspects for compounds of Formula (I), R3is -NH-L'-N(R RI!.

[0062] In one embodiment, L1is propylene, and RAand RBare each methyl.

[0063] In one embodiment, L1is propylene, and RAand RBare each ethyl.

[0064] In certain aspects for compounds of Formula (I), R3is -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R\

[0065] In one embodiment, L2is a bond, and the heterocyclyl is pyrrolidinyl optionally substituted with one or more R5.

[0066] In one embodiment, the pyrrolidinyl is substituted with one R\ wherein R3is ethyl or isopropyl. In another embodiment, L2is a C1-C4 alkylene, and the heterocyclyl is pyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl, each optionally substituted with one or more R5.

[0067] In one embodiment, L2is ethylene or propylene.

[0068] In one embodiment, the heterocyclyl is piperazinyl optionally substituted with one or more R3, wherein R5is methyl.

[0069] In another embodiment, L2is dimethylethylene, and the heterocyclyl is pyrrolidinyl, piperidinyl, or morpholinyl.

[0070] In another embodiment, L2is dimethylethylene, and the heterocyclyl is piperazinyl optionally substituted with one or more R5, wherein R5is methyl.In certain aspects for compounds of Formula (I), the compound of Formula (I) is selected from the group consisting of:Fpharmaceutically acceptable salts thereof.

[0071] The compounds of Formula (I) and pharmaceutically acceptable salts thereof may be formulated into pharmaceutical compositions.

[0072] The compounds disclosed herein include all pharmaceutically acceptable isotopically-labeled compounds, in which one or more atoms of the compounds disclosed herein are replaced by atoms having the same atomic number and an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can beincorporated into the compounds disclosed herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, chlorine, and iodine, such as2H,3H,nC,13C,14C,13N,15N,15O,17O,31P,32P,33S,18F,36C1,123I, and123I. Such radiolabelled compounds could help determine or measure the effectiveness of the compounds, for example, by characterizing binding affinities and kinetics of binding to PLK1 and other modes of action. Certain compounds of the disclosure, when labeled with radioactive isotopes, such as tritium,3H, and carbon-14,14C, could also help characterize drug distribution and in vivo tissue expression of PLK1.

[0073] When labeled with heavier isotopes such as deuterium,2H, compounds disclosed herein may exhibit therapeutic advantages such as greater metabolic stability, resulting in longer in-vivo half-lives, which in turn could reduce the therapeutic dosing requirements.

[0074] In order to better characterize substrate receptor occupancy or tissue expression in live specimens statically or longitudinally, replacing atoms of compounds disclosed herein with positron emitting isotopes, such asnC,14F,15O, and13N, may allow for enabling Positron Emission Tomography (PET) studies.

[0075] Isotopically-labeled compounds, as disclosed herein, can be prepared by conventional techniques known to those skilled in the art or by processes that are analogous to those described in the associated examples and schemes using an appropriate isotopically-labeled reagent in place of the non-labeled reagent previously employed.

[0076] Some of the compounds disclosed herein may exist as stereoisomers. The compounds disclosed herein include all stereoisomers, both as pure individual stereoisomer preparations as well as preparations enriched of each, and both the racemic mixtures of such stereoisomers and the individual diastereomers and enantiomers that may be separated according to methods known to those skilled in the art. Additionally, the compounds disclosed herein include all individual tautomeric states of the compounds and mixtures thereof.

[0077] To emphasize, unless explicitly indicated otherwise, the compounds of the present invention include all stereoisomers, including enantiomers, diastereomers, and racemates, as well as all tautomeric forms thereof. The present invention also encompasses all pharmaceutically acceptable salts, solvates, hydrates, polymorphs, and isotopically labeled derivatives (e.g., suchas those enriched with deuterium or other stable or radioactive isotopes) of the disclosed compoundsPHARMACEUTICAL COMPOSITIONS

[0078] In another aspect, the invention provides pharmaceutical compositions comprising a PLK1 inhibitor according to the invention and a pharmaceutically acceptable earner, excipient, or diluent. Compounds of the invention may be formulated by any method well-known in the art. They may be prepared for administration by any route, including, without limitation, parenteral, oral, sublingual, transdermal, topical, subcutaneous, intranasal, intratracheal, or intrarectal. In certain embodiments, compounds of the invention are administered intravenously in a hospital setting. In certain other embodiments, administration may preferably be done by oral route.

[0079] The characteristics of the carrier depend on the route of administration. As used herei n, the term “pharmaceutically acceptable” means a non-toxic material compatible with a biological system such as a cell, cell culture, tissue, or organism, and that does not interfere with the effectiveness of the biological activity of the active ingredient(s). Thus, compositions according to the invention may contain, in addition to the inhibitor, diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials well-known in the art. The preparation of pharmaceutically acceptable formulations is described in, e.g., Remington’s Pharmaceutical Sciences, 18thEdition, ed. A. Gennaro, Mack Publishing Co., Easton, Pa., 1990.

[0080] As used herein, the term pharmaceutically acceptable salts refers to salts that retain the desired biological activity of the above-identified compounds and exhibit minimal or no undesired toxicological effects. Examples of such salts include but are not limited to acid addition salts formed with inorganic acids (for example, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like), and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, naphthalenedi sulfonic acid, and polygalacturonic acid. The compounds can also be administered as pharmaceutically acceptable quaternary salts known by those skilled in the art, which specifically include the quaternary ammonium salt of the Formula NRhZ', wherein R ishydrogen, alkyl, or benzyl, and Z is a counterion, including chloride, bromide, iodide, O-alkyl, toluenesulfonate, methyl sulfonate, sulfonate, phosphate, or carboxylate (such as benzoate, succinate, acetate, glycolate, maleate, malate, citrate, tartrate, ascorbate, benzoate, cinnamoate, mandeloate, benzyloate, and diphenylacetate).

[0081] The active compound is included in the pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver to a patient a therapeutically effective amount without causing serious toxic effects in the patient treated. A dose of the active compound for all of the above-mentioned conditions ranges from about 0.01 to 300 rag / kg, preferably 0.1 to 100 rag / kg per day, more generally 0.5 to about 25 mg per kilogram body weight of the recipient per day.

[0082] A typical topical dosage will range from 0.01-3 % wt / wt in a suitable carrier. The effective dosage range of the pharmaceutically acceptable derivatives can be calculated based on the weight of the parent compound to be delivered. If the derivative exhibits activity in itself, the effective dosage can be estimated as above using the weight of the derivative or by other means known to those skilled in the art.

[0083] The pharmaceutical compositions comprising compounds of the present invention may be used in the methods described herein.METHODS OF USE

[0084] In yet another aspect, the invention provides methods for inhibiting PLK1 activity in a cell, comprising contacting the cell in which inhibition of PLK1 activity is desired with a therapeutically effective amount of a compound of Formula (I), pharmaceutically acceptable salts thereof or pharmaceutical compositions containing the compound or pharmaceutically acceptable salt thereof.

[0085] The compositions and methods provided herein are deemed particularly useful for inhibiting PLK1 activity in a cell, including cells that overexpress PLK1. In one embodiment, a cell in which inhibition of PLK1 activity is desired is contacted with an adequate amount of a compound of Formula (I) to modulate the activity of a PLK1 kinase negatively. In other embodiments, a therapeutically effective amount of pharmaceutically acceptable salt or pharmaceutical compositions containing the compound of Formula (I) may be used. In certainembodiments, contacting the cell with an adequate amount or a therapeutically effective amount of a compound of Formula (I) occurs in vivo. In certain embodiments, contacting the cell with an adequate amount or a therapeutically effective amount of a compound of Formula (I) occurs in vitro. In one embodiment, the therapeutically effective amount of Formula (I) compound is between about 0.01 to 300 mg / kg per day. In one embodiment, the therapeutically effective amount of the compound of Formula (I) is between about 0.1 to 100 mg / kg per day.

[0086] In one embodiment, methods for treating a patient having PLKl-mediated disease or disorder, comprising administering to the patient a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, alone or combined with a pharmaceutically acceptable carrier, excipient or diluents are provided.

[0087] In one embodiment, the PLKl-mediated disease or disorder results from overexpression of PLK1. In one embodiment, the PLKl-mediated disease or disorder is cancer. In one embodiment, the PLKl-mediated cancer is a TP 53 - or Ras-mutated cancer. In one embodiment, PLKl-mediated cancer is melanoma, non-small cell lung cancer (NSCLC), carcinomas of the head and neck, esophageal, pharynx, breast, liver, endometrium, colorectum, ovary, pancreas or prostate.

[0088] By negatively modulating the activity of PLK1, particularly in cases of cells overexpressing the PLK1 enzyme, the methods are designed to modulate the activity of PLK1, in some embodiments, to treat certain PLKl-mediated diseases or disorders, such as cancer. The cells / patient may be contacted in a single dose or multiple doses in accordance with a particular treatment regimen to induce the desired negative modulation of PLK1. For example, hematological or solid cancers may be monitored using well-known methods, including blood draws, X-rays, or other imaging techniques, such as CT scans or MRI scans, to assess the effectiveness of treatment, and dosages may be adjusted accordingly by the attending medical practitioner.

[0089] The concentration and route of administration to the patient may vary' depending on the severity of the disease. The compounds, pharmaceutically acceptable salts thereof, and pharmaceutical compositions comprising such compounds and salts also may be co-administered with other compounds or used in combination with other treatments, such as surgicalintervention, either as an adjuvant prior to surgery or post-operatively The degree of PLK1 inhibition may be monitored in the patient using well-known assay methods, including those in Example A, to assess the effectiveness of treatment.GENERAL REACTION SCHEMES AND EXAMPLES

[0090] The compounds of the present invention may be prepared using commercially available reagents using the synthetic methods and reaction schemes described herein or using other reagents and conventional methods well known to those skilled in the art.

[0091] For instance, intermediates for preparing compounds and compounds of Formula (I) of the present invention may be prepared according to General Reaction Schemes I - XIII:GENERAL REACTION SCHEME ISuzuki Oxidative Olefin Cleavage

[0092] Compounds of Formula (I) wherein R1is hydrogen, halogen, hydroxy, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more R4, and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R3may be prepared according to General Reaction Scheme I. Compound 7 is an example of Formula (I) wherein R1and R2is H and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclylis optionally substituted with one or more R?and R6is OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3. A dihalopyridopyrimidine 1 is treated with an aniline 2, for example, under SₙAr conditions at elevated temperature, to form pyridopyrimidyl aniline 3, which is subjected to Pd catalyzed cross-coupling conditions, for example, Heck conditions with pinacolvinylboronate to afford the anilinopyridopyrimidyl styrene 4. Pyridopyrimidine styrene 4 is subjected to oxidative cleavage conditions, for example, osmate and permanganate, to furnish anilinopyridopyrimidine aldehyde 5. Aldehyde 5 was then subjected to olefination conditions, for example, with a phosphorylsulfonamidyl ylide to generate vinyl sulfonamide 6, which is then subjected to acidic deprotection conditions, for example, trifluoroacetic acid to provide the desired compound 7 of Formula (I).GENERAL REACTION SCHEME II

[0093] Compounds of Formula (I) wherein R1is hydrogen, halogen, hydroxy, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more R4, and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5may be prepared according to General Reaction Scheme II. Compound 7 is anexample of Formula (T) wherein R1and R2are H and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5and R6is OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3. 1 is reacted with 8 in S\Ar conditions in the presence of Bronstead-Lowry acid, for example, TFAA, to provide halide 9, which was reacted with a vinylboronic ester under Suzuki conditions to provide olefin 10. The carbon-carbon double bond of olefin 10 was subjected to oxidative cleavage conditions, for example, with Osmate and Magnesium Oxide to give aldehyde 11. Aldehyde 11 was subjected to Wittig conditions, for example, with a phosphorous ylide and base to generate vinyl sulfonamide 12, which was deprotected with an acid such as HCl(aq) or TFAA to provide free-amine 13, which was alkylated via reductive amination with aldehyde or ketone to give the desired product 7 of Formula (I).General Reaction Scheme III

[0094] Compounds of Formula (I) wherein R1is hydrogen, halogen, hydroxy, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more R4, and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one ormore R5may be prepared according to General Reaction Scheme TIT. Compound 7 is an example of Formula (I) wherein R1and R2are H and R3is heterocyclyl optionallysubstituted with one or more R4, -NH-LT-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5and R6is OCF3, SCH3,cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3. Dihalo-aza- quinazoline 1 was reacted with diaminobenzene 14 under SₙAr conditions in the presence of a Bronstead-Lowry acid, such as TFAA, to provide halide 15, which was coupled with avinyl boronic acid pinacol ester to generate olefin 16. The carbon-carbon double bond of olefin is oxidatively cleaved to aldehyde 17, for example, with osmate and metal oxide.Aldehyde 17 was exposed to Wittig conditions, for example, with a phosphorous ylide to generate vinyl Boc sulfonamide 18, which was immediately deprotected with protic acid to vinyl sulfonamide aniline 19, which was exposed to reductive amination conditions, such as carbonyl reactant and borohydride to grant 7 of Formula (I).General Reaction Scheme IVCompounds of Formula (I) wherein R1is hydrogen, halogen, hydroxy, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more R4, and R3is heterocyclyl optionally substituted with one or more R4, -NH-Ll-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionallysubstituted with one or more R5may be prepared according to General Reaction Scheme TV.Compound 7 is an example of Formula (I) wherein R1and R2are H and R3is heterocyclyl optionally substituted with one or more R4, -NH-Ll-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5and R6is OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3. Weinreb amide 9 underwent S\ Ar substitution with aniline 8 in the presence of Bronstead-Lowry acid, for example, TFAA, to synthesize Boc-protected 20, which was deprotected with an acid, for example, HC1, generating free amine 21, which was alkylated via reductive amination, for instance with alkyl carbonyl and borohydride, to provide 22. Weinreb amide was reduced, for example, with DiBAL-H to give aldehyde 23, which was reacted with phosphorous ylide to bequeath Boc vinyl sulfonamide 24, which was deprotected with acid, for example, HC1 to provide desired product 7 of Formula (I).General Reaction Scheme V

[0096] Compounds of Formula (I) wherein R1is hydrogen, halogen, hydroxy, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more R4, and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5may be prepared according to General Reaction Scheme V. Compound 7 is an example of Formula (I) wherein R1and R2is H and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5and R6is OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3. Dihalopyridopyrimidine 9 wasreacted under SNAr conditions with Boc piprizylaniline 25, for example, in a polar aprotic solvent in the presence of a Bronstead-Lowry acid to impart TFA anilinopyridopyrimidyl halide 26, which underwent Pd-catalyzed cross-coupling conditions, for example, Heck conditions with pinacolvinylboronate to afford TFA anilinopyridopyrimidyl styrene 26. Olefin cleavage conditions, such as osmate and permanganate, were imposed upon styrene 27 to generate TFA anilinopyridopyrimidyl aldehyde 27. Subsequent Wittig conditions, for example, with a phosphorylsulfonamidyl ylide to generate vinyl sulfonamide 24, were then subjected to acidic deprotection conditions, for example, trifluoroacetic acid to provide the desired compound 7 of Formula (I).25 SNAr-Core Oxidative OlefinCleavage

[0097] Compounds of Formula (I) wherein R1is hydrogen, halogen, hydroxy, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more R4, and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5may be prepared according to General Reaction Scheme VI. Compound 7 is an example of Formula (I) wherein R1and R2are H and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5and R6is OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3. Fuseddichlorobicycle 1 was reacted with aniline 25 under SₙAr conditions, for example, in the presence of TFAA and a polar aprotic solvent to generate coupled product 28, which underwent Suzuki coupling with vinylboronic pinacol ester to provide olefin 29. Olefin 29 was exposed to oxidative cleavage conditions, for example, Osmate and Magnesium Oxide to generate olefin 30, which was olefinated with, for example, phosphorous ylide to generate Boc vinylsulfonamide 31, which was deprotected with acid, for example, TFAA to generate desired product 7 of Formula (I).General Reaction Scheme VII

[0098] Compounds of Formula (I) wherein R1is hydrogen, halogen, hydroxy, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more R4, and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5may be prepared according to General Reaction Scheme VII. Compound 7 is an example of Formula (I) wherein R1and R2are H and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5and R6is OCF3, SCH3,cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3. Weinreb amide 9 was coupled to aniline 32 under SNAC conditions for example with polar protic solvent, for example, t-BuOH to generate coupled Weinreb amide 33 which was reduced under hydride conditions for example with DiBAL-H to furnish aldehyde 34 which was olefinated under Wittig conditions for example with a phosphorous sulfonamide ylide to provide vinylsulfonamide 35. Reductive amination of free amine 35 provides protected vinyl sulfonamide 36, which was deprotected with Bronsted-Lowrey acid, for example, HC1, to generate the desired product 7 of Formula (I).General Reaction Scheme VIIISNAr— F

[0099] Compounds of Formula (I) wherein R1is hydrogen, halogen, hydroxy, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more R4, and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5may be prepared according to General Reaction Scheme VIII. Compound 7 is an example of Formula (I) wherein R1and R2are H and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5and R6is OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3. Protected chloropyridino Boc sulfonamide 36 was subjected to SₙAr conditions, for example, with KF, DABCO, and DMSO, to furnish fluoropyridyl 37, which was deprotected with acid, for example, HC1 or TFAA, to generate the desired product 7 of Formula (I).General Reaction Scheme IXX XReductive Amination Boc Deprotection

[0100] Compounds of Formula (I) wherein R1is hydrogen, halogen, hydroxy, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more R4, and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5may be prepared according to General Reaction Scheme IX. Compound 7 is an example of Formula (I) wherein R1and R2are H and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5and R6is OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3. Free amine 38 underwent reductive amination with an alkyl carbonyl and borohydride to provide Boc-protected vinyl sulfonamide 36, which was deprotected with protic acid, for example, HC1 or TFAA, to generate the desired product 7 of Formula (I).General Reaction Scheme X

[0101] Compounds of Formula (I) wherein R1is hydrogen, halogen, hydroxy, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more R4, and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5may be prepared according to General Reaction Scheme X. Compound 7 is an example of Formula (I) wherein R1and R2is H and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5and R6is OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3. 6,6-Fused bicyclic dichloride 1 was coupled to aniline 10 under SₙAr conditions, for example, with a polar protic solvent such as t-BuOH to provide coupled product 27, which was coupled to vinyl boronic acid ester under Suzuki Coupling conditions, furnishing olefin 28, which was oxidatively cleaved with Osmate and Magnesium to generate olefin 29, which was olefinated with phosphorous ylide to form Boc-protected vinyl sulfonamide 30, which was deprotected to vinylsulfonamide 31, and underwent reductive amination with aldehyde or ketone to give compound 7 of Formula (I).General Reaction Scheme XISNAr-Ccre Suzuki Coupling

[0102] Compounds of Formula (I) wherein R1is hydrogen, halogen, hydroxy, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more R4, and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5may be prepared according to General Reaction Scheme XI. Compound 7 is an example of Formula (I) wherein R1and R2are H and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5and R6is OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3.Dichloropryidopyrimidine 1 was coupled with aniline 32 under SₙAr conditions, for example, with polar protic solvent t-BuOH to furnish coupled biaryl amine 33, which was subjected to Suzuki coupling conditions with vinyl boronic pinacol ester to give olefin 34, which was alkylated via reductive amination with alkyl carbonyl and borohydride to produce olefin 4, which was oxidatively cleaved via Osmate and KMnO4 to generatealdehyde 5. Aldehyde 5 underwent Wittig olefination with a phosphorous ylide to generate Boc-protected olefin 6, which was deprotected to the desired product 7 of Formula (I).General Reaction Scheme XIIOH MeSuzuki Coupling Boc Deprotection

[0103] Compounds of Formula (I) wherein R1is hydrogen, halogen, hydroxy, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more R4, and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5may be prepared according to General Reaction Scheme XII. Compound 7 is an example of Formula (I) wherein R1and R2are H and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5and R6is OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3. Halo-pyrido Boc-protected vinylsulfonamide 26 was subjected to Suzuki Coupling conditions with vinyl boronic pinacol ester to provide Boc-protected vinyl sulfonamide 35, which was deprotected with protic acid, for example, HC1, to generate desired product 7 of Formula (I).General Reaction Scheme XIII

[0104] Compounds of Formula (I) wherein R1is hydrogen, halogen, hydroxy, C1-C6 alkyl, cycloalkyl, haloalkyl, aniline, R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more R4, and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1- N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5may be prepared according to General Reaction Scheme XIII. Compound 7 is an example of Formula (I) wherein R1and R2are H and R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), or -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5and R6is OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3. Dihalo pyridopyrimidine was coupled to aniline 36 under SₙAr conditions, for example, with heat and t-BuOH to generate coupled halopyrodopyrimidine free amine 37, which was immediately alkylated with aldehyde or ketone and borohydride to furnish 3° amine 38. Chloropyridopyrimidine 38 was olefinated under Suzuki conditions with the vinyl boronic ester to provide olefin 39, which underwent oxidative cleavage with Osmate and KMnO4 to give aldehyde 40, which was reacted with phosphorous ylide to furnish vinylsulfonamide 41, which was deprotected with protic acid to give the desired product 7 of Formula (I).INTERMEDIATES

[0105] The following intermediate may be used in the preparation of compounds in Formula (I).INTERMEDIATE ACl O Cl~)— Cl N-C=O THF, rt, 1 h; NH3, POCh, DIPEA, MeOH, rt, 3 h, 88% - Toluene, 42% step 1 step 2A-1 A-2 Pd(PPh3)2CI2, SnBu3H, DCM, 0 °C, 2 h jj 'N step sIntermediate AStep 1: The procedure was quoted from patent WO2020 / 146613. To a solution of ethyl 4-amino-6-chloro-pyridine-3 -carboxylate (14.0 g, 69.8 mmol) in THF (280 mL), 2, 2, 2-trichloroacetylisocyanate (26.3 g, 140 mmol) was added at room temperature. The mixture was stirred for 1 h and concentrated under vacuum. The residue was triturated with petroleum ether (50 mL) to afford a white solid. The solids were then dissolved in MeOH (400 mL), followed by a dropwise addition of methylamino (40% solution in MeOH; 18 mL) at 0 °C. After stirring at room temperature for 3 h, the reaction mixture was concentrated under reduced pressure, and the residue was triturated in ethyl acetate. Then, the mixture was filtered, and the filter cake was collected to give ethyl 7-chloropyrido[4,3-d]pyrimidine-2,4(1H,3H)-dione as a white solid (12.1 g, 88 % yield), m / z (ESI), calcd. for C7H4ClN3O2: 197.0, found: 198.0, [M+H]+.1H NMR (300 MHz, DMSO-d6) δ 8.64 (s, 1H), 6.98 (s, 1H).Step 2: The procedure was quoted from patent W02020 / 146613. 7-chloropyrido[4,3-d]pyrimidine-2,4-diol (2.4 g, 12.1 mmol.) was dissolved in toluene (40mL), phosphorus oxychloride (9.3 g, 60.7 mmol) and diisopropylethylamine (7.8 g, 60.7 mmol) were added sequentially. The mixture was stirred at 110 °C for 2 h, cooled to room temperature, and concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography (C18, column; mobile phase: MeCN and 0.05% FA aqueous solution; gradient: 5% CAN to 100% CAN in 30 min) to obtain 2,4,7-trichloropyrido[4,3-dJpyrimidine as a white solid (1.2 g, 42% yield), m / z (ESI), calcd. for C7H2Cl3N3: 232.9, found: 234, [M+H]+.Step 3: 2,4,6-trichloropyrido[3,4-d]pyrimidine (1.1 g, 4.7 mmol) and Pd(PPh3)2Cl2(340 mg, 0.47 mmol) wereloaded into a round-bottle flask equipped with a stirring bar, then the flask was sealed with a rubber stopper followed by evacuating-N2-backfill round for 3 times. Afterward, DCM (20 mL) was introduced, and the mixture was cooled to 0 °C before adding tributyltin hydride (2.7 g, 9.41 mmol). After stirring at 0 °C for 2 h, saturated potassium fluoride aqueous solution was added, and the mixture was extracted with ethyl acetate (100 mL x 3 times). The combined organic layer was dried over anhydrous MgSO4and concentrated under reduced pressure. The residue was eluted with PE / EA (10 / 1, v / v) from a silica gel column to yield 2,7-dichloropyrido[4,3-d]pyrimidine as a white solid. (400.2 g, 42.5% yield), m / z (ESI), calcd. for CsHsCLNsO: 198.9, found: 200.0, [M+H]+. ’H NMR (400 MHz, DMSO-d6) 59.86 (s, 1H), 9.54 (s, 1H), 8.14 (s, 1H).INTERMEDIATE BSeO2, 1,4-dioxane,NHBoc 100 °C, 24 h, DBU, DCM, 0 °C → 90% crude oil rt, O / N, 45% Step 1 Step 2 7234-25-5POCI3, 100 °C, MeNH(OMe), Me3AI, 2 h. 80% THF, 0 °C, 3h, 50%rStep 3 Step 4B-3 PHRT-010085-5SO2CI2, MeCN / DCM, - 0 °C, 0.5h, - - 94% Step 5B-4 Intermediate BPHRT-010108-11 PHRT-010108-12Step 1: 4-bromo-2-nitro-1-(trifluoromethoxy)benzene (1.0 g, 3.49 mmol) and t-BuONa (674.0 mg, 7.01 mmol) were added to a 10 mL microwave tube at room temperature. tert-Butyl piperazine- 1 -carboxylate (980.1 mg, 5.26 mmol) in toluene (15 mL) was added to the mixture under a nitrogen atmosphere at room temperature. The mixture was stirred for 1 h at 100 °C under a nitrogen atmosphere. The desired product could be detected by LCMS. The mixture wasallowed to cool to room temperature. Water (100 mL) was added to the mixture. The mixture was extracted with EtOAc (100 mL x 3 times). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: EtOAc / PE = 1 / 14, v / v) to afford tert-butyl 4-[3-nitro-4-(trifluoromethoxy)phenyl]piperazine-l-carboxylate (900.0 mg, 62.5% yield) as a yellow oil. m / z (ESI), calcd. for C16H20F3N3O5: 391.1, found: 336.1 [M+H-tBu]+.Step 2: tert-Butyl 4-[3-nitro-4-(trifluoromethoxy)phenyl]piperazine-1-carboxylate (2.7 g, 6.89 mmol), t-BuOH (60 mL), hydrogen chloride (252.1 mg, 1.52 mmol), and Pd / C (2.7 g, 25.37 mmol) were added to a 250 mL round-bottom flask at room temperature. The resulting mixture was stirred for 1.5 h at room temperature under a hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with EtOAc (60 mL). The organic layer was basified to a pH of 8 using NH3·H2O. After separating the phases, the aqueous layer was extracted with EtOAc (200 mL x 3 times). The combined organic extracts were washed with brine (200 mL), dried over anhydrous Na2SO4, and concentrated under a vacuum to afford tert-butyl 4-[3-amino-4-(trifluoromethoxy)phenyl]piperazine-1-carboxylate (2.3 g, 92.3% yield) as a brown solid, m / z (ESI), calcd. for C16H22F3N3O3: 361.2, found: 362.1 [M+H]+.INTERMEDIATE C-lStep 1: Into a 10 mL microwave tube were added 4-bromo-2-nitro-l -(trifluoromethoxy )benzene (1.0 g, 3.49 mmol) and t-BuONa (674.0 mg, 7.01 mmol) at room temperature. Then, the mixture was added to tert-butyl piperazine- 1 -carboxylate (980.1 mg, 5.26 mmol) in toluene (15 mL) under a nitrogen atmosphere at room temperature. The mixture was stirred for 1 h at 100 °C under a nitrogen atmosphere. The desired product could be detected by LCMS. The mixture was allowed to cool down to room temperature. Water (100 mL) was added to the mixture. The mixture wasextracted with EtOAc (100 mL x 3 times). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SC>4, and concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: EtOAc / PE = 1 / 14, v / v) to afford tert-butyl 4-[3-nitro-4-(trifluoromethoxy)phenyl]piperazine-1-carboxylate (900.0 mg, 62.5% yield) as a yellow oil. m / z (ESI) calcd. for C16H20F3N3O5: 391.1, found: 336.1 [M+H-tBu]+.Step 2: Into a 250 mL round-bottom flask were added tert-butyl 4-[3-nitro-4-(trifluoromethoxy)phenyl]piperazine-1-carboxylate (2.7 g, 6.89 mmol), t-BuOH (60 mL), hydrogen chloride (252.1 mg, 1.52 mmol) and Pd / C (2.7 g, 25.37 mmol) at room temperature. The resulting mixture was stirred for 1.5 h at room temperature under a hydrogen atmosphere. The resulting mixture was filtered, and the filter cake was washed with EtOAc (60 mL). The organic layer was basified to pH 8 with NH3·H2O. After separating phases, the aqueous layer was extracted with EtOAc (200 mL x 3 times). The combined organic extracts were washed with brine (200 mL), dried over anhydrous Na2SO4, and concentrated under a vacuum to afford tert-butyl 4-[3-amino-4-(trifluoromethoxy)phenyl]piperazine-1-carboxylate (2.3 g, 92.3% yield) as a brown solid, m / z (ESI) calcd. for C16H22F3N3O3: 361.2, found: 362.1 [M+H]+.The Intermediates C-2 to C-8 shown in Table A were prepared following the teachings of the General Reaction Schemes and the method to prepare INTERMEDIATE C-l.TABLE AT|—0 S?tructure intermediate No. Spectral Data 04°" / _ C-l 362.1 [M+H]+°7 ~\C-2 237.0 [M+H-tBu] +F\A0 NH2.F^00 0 C-3 376.2 [M+H] +U.NAOAH NH,A*, HV C-4 251.1 [M+H] +NH2¥F0 '^X.'NX\ A 0 C-5 376.1 [M+H] +ATAAoLO=o Table A contStructure intermediate No. Spectral Data NHo:A i..OA. NTO C-6 376.2 [M+H] + °'x[\NH?AVS r-OFA, AN.'ASC-7 330.2 [M+H] +NH,C-8 334.2 [M+H] + HINTERMEDIATE D-lH2N N NBoc Ephos Pd, Ephos, CS2CO3, dioxane, TFAA, pyridine, 100 °C " FA. DCM, it. 1 h rt, 1 h Step 1 Step 2 Step 3 BrB2(OH)4DMF, rip 4-bipyridine Step 4INTERMEDIATE D-1Step 1: To a solution of 4-bromo-2-nitro-l-(trifluorom ethoxy )benzene (2.1 g, 7.11 mmol) in 1,4-dioxane (100 mL) were sequentially added tert-butyl 4-(4-aminophenyl)piperazine-1-carboxylate (2.3 g, 8.37 mmol), Ephos (373 mg, 0.71 mmol), Ephos Pd (642.2 mg, 0.71 mmol), and cesium carbonate (6.8 g, 20.91 mmol). Then, the mixture was bubbled with N2 flow for 5 minutes to expel air gas in the flask, after which the mixture was stirred at 100 °C under N2 atmosphere for 16 h. After cooling to room temperature, insoluble solids were removed by filtration, and the filtrate was diluted with water (250 mL) and then extracted with ethyl acetate (3^200 mL). Organic phases were combined and dried over anhydrous MgSO4, followed by concentration under reduced pressure. The resultant residue was purified with a silica-gel column (eluent: PE / EA = 1 / 1, v / v) to yield tert-butyl 4-(4-((3-nitro-4-(trifluoromethoxy)phenyl)amino)phenyl)piperazine-1-carboxylate as yellow oil (2.1 g, 60% yield), m / z (ESI) calcd. for C22H25F3N4O5: 482.18, found: 483.18, [M+H]+.Step 2: T ert-butyl 4-(4-((3 -nitro-4-(trifluoromethoxy )phenyl )amino)phenyl )piperazine- 1 -carboxylate (2.1 g, 4.14 mmol) was dissolved in DCM (60 mL), then trifluoroacetic acid (20 mL) was added to that. The reaction mixture was stirred at room temperature for 0.5 h, then concentrated under reduced pressure to obtain trifluoroacetate, 3-nitro-N-(4-(piperazin- l-yl)phenyl)-4-(trifluoromethoxy)aniline as yellow solids (2.5 g), which was used in the next step without further purification, m / z (ESI) calcd. for C17H17F3N4O3: 382.13, found: 383.13, [M+H]+.Step 3: The above trifluoroacetate, 3-nitro-N-(4-(piperazin-l-yl)phenyl)-4-(trifluoromethoxy)aniline (2.5 g, 4.14 mmol) was dissolved in pyridine (50 mL), then trifluoroacetic anhydride (3.1 g, 14.51 mmol) was added to that. The mixture was stirred at room temperature for 1 hour, then concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: PE / EA = 1 / 1, v / v) to yield 2,2,2-trifluoro-N-(3-nitro-4-(trifluoromethoxy)phenyl)-N-(4-(4-(2,2,2-trifluoroacetyl)piperazin-l-yl)phenyl)acetamide as yellow solid (2.0 g, 84.0% overall yield of two steps), m / z (ESI) cal cd. for C21H15F9N4O5: 574.09, found: 575.09, [M+H]+.Step 4: 2,2,2-trifluoro-N-(3-nitro-4-(trifluoromethoxy)phenyl)-N-(4-(4-(2,2,2-trifluoroacetyl)piperazin-l-yl)phenyl)acetamide (2.0 g, 3.51 mmol) and B2(OH)4(1.6 g, 17.71 mmol) were dissolved in DMF (N, N-dimethylamine; 40 mL), which was then cooled to 0 °C, followed by addition of 1,4-bipyridine (55.2 mg, 0.35 mmol) dissolved in DMF (1 mL). The mixture was stirred at room temperature for 30 minutes. Then diluted with water and extracted with ethyl acetate (550 mL x 3 times). The combined organic layer was dried over anhydrous MgSO4 and concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: EA / PE = 1 / 2, v / v) to yield N-(3-amino-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-(4-(4-(2,2,2-trifluoroacetyl)piperazin-l-yl)phenyl)acetamide Intermediate D-l as yellow solid (1.3 g, 70% yield), m / z (ESI) calcd. for C21H17F9N4O3: 544.12, found: 545.12, [M+H]+.The Intermediates D-2 to D-4 shown in Table B were prepared following the teachings of the General Reaction Schemes and the method to prepare INTERMEDIATE D-l.TABLE BStructure Intermediate No. Spectral Data0NH,D-l 545.12, [M+H] +T FNH2o%''' N D-2 468.29 [M+H] +T FO'^ z^pFFNH29cF bs, Jk ~XwirD-3 524, [M+H]+F. TFV> ONH2D-4 452.05 [M-H]+I F- >^FFrINTERMEDIATE E-lEphos Pd G4 / Ephos, Cs2CO3, 1 4- HCOOH, Tol STAB, DCM dicxane, 90 °C, 1 5 h rt, 1 h Step 2 Step 3TFAA, pyridine, B2(OH)4, 4,4-bipyridine rt, 1 h DMF, rt, 20 min Step 4 Step 5INTERMEDIATE E-1Step 1: To a solution of 4-bromo-2-nitro-l-(trifluorom ethoxy )benzene (2.0 g, 7.00 mmol) in 1,4-dioxane (200 mL) was added l,4-dioxaspiro[4.5]decan-8-amine (2.2 g, 13.99 mmol), Ephos PdG4 (640.0 mg, 0.70 mmol), Ephos (370.0 mg, 0.70 mmol ), and cesium carbonate (6.8 g, 20.98 mmol). The mixture was stirred at 90 °C for 1.5 h. After being cooled to room temperature, insoluble solids were removed by filtration, and the filtrate was diluted with water and then extracted with ethyl acetate (200 mL × 3 times). Organic phases were combined and dried over anhydrous MgSO4, followed by concentration under reduced pressure. The resultant residue was purified with a silica-gel column (eluent: PE / EA = 10 / 1, v / v) to N-(3-nitro-4-(trifluoromethoxy)phenyl)-l,4-dioxaspiro[4.5]decan-8-amine (1.4 g, 56% yield) as yellow oil. m / z (ESI) calcd. for C15H17F3N2O5: 362.1, found: 363.1, [M+H]+.Step 2: N-(3-nitro-4-(trifluoromethoxv)phenvl)-l,4-dioxaspiror4.5]decan-8-amine (1.4 g, 3.87 mmol) and HCOOH / Toluene (v / v =1 / 3; 120 mL) and stirred at 110 °C for 1 h, after being cooled to room temperature. The resultant residue was purified with a silica-gel column (eluent: PE / EA = 10 / 1, v / v) to 4-((3-nitro-4-(trifluoromethoxy)phenyl)amino)cyclohexan-l-one (1.4 g, 80% yield) as yellow oil. m / z (ESI) calcd. for C13H13F3N2O4: 318.1, found: 319.1, [M+H]+.Step 3: A solution of 4-((3-nitro-4-(trifluoromethoxy)phenyl)amino)cyclohexan-l-one (900.0 mg, 2.83 mmol) and morpholine (739.1 mg, 8.48 mmol) were dissolved in CH2CI2 (90 mL), then AcOH (50 uL) was added. The mixture was stirred at room temperature for 10 min before adding sodium triacetoxyborohydride (1.8 g, 8.48 mmol) in portions. After stirring for another 1 h, the reaction mixture was diluted with water and extracted with ethyl acetate (100 mL x 3 times). The combined organic layers dried over anhydrous MgSO4 and concentrated under reduced pressure to yield N-(4-morpholinocyclohexyl)-3-nitro-4-(trifluoromethoxy)aniline as yellow solids (1.0 g, 91% yield), which was used in the next step without further purification, m / z (ESI) calcd. for C17H22F3N3O4: 389.2, found: 390.1, [M+H]+Step 4: N-(4-morpholinocvclohexvl)-3-nitro-4-(trifluoromethoxy)aniline (1.0 g, 2.57 mmol) was dissolved in pyridine (100 mL), then trifluoroacetic anhydride (1.08 g, 5.14 mmol) was added to it. The mixture was stirred at room temperature for 1 h and concentrated under reduced pressure afterward. The residue was purified with a silica-gel column (eluent: PE / EA = 1 / 1, v / v) to yield 2,2,2-trifluoro-N-(4-morpholinocyclohexyl)-N-(3-nitro-4-(trifluoromethoxy)phenyl)acetamide (1.2 g, 94% yield) as yellow solids, m / z (ESI) calcd. for C19H21F6N3O5: 485.1, found: 486.1, [M+H]+.Step 5: 2.2.2-trifluoro-N-('4-morpholinocvclohexvl)-N-(3-nitro-4-(trifluoromethoxy)phenyl)acetamide (1.2 g, 2.47 mmol) was dissolved in DMF (N, N-dimethylformamide; 60 mL), followed by addition of B2(OH)4 (660.0 mg, 7.42 mmol). The mixture was cooled to 0 °C before slowly adding 1,4-bipyridine (3.86 mg, 0.03 mmol) dissolved in DMF (1 mL). After being stirred at room temperature for 20 minutes, the reaction solution was diluted with water and extracted with ethyl acetate (500 mL × 3 times). The combined organic layers were dried over anhydrous MgSO4 and concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: DCM / MeOH = 10 / 1, v / v) to yield N-(3-amino-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-(4-morpholinocyclohexyl)acetamide (793.0 mg, 72% yield) Intermediate E-l as yellow solids, m / z (ESI) calcd. for C19H23F6N3O3: 455.2, found: 456.1, [M+H]+.The Intermediates E-2 to E-7 shown in Table C were prepared following the teachings of the General Reaction Schemes and the method to prepare INTERMEDIATE E-l.TABLE CStructure Intermediate No. Spectral Data NH2. CT' E-l 456.1, [M+H]+F\FA * orFNH2N^'X-FF UlNXj E-2 497.2, [M+H]+YFo' AFrE-3 531.0, [M+H]+NH2f^OQ ‘’FE-4 484.1, [M+H]+ bAA QrF, GJJbb-b ^ E-5 482.48, [M+H] + oo^ LL. LLLL- IX u.F?H2r\TbkOJ F E-6 440.4, [M+H]+ °>FF r"\TtX cr- E-7 440.4, [M+H]+7 F°>rINTERMEDIATE F-l196613-57-7 Ruphos, Ruphos TFA, DCM, TFAA, Pd,100°C,8h rt, 0.5 hypyridine, rt, 1 h Step 1 Step 2 Step 395668-20-5INTERMEDIATE F-1Step 1: To a solution of 4-bromo-2-nitro-l-(trifluorom ethoxy )benzene (2.0 g, 6.99 mmol) in toluene (200 mL) were sequentially added tert-butyl 4-aminoazepane-l -carboxylate (2.2 g, 10.41 mmol), Ruphos Pd G4 (585.2 mg, 0.70 mmol), Ruphos (326.1 mg, 0.70 mmol), and cesium carbonate (4.5 g, 13.90 mmol). Then the mixture was bubbled with N2 flow for 5 minutes to expel air gas in the flask, after which the mixture was stirred at 100 °C under N2 atmosphere for 4 h. After being cooled to room temperature, insoluble solids were removed by filtration, and the filtrate was diluted with water (250 mL), then extracted with ethyl acetate (3x500 mL). Organic phases were combined and dried over anhydrous MgSO4, followed by concentration under reduced pressure. The resultant residue was purified with a silica-gel column (eluent: PE / EA = 2 / 1, v / v) to yield tert-butyl 4-((3-nitro-4-(trifluoromethoxy)phenyl)amino)azepane-l -carboxylate as yellow oil (2.1 g, 70.6% yield), m / z (ESI) calcd. for C18H24F3N3O5: 419.17, found: 420.17, [M+H]+.Step 2: Tert-butyl 4-((3-nitro-4-(trifluoromethoxy)phenyl)amino)azepane-l-carboxylate (2.1 g, 5.01 mmol) was dissolved in DCM (dichloromethane; 30 mL), then trifluoroacetic acid (10 mL) was added to that. The reaction mixture was stirred at room temperature for 0.5 h, followed by concentration under reduced pressure to afford trifluoroacetate, N-(3-nitro-4-(trifluoromethoxy)phenyl)azepan-4-amine as brown semi-solids (2.6 g), which was used in next step without further purification. m / z(ESI), calcd. for C13H16F3N3O3: 319.1, found 320.0,[M+H]+.Step 3: The above-obtained trifluoroacetate, N-(3-nitro-4-(trifluoromethoxy)phenyl)azepan-4-amine (2.6 g, 5.01 mmol) was dissolved in pyridine (30 mL), then TFAA (trifluoroacetic anhydride; 5.3 g, 25.07 mmol) were added to it. The mixture was stirred at rt for 1 h, then concentrated under reduced pressure to yield 2,2,2-trifluoro-N-(3 -nitro-4-(trifluorom ethoxy )phenyl)-N-(l -(2,2,2-trifluoroacetyl)azepan-4-yl)acetamide yellow solid (3.6 g), which was used in next step without further purification. m / z(ESI) calcd. for C17H14F9N3O5: 511.1, found: 512.1, [M+H]+.Step 4: 2,2,2-trifluoro-N-(3-nitro-4-(trifluoromethoxy)phenyl)-N-(l-(2,2,2-trifluoroacetyl)azepan-4-yl)acetamide (3.6 g, 5.01 mmol) and B2(OH)4 (1.9 g, 20.5 mmol) were dissolved in DMF (N, N-dimethylformamide; 30 mL), which was then cooled to 0 °C before slow addition of 1,4-bipyridine (7.8 mg, 0.05 mmol) dissolved in DMF (0.5 mL). The mixture was stirred at room temperature for 20 minutes, then diluted with water and extracted with ethyl acetate (550 mL x 3 times). The combined organic layers were dried over anhydrous MgSO4 and concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: CH2Cl2 / MeOH = 10 / l, v / v) to afford N-(3-amino-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-(l-(2,2,2-trifluoroacetyl)azepan-4-yl)acetamide Intermediate F-l as yellow solid (1.7 g, 70.6% overall yield of first three steps). m / z(ESI), calcd. for C17H16F9N3O3: 481.1, found: 482.1 [M+H]+.The Intermediates F-2 to F-9 shown in Table D were prepared following the teachings of the General Reaction Schemes and the method to prepare INTERMEDIATE F-l.TABLE DX( -npS V V z ■n—tructure Intermediate No. Spectral Data NKHr~, ■ - > oF-7 416.2, [M+H]+ F. T IQ-v°F-8 402.1, [M+H] +NH2F-9 400.2, [M+H]+ E T FV°F-10 386.3, [M+H]+oLL. LL X11-"INTERMEDIATE G-l£>— OH Ruphos, Ruphos K2C03, DMF, Rd, Cs2CO3, Tol., TFA, DCM, 50°C, 20 h, 100 °C, 16 h. rt, 0.5 h Step 1 Step 2 Step 3 Br 364-73-8B2(0H)2, 4,4- bipyridine, TFAA, pyridine, DMF, rt, 1 h_ rt 1 h Step 4 Step 5Intermediate G-1Step 1: To a solution of 4-bromo-2-fluoro-l -nitrobenzene (5.0 g, 22.72 mmol) in DMF (30 mL) were added cyclopropanol (1.6 g, 27.26 mmol), and potassium carbonate (9.4 g, 68.16 mmol).Then the mixture was stirred at 50 °C for 20 h. After being cooled to room temperature,insoluble solids were removed by fdtration, and the filtrate was diluted with water, then extracted with ethyl acetate (200 mL x 3 times). Organic phases were combined and dried over anhydrous MgSO4, followed by concentration under reduced pressure to 4-bromo-l-cyclopropoxy-2-nitrobenzene (4.7 g, 51.0% yield) as yellow oil. 1HNMR (300 MHz, Chloroform-d) 87.95 (d, J= 2.4 Hz, 1H), 7.64 (dd, J = 9.0, 2.5 Hz, 1H), 7.37 (d, J = 9.0 Hz, 1H), 3.87 (p, J = 4.5 Hz, 1H),0.89 (s, 4H).Step 2: To a solution of 4-bromo-2-nitro-l-(trifluorom ethoxy )benzene (2.5 g, 8.74 mmol) in 1,4-dioxane (250 mL) were sequentially added ert-butyl 4-aminoazepane-l -carboxylate (2.1 g, 9.69 mmol), RuPhos Pd G4 (710.0 mg, 0.97 mmol), RuPhos (450.0 mg, 0.97 mmol), and cesiumcarbonate (6.3 g, 19.37 mmol). Then, the mixture was bubbled with N2 flow for 5 minutes toexpel air gas in the flask, after which the mixture was stirred at 100 °C under N2 atmosphere for16 h. After cooling to room temperature, insoluble solids were removed by filtration, and thefiltrate was diluted with water (250 mL) and then extracted with ethyl acetate (3 * 200 mL).Organic phases were combined and dried over anhydrous MgSO4, followed by concentrationunder reduced pressure. The resultant residue was purified with a silica-gel column (eluent: PE / EA = 1 / 1, v / v) to yield tert-butyl 4-((4-cyclopropoxy-3-nitrophenyl)amino)azepane-l-carboxylate as yellow oil (2.3 g, 60% yield), m / z (ESI) calcd. for C20H29N3O5: 391.2, found: 414.2 [M+Na]+.Step 3: Tert-butyl 4-((4-cyclopropoxy-3-nitrophenyl)amino)azepane-l-carboxylate( 2.6 g, 6.60 mmol) was dissolved in DCM (200 m ), then trifluoroacetic acid (40 mL) was added to it. The reaction mixture was stirred at room temperature for 1 h, followed by concentration by a rotary evaporator to yield a crude product of N-(4-cyclopropoxy-3-nitrophenyl)azepan-4-amine (1.8 g, 95.0% yield) as yellow semi-solids, which was used for the next step without further purification, m / z (ESI) calcd. for C15H21N3O3: 291.2, found: 292.1 [M+H]+.Step 4: N-(4-cyclopropoxy-3-nitrophenyl)azepan-4-amine (1.8 g, 6.20 mmol) was dissolved in pyridine (150 mL), then trifluoroacetic anhydride (2.6 g, 12.40 mmol) was added to that. The mixture was stirred at room temperature for 1 h, then concentrated under reduced pressure; the resultant residue was purified with a silica-gel column (eluent: EA / PE = 1 / 1, v / v) to yield N-(4-cyclopropoxy-3-nitrophenyl)-2,2,2-trifluoro-N-(l-(2,2,2-trifluoroacetyl)azepan-4-yl)acetamide as yellow solid (2.5 g, 83% yield), m / z (ESI) calcd. for C19H19F6N3O5: 483.1, found LCMS: 506.0, [M+Na]+.Step 5: N-(4-cyclopropoxy-3-nitrophenyl)-2,2,2-trifluoro-N-(l-(2,2,2-trifluoroacetyl)azepan-4-yl)acetamide (1.8 g, 3.70 mmol) was dissolved in DMF (N, N-dimethylformamide; 40 mL), followed by addition of B2(OH)4 (1.0 g, 11.20 mmol). The mixture was cooled to 0 °C before slowly adding 1,4-bipyridine (11.6 mg, 0.07 mmol) dissolved in DMF (1 mL). After being stirred at room temperature for 20 minutes, the reaction solution was diluted with water and extracted with ethyl acetate (500 mL × 3 times). The combined organic layers were dried over anhydrous MgSO₄ and concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: DCM / MeOH = 10 / 1, v / v) to yield N-(3-amino-4-cyclopropoxyphenyl)-2,2,2-trifluoro-N-(l-(2,2,2-trifluoroacetyl)azepan-4-yl)acetamide Intermediate G-l as yellow solids (2.0 g, 83.5% yield), m / z (ESI) calcd. for C19H21F6N3O3: 453.1, found: 454.1, [M+H]+.The Intermediates G-2 shown in Table E were prepared following the teachings of the General Reaction Schemes and the method to prepare INTERMEDIATE G-l.TABLE EStructure Intermediate No. Spectral DataG-l 454.1, [M+H] +NH20G-2 440.2, [M+H] +7 FG-3 334.2, [M+H] +0LL LL INTERMEDIATE H-lO3 Ll_ / HN'^yXX'OH / < 0= A^NBoc yjz~ tBuXphos Pd 0 G3, NaOPh, TBSCi, imidozale,To!, 100 °C, 4 hrPCM, rt, 16 hrStep 1 Step 2B2(OH)4, 1,4- bipyridine, DMF, rt, 1 h Step 3INTERMEDIATE H-1Step 1: To a solution of 4-bromo-2-nitro-l-(trifluoromethoxy)benzene (3.0 g, 10.49 mmol) in toluene (150 mb) were sequentially added tert-butyl 2-(hydroxymethyl)piperazine-l -carboxylate (3.4 g, 15.73 mmol), t-BuXPhos Pd G3 (832.8 mg, 1.05 mmol), t-BuXPhos (445.6 mg, 1.05 mmol), and sodium benzenolate (3.6 g, 31.47 mmol). Then the mixture was bubbled with N2 flow for 5 minutes to expel air in the flask, after which the mixture was stirred at 100 °C underN2 atmosphere for 4 h. After being cooled to room temperature, insoluble solids were removed by fdtration, and the filtrate was diluted with water then extracted with ethyl acetate (200 mb x 3 times). Organic phases were combined and dried over anhydrous MgSO4, followed by concentration under reduced pressure. The resultant residue was purified with a silica-gel column (eluent: PE / EA = 1 / 1, v / v) to yield tert-butyl 2-(hydroxymethyl)-4-(3-nitro-4-(trifluoromethoxy)phenyl)piperazine-l -carboxylate as yellow solid (1.2 g, 27% yield), m / z (ESI) calcd. for C17H22F3N3O6: 421.1, found: 422.1, [M+H]+.Step 2: Tert-butyl 2-(hydroxymethyl)-4-(3-nitro-4-(trifluoromethoxy)phenyl)piperazine-l-carboxylate (1.2 g, 2.85 mmol) and imidazole (600.6 mg, 8.54 mmol) was dissolved in DCM (150 mL), then tert-butylchlorodimethylsilane (860.3 mg, 5.69 mmol) was added to it. The mixture was stirred at room temperature for 16 h, followed by concentration under reduced pressure; the resultant residue was purified with a silica-gel column (eluent: PE / EA = 1 / 1, v / v) to yield tert-butyl 2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(3-nitro-4-(trifluoromethoxy)phenyl)piperazine-l -carboxylate as yellow solid (1.3 g, 86 % yield), m / z (ESI) calcd. for C20H17F6N3O5: 535.2, found: 536.2, [M+H]+.Step 3: Tert-butyl 2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(3-nitro-4-(trifluoromethoxy)phenyl)piperazine-l -carboxylate (1.3 g, 2.43 mmol) andB2(OH)4 (650.0 mg, 7.281 mmol) were dissolved in DMF (N, N-dimethylformamide; 55 mL), followed by slow addition of 1,4-bipyridine (1.9 mg, 0.01 mmol) dissolved in DMF (0.5 mL). The mixture was stirred at room temperature for 1 hour, then diluted with water and extracted with ethyl acetate (550 mL x 3 times). The combined organic layers were dried over anhydrous MgSO₄ and concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: DCM / MeOH = 10 / 1, v / v) to afford tert-butyl 4-(3-amino-4-(trifluoromethoxy)phenyl)-2-(((tert-butyldimethylsilyl)oxy)methyl)piperazine-l -carboxylate as yellow solid (1.1 g, 83% yield), m / z (ESI), calcd. for C23H38F3N3O4Si: 505.2, found: 506.2, [M+H]+.INTERMEDIATE l-lEphos Pd, EPhos, CS2CO3.To!., 100 °C, 4 h Step 1B2(OH)4, 1,4- bipyridine, DMF, rt, 1 h > Step 4INTERMEDIATE 1-1Step 1: To a solution of 4-bromo-2-nitro-l-(trifluorom ethoxy )benzene (5.1 g, 17.83 mmol) in toluene (250 mL) were sequentially added tert-butyl 4-aminopiperidine-l -carboxylate (2.4 g, 21.34 mmol), Ephos Pd G3 (813.3 mg, 0.89 mmol), Ephos (441.4 mg, 0.89 mmol), CS2CO3 (17.4 g, 53.49 mmol). Then, the mixture was bubbled with N2 flow for 5 minutes to expel air gas in the flask, after which the mixture was stirred at 100 °C under N2 atmosphere for 4 h. After cooling to room temperature, insoluble solids were removed by filtration. The filtrate was diluted with water and then extracted with ethyl acetate (250 mL x 3 times). Organic phases were combined and dried over anhydrous MgSO4, followed by concentration under reduced pressure. The resultant residue was purified with a silica-gel column (eluent: PE / EA= 1 / 1, v / v) to yield tertbutyl 4-((4-cyclopropyl-3-nitrophenyl)amino)piperidine-l-carboxylate(7.9 g, 80.0% yield) as yellow oil. m / z (ESI) calcd. for C19H27N3O4: 361.2, found: 362.2, [M+H]+.Step 2: Tert-butyl 4-((4-cyclopropyl-3-nitrophenyl)amino)piperidine-l-arboxylate (7.9 g, 21.88 mmol) was dissolved in DCM (120 mL), then trifluoroacetic acid (24 mL) was added to it. The reaction mixture was stirred at room temperature for 1 h, followed by concentration by a rotary evaporator to yield trifluoroacetate of N-(4-cyclopropyl-3-nitrophenyl)piperidin-4-amine as yellow semi-solids (9.6 g), which was used for the next step without further purification, m / z (ESI) calcd. for C14H19N3O2: 261.2, found: 262.2, [M+H]+.Step 3: The above-obtained trifluoroacetate of N-(4-cyclopropyl-3-nitrophenyl)piperidin-4-amine (9.6 g) was dissolved in pyridine (200 mL), then trifluoroacetic anhydride (30.9 g, 147.20 mmol) was added to it. The mixture was stirred at room temperature for 1 h, then concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: DCM / MeOH = 10 / 1, v / v) to yield N-(4-cyclopropyl-3-nitrophenyl)-2,2,2-trifluoro-N-(l -(2,2,2-trifluoroacetyl)piperidin-4-yl)acetamide as yellow solids (2.5 g, 24.0% overall yield of two steps), m / z (ESI) calcd. for C18H17F6N3O4: 453.2, found: 454.2, [M+H]+.Step 4: N-(4-cyclopropyl-3-nitrophenyl)-2,2,2-trifluoro-N-(l-(2,2,2-trifluoroacetyl)piperidin-4-yl)acetamide (2.5 g, 5.52 mmol) was dissolved in DMF (N, N-dimethylformamide; 50 mL), followed by addition of B2(OH)4 (2.4 g, 27.60 mmol). The mixture was cooled to 0 °C before slowly adding 1,4-bipyridine (83.6 mg, 0.55 mmol) dissolved in DMF (2 mL). After being stirred at room temperature for 20 minutes, the reaction solution was diluted with water and extracted with ethyl acetate (500 mL x 3 times). The combined organic layers were dried over anhydrous MgSO4 and concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: DCM / MeOH = 10 / 1, v / v) to yield N-(3-amino-4-cyclopropylphenyl)-2,2,2-trifluoro-N-(l-(2,2,2-trifluoroacetyl)piperidin-4-yl)acetamide as yellow solid (1.1 g, 50% yield), m / z (ESI) calcd. for C18H19F6N3O2: 423.2, found: 424.2, [M+H]+.The Intermediate 1-2 shown in Table F was prepared following the teachings of the General Reaction Schemes and the method to prepare INTERMEDIATE 1-1.TABLE FStructure Intermediate No. Spectral DataO F NH,y FO' A. l-l 454.39, [M+H] +FFNH20I-2 440.36, [M+H] +I F0>FINTERMEDIATE J-l>-BF3-K? Ephos Pd NO-: B2(OH)4, 1,4- Pd(AcO;2, K3PO4, EPhos Cs2CO3, bipyridine, Tol. H?O. 80CC, 16 h To!. 100 C OMF, rt. 1 htStep 1 Step 2 Step 3INTERMEDIATE J-1 Step 1: To a solution of l-bromo-4-chloro-2-nitrobenzene (5.0 g, 15.20 mmol) in toluene (150 mb) were added potassium cyclopropyltrifluoroborate (4.1 g, 19.80 mmol), tricyclohexyl phosphine (593.0 mg, 1.52 mmol), palladium acetate (238.0 mg, 0.76 mmol ), and potassium phosphate tribasic (18.0 g, 60.80 mmol). The mixture was bubbled with N2 flow for 5 minutes to expel air gas in the flask, after which the mixture was stirred at 80 °C under N2 atmosphere for 16 h. After being cooled to room temperature, insoluble solids were removed by filtration, and the filtrate was diluted with water and then extracted with ethyl acetate (200 mL x 3 times). Organic phases were combined and dried over anhydrous MgSO4, followed by concentration under reduced pressure to 4-chloro-l-cyclopropyl-2-nitrobenzene (4.7 g, 51.0% yield) as yellow oil. m / z (ESI) calcd. for: C9H8CINO2: 197.0, found: 198.0, [M+H]+.Step 2: To a solution of 4-bromo-l-cyclopropoxy-2-nitrobenzene (4.0 g, 20.30 mmol) in 1,4-dioxane (150 mL) were added tert-butyl piperazine- 1 -carboxylate (5.7 g, 30.45 mmol), EPhos PdG4 (1.3 g, 1.41 mmol), EPhos (760.0 mg, 1.42 mmol), and cesium carbonate (20.0 g, 60.90 mmol). Then, the mixture was bubbled with N2 flow for 5 minutes to expel air gas in the flask, after which the mixture was stirred at 100 °C under N2 atmosphere for 16 h. After being cooled to room temperature, insoluble solids were removed by filtration, and the filtrate was diluted with water and then extracted with ethyl acetate (200 mL x 3 times). Organic phases were combined and dried over anhydrous MgSO4, followed by concentration under reduced pressure. The resultant residue was purified with a silica-gel column (eluent: PE / EA = 10 / 1, v / v) to yield tert-butyl 4-(4-cyclopropyl-3-nitrophenyl)piperazine-l -carboxylate as yellow oil (3.6 g, 50.0% yield), m / z (ESI) calcd. for: C18H25N3O4: 347.2, found: 348.2, [M+H]+.Step 3: Tert-butyl 4-(4-cyclopropyl-3-nitrophenyl)piperazine-l-carboxylate (3.5 g, 10.05 mmol) was dissolved in DMF (N, N-dimethylformamide; 50 mL), followed by addition of B2(OH)4 (2.7 g, 30.17 mmol). The mixture was cooled to 0 °C before slowly adding 1,4-bipyridine (16.0 mg, 0.10 mmol) dissolved in DMF (0.5 mL). After being stirred at room temperature for 20 minutes, the reaction solution was diluted with water and extracted with ethyl acetate (500 mL x 3 times). The combined organic layers were dried over anhydrous MgSO₄ and concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: DCM / MeOH = 10 / 1, v / v) to yield tert-butyl 4-(3-amino-4-cyclopropylphenyl)piperazine-l -carboxylate as yellow solids (6.0 g, 85% yield), m / z (ESI) calcd. for C18H27N3O2: 317.2, found: 318.2, [M+H]+INTERMEDIATE K-lStep 1: To a solution of 4-bromo-2-fluoro-l -nitrobenzene (5.0 g, 22.72 mmol) in DMF (30 mL) were added cyclopropanol (1.6 g, 27.26 mmol), and potassium carbonate (9.4 g, 68.16 mmol). Then the mixture was stirred at 50 °C for 20 h. After being cooled to room temperature, insoluble solids were removed by filtration, and the filtrate was diluted with water, then extracted with ethyl acetate (200 mL x 3 times). Organic phases were combined and dried over anhydrous MgSO4, followed by concentration under reduced pressure to 4-bromo-l-cyclopropoxy-2-nitrobenzene (4.7 g, 51.0% yield) as yellow oil. 1HNMR (300 MHz, Chloroform-d) 87.95 (d, J = 2.4 Hz, 1H), 7.64 (dd, J = 9.0, 2.5 Hz, 1H), 7.37 (d, J = 9.0 Hz, 1H), 3.87 (p, J = 4.5 Hz, 1H), 0.89 (s, 4H).Step 2: N- 4-cvclopropoxv-3-nitrophenyl)-l-isopropylpyrrolidin-3-amine (600.0 mg, 1.96 mmol) was dissolved in pyridine (30 mL), then trifluoroacetic anhydride (1.2 g, 5.90 mmol) was added thereto. The mixture was stirred at room temperature for 1 h, then concentrated under reduced pressure to yield N-(4-cyclopropoxy-3-nitrophenyl)-2,2,2-trifluoro-N-(l-isopropylpyrrolidin-3-yl)acetamide as yellow solids (790.0 mg), which was used in the next step without further purification, m / z (ESI) calcd. for C18H22F3N3O4: 401.2, found: 402.1, [M+H]+.Step 3: N-(4-cvclopropoxv-3-nitrophenyl)-2.2.2-trifluoro-N-Cl-isopropylpyrrolidin-3-yl)acetamide (790.1 mg, 1.96 mmol) and B2(OH)4(675.0 mg, 7.50 mmol) were dissolved in DMF (N, N-dimethylformamide; 40 mL), which was cooled to 0 °C, followed by addition of 1,4-bipyridine (23.4 mg, 0.15 mmol) dissolved in DMF (0.5 mL). The mixture was stirred at room temperature for 20 minutes then diluted with water and extracted with ethyl acetate (550 mL x 3 times). The combined organic layers were dried over anhydrous MgSO4and concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: CH2Cl2 / MeOH =10 / 1, v / v) to afford N-(3-amino-4-cyclopropoxyphenyl)-2,2,2-trifluoro-N-(l-isopropylpyrrolidin-3-yl)acetamide as yellow solids (600.0 mg, 82% overall yield of two steps). m / z(ESI), calcd. for C18H24F3N3O2: 371.2, found: 372.3, [M+H]+.TABLE GStructure Intermediate No. Spectral DataNH2XZYS ruK-l 372.3, [M+H] +T FK-2 372.2, [M+H] +NH2K-3 360.2, [M+H] +T Fo>FNH2K-4 374.2, [M+H] +F. i AAzQ |o / AEXAMPLESo

[0106] The following examples are intended to illustrate specific embodiments of the invention further and are not intended to limit the scope of the invention.EXAMPLE 1

[0107] Step 1: To a mixture of methyl N-[3-amino-4-(trifluoromethoxy)phenyl]carbamate (813.0 mg, 3.25 mmol), 2,7-dichloropyrido[4,3-d]pyrimidine (500.0 mg, 2.50 mmol) in DMSO (10 mL) was added DIEA (323.0 mg, 2.50 mmol). The mixture was stirred for 22 h at 100 °C. EtOAc (20 mL) and H2O (20 mL) were added to the mixture. After separating phases, the aqueous phase was extracted with EtOAc (20 mL x 2 times). The combined organic layers were washed with brine (50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: EtOAc / PE = 1 / 5, v / v) to afford methyl N-[3-({7-chloropyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]carbamate (170.0 mg, 12.3% yield) as ayellow solid, m / z (ESI) calcd. for C16H11ClF3N5O3: 413.1, found: 414.1 [M+H]+.

[0108] Step 2: To a mixture of methyl N,-[3-({7-chloropyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]carbamate (320.0 mg, 0.77 mmol) in 1,4-dioxane (4 mL) and H2O (1 mL) were added Pd(dppf)C12 (100.0 mg, 0.16 mmol), 2-ethenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (476.4 mg, 3.09 mmol) and K3PO4 (328.0 mg, 1.55 mmol). The mixture was stirred at 80 °C for 2 h under a nitrogen atmosphere. EtOAc (20 mL) and H2O (20 mL) were added to the mixture. After separating phases, the aqueous phase was extracted with EtOAc (20 mL x 2 times). The combined organic layers were washed with brine (50 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified with a silica-gel column(eluent: EtOAc / PE = 1 / 5, v / v) to afford methyl N-[3-({7-ethenylpyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]carbamate (190.0 mg, 53.9% yield) as a yellow solid. m / z (ESI) calcd. for C18H14F3N5O3: 405.1, found: 406.1 [M+H]+.

[0109] Step 3: To a solution of methyl N-[3-({7-ethenylpyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]carbamate (180.0 mg, 0.44 mmol) in THF (9 mL) was added NaIO4 (569.0 mg, 2.66 mmol) and OsO4 (22.0 mg, 0.09 mmol). The mixture was stirred at room temperature for 3 h. H2O (20 mL) was added to the mixture. The resulting mixture was extracted with DCM (20 mL x 3 times). The combined organic layers were washed with brine (30 mL) and dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure to provide methyl (3-((7-formylpyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)carbamate (150.0 mg, 83.3% yield) as a yellow solid, m / z (ESI) calcd. for C17H12F3N5O4: 407.1, found: 408.1 [M+l]+.

[0110] Step 4: A solution of tert-butyl N-(diphenylphosphoroso)methanesulfonylcarbamate (75.7 mg, 0.19 mmol) in DMF (4 mL) was added NaH (11.1 mg, 0.48 mmol, 60%wt) in portions at 0 °C. The resulting mixture was stirred for 30 min at room temperature. Then methyl N-[3-({7-formylpyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]carbamate (78.1 mg, 0.19 mmol) was added and the mixture was stirred for 6 h at room temperature. The reaction mixture was eluted with ACN / H2O (0.05%TFA) (from 50% to 100% in 30 min) from a C18 column to afford methyl N-[3-({7-[(Z)-2-[(tert-butoxycarbonyl)aminosulfonyl]ethenyl]pyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]carbamate (30.0 mg, 26.0% yield) as a yellow solid, m / z (ESI) calcd. for C23H23F3N6O7S: 584.1, found: 485.1 [M+H-Boc]+.Step 5: To the mixture of methyl N-[3-({7-[(E)-2-[(tert-butoxycarbonyl)aminosulfonyl]ethenyl]pyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]carbamate (25.0 mg, 0.04 mmol) in DCM (1.5 mL) was added TFA (0.3 mL). The mixture was stirred at rt for 2h. The desired product could be detected by LCMS. The mixture was concentrated. Prep-HPLC purified the crude product with the following conditions (Column: XSelect CSH Cl 8 Column, 19*250mm, 5pm; Mobile Phase A: Water(0.1% FA), Mobile Phase B: ACN; Flow rate: 25 mL / min mL / min; Gradient:32%B to 52%B in 10 min; Wave Length: 254nm / 220nm; RT: 9.17 min) to afford methyl N-[3-({7-[(E)-2-sulfamoylethenyl]pyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]carbamate (2.5 mg, 12.0% yield) as a white solid, m / z (ESI) calcd. for C18H15F3N6O5S: 484.1, found: 485.1 [M+H]+. 1HNMR (300 MHz, DMSO-d6) 59.42 (s, 1H), 9.18 (s, 1H), 8.01 (s, 1H), 7.67 (s, 1H), 7.57 (d, J = 15.0 Hz, 1H), 7.49 -7.26 (m, 3H), 3.67 (s, 3H). 19F NMR (282 MHz, DMSO) 5 -56.76.Following the teachings of the General Reaction Schemes and the synthesis procedure for Example 1 were prepared as shown in Table I:TABLE IExample # Structure Name NMR LCMS tert-butyl (E)-4-(3-((7-(2- 1H NMR (400 MHz, DMSO-d6) 6981 sulfamoylvinyl)pyrido[4,3- (s, 1H), 9.44 (s, 1H), 9.18 (s, 1H), 7.76 d]pyrimidin-2-yl)amino)-4- (s, 1H), 760 - 7.47 (m, 3H), 7.32 -7.232 596.1 [M+H]+(trifluoromethoxy)phenyl)piperazin (m, 3H), 6.90 - 6.87 (m, 1H), 3.50 - 3.46e-l-carboxylate (m, 4H), 3.18 - 3.16 (m, 4H), 1.43 (s,9H). 19F NMR (376 MHz, DMS0-d6) 5 - (E)-2-(2-((5-(4-aminopiperidin-l-yl)- 1H NMR (400 MHz, DMSO-d6) 59.442- (s, 1H), 9.20 (s, 1H), 7.69 (s, 1H), 7.60 (trifluoromethoxy)phenyl)amino)py (d, J = 15.0 Hz, 1H), 7.50 - 7.46 (m, 2H), rido[4,3-d]pyrimidin-7-yl)ethene-l- 7.30 - 7.28 (m, 1H), 6.93 - 6.90 (m, 1H),3 sulfonamide 3.86 (s, 2H), 3.31 -3.21 (m, 1H), 2.88 510.1 [M+H]+(t, J = 12.3 Hz, 2H), 1.99 (d, J = 12.1 Hz,2H), 1.69-1.56 (m, 2H) 19F NMR (376MHz, DMSO-d6) 6 -56.8(E)-2-(2-((5-((2-methyl-l- 1H NMR (400 MHz, DMSO-d6) 59.46 morpholinopropan-2-yl)amino)-2- (s, 1H), 9.23 (s, 1H), 7.68 -7.56 (m, (trifluoromethoxy)phenyl)amino)py 2H), 7.49 (d, J = 15.0 Hz, 1H), 7.38 (s,4 568.2, [M+H]+ * 0x0 rido[4,3-d] py rimidin-7-yl)ethene-l- 1H), 7.21 (d, J = 8.6 Hz, 1H), 6.76 (d, J =sulfonamide 11.4 Hz, 1H), 3.49 (s, 4H), 3.49 (s, 6H),1.46 (s, 6H).(E)-2-(2-((5-(((ls,4s)-4- 'H NMR (300 MHz, Acetonitrile-d3) 9 32 morpholinocyclohexyl)amino)-2- (s, 1H), 9.12 (s, 1H), 7.77 (d, J = 3.0 Hz, (trifluoromethoxy)phenyl)amino)py 1H), 7.66 - 7.45 (m, 3H), 7.11 (dd, J =rido[4,3-d]pyrimidin-7-yl)ethene-l- 8.8, 1.8 Hz, 1H), 6.40 (dd, J = 8.9, 2.8 sulfonamide Hz, 1H), 3.68 - 3.53 (m, 4H), 3.20 (t, J5 594.3 [M+H]+= 11.3 Hz, 1H), 2 53 (d, J = 4.6 Hz, 4H),2.26 (dd, J = 24.3, 12.0 Hz, 3H), 2.01 (s,2H), 1.30 (dt, J = 40.1, 12.2 Hz, 5H).19F NMR (282 MHz, Acetonitrile-d3) 6 -59.06.Example # Structure Name NMR LCMS (E)-2-(2-((5-(((lr,4r)-4- ’H NMR (500 MHz, DMSO-d6) 9 43 (s, morpholinocyclohexyl)amino)-2- 1H), 9.18 (s, 1H), 767 (s, 1H), 7.57 (d, J (trifluoromethoxy)phenyl)amino)py = 15.0 Hz, 1H), 746 (d, J = 149 Hz, 1H), rido[4,3-d]pyrimidin-7-yl)ethene-l- 7.13 (d, J = 8.7 Hz, 1H), 7.05 (d, J = 5.8 sulfonamide Hz, 1H), 6.49 (dd, J = 8.9, 2.9 Hz, 1H),4.01 (d, J = 12.5 Hz, 2H), 3.69 (d, J =6 ^cyk *' "N:1?594.3, [M+H]+12.2 Hz, 2H), 3.41 (d, J = 12.2 Hz, 2H), f \ _! 3.28 - 308 (m, 5H), 2.14 (q, J = 15.8,13.7 Hz, 4H), 1.89 (s, 1H), 1.62 - 1.52(m, 2H), 1.24 (q, J = 12.0, 11.3 Hz, 2H).19F NMR (471 MHz, Methanol-d4) 6 - 60.02, -77.05.(E)-2-(2-((5-(((lr,4r)-4-(pyrrolidin-l 1H NMR (400 MHz, Methanol-d4) 6 yl)cyclohexyl)amino)-2- 9.26 (s, 1H), 9.03 (s, 1H), 7.60 (s, 1H), (trifluoromethoxy)phenyl)amino)py 7.56 (d, J = 14.9 Hz, 1H), 7.51 (s, 1H),rido[4,3-d]pyrimidin-7-yl)ethene-l- 7.42 (d, J = 15.0 Hz, 1H), 7.02 (d, J = 8.8 sulfonamide Hz, 1H), 6.41 (dd, J = 8.9, 2.8 Hz, 1H),7 578.2 [M+H]+3.58-3.54 (m, 4H), 3.15 -2.97 (m,4H), 2.09-2.01 (m, 5H), 197 -1.88(m, 5H), 1.80- 1.67 (m, 4H). 19F NMR(376 MHz, Methanol-d4) 6 -60.01, - 76.96.(E)-2-(2-((5-(((ls,4s)-4-(4- ’H NMR (500 MHz, Methanol-d4) 9.37 phenylpiperazin-1- (s, 1H), 9.15 (s, 1H), 7.81 - 7.77 (m, yl)cyclohexyl)amino)-2- 1H), 7.71 - 7.62 (m, 2H), 7.54 (d, J = (trifluoromethoxy)phenyl)amino)py 14.9 Hz, 1H), 7.30 - 7.22 (m, 2H), 7.10 rido[4,3-d]pyrimidin-7-yl)ethene-l- (dt, J =8.7, 1.5 Hz, 1H), 7.05 - 7.00 (m,W sulfonamide 2H), 6.87 (t, J = 7.3 Hz, 1H), 6.47 (dd, J =8 8.9, 2.8 Hz, 1H), 3.36 (s, 1H), 3.24 (t, J = 602.1, [M+H]+ w i 5.0 Hz, 4H), 2.84 (t, J = 5.0 Hz, 4H), 2.49 ^K,‘" O 'NC: N^O - 2 40 (m, 1H), 2.34 (d, J = 12.6 Hz,2H), 2.17 (d, J = 11.9 Hz, 2H), 1.58 - 1.47 (m, 2H), 1.39 - 1.25 (m, 4H) 19FNMR (471 MHz, Methanol-d4) 6 -59.99(E)-2-(2-((5-(((lR,4r)-4-((2S,6R)-2,6 1H NMR (500 MHz, Methanol-d4) 6 dimethylmorpholino)cyclohexyl)ami 9.36 (s, 1H), 9.14 (s, 1H), 7.70 - 7.59no)-2- (m, 3H), 7.53 (d, J = 14.9 Hz, 1H), 7.10 (trifluoromethoxy)phenyl)amino)py (dd, J = 8.9, 1.5 Hz, 1H), 6.46 (dd, J =rido[4,3-d]pyrimidin-7-yl)ethene-l- 9.0, 2.8 Hz, 1H), 3.70 (dqd, J = 12.5, sulfonamide 6.1, 1.9 Hz, 2H), 3.23 (tt, J = 11.1, 3.89 3 >H^ 611.2 [M+H]+Hz, 1H), 2.88 (d, J = 11.2 Hz, 2H), 2.38- 2.27 (m, 3H), 2.09 (d, J = 12.4 Hz, 2H),1.95 (q, J = 10.7 Hz, 2H), 1.51 - 1.40 (m,2H), 1.33 - 1.27 (m, 2H), 1 17 (d, J = 6.3Hz, 6H). 19F NMR (471 MHz, Methanol-d4) 6 -5998.Example # Structure Name NMR LCMS (E)-2-(2-((5-(((lR,4s)-4-((2SR,6SR)- 1H NMR (500 MHz, Methanol-d4) 52,6- 9.38 (s, 1H), 9.15 (s, 1H), 7.73 (s, 1H),A dimethylmorpholino)cyclohexyl)ami 7.68 (d, J = 14.9 Hz, 1H), 7.62 (s, 1H), Q no)-2- 7.53 (d, J = 14.9 Hz, 1H), 7.14 (dt, J = (trifluoromethoxy)phenyl)amino)py 9.0, 1.4 Hz, 1H), 6.54 (dd, J = 8.9, 2.8rido[4,3-d]pyrimidin-7-yl)ethene-l- Hz, 1H), 3.90 (ddd, J = 11.0, 6.2, 1.8 Hz,10 sulfonamide 2H), 3.73 (t, J = 3.1 Hz, 1H), 3.49 (d, J = 611.2 [M+H]+12.1 Hz, 3H), 2.82 (t, J = 11.6 Hz, 2H),2.27 (d, J = 13.8 Hz, 2H), 2.03 (d, J =11.8 Hz, 2H), 1.95 (q, J = 12.1 Hz, 2H),1.81 (t, J = 13.6 Hz, 2H), 1.29 (d, J = 6.2Hz, 6H). 19F NMR (471 MHz, Methanol- d4) 6 -6003, -77.04.(S, E)-2-(2-((5-(2-(pyrrolidin-l- 1H NMR (400 MHz, Methanol-d4) 5 ylmethyl)pyrrolidin-l-yl)-2- 9.38 (s, 1H), 9.15 (s, 1H), 7.73 - 7.64 (trifluoromethoxy)phenyl)amino)py (m, 3H), 7.54 (d, J = 15.0 Hz, 1H), 7.28- rido[4,3-d]pyrimidin-7-yl)ethene-l- 7.22 (m, 1H), 6.55 (dd, J = 9.1, 3.0 Hz,p sulfonamide 1H), 4.29-4.16 (m, 1H), 390 (dd, J = 11 13.5, 6.7 Hz, 1H), 3.72 (d, J = 11.8 Hz, 564.30, [M+H]+1H), 3.62 (d, J = 8.4 Hz, 1H), 3.34 (s,1H), 3.24-3.11 (m, 2H), 2 18 (dq, J =18.6, 11.9, 9.1 Hz, 5H), 2.08 - 1.96 (m,3H). 19F NMR (376 MHz, Methanol-d4)6-59.99, -77.14.(S, E)-2-(2-((5-(3-(hydroxymethyl)-4 ’H NMR (300 MHz, Methanol-d4) 9.37 methylpiperazin-l-yl)-2- (s, J = 0.8 Hz, 1H), 9.13 (s, 1H), 8.17 (s, (trifluoromethoxy)phenyl)amino)py 1H), 7.72 - 7.61 (m, 2H), 7.54 (d, J =AW rido[4,3-d]pyrimidin-7-yl)ethene-l- 15.0 Hz, 1H), 7.25 (dd, J = 9.0, 1.6 Hz, sulfonamide 1H), 6.82 (dd, J =9.1, 3.0 Hz, 1H), 3.9412 540.2, [M+H]+ f Xx O A - 3 82 (m, 2H), 3.61 (dd, J = 11.2, 7.1z<< ■• Hz, 2H), 2.96 (t, J = 10.3 Hz, 2H), 2.82- 2 65 (m, 2H), 2.59 - 2.45 (m, 2H),2.42 (s, 3H). 19F NMR (282 MHz,Methanol-d4) 6 -59.76.(E)-2-(2-((5-(((ls,4s)-4-(pyrrolidin- 1H NMR (400 MHz, Methanol-d4) 8AA^ 'P l-yl)cyclohexyl)amino)-2- 9.35 (s, 1H), 9.13 (s, 1H), 7.70 - 7.60 (trifluoromethoxy)phenyl)amino)py (m, 3H), 7.51 (d, J = 15.0 Hz, 1H), 7.11rido[4,3-d]pyrimidin-7-yl)ethene-l- (d, J = 8.9 Hz, 1H), 6.51 (dd, J = 9.0, 2.8 X o sulfonamide Hz, 1H), 3.71 -3 60 (m, 4H), 3.23 - 13 578.2 [M+H]+3.08 (m, 4H), 2.19 -2.13 (m, 5H), 1.97-2.05 (m, 5H), 1.95 - 1.88 (m, 2H),1.81 (q, J = 13.0, 11.8 Hz, 2H). 19F NMR(376 MHz, Methanol-d4) 6 -59.99, - 76.93(E)-2-(2-((5-((2-(piperidin-l- 1H NMR (400 MHz, Methanol-d4) 6 yl)ethyl)amino)-2- 9.37 (s, 1H), 9.14 (s, 1H), 7.78 (s, 1H), (trifluoromethoxy)phenyl)amino)py 7.66 (d, J = 15.0 Hz, 2H), 7.52 (d, J =rido[4,3-d]pyrimidin-7-yl)ethene-l- 14.8 Hz, 1H), 7.19 (d, J = 10.1 Hz, 1H), sulfonamide 6.55 (dd, J = 8.9, 2.8 Hz, 1H), 3.62 (dd, J14 538.2, [M+H]+= 12.0, 5.5 Hz, 4H), 3.38 (t, J = 5.9 Hz,2H), 3.09-2.99 (m, 2H), 199 (d, J =15.8 Hz, 2H), 1.81 (d, J = 15.7 Hz, 2H),1.62 - 1.52 (m, 2H). 19F NMR (376MHz, Methanol-d4) 6 -60.01, -77.00.Example # Structure Name NMR LCMS 0 o (E)-2-(2-((5-(((ls,4s)-4-(4,4- 1H NMR (300 MHz, Methanol-d4) 5dimethylpiperidin-1- 9.38 (s, 1H), 9.15 (s, 1H), 7.74 - 7.62 yl)cyclohexyl)amino)-2- (m, 3H), 7.53 (d, J = 15.0 Hz, 1H), 7.13 (trifluoromethoxy)phenyl)amino)py (dd, J = 8.9, 1.5 Hz, 1H), 6.50 (dd, J =rido[4,3-d]pyrimidin-7-yl)ethene-l- 8.9, 2.8 Hz, 1H), 3.49 - 3.38 (m, 2H), 15 5 o°* z sulfonamide 3.27-3.15 (m, 3H), 2.40 (d, J = 13.0 Hz, 620.2, [M+H]+ \ C 2H), 2.27 (d, J = 12.3 Hz, 2H), 1.80- 1.66 (m, 6H), 1.47-1.31 (m, 3H), 1.10(d, J = 6.6 Hz, 6H). 19F NMR (282 MHz, Methanol-d4) 6 -60.00, -77.09.(E)-2-(2-((5-(((lr,4r)-4-(4,4- 1H NMR (300 MHz, Methanol-d4) 5dimethy lpiperidin-1- 9.38 (s, 1H), 9.15 (s, 1H), 7.74 (s, 1H), yl)cyclohexyl)amino)-2- 7.72-7.61 (m, 2 H ),7.53 (d, J = 14.9 Hz, (trifluoromethoxy)phenyl)amino)py 1H), 7.14 (d, J = 8.9 Hz, 1H),6.53 (d, J =rido[4,3-d]pyrimidin-7-yl)ethene-l- 9.0 Hz, 2H), 3.70 (d, J = 20.0 Hz, 2H),16 sulfonamide 3.48-3.41 (m, 2H), 3.25-3.14 (m, 2H), 620.2, [M+H]+3.12-3.05 (m, 1H), 2.26 (d, J = 13.5 Hz,2H), 2.10-2.00 (m, 4H), 183 (d, J =12.7 Hz, 2H), 1.74 (s, 4H), 1.32 (s, 2H),1.10 (d, J = 6.5 Hz, 6H). 19F NMR (282MHz Mpthannl-d4) 6-60 0? -7706(E)-2-(2-((5-(((ls,4s)-4-(4- 1H NMR (300 MHz, Methanol-d4) 5 acetylpiperazin-1- 9.37 (d, J = 2.1 Hz, 1H), 9.14 (d, J =3.4 yl)cyclohexyl)amino)-2- Hz, 1H), 7.82 (s, 1H), 7.70 - 7.50 (m, (trifluoromethoxy)phenyl)amino)py 3H), 7.13-7.07 (m, 1H), 648 (ddd, J =rido[4,3-d]pyrimidin-7-yl)ethene-l- 19.3, 8.9, 2 7 Hz, 1H), 4.86 (dd, J = 5.6, sulfonamide 3.0 Hz, 1H), 3.65 -3.55 (m, 5H), 3 29 - 17 3.18 (m, 1H), 2.66 (dt, J = 19.5, 5.2 Hz, 635.2, [M+H]+ Gc'rS _> i °‘1H \ — f \ — / o 4H), 2.51-2.41 (m, 1H), 2 36 -2.29 < ix ^ / (m, 2H), 2.12 (d, J = 5.1 Hz, 3H), 2.07 (d,J = 10.5 Hz, 2H), 1.79 - 1.70 (m, 2H), 0 • 1.58-1.44 (m, 2H), 1.38-1.27 (m,3H). 19F NMR (282 MHz, Methanol-d4)6-59.99(E)-2-(8-methoxy-2-((5-(4-1H NMR (300 MHz, Methanol-d4) 9.36 methylpiperazin-l-yl)-2- (s, 1H), 889 (s, 1H), 8.18 (s, 1H), 7.90 (trifluoromethoxy)phenyl)amino)py (d, J = 15.1 Hz, 1H), 7.62 (d, J = 15.1 Hz, rido[4,3-d]pyrimidin-7-yl)ethene-l- 1H), 7.33 - 7.20 (m, 1H), 6.82 (dd, J =18 540.1, [M+H]+ 0 sulfonamide 9.1, 3.0 Hz, 1H), 4.15 (s, 3H), 3.33 (s,4H), 2.66 (t, J = 5.0 Hz, 4H), 2.38 (s,3H). 19F NMR (282 MHz, Methanol-d4)6-59.79.(E)-2-(2-((5-((2-(pyrrolidin-l- 1H NMR (400 MHz, Methanol-d4) 6 yl)ethyl)amino)-2- 9.26 (d, J = 0.7 Hz, 1H), 9.03 (s, 1H), (trifluoromethoxy)phenyl)amino)py 7.73 -7.67 (m, 1H), 7.61 -7.56 (m,rido[4,3-d]pyrimidin-7-yl)ethene-l- 2H), 7.54-7.50 (m, 1H), 747 -7.39 sulfonamide (m, 2H), 7.04 - 7.00 (m, 1H), 6.37 (dd, J19 = 8.9, 2.8 Hz, 1H), 4.00 (dt, J = 14.1, 7.1 524.2, [M+H]+Hz, 1H), 3.27-3 23 (m, 2H), 2.70 (t, J =7.0 Hz, 2H), 2.57 (t, J = 5.2 Hz, 4H), 1.80-1.70 (m, 4H), 1.28 (t, J = 7.0 Hz, 1H),1.19 (s, 1H). 19F NMR (376 MHz,Methanol-d4) 6 -59.98. -76.92.EXAMPLE 20Intermediate A DIEA. DMSO, 100 °CUSO4. NalOd, THE H2O, rt, 1 h Step 3HCHO STAB, ft Step 6Example 20Step 1; To a stirred mixture of tert-butyl 4-[3-amino-4-(trifluoromethoxy)phenyl]piperazine-l-carboxylate (1.0g, 2.89 mmol) and 2,7-dichloropyrido[4,3-d]pyrimidine (720.1 mg, 3.60 mmol) in DMSO (8 mL) was added DIPEA (933.1 mg, 7.22 mmol) in portions at room temperature under nitrogen atmosphere. The resulting mixture was stirred for 2 h at 100 °C. The desired product could be detected by LCMS. The resulting mixture was diluted with water (50 mL). The resulting mixture was extracted with EtOAc (50 mL x 3 times). The combined organic layers were washed with brine (150 mL) and dried over anhydrous Na2SO4. The fdtrate was concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: EtOAc / PE = 1 / 6, v / v) to afford tert-butyl 4-[3-({7-chloropyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]piperazine-l -carboxylate as a yellow oil (400.0 mg, 26.4% yield), m / z (ESI) calcd. for C23H24ClF3N6O3: 524.2, found: 525.1 [M+H]+.Step 2: To a mixture of tert-butyl 4-[3-({7-chloropyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]piperazine-l -carboxylate (230.1 mg, 0.44 mmol), Pd(dppf)C12 (57.0 mg, 0.09 mmol), K3PO4 (186.0 mg, 0.88 mmol) was added 2-ethenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (270.0 mg, 1.75 mmol) in dioxane (9.2 mL) and H2O (2.3 mL) dropwise at roomtemperature under nitrogen atmosphere. The resulting mixture was stirred overnight at 80 °C under a nitrogen atmosphere. The desired product could be detected by LCMS. The reaction was quenched with Water at room temperature. The resulting mixture was extracted with CH2CI2 (200 mL x 3 times). The combined organic layers were washed with brine (500 mL), dried over anhydrous Na2SC>4, and concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: EtOAc / PE = 1 / 1, v / v) to afford tert-butyl 4-[3-({7-ethenylpyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]piperazine-l -carboxylate as a yellow oil (180.0 mg, 75.6% yield), m / z (ESI) calcd. for C25H27F3N6O3: 516.2, found: 517.2 [M+H]+.Step 3: Into an 8 mL vial were added tert-butyl 4-[3-({7-ethenylpyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]piperazine-l -carboxylate (350.0 mg, 0.68 mmol), THF (7 mL) and H2O (2.3 mL) at room temperature. Then the above mixture was added OsCL (690 uL, 0.14 mmol) and NalCL (870.0 mg, 4.07 mmol) at 0 °C. The final reaction mixture was stirred for 1 h at room temperature. The desired product could be detected by LCMS. The resulting mixture was diluted with ice water (100 mL). The resulting mixture was extracted with CHCI3 (100 mL x 3 times). The combined organic layers were washed with brine (300 mL), dried over anhydrous Na2SC>4, and concentrated under reduced pressure. The residue was eluted with ACN / H2O (0.05%TFA) (from 10% to 50% in 30 min) from a C18 column to afford tert-butyl 4-[3-({7-formylpyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]piperazine-l-carboxylate as a brown oil (220.0 mg, 50.1% yield), m / z (ESI) calcd. for C24H25F3N6O4: 518.2, found: 519.3 [M+H]+.Step 4; Into a 40 mL vial were added tert-butyl N-(diphenylphosphoroso) methanesulfonylcarbamate (160.0 mg, 0.41 mmol) and DMF (10 mL) at room temperature. NaH (41.0 mg, 1.71 mmol) was added to the above mixture at 0 °C. The mixture was stirred for 30 min at room temperature under a nitrogen atmosphere. Then to the stirred mixture was added tert-butyl 4-[3-({7-formylpyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]piperazine-l -carboxylate (210.1 mg, 0.41 mmol). The final reaction mixture was stirred for 2 h at room temperature. The desired product could be detected by LCMS. The resulting mixture was diluted with water (100 mL). The resulting mixture was extracted with EtOAc (100 mL x 3 times). The combined organic layers were washed with brine (200 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residuewas eluted with ACN / H2O (0.05%TFA) (from 50% to 90% in 30 min) from a C18 column to afford tert-butyl 4-[3-({7-[(E)-2-[(tert-butoxycarbonyl)aminosulfonyl]ethenyl]pyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]piperazine-l -carboxylate (190.0 mg, 64.7% yield) as a yellow powder, m / z (ESI) calcd. for C30H36F3N7O7S: 695.2, found: 696.3 [M+H]+.Step 5: To the mixture of tert-butyl 4, -[3 -({7-[(E)-2- [(tertbutoxy carbonyl)aminosulfonyl]ethenyl]pyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]piperazine-l -carboxylate (100.0 mg, 0.14 mmol) in DCM (3 mL) was added TFA (0.6 mL). The mixture was stirred at room temperature for Ih. The desired product could be detected by LCMS. The mixture was concentrated. The residue was dissolved by DCM (15 mL) and washed by sat. aq. NaHCO₃. The H2O layer was extracted by CHCl₃ / iPrOH (3 / 1; 10 mL x 3 times). The organic layer was combined, dried over Na2SC>4, filtered, and concentrated to afford (E)-2-(2-{[5-(piperazin-l-yl)-2-(trifluoromethoxy)phenyl]amino}pyrido[4,3-d]pyrimidin-7-yl)ethenesulfonamide (70.0 mg, 98.3% yield) as yellow solid, m / z (ESI) calcd. for C20H20F3N7O3S: 495.1, found: 496.1 [M+H]+.Step 6; To the mixture of (E)-2-(2- {[5 -(piperazin- 1 -yl)-2-(trifluoromethoxy)phenyl]amino}pyrido[4,3-d]pyrimidin-7-yl)ethenesulfonamide (40.0 mg, 0.08 mmol) in CH2CI2 (3 mL) was added HCHO (5.0 mg, 0.16 mmol), Sodium triacetoxyborohydride (26.0 mg, 0.12 mmol) and CH3COOH (0.01 mL, 0.16 mmol). The mixture was stirred at room temperature for 2 h. The desired product could be detected by LCMS. The mixture was added with H2O (10 mL) and extracted by DCM (5 mL x 3 times). The organic layer was combined, dried over Na2SO4, filtered, and concentrated. Prep-HPLC purified the crude product with the following conditions (Column: XSelect CSH C18 Column, 19*250mm, 5pm; Mobile Phase A: 0.05% TFA aqueous solution, Mobile Phase B: ACN; Flow rate: 25 mL / min; Gradient: 13%B to 33%B in 10 min; Wave Length: 254nm / 220nm nm; RT: 9.63 min) to afford (E)-2-(2-{[5-(4-methylpiperazin-l-yl)-2-(trifluoromethoxy)phenyl]amino}pyrido[4,3-d]pyrimidin-7-yl)ethenesulfonamide (5.8 mg, 13.0% yield) as a yellow solid, m / z (ESI) calcd. for C21H22F3N7O3S: 509.1, found: 510.1 [M+H]+.1HNMR(400 MHz, DMSO-d6) 89.42 (s, IH), 9.18 (s, IH), 7.68 (s, IH), 7.59 - 7.44 (m, 3H), 7.33 - 7.31 (m, IH), 6.95 - 6.92 (m, IH), 3.86 -3.81 (m, 2H), 3.52 (d, J = 12.0 Hz, 2H), 3.17 (t, J = 11.9 Hz, 2H), 3.02 (t, J = 12.6 Hz, 2H), 2.85 (s, 3H).19F NMR (376 MHz, DMSO-d6) 8 -56.8.Following the teachings of the General Reaction Schemes and the synthesis procedure for Example 20 were prepared as shown in Table II:TABLE IIExample # Structure Name NMR LCMS (E)-2-(2-((5-(4-(2- 1H NMR (300 MHz, DMSO-d6) 69.90 hydroxyethyl)piperazin-l-yl)-2- (s, 1H), 9.75 (s, 1H), 9.46 (s, 1H), 9.20 (trifluoromethoxy)phenyl)amino)py (s, 1H), 7.74 (s, 1H), 7.55 - 7.50 (m,rido[4,3-d]pyrimidin-7-yl)ethene-l- 2H), 7.47 (d, J = 15.0 Hz, 1H), 7.37 - 21 ^6 sulfonamide 7.28 (m, 1H), 7.27 (s, 2H), 6.98 - 6.91 540.1 [M+H] +(m, 1H), 5.43 (s, 1H), 3.93 - 3.73 (m,4H), 3.61 (s, 2H), 3.31 -3.06 (m, 6H).19F NMR (376 MHz, DMSO-d6) 6 -56.8.( E)-2-(2-( (5-(4-ethy Ipiperazin- 1-yl)- 1H NMR (400 MHz, DMSO-d6) 89.442- (d, J = 3.7 Hz, 1H), 9.19 (d, J = 2.2 Hz,ppi (trifluoromethoxy)phenyl)amino)py 1H), 7.73-7.67 (m, 1H), 7.60- 7.55 (m,rido[4,3-d]pyrimidin-7-yl)ethene-l- 2H), 7.47 - 7.44 (m, 1H), 7.34- 7.32 (m,22 sulfonamide 1H), 6.96 - 6.93 (m, 1H), 3.90- 3.76 (m, 524.0 [M+H]+4H), 3.21 - 3.00 (m, 6H), 1.26 (t, J = 7.3Hz, 3H). 19F NMR (376 MHz, DMSO-d6)6 -56.8.(E)-2-(2-((5-(4-methyl-2- 1H NMR (400 MHz, DMSO-d6) 89.46oxopiperazin-l-yl)-2- (s, 1 H), 9.20 (s, 1 H), 7.87 (d, 1 H), 7.73 (trifluoromethoxy)phenyl)amino)py (s, 1 H), 7.62 (s, 1 H), 7.43 (m, 2 H),23 524.3 [M+H]+FVos-^- rido[4,3-d]pyrimidin-7-yl)ethene-l- 7.32 (m, 1 H), 3.75 - 3.68 (m, 2 H), 3.14FA L 1 sulfonamide (s, 2 H), 2.75 (m, 2 H), 2.29 (s, 3 H). 19FNMR (376 MHz, DMSO-d6) 6 -56.42.1H NMR (300 MHz, DMSO-d6) 89.42 (E)-2-(2-((5-(4-methyl-l,4-diazepanl-yl)-2- (s, 1H), 9.12 (s, 1H), 7.66 (s, 1H), 7.61 - (trifluoromethoxy)phenyl)amino)py 7.49 (m, 2H), 7.21 - 7.17 (m, 2H), 6.62 - rido[4,3-d]pyrimidin-7-yl)ethene-l- 6.58 (m, 1H), 3.60 - 3.51 (m, 2H), 2.47 - sulfonamide 3.43 (m, 2H), 2.65 - 2.62 (m, 2H), 2.48 - 24 524.1 [M+H]+ %d'b2.46 (m, 2H), 2.26 (s, 3H), 1.91 (t, J = \--N 8.4 Hz, 2H). 19F NMR (282 MHz, DMSO- d6) 8-56.95. 19F NMR(376 MHz,Methanol-d4) 6-59.74.Example 25Step 1: To a solution of 4-bromo-2-nitro-l-(trifluorom ethoxy )benzene (4.1 g, 14.01 mmol) in 1,4-di oxane (200 mL) were sequentially added tert-butyl 4- (3 -aminopropyl) piperazine- 1-carboxylate (5.0 g, 20.52 mmol), Ruphos PdG4 (584.2 mg, 0.71 mmol), Ruphos (326.1 mg, 0.71 mmol), and cesium carbonate (9.1 g, 27.72 mmol). Then the mixture was bubbled with N2 flow for 5 minutes to expel air gas in the flask, after which the mixture was stirred at 90 °C under N2 atmosphere for 16 h. After being cooled to room temperature, insoluble solids were removed by filtration, and the filtrate was diluted with water (250 mL) then extracted with ethyl acetate (3x500 mL). Organic phases were combined and dried over anhydrous MgSO4, followed by concentration under reduced pressure. The resultant residue was purified with a silica-gel column (eluent: DCM / MeOH = 10 / 1, v / v) to yield tert-butyl 4-(3-((3-nitro-4-(trifluoromethoxy)phenyl)amino)propyl)piperazine-l -carboxylate as yellow oil (4.4 g, 70% yield), m / z (ESI) calcd. for C19H27F3N4O5: 448.19, found: 449.19, [M+H]+.Step 2; tert-butyl 4-(3-((3-nitro-4-(trifluoromethoxy)phenyl)amino)propyl)piperazine-l-carboxylate (4.4 g, 9.81 mmol) was dissolved in DCM (150 mL), then trifluoroacetic acid (30mL) was added thereto. The reaction mixture was stirred at room temperature for 0.5 h, when the desired product was almost quantitatively generated as per LCMS analysis. Thereafter, the reaction solution was concentrated under reduced pressure to afford trifluoroacetate, (E)-2-((5-(piperazin-l-yl)-2-(trifluoromethoxy)phenyl)amino)-7-(2-sulfamoylvinyl)pyrido[4,3-d]pyrimidine 6-oxide as yellow solids (5.4 g), which was used in the next step without further purification, m / z (ESI) calcd. for C14H19F3N4O3: 348.14, found: 349.14, [M+H]+.Step 3; The above obtained trifluoroacetate, (E)-2-((5 -(piperazin- l-yl)-2-(trifluoromethoxy)phenyl)amino)-7-(2-sulfamoylvinyl)pyrido[4,3-d]pyrimidine 6-oxide (5.4 g, 9.81 mmol)was dissolved in pyridine (120 mL), then trifluoroacetic anhydride (7.2 g, 34.11 mmol) was added thereto. The mixture was stirred at room temperature for 1 hour, then concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: PE / EA= 1 / 1, v / v) to yield 2,2,2-trifluoro-N-(3-nitro-4-(trifluoromethoxy)phenyl)-N-(3-(4-(2,2,2-trifluoroacetyl)piperazin-l-yl)propyl)acetamide as yellow solid (2.3 g, 43% overall yield of two steps), m / z (ESI) calcd. for C18H17F9N4O5: 540.11, found: 541.11, [M+H]+.Step 4: 2,2, 2-trifhioro-N-(3-nitro-4-(tri fluoromethoxy )phenyl)-N-(3-(4-(2, 2,2-trifhioroacetyl)piperazin-l-yl)propyl)acetamide (2.2 g, 4.16 mmol) and B2(OH)4(1.8 g, 20.81 mmol) were dissolved in DMF (N, N-dimethylamine; 50 mL), which was then cooled to 0 °C before slow addition of DMF solution (0.5 mL) of 1,4-bipyridine (6.5 mg, 0.04 mmol). The mixture was stirred at room temperature for 30 minutes, then diluted with water and extracted with ethyl acetate (550 mL x 3 times). The combined organic layer was dried over anhydrous MgSO4and concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: CH2C12 / EA= 10 / 1, v / v) to yield N-(3-amino-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-(3-(4-(2,2,2-trifluoroacetyl)piperazin-l-yl)propyl)acetamide as yellow solid (1.1 g, 50% yield), m / z (ESI) calcd. for C18H19F9N4O3: 510.13, found: 511.13, [M+H]+.Step 5: The mixture of N-(3-amino-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-(3-(4-(2,2,2-trifluoroacetyl)piperazin-l-yl)propyl)acetamide (860.1 mg, 1.68 mmol), 2,7-dichloropyrido [4,3-d] pyridine (671.3 mg, 3.37 mmol) and z-PrOH (20 mL) was stirred at 120 °C overnight. After being cooled to room temperature, the mixture was concentrated under reduced pressure, and the residue was purified with a silica gel column (eluent: CH2CI2 / EA = 2 / 1, v / v) to yield N-(3-((7-chloropyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-(3-(4-(2,2,2-trifluoroacetyl)piperazin-l-yl)propyl)acetamide as yellow solid (600.1 mg, 52% yield), m / z (ESI) calcd. for C25H21CIF9N7O3: 673.13, found: 674.13, [M+H]+.Step 6 N-(3-((7-chloropyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-(3-(4-(2,2,2-trifluoroacetyl)piperazin-l-yl)propyl)acetamide (600.1 mg, 0.89 mmol), Pd(dppf)Ch (72.2 mg, 0.08 mmol) and K3PO4 (253.8 mg, 1.78 mmol) were loaded into a singleneck round-bottom flask, followed by addition of solvent mixture of 1,4-dioxane and H₂O (20 mL, v / v = 5 / 1). The flask was capped by a septum, and the mixture was bubbled with N2 flow through a needle for 10 minutes, after which 4,4,5,5-tetramethyl-2-vinyl-l,3,2-dioxaborolane (411.3 mg, 2.67 mmol) was injected using a syringe. After being stirred at 90 °C for 3 h, the reaction mixture was diluted with water and extracted with ethyl acetate (100 mL x 3 times). The combined organic layer was dried over anhydrous MgSO₄ and concentrated under reduced pressure; and the residue was purified with a silica-gel column (eluent: DCM / MeOH = 10 / 1, v / v) to yield N1-(3-(piperazin-l-yl)propyl)-4-(trifluoromethoxy)-N3-(7-vinylpyrido[4,3-d]pyrimidin-2-yl)benzene-l,3-diamine as yellow solid (180.3 mg, 30% yield), m / z (ESI) calcd. for C23H26F3N7O: 473.22, found: 474.22, [M+H]+.Step 7: N1-(3-(piperazin-l-yl)propyl)-4-(trifluoromethoxy)-N3-(7-vinylpyrido[4,3-d]pyrimidin-2-yl)benzene-l,3-diamine (170.2 mg, 0.25 mmol) was dissolved in THF (tetrahydrofuran; 4 mL), then OsO4(14.8 mg, 0.05 mmol) solution in THF (1 mL) was added thereto, followed by addition of water (5 mL) and sodium periodate (218.1 mg, 1.01 mmol). The mixture was stirred at room for 1 hour and subsequently extracted with ethyl acetate (100 mL x 3 times). The combined organic layer was dried over anhydrous MgSCU and concentrated under reduced pressure to yield 2-((5-((3-(piperazin-l-yl)propyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidine-7-carbaldehyde as yellow solid (68.3 mg, 40% yield), which was used in the next step without further purification, m / z (ESI) calcd. for C22H24F3N7O2: 475.19, found: 476.19, [M+H]+.Step 8: Tert-butyl (((diphenylphosphoryl)methyl)sulfonyl)carbamate (53.2 mg, 0.13 mmol) was dissolved in DMF (5 mL), and NaH (60wt%; 21.6 mg, 0.91 mmol) was then added. The mixture was stirred at room temperature for 30 minutes, after which 2-((5-((3-(piperazin-l-yl)propyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidine-7-carbaldehyde (60.1 mg, 0.08 mmol) was introduced thereto, followed by additional 1-hour stirring at room temperature. Afterward, the reaction mixture was quenched by slow addition of water (0.5 mL), then extracted with ethyl acetate (50 mL * 3 times). The combined organic layer was dried over anhydrous MgSO4and concentrated under reduced pressure, and the residue was purified with a silica-gel column (eluent: DCM / MeOH = 10 / 1, v / v) to yield tert-butyl (E)-((2-(2-((5-((3-(piperazin-l-yl)propyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (40.2 mg, 68% yield) as yellow solid, which was used in the next step without further purification, m / z (ESI) calcd. for C28H35F3N8O5S: 652.24, found: 653.24, [M+H]+.Step 9; Tert-butyl (E)-((2-(2-((5-((3-(piperazin-l-yl)propyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (40.1 mg, 0.06 mmol) was dissolved in DMF (4 mL), followed by addition of formaldehyde (40% aqueous solution; 9.5 mg, 0.27 mmol). After being stirred for 10 minutes, NaBH(AcO)3 (42.5 mg, 0.20 mmol) was added thereto. The mixture was stirred at room temperature for about 0.5 h then diluted with water and extracted with DCM / MeOH (10 / 1, v / v). The combined organic phase was dried over anhydrous MgSO4 and concentrated under reduced pressure, the residue was purified with a silica-gel column (eluent: DCM / MeOH = 4 / 1, v / v) to yield tert-butyl (E)-((2-(2-((5-((3-(4-methylpiperazin-l-yl)propyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (30.1 mg, 75% yield) as yellow solid, m / z (ESI) calcd. for C29H37F3N8O5S: 666.72, found: 667.72, [M+H]+.Step 10: Tert-butyl (E)-((2-(2-((5-((3-(4-methylpiperazin-l-yl)propyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (30.1 mg, 0.05 mmol) was dissolved in DCM (dichloromethane; 2.5 mL), then trifluoroacetic acid (0.5 mL) was added thereto. The reaction mixture was stirred at room temperature for 0.5 h, then concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography (column: Xselect CSH C18 OBD column, 30*150 mm; mobile phase A: 0.05% TFA aqueous solution, mobile phase B: ACN; flow rate: 60 mL / min; gradient: 3% B to 32% B in 11 min; wave length: 254 nm; RT: 9.5 min) to obtain trifluoroacetate, (E)-2-(2-((5-((3-(4-methylpiperazin-l-yl)propyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)ethene-l -sulfonamide as yellow solids (1.1 mg, 4.4% yield), m / z (EST) calcd. for C24H29F3N8O3S: 566.72, found: 567.72, [M+H]+.1H NMR (300 MHz, Methanol-d4) δ 9.38 (s, 1H), 9.16 (s, 1H), 7.75-7.61 (m, 3H), 7.54 (d, J = 15.0 Hz, 1H), 7.15 (d, J = 8.5 Hz, 1H), 6.51 (dd, J = 8.9, 2.7 Hz, 1H), 3.67 (s, 2H), 3.26 (d, J = 15.4 Hz, 6H), 2.94-2.82 (m, 4H), 2.80 (s, 3H), 1.98 (s, 2H).19F NMR (376 MHz, Methanol-d4) 8 -59.99, -77.06.Example 26A I STAB, CH2CI2, AcOH, rt Step 6Example 26Step 1: To a stirred mixture of tert-butyl N-[3-amino-4-(trifluoromethoxy)phenyl]carbamate (200.0 mg, 0.68 mmol) and 2,7-dichloropyrido[4,3-d]pyrimidine (136.8 mg, 0.68 mmol) in 1,4-dioxane (5 mL) was added TsCl (156.5 mg, 0.82 mmol) dropwise at room temperature under nitrogen atmosphere. The resulting mixture was stirred at 50 °C for 2 h under nitrogen atmosphere. The reaction was monitored by LCMS. Desired product could be detected by LCMS. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with EtOAc (10 mL x 3 times). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SC>4. After fdtration, the filtrate was concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: EtOAc / PE = 1 / 5, v / v) to afford tert-butyl N-[3-({7-chloropyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]carbamate (90.0 mg, 28.9% yield) as a white solid, m / z (ESI) calcd. for C19H17CIF3N5O3: 455.1, found: 456.0 [M+H]+.Step 2; To a stirred solution of tert-butyl N-[3-({7-chloropyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]carbamate (80.0 mg, 0.18 mmol) and 2-ethenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (32.4 mg, 0.21 mmol) in 1,4-dioxane (4 mL) and H2O (1 mL) were added Pd(dppf)C12 (11.4 mg, 0.02 mmol) and K3PO4 (111.7 mg, 0.53 mmol) in portions at roomtemperature under nitrogen atmosphere. The resulting mixture was stirred at 80 °C for 2 h under nitrogen atmosphere. The reaction was monitored by LCMS. Desired product could be detected by LCMS. The resulting mixture was diluted with water (10 mL). The resulting mixture was extracted with EtOAc (10 mL x 3 times). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: EtOAc / PE = 1 / 5, v / v) to afford tert-butyl N-[3-({7-ethenylpyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]carbamate (40.0 mg, 50.6% yield) as a yellow oil. m / z (ESI) calcd. for C21H20F3N5O3: 447.1, found: 448.2 [M+H]+.Step 3: To a stirred solution of tert-butyl N-[3-({7-ethenylpyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]carbamate (540.0 mg, 1.21 mmol) in tetrahydrofuran (10 mL), water (3.3 mL) was added sodium periodate (1548.9 mg, 7.24 mmol) and OsO4 (61.4 mg, 0.24 mmol) in portions at 0 °C under air atmosphere. The resulting mixture was stirred for 2 h at room temperature under air atmosphere. The reaction was monitored by LCMS. Desired product could be detected by LCMS. EtOAc (20 mL) and H2O (20 mL) were added to the mixture. After separation of phases, the aqueous phase was extracted with EtOAc (20 mL x 2 times). The combined organic phases were washed with brine (50 mL x 2 times), dried with Na2SO4, filtered and concentrated to afford tert-butyl N-[3-({7-formylpyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]carbamate (500.0 mg, 92.6% yield) as a yellow solid crude, m / z (ESI) calcd. for C20H18F3N5O4: 449.1, found: 450.1 [M+H]+.Step 4; To a stirred mixture of tert-butyl N-(diphenylphosphoroso)methanesulfonylcarbamate (439.9 mg, 1.11 mmol) in DMF (20 mL) was added NaH (111.2 mg, 2.78 mmol, 60%wt) and the mixture was stirred for 30 min at room temperature. To the above mixture was added tert-butyl N-[3-({7-formylpyrido[4,3-d]pyrimidin-2-yl}amino)-4-(trifluoromethoxy)phenyl]carbamate (500.0 mg, 1.11 mmol) and the mixture was stirred for 4 h at room temperature. The reaction was monitored by LCMS. Desired product could be detected by LCMS. DCM (20 mL) and H2O (20 mL) were added to the mixture. After separation of phases, the aqueous phase was extracted with DCM (20 mL x 2 times). The combined organic layers were washed with brine (50 mL x 3 times), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was eluted with ACN / H2O (0.05%TFA) (from 50% to 100% in 30 min)from a Cl 8 column to afford tert-butyl N-[(E)-2-[2-({5-[(tert-butoxycarbonyl)amino]-2-(trifluoromethoxy)phenyl}amino)pyrido[4,3-d]pyrimidin-7-yl]ethenesulfonyl]carbamate (600.0 mg, 86.0% yield) as a yellow solid, m / z (ESI) calcd. for C26H29F3N6O7S: 626.2, found: 627.1 [M+H]+.Step 5: To a stirred solution of tert-butyl N-[(E)-2-[2-({5-[(tert-butoxycarbonyl)amino]-2-(trifluoromethoxy)phenyl}amino)pyrido[4,3-d]pyrimidin-7-yl]ethenesulfonyl]carbamate (600.0 mg, 0.96 mmol) in DCM (10 mL) was added TFA (5 mL) drop wise and the mixture was stirred for 3 h at room temperature. The reaction was monitored by LCMS. Desired product could be detected by LCMS. The mixture was basified to pH 8 with NH3·H2O. The resulting mixture was extracted with CHCI3 (50 mL x 3 times). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The resulting mixture was concentrated under reduced pressure to afford (E)-2-(2-{[5-amino-2-(trifluoromethoxy)phenyl]amino}pyrido[4,3-d]pyrimidin-7-yl)ethenesulfonamide (360.0 mg, 88.2% yield) as a yellow solid, m / z (ESI) calcd. for C16H13F3N6O4S: 426.1, found: 427.1 [M+H]+.Step 6; To a stirred solution of (E)-2-(2-{[5-amino-2-(trifluoromethoxy)phenyl]amino}pyrido[4,3-d]pyrimidin-7-yl)ethenesulfonamide (25.0 mg, 0.06 mmol) and 3-(dimethylamino)-2,2-dimethylpropanal (11.3 mg, 0.09 mmol) in DCM (1 mL) were added STAB (24.8 mg, 0.12 mmol) and AcOH (20 uL, 0.35 mmol) dropwise at room temperature. The resulting mixture was stirred for 2 h at room temperature. The reaction was monitored by LCMS. The resulting mixture was diluted with water (2mL). The resulting mixture was extracted with CH2CI2 (5 mL x 3 times). The combined organic layers were washed with brine (15 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The crude product was purified by Prep-HPLC with the following conditions (Column: XSelect CSH C18 Column, 19*250mm, 5pm; Mobile Phase A: 0.05% TFA aqueous solution, Mobile Phase B: ACN; Flow rate: 25 mL / min; Gradient: 20%B to 40%B in 9 min; Wave Length: 254nm / 220nm; RT: 7.63 min) to afford (E)-2-{2-[(5-{[3-(dimethylamino)-2,2-dimethylpropyl]amino}-2-(trifluoromethoxy)phenyl)amino]pyrido[4,3-d]pyrimidin-7-yl} ethenesulfonamide (2.4 mg, 7.3% yield) as an off-white solid, m / z (ESI) calcd. for C23H28F3N7O3S: 539.2, found: 540.1 [M+H]+.1HNMR(300 MHz, DMSO-d6) 89.38 (s, 1H),9.15 (s, 1H), 7.73 – 7.33 (m, 3H), 7.18 - 7.10 (m, 2H), 6.64 – 6.54 (m, 1H), 3.13 (s, 2H), 2.98 (s, 2H), 2.82 (s, 6H), 1.08 (s, 6H).19F NMR (282 MHz, DMSO-d6) 8 -56.8.AFollowing the teachings of the General Reaction Schemes and the synthesis procedure for Example 26 were prepared as shown in Table III:TABLE IIIExample # Structure Name NMR LCMS (E)-2-(2-((5-((3-(pyrrolidin-l- 1H NMR (400 MHz, DMSO-d6) 59.68 yl)propyl)amino)-2- (s, 1H), 9.41 (s, 1H), 9.16 (s, 1H), 7.69 (trifluoromethoxy)phenyl)amino)py (s, 1H), 7.56 (d, J = 14.9 Hz, 1H), 7.45rido[4,3-d]pyrimidin-7-yl)ethene-l- (d, J = 14.9 Hz, 1H), 7.23 (d, J = 13.8 Hz,sulfonamide 2H), 7.11 (d, J = 9.6 Hz, 1H), 6.96 (s,27 538.3, [M+H] +1H), 6.45 (dd, J = 9.0, 2.8 Hz, 1H), 5.99(t, J =5.3 Hz, 1H), 3.08 -3.03 (m, 2H),2.42 -2.30 (m, 6H), 1.75 -1.70 (m,2H), 1.64 (s, 4H). 19F NMR(376 MHz,DMS0-d6) 8-56.98.(E)-2-(2-((5-((3- 1H NMR (400 MHz, Acetonitrile-d3) 8 morpholinopropyl)amino)-2- 9.33 (s, 1H), 9.13 (s, 1H), 7.80-7.72 (trifluoromethoxy)phenyl)amino)py (m, 1H), 7.66 (s, 1H), 7.63 (d, J = 15.0rido[4,3-d]pyrimidin-7-yl)ethene-l- Hz, 1H), 7.54 (d, J = 14.9 Hz, 1H), 7.17sulfonamide (d, J = 10.2 Hz, 1H), 6.48 (s, 1H), 4.07 - 28 3.92 (m, 2H), 3.80 (d, J = 12.4 Hz, 2H), 554.17, [M+H]+ p J 3.59 (d, J =2.6 Hz, 1H), 3.45 (d, J = 12.1Q Hz, 2H), 3.29-3.17 (m, 4H), 3.06 (d, J =9.9 Hz, 2H), 2.08 (dt, J = 14.5, 6.6 Hz,2H). 19F NMR (377 MHz, Acetonitrile- d3) 8-58.98, -75.85.(E)-2-(2-((5-((l-methylpiperidin-4- 1H NMR (300 MHz, DMSO-d6) 89.41yl)amino)-2- (s, 1 H), 9.17 (s, 1 H), 7.65 (s, 1 H), 7.56 (trifluoromethoxy)phenyl)amino)py (m, 1 H), 7.45 (m, 1 H), 7.10 (m, 1 H),29 rido[4,3-d]pyrimidin-7-yl)ethene-l- 7.03 (s, 1H), 6.48 (m, 1 H), 3.14 (s, 1 H), 524.2 [M+H]+ sulfonamide 2.74 (s, 2 H), 2.16 (s, 3 H), 2.09 - 1.87(m, 4 H), 1.45 (m, 2 H). 19F NMR (376MHz, DMSO-d6) 8 -57.0.( E)-2-(2-((5-(( 1-ethy lpiperidin-4- 1H NMR (300 MHz, DMSO-d6) 89.41yl)amino)-2- (s, 1 H), 9.17 (s, 1 H), 7.65 (s, 1 H), 7.56 (trifluoromethoxy)phenyl)amino)py (m, 1 H), 7.45 (m, 1 H), 7.15 (m, 1 H),rido[4,3-d]pyrimidin-7-yl)ethene-l- 7.00 (m, 1 H), 6.48 (m, 1 H), 3.15 (s, 130 538.1 [M+H]+ sulfonamide H), 2.84 (m, 2 H), 2.38 - 2.27 (m, 2 H),2.06 - 1.89 (m, 4 H), 1.45 - 1.31 (m, 2H), 0.99 (m, 3 H). 19F NMR(376 MHz,DMSO-d6) 8-57.0.(E)-2-(2-((5-((l-isopropylpiperidin- 1H NMR (400 MHz, DMSO-d6) 89.414-yl)amino)-2- (s, 1 H), 9.17 (s, 1 H), 7.64 (s, 1 H), 7.56 (trifluoromethoxy)phenyl)amino)py (m, 1 H), 7.44 (m, 1 H), 7.26 (s, 1 H),NH, rido[4,3-d]pyrimidin-7-yl)ethene-l- 7.10 (m, 1 H), 7.03 (s, 1 H), 6.47 (m, 131FYT1 / -A sulfonamide H), 3.16 (m, 1 H), 2.82 - 2.74 (m, 2 H), 552.2 [M+H]+2.72 - 2.65 (m, 1 H), 2.27 - 2.16 (m, 2H), 1.95 (m, 2 H), 1.44 - 1.30 (m, 2 H),0.96 (m, 6 H). 19F NMR (376 MHz,DMSO-d6) 6-57.01.Example # Structure Name NMR LCMS (E)-2-(2-((5-(4-(3- 1H NMR (300 MHz, DMSO-d6) 89.45 (dimethylamino)propyl)piperazin-l- (s, 1H), 9.20 (s, 1H), 7.72 (s, 1H), 7.58yl)-2- (d, J = 14.8 Hz, 2H), 7.48 (s, 1H), 7.34 (trifluoromethoxy)phenyl)amino)py (d, J =8.9 Hz, 1H), 6.96 (dd, J = 9.1, 2.932F VF°^NQ rido[4,3-d]pyrimidin-7-yl)ethene-l- Hz, 1H), 3.88 (s, 2H), 3.61 (s, 2H) 3.18 581.25 [M+H]+J sulfonamide (dt, J = 27.3, 8.0 Hz, 8H), 2.82 (s, 6H),2.10 (dt, J = 15.8, 6.3 Hz, 2H), 1.23 (s,1H). 19F NMR (282 MHz, DMSO-d6) 8 - 56.79, -74.25.(E)-2-(2-((5-((l-benzylpiperidin-4- 1H NMR (300 MHz, DMSO-d6) 89.41yl)amino)-2- (s, 1 H), 9.16 (s, 1 H), 7.64 (s, 1 H), 7.57 (trifluoromethoxy)phenyl)amino)py 7.45 (m, 2 H), 7.38 - 7.10 (m, 5 H), 7.03rido[4,3-d]pyrimidin-7-yl)ethene-l- (s, 2 H), 6.47 (m, 1 H), 3.48 (s, 2 H),33 600.1 [M+H]+ bb ^ / sulfonamide 3.18 (m, 1 H), 2.80 (m, 2 H), 2.09 (m, 2H), 1.93 (m, 2 H), 1.43 (m, 2 H). 19Fb NMR (376 MHz, DMSO-d6) 8 -56.8.(E)-2-(2-((5-(((lr,4r)-4-(piperidin-l- ’H NMR (500 MHz, DMSO-d6) 9.65 (s, yl)cyclohexyl)amino)-2- 1H), 9.42 (s, 1H), 9.17 (s, 1H), 7.66 (s, (trifluoromethoxy)phenyl)amino)py 1H), 7.57 (d, J = 14.9 Hz, 1H), 7.44 (d, Jrido|4,3-d]pyrimidin-7-yl)ethene-l- = 14.9 Hz, 1H), 7.24 (s, 2H), 7.09 (d, J = sulfonamide 8.9 Hz, 1H), 6.98 (s, 1H), 6.45 (dd, J =9.1, 2.8 Hz, 1H), 5.77 (d, J = 7.8 Hz, 1H),34 3.51 (s, 2H), 2.45 (s, 4H), 2.29 (d, J = 592.3, [M+H]+.bb >-N A ) 20.8 Hz, 1H), 2.07 (d, J = 12.0 Hz, 2H),1.78 (d, J = 12.2 Hz, 2H), 1.47 (s, 2H),1.36 (d, J = 11.1 Hz, 2H), 1.24 (s, 2H),1.16 (q, J = 12.1 Hz, 2H), 0.85 (d, J = 7.0Hz, 1H). 19F NMR (471 MHz, DMSO- d6) 8-56.96.(E)-2-(2-((5-((( ls,4s)-4-(piperidin-l- ’H NMR (400 MHz, Acetonitrile-d3) 9.33rf 5yl)cyclohexyl)amino)-2- (s, 1H), 9.12 (s, 1H), 7.69 (s, 1H), 7.65p b (trifluoromethoxy)phenyl)amino)py — 7.30 (m, 3H), 7.14 (d, J =8.9 Hz, 1H),rido[4,3-d]pyrimidin-7-yl)ethene-l- 6.51 (dd, J = 9.0, 3.0 Hz, 1H), 3.59 (s,35 b 592.3, [M+H] + AYS sulfonamide 2H), 2.41 (s, 1H), 1.95 - 1.43 (m, 11H), O 1.27 (s, 2H), 0.87 (d, J = 7.3 Hz, 1H).19F NMR (376 MHz, Acetonitrile-d3) 8 - 58.98.(E)-2-(2-((5-(((lr,4r)-4- 1H NMR (400 MHz, DMSO-d6) 69.42 (dimethylamino)cyclohexyl)amino)- (s, 1 H), 9.18 (s, 1 H), 7.65 (s, 1 H), 7.602- -7.52 (m, 1 H), 7.48 -7.42 (m, 1 H), (trifluoromethoxy)phenyl)amino)py 7.14 - 7.06 (m, 2 H), 6.46 -6.43 (m, 136 rido[4,3-d]pyrimidin-7-yl)ethene-l- H), 3.14- 3.09 (m, 1 H), 2.17 (s, 6 H), 552.2 [M+H]+ sulfonamide 2.10-2.07 (m, 2 H), 1.89 -1.82 (m, 2H), 1.78-1.70 (m, 1 H), 1.25-1.22 (m,4 H). 19F NMR (376 MHz, DMSO-d6) 6 - 57.03.(E)-2-(2-((5-(((ls,4s)-4- 1H NMR (400 MHz, DMSO-d6) 89.42 (dimethylamino)cyclohexyl)amino)- (s, 1 H), 9.17 (s, 1 H), 7.66 - 7.58 (m, 22- H), 7.60-7.55 (m, 1 H), 7.47 - 7.03 (m,37 bb (trifluoromethoxy)phenyl)amino)py 2 H), 6.56- 6.51 (m, 1 H), 3.44 -3.38 552.2 [M+H]+ rido[4,3-d]pyrimidin-7-yl)ethene-l- (m, 1 H), 2.18 -2.11 (m, 7 H), 1.76 - sulfonamide 1.51 (m, 8 H). 19F NMR (376 MHz,DMSO-d6) 6-57.03.Examples 38 & 39Step 1; To the mixture of (E)-2-(2-{[5-(piperazin-l-yl)-2-(trifluoromethoxy)phenyl]amino}pyrido[4,3-d]pyrimidin-7-yl)ethenesulfonamide (120.2 mg, 0.24 mmol) and tert-butyl N-(4-oxocyclohexyl)carbamate (103.3 mg, 0.48 mmol) in CH2CI2 (5 mL) was added STAB (76.9 mg, 0.36 mmol) and AcOH (20 uL). The mixture was stirred at rt for 2 h. The desired product could be detected by LCMS The mixture was added with H2O (20 mL) and extracted by DCM (10 mL x 3 times). The organic layer was combined, dried over Na2SC>4, filtered, and concentrated. The residue was eluted with ACN / H2O (0.05%TFA) (from 50% to 90% in 30 min) from a C18 column to afford tert-butyl N-(4-{4-[3-({7-[(E)-2-sulfamoylethenyl]pyrido[4, 3-d]pyrimidin-2-yl}amino)-4-(tri fluoromethoxy )phenyl]piperazin-l -yl}cyclohexyl)carbamate (100.0 mg, 59.6% yield) as yellow solid, m / z (ESI) calcd. for C31H39F3N8O5S: 692.3, found: 693.1 [M+H]+.Step 2: To the mixture of (E)-2-[2-({5-[4-(4-aminocyclohexyl)piperazin-l-yl]-2-(trifluoromethoxy)phenyl}amino)pyrido[4,3-d]pyrimidin-7-yl]ethenesulfonamide (100.2 mg, 0.17 mmol) in CH2CI2 (5 mL) was added TFA (1 mL). The mixture was stirred at rt for Ih. The desired product could be detected by LCMS. The mixture was concentrated. The crude product was purified by Prep-HPLC with the following conditions (Column: YMC-Actus Triart Cl 8 ExRS30*150 mm; Mobile Phase A: Water(10mmol / L NH4HC03+0.05%NH3H2O), Mobile Phase B: ACN; Flow rate: 60 mL / min mL / min; Gradient: 18% B to 34% B in 12 min; Wave Length: 254nm / 220nm; RTl(min): 10.18, 11.77(min): ) to afford assumed trans (E)-2-{2-[(5-{4-[(lr,4r)-4-aminocyclohexyl]piperazin-l-yl}-2-(trifluoromethoxy)phenyl)amino]pyrido[4,3-d]pyrimidin-7-yl } ethenesulfonamide (the first peak, 10.0 mg, 9.89% yield) as a yellow solid and assumed trans (E)-2-{2-[(5-{4-[(ls,4s)-4-aminocyclohexyl]piperazin-l-yl}-2-(trifluoromethoxy )phenyl)amino]pyrido[4,3-d]pyrimidin-7-yl (ethenesulfonamide (the second peak, 10.1 mg, 9.9% yield) as a yellow solid.Pl: m / z (ESI) calcd. for C₂₆H₃₁F₃N₈O₃S: 592.2, found: 593.1 [M+H]+.. 'H NMR (400 MHz, DMSO-d6) 89.43 (s, 1H), 9.17 (s, 1H), 7.72 (s, 1H), 7.59 - 7.47 (m, 2H), 7.43 (s, 1H), 7.24 (d, J = 8.9 Hz, 1H), 6.86 - 6.83 (m, 1H), 3.15 (t, J = 4.6 Hz, 4H), 2.63 (t, J = 4.9 Hz, 4H), 2.53 (s, 1H), 2.23 (t, J = 11.4 Hz, 1H), 1.81 (d, J = 11.3 Hz, 4H), 1.26 - 1.02 (m, 4H).19F NMR (376 MHz, DMSO-d6) 8 -56.8.P2: m / z (ESI) calcd. for C₂₆H₃₁F₃N₈O₃S: 592.2, found: 593.1 [M+H]+. 'H NMR (400 MHz, DMSO-d6) 89.43 (s, 1H), 9.17 (s, 1H), 7.73 (s, 1H), 7.59 - 7.51 (m, 2H), 7.47 - 7.44(m, 1H), 7.25 (d, J = 9.0 Hz, 1H), 6.86 - 6.84 (m, 1H), 3.17 (t, J = 4.9 Hz, 4H), 2.86 (s, 1H), 2.62 (t, J = 4.9 Hz, 4H), 2.18 (t, J = 8.9 Hz, 1H), 1.76 - 1.71 (m, 2H), 1.52 -1.46 (m, 6H).19F NMR(376 MHz, DMSO-d6) 8 -56.8.Following the teachings of the General Reaction Schemes and the synthesis procedure for Examples 38 & 39 were prepared as shown in Table IV:TABLE IVExample # Structure Name NMR LCMS (E)-2-(5-chloro-2-((5-(4- 1H NMR (300 MHz, DMSO-d6) 89.42methylpiperazin-l-yl)-2- (s, 1H), 9.12 (s, 1H), 7.66 (s, 1H), 7.61 - (trifluoromethoxy)phenyl)amino)py 7.49 (m, 2H), 7.21 - 7.17 (m, 2H), 6.62 - rido[4,3-d]pyrimidin-7-yl)ethene-l- 6.58 (m, 1H), 3.60 - 3.51 (m, 2H), 2.47 - 40 sulfonamide 3.43 (m, 2H), 2.65 - 2.62 (m, 2H), 2.48 - 544.1, [M+H] +2.46 (m, 2H), 2.26 (s, 3H), 1.91 (t, J =8.4 Hz, 2H). 19F NMR (282 MHz, DMSO- d6) 8-56.95. 19F NMR(376 MHz,Methanol-d4) 8-59.74.(E)-2-(2-((2-cyclopropyl-5-(4- 1H NMR (400 MHz, DMSO-d6) 89.42methylpiperazin-1- (d, J = 53.3 Hz, 2H), 7.54 (d, J = 14.9 Hz, yl)phenyl)amino)-5- 2H), 7.41 (d, J = 14.9 Hz, 1H), 7.27 (s, methylpyrido[4,3-d]pyrimidin-7- 2H), 6.90 (d, J = 8.6 Hz, 1H), 6.73 (dd, J41 yl)ethene-l-sulfonamide = 8.6, 2.5 Hz, 1H), 3.13 - 3.05 (m, 4H), 480.2, [M+H] +2.84 (s, 3H), 2.48 - 2.42 (m, 4H), 2.22(s, 3H), 1.88 (ddd, J = 13.7, 8.4, 5.4 Hz,1H), 1.23 (s, 1H), 0.78 -0.70 (m, 2H),0.53 -0.44 (m, 2H).Example # Structure Name NMR LCMS (E)-2-(5-chloro-2-((2-cyclopropyl-5- 1H NMR (400 MHz, DMSO-d6) 89.79(4-methylpiperazin-l- (s, 1H), 9.40 (s, 1H), 7.70 (s, 1H), 7.51 - yl)phenyl)amino)pyrido[4,3- 7.38 (m, 2H), 7.25 (s, 1H), 7.13 (s, 1H),t o d]pyrimidin-7-yl)ethene-l- 6.90 (d, J =8.6 Hz, 1H), 6.77 (d, J = 10.6 42 500.1, [M+H] + sulfonamide Hz, 1H), 3.10 (s, 4H), 2.44 (s, 4H), 2.21t f p (s, 3H), 1.87 (dt, J = 13.5, 5.6 Hz, 1H),1.23 (s, 1H), 0.74 (d, J =8.3 Hz, 2H),0.50 (d, J =4.4 Hz, 2H).rac-(R, E)-2-(5-chloro-2-((5-((l- 1H NMR (400 MHz, DMSO-d6) 610.00 methylazepan-4-yl)amino)-2- (s, 1H), 9.43 (s, 1H), 7.71 (s, 1H), 7.54 - (trifluoromethoxy)phenyl)amino)py 7.36 (m, 2H), 7.28 (s, 2H), 7.11 (d, J =rido[4,3-d]pyrimidin-7-yl)ethene-l- 8.8 Hz, 1H), 6.84 (s, 1H), 6.45 (d, J = 8.943 sulfonamide Hz, 1H), 5.88 (d, J = 7.8 Hz, 1H), 3.47 572.2, [M+H] +(tt, J = 8.6, 3.8 Hz, 1H), 2.68 -2.52 (m,2H), 2.48 - 2.34 (m, 2H), 2.24 (s, 3H),2.01 -1.85 (m, 2H), 1.77 -1.44 (m,4HJ.19F NMR (376 MHz, DMS0-d6) 8 -Example 4444-3 44-4 44-5Example 44Step 1: The mixture of tert-butyl 4-(3-amino-4-cyclopropoxyphenyl)piperazine-l -carboxylate (1.1 g, 3.30 mmol), methyl 2,5-dichloropyrido[4,3-d]pyrimidine-7-carboxylate (852.2 mg, 3.30 mmol) and z-PrOH (42 mL) was stirred at 80 °C for about 1 hour. After being cooled to room temperature, the mixture was concentrated under reduced pressure, and the residue was purified with a Cl 8 column to yield methyl 2-((5-(4-(tert-butoxycarbonyl)piperazin-l-yl)-2- cyclopropoxyphenyl)amino)-5-chloropyrido[4,3-d]pyrimidine-7-carboxylate as yellow solids (610.0 mg, 33.0% yield). m / z(ESI), calcd. for C27H31CIN6O5: 554.2, found: 555.3, [M+H]+.Step 2; Methyl 2-((5-(4-(tert-butoxycarbonyl)piperazin- 1 -yl)-2-cyclopropoxyphenyl)amino)-5- chloropyrido[4,3-d]pyrimidine-7-carboxylate (760.0 mg, 1.36 mmol) was dissolved in ACN (15 mL), then AICI3 (546.5 mg, 4.08 mmol) was added thereto. The mixture was stirred at room temperature for one hour, then diluted with water and extracted with dichloromethane (50 mL x 3 times). The combined organic layers were dried over anhydrous MgSO4and concentrated under reduced pressure to yield methyl 5-chloro-2-((2-cyclopropoxy-5-(piperazin-l- yl)phenyl)amino)pyrido[4,3-d]pyrimidine-7-carboxylate (320.0 mg, 51.0% yield) as yellowsemi-solids, which was used for the next step without further purification. m / z(ESI), calcd. for C22H23CIN6O3: 454.2, found: 455.2, [M+H]+.Step 3; Methyl 5-chloro-2-((2-cyclopropoxy-5-(piperazin-l-yl)phenyl)amino)pyrido[4,3-d]pyrimidine-7-carboxylate (320.0 mg, 0.70 mmol) and HCHO (263.7 g, 3.50 mmol) were dissolved in DCM (dichloromethane; 30 mL), followed by addition of NaBH(AcO)3 (1.3 g, 2.10 mmol). The mixture was stirred at room temperature for 30 minutes, then diluted with water and extracted with ethyl acetate (50 mL x 3 times). The combined organic layers were dried over anhydrous MgSO4and concentrated under reduced pressure. The residue was purified with a Cl 8 column to afford methyl 5-chloro-2-((2-cyclopropoxy-5-(4-methylpiperazin-l-yl)phenyl)amino)pyrido[4,3-d]pyrimidine-7-carboxylate as yellow solids (120.0 mg, 44.0% yield). m / z(ESI), calcd. for C23H25ClN6O3:468.2, found: 469.2, [M+H]+.Step 4: To a solution of morpholine (75.8 mg, 0.84 mmol) in THF (tetrahydrofuran; 8mL) under nitrogen atmosphere was added diisobutylaluminium hydride (DIBALH; IM in THF, 0.84 mL, 0.84 mmol) at 0 °C, and the mixture was stirred at 0 °C for 30 minutes. Afterward, the mixture was cooled to -20 °C, and methyl 5-chloro-2-((2-cyclopropoxy-5-(4-methylpiperazin-l-yl)phenyl)amino)pyrido[4,3-d]pyrimidine-7-carboxylate (100.0 mg, 0.21 mmol) solution in dry THF (2 mL) was introduced via a syringe. The solution was stirred at -20 °C for 10 minutes, quenched with H2O, and extracted with ethyl acetate (50 mL x 3 times). The combined organic layer was dried over anhydrous MgSCU and concentrated under reduced pressure. The residue was purified with a C18 column to afford 5-chloro-2-((2-cyclopropoxy-5-(4-methylpiperazin-l-yl)phenyl)amino)pyrido[4,3-d]pyrimidine-7-carbaldehyde as yellow solids (45.2 mg, 50.0% yield). m / z(ESI), calcd. for C22H23ClN6O2:438.2, found:439.1, [M-H]+.Step 5; To a solution of tert-butyl (((diphenylphosphoryl)methyl)sulfonyl)carbamate (72.1 mg, 0.18 mmol) in DMF (N, N-dimethylformamide; 5 mL) was added NaH (60wt%; 55.0 mg, 0.54 mmol), and the mixture was stirred at room temperature for 30 min. Then 5-chloro-2-((2-cyclopropoxy-5-(4-methylpiperazin-l-yl)phenyl)amino)pyrido[4,3-d]pyrimidine-7-carbaldehyde (40.0 mg, 0.09 mmol) was added to the above solution before being further stirred for another 1 hour. Afterward, the reaction mixture was quenched by slowly adding water (0.5 mL), then extracted with ethyl acetate (50 mL x 3 times). The combined organic layer was dried overanhydrous MgS04 and concentrated under reduced pressure to yield tert-butyl (E)-((2-(5-chloro-2-((2-cyclopropoxy-5-(4-methylpiperazin-l-yl)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate as yellow solids (55.0 mg, 87.0% yield), which was used in the next step without further purification, m / z (ESI), calcd. for C28H34ClN7O5S:615.2, found:616, [M+H]+.Step 6; Tert-butyl (E)-((2-(5-chloro-2-((2-cyclopropoxy-5-(4-methylpiperazin-l-yl)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (50.0 mg, 0.17 mmol) was dissolved in DCM (3 mL), then trifluoroacetic acid (1 mL) was added to it. The reaction mixture was stirred at room temperature for 0.5 h and then concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography (column: X Bridge Prep Cl 8 OBD, 30*150 mm, 5 pm; mobile phase A: 10 mM aqueous solution of NH4HCO3, mobile phase B: ACN; flow rate: 60 mL / min; gradient: 33% B to 49% B in 10 min; wavelength: 254nm; RT: 9.95 min) to obtain (E)-2-(5-chloro-2-((2-cyclopropoxy-5-(4-methylpiperazin-l-yl)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)ethene-l-sulfonamide (2.4 mg, 5.7% yield) as yellow solids, m / z (ESI), calcd. for C23H26CIN7O3S: 515.2, found: 516.2, [M+H]+. 'H NMR (400 MHz, DMSO-rL) 89.40 (s, 1H), 9.19 (s, 1H), 7.76 (s, 1H), 7.59 (s, 1H), 7.48 (s, 2H), 7.25 (d, J = 18.4 Hz, 2H), 7.20 (s, 1H), 6.79 (dd, J= 8.9, 2.8 Hz, 1H), 3.85 (dt, J= 5.6, 3.1 Hz, 1H), 3.11 -3.03 (m, 4H), 2.48 (d, J= 4.6 Hz, 2H), 2.23 (s, 3H), 1.23 (s, 2H), 0.70 (d, J= 5.3 Hz, 2H), 0.62 (s, 2H).Following the teachings of the General Reaction Schemes and the synthesis procedure for Example 44 were prepared as shown in Table V:TABLE VExample # Structure Name NMR LCMS (E)-2-(5-chloro-2-((2-cyclopropoxy- 1H NMR (300 MHz, DMSO-d6) 89.385-((3-(dimethylamino)-2,2- (s, 1H), 9.14 (s, 1H), 7.71 (s, 1H), 7.53 - dimethylpropyl)amino)phenyl)amin 7.37 (m, 2H), 7.33 - 7.07 (m, 4H), 6.44o)pyrido[4,3-d]pyrimidin-7- (dd, J = 8.7, 2.4 Hz, 1H), 5.07 (s, 1H),45 546.2, [M+H] + yl)ethene-l-sulfonamide 3.83 -3.75 (m, 1H), 3.16 -3.03 (m,2H), 2.47 (d, J = 6.8 Hz, 2H), 2.38 (s,4H), 1.56-1.45 (m, 4H), 1.43 - 1.33(m, 2H), 0.69 -0.56 (m, 4H).Example # Structure Name NMR LCMS (S, E)-2-(5-chloro-2-((2- 1H NMR (400 MHz, DMSO-d6) 59.38cyclopropoxy-5-((l- (s, 1H), 9.14 (s, 1H), 8.19 (s, 1H), 7.70 isopropylpyrrolidin-3- (s, 1H), 7.49 -7.36 (m, 2H), 7.28 (s, yl)amino)phenyl)amino)pyrido[4,3- 2H), 7.22 (s, 1H), 7.11 (d, J = 8.8 Hz,d]pyrimidin-7-yl)ethene-l- 1H), 6.41 (dd, J = 8.8, 2.7 Hz, 1H), 3.86sulfonamide (s, 1H), 3.78 (tt, J = 6.2, 3.0 Hz, 1H),46 3.00 (dd, J = 9.7, 6.9 Hz, 1H), 2.80 (q, J = 544.20, [M+H]+8.0 Hz, 1H), 2.64 (q, J = 7.9, 7.3 Hz, 1H),2.57 -2.51 (m, 2H), 2.21 (dq, J = 13.7,7.6 Hz, 1H), 1.65 (dq, J = 12.9, 6.8 Hz,1H), 1.07 (t, J = 6.1 Hz, 6H), 0.75 -0.63(m, 2H), 0.62 -0.58 m, 2H).(E)-2-(5-chloro-2-((2-cyclopropoxy- ’H NMR (300 MHz, DMSO-d6) 9.38 (s,5-((2-(piperidin-l- 1H), 9.14 (s, 1H), 7.71 (s, 1H), 7.53 - yl)ethyl)amino)phenyl)amino)pyrid 7.37 (m, 2H), 7.33 - 7.07 (m, 4H), 6.44o[4,3-d]pyrimidin-7-yl)ethene-l- (dd, J = 8.7, 2.4 Hz, 1H), 5.07 (s, 1H),47 544.2, [M+H]+.sulfonamide 3.83 — 3.75 (m, 1H), 3.16 — 3.03 (m,2H), 2.47 (d, J = 6.8 Hz, 2H), 2.38 (s,4H), 1.56 - 1.45 (m, 4H), 1.43 - 1.33(m, 2H), 0.69 - 0.56 (m, 4H).(E)-2-(5-chloro-2-((2-cyclopropoxy- 'H NMR (300 MHz, DMSO-d6) 9.39 (s,5-((2- 1H), 9.15 (s, 1H), 7.71 (s, 1H), 7.54 — (diethylamino)ethyl)amino)phenyl) 7.42 (m, 2H), 7.41 - 7.22 (m, 3H), 7.11 amino)pyrido[4,3-d]pyrimidin-7- (d, J = 8.7 Hz, 1H), 6.44 (dd, J = 8.8, 2.448 HNAA^1''**XKH2yl)ethene-l-sulfonamide Hz, 1H), 5.07 (s, 1H), 3.86 - 3.69 (m, 532.2, [M+H]+.1H), 3.07 (t, J = 6.4 Hz, 2H), 2.61 (t, J =■'N k / 6.8 Hz, 2H), 2.55 (d, J = 7.1 Hz, 4H),1.27 (d, J = 23.3 Hz, 1H), 0.97 (d, J =14.1 Hz, 6H), 0.71 - 0.56 (m, 4H).Example 49Example 49 Step 1: The mixture of N-(3-amino-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-(4-(morpholinomethyl)phenyl)acetamide (420.0 mg, 0.91 mmol), 2,7-dichloropyrido[4,3-d]pyrimidine (217.5 mg, 1.09 mmol) and i-PrOH (42 mL) was stirred at 120 °C for about 20 hours. After being cooled to room temperature, the mixture was concentrated under reduced pressure, and the residue was purified with a silica-gel column (eluent: CLLCh / MeOH = 10 / 1, v / v) to yield N-(3-((7-chloropyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-(4-(morpholinomethyl)phenyl)acetamide as yellow solid (425.0 mg, 74.8% yield), m / z (ESI) calcd. for C27H21ClF6N6O3:626.1, found:627.2, [M+H]+.Step 2; N-(3-((7-chloropvridor4.3-d1pvrimidin-2-vl)amino)-4-(trifluoromethoxv)phenvl)-2.2.2-trifluoro-N-(4-(morpholinomethyl)phenyl)acetamide (300.0 mg, 0.48 mmol), Pd(dppf)C12 (35.1 mg, 0.1 mmol) and K3PO4 (304.7 mg, 1.44 mmol) were loaded into a single-neck round-bottom flask, followed by addition of solvent mixture of 1,4-di oxane and H2O (30 mL, v / v = 5 / 1). A septum capped the flask, and the mixture was bubbled with N2 flow through a needle for 10 minutes, after which 2-ethenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (221.1 mg, 1.44 mmol) was injected via a syringe. After stirring at 90 °C for 3 h, the reaction mixture was diluted with water and extracted with ethyl acetate (100 mL x 3 times). The combined organic layer was dried over anhydrous MgSO4and concentrated under reduced pressure. The residue was elutedwith CH₂Cl₂ / MeOH (v / v = 10 / 1) from a silica gel column to yield N'-(4-(morpholinomethyl)phenyl)-4-(trifluoromethoxy)-N3-(7-vinylpyrido[4,3-d]pyrimidin-2-yl)benzene-l,3-diamine as yellow solid (249.7 mg, 60% yield), m / z (ESI) calcd. for C27H25F3N6O2: 522.2, found: 523.2, [M+H]+.Step 3: N1-(4-(morpholinomethyl)phenyl)-4-(trifluoromethoxy)-N3-(7-vinylpyrido[4,3-d]pyrimidin-2-yl)benzene-l,3-diamine (180.0 mg, 0.34 mmol) and morpholine (29.9 mg, 0.34 mmol) were dissolved in THF (tetrahydrofuran; 8 mL), then OsO4 (17.5 mg, 0.07 mmol) solution in THF (1 mL) was added to that, followed by addition of water (9 mL) and NaIO₄ (186.7 mg, 0.87 mmol). The mixture was stirred for 1 hour and extracted with ethyl acetate (50 mL * 3 times). The combined organic layer was dried over anhydrous MgSO₄ and concentrated under reduced pressure to yield crude product of 2-((5-((4-(morpholinomethyl)phenyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidine-7-carbaldehyde as yellow solid (170.2 mg, 94.4% yield), which was used in the next step without further purification. m / z( ESI) calcd. for C26H23F3N6O3: 524.2, found:525.2, [M+H]+.Step 4: tert-butyl (((diphenylphosphoryl)methyl)sulfonyl)carbamate (113.1 mg, 0.29 mmol) was dissolved in DMF (15 mL), and NaH (60wt%; 45.6 mg, 1.14 mmol) was then added. The mixture was stirred at room temperature for 30 minutes, after which 2-((5-((4-(morpholinomethyl)phenyl)amino)-2-(tri fluoromethoxy )phenyl)amino)pyrido[4,3-d]pyrimidine-7-carbaldehyde (150.0 mg, 0.29 mmol) was introduced to it, followed by additional 1-hour stirring at room temperature. Afterward, the reaction mixture was quenched by slowly adding water (0.5 mL), then extracted with ethyl acetate (50 mL x 3 times). The combined organic layer was dried over anhydrous MgSCU and concentrated under reduced pressure to yield tert-butyl (E)-((2-(2-((5-((4-(morpholinomethyl)phenyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate as yellow solids (128.0 mg, 64% yield), which was used in the next step without further purification, m / z (ESI) calcd. for C₃₂H₃₄F₃N₇O₆S: 701.2, found: 702.2, [M+H]+.Step 5: tert-butyl (E)-((2-(2-((5-((4-(morpholinomethyl)phenyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (100.0 mg, 0.15 mmol) was dissolved in DCM (dichloromethane; 10 mL), then trifluoroacetic acid (2mL) was added to that. The reaction mixture was stirred at room temperature for 0.5 h, followed by concentration under reduced pressure; the residue was purified by high-pressure preparation HPLC (X Bridge) Shield RP 18 OBD Column, 19*250 mm, 5pm; mobile phase A: 0.1% FA d;aqueous solution, mobile phase B: ACN; flow rate: 25 mL / min; gradient: 21% B to 51% B in 7A)min; wavelength: 254 nm) to obtain (E)-2-(2-((5-((4-(morpholinomethyl)phenyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)ethene-l -sulfonamide (9.0 mg) as yellow solids. m / z( ESI) calcd. for C27H26F3N7O4S: 601.2, found LCMS: 602.1 [M+H]+. 'H NMR (500 MHz, DMSO-d₆) 59.44 (s, 1H), 9.19 (s, 1H), 7.73 - 7.58 (m, 3H), 7.56 (s, 1H), 7.47 (d, J= 15.0 Hz, 1H), 7.22 (d, J = 8.0 Hz, 2H), 7.14 (d, J= 8.0 Hz, 2H), 6.93 - 6.87 (m, 1H), 3.38 (s, 2H), 2.32 (s, 4H).Following the teachings of the General Reaction Schemes and the synthesis procedure for Example 49 were prepared as shown in Table VI:TABLE VIExample # Structure Name NMR LCMS(E)-2-(2-((5-((3-(piperidin-l- 1H NMR (400 MHz, Methanol-d4) 6yl)propyl)amino)-2- 9.34 (s, 1H), 9.12 (s, 1H), 7.71 -7.60 (trifluoromethoxy)phenyl)amino)py (m, 3H), 7.52 (d, J = 15.0 Hz, 1H), 7.10AiAzy*- rido[4,3-d]pyrimidin-7-yl)ethene-l- (dt, J = 8.9, 1.4 Hz, 1H), 6.45 (dd, J =50FVxji0552.30 [M+H]+ sulfonamide 8.9, 2.8 Hz, 1H), 3.19 (t, J = 6.8 Hz, 2H),Hu 2 61 - 2 39 (m, 6H), 1 93 - 1 82 (m,2H), 1.62 (p, J = 5.6 Hz, 4H), 1.48 (s,2H). 19F NMR (376 MHz, Methanol-d4)(E)-2-(2-((5-((2- 1H NMR (400 MHz, Methanol-d4) 6 morpholinoethyl)amino)-2- 9.35 (s, 1H), 9.12 (s, 1H), 7.65 (d, J = (trifluoromethoxy)phenyl)amino)py 16.9 Hz, 3H), 7.53 (d, J = 14.9 Hz, 1H),51 rido[4,3-d]pyrimidin-7-yl)ethene-l- 7 11 (d, J = 89 Hz, 1H), 6 47 (dd, J = 89, 540 20, [M+H]+ sulfonamide 2.8 Hz, 1H), 3.77 - 3.63 (m, 4H), 3.33(s, 2H), 2.67 (t, J = 6.6 Hz, 2H), 2.56 (s,4H). 19F NMR (376 MHz, Methanol-d4)(E)-2-(2-((5-((2- 1H NMR (400 MHz, Methanol-d4) 6 (diisopropylamino)ethyl)amino)-2- 9.37 (s, 1H), 9.15 (s, 1H), 7.78 (d, J = (trifluoromethoxy)phenyl)amino)py 2.1 Hz, 1H), 7.66 (d, J = 14.9 Hz, 2H),rido[4,3-d]pyrimidin-7-yl)ethene-l- 7 56 - 7 48 (m, 1H), 7 21 -7 17 (m,52 sulfonamide 1H), 6.55 (dd, J = 8.9, 2.8 Hz, 1H), 3.80 554.3, [M+H]+P ' r (p, J = 6.6 Hz, 2H), 3.58 (q, J = 6.2, 5.7Hz, 2H), 3.39 (t, J = 6.6 Hz, 2H), 1.40 (d,J = 6.6 Hz, 12H). 19F NMR (376 MHz,Methanol-d4) 6 -60.10 (d, J = 81.3 Hz), -Example# Structure Name IMMR LCMS (E)-2-(2-((5-((l-(tetrahydro-2H- ’H NMR (300 MHz, Methanol-d4) 9.37pyran-4-yl)piperidin-4-yl)amino)-2- (s, 1H), 9.15 (s, 1H), 7.78 (s, 1H), 7.70 (trifluoromethoxy)phenyl)amino)py — 748 (m, 3H), 7 11 (d, J = 9 0 Hz, 1H),rido[4,3-d]pynmidin-7-yl)ethene-l- 6.49 (dd, J = 9.0, 2.6 Hz, 1H), 4.60 (d, J = sulfonamide 1.7 Hz, 1H), 4.05 (d, J = 11.4 Hz, 2H),53 3.45 (t, J = 11.7 Hz, 2H), 3.17 — 3.06 594.2, [M+H]+FvX', / XX^' t ULr. Q^2>NHl(m, 2H), 2.64 — 2.54 (m, 1H), 2.44 (t, J= 11.4 Hz, 2H), 2.21 (d, J = 13.0 Hz, 2H),1.89 (d, J = 14.0 Hz, 2H), 1.69 — 1.51(m, 4H). 19F NMR (282 MHz, Methanol- d4) 6 -59.99(E)-2-(2-((5-((l,2,2,6;6- 1H NMR (400 MHz, DMSO-d6) S 9.71 pentamethylpiperidin-4-yl)amino)-2 (s, 1H), 9.42 (s, 1H), 9.17 (s, 1H), 7.67 (trifluoromethoxy)phenyl)amino)py (s, 1H), 7.56 (d, J = 14.9 Hz, 1H), 7.42rido[4,3-d]pyrimidin-7-yl)ethene-l- (d, J = 14.9 Hz, 1H), 7.27 (s, 2H), 7.11sulfonamide (d, J = 8.7 Hz, 1H), 6.97 (d, J = 2.1 Hz,54 580.3, [M+H]+ M4 1H), 6.48 (dd, J = 8.9, 2.6 Hz, 1H), 5.76(d, J = 7.7 Hz, 1H), 3.51 (s, 1H), 2.20 (s,3H), 1.87 (d, J = 13.5 Hz, 2H), 1.25 (t, J =12.0 Hz, 2H), 1.11 (s, 6H), 1.05 (s, 6H).19F NMR (376 MHz, Methanol-d4) 6 - (E)-2-(2-((5-((4- 1H NMR (400 MHz, DMSO-d6) 69.65 (diethylamino)butyl)amino)-2- (s, 1H), 9.41 (s, 1H), 9.16 (s, 1H), 7.69 (trifluoromethoxy)phenyl)amino)py (s, 1H), 7.56 (d, J = 14.9 Hz, 1H), 7.45N^Y^N rido[4,3-d]pyrimidin-7-yl)ethene-l- (d, J = 14 9 Hz, 1H), 7 24 (s, 2H), 7 1155 v°>«A > sulfonamide (d, J = 8.8 Hz, 1H), 6.96 (s, 1H), 6.46 554.2, [M+H]+ a,^uki (dd, J = 8.9, 2.8 Hz, 1H), 5.96 (t, J = 5.3Hz, 1H), 3.01 (q, J = 6.5 Hz, 2H), 2.45 - 2.32 (m, 6H), 1.61 - 1.40 (m, 4H), 0.91(t, J = 7.1 Hz, 6H).Example 56NH2O ’S Al / 'CF;,iFhOH, 120 °C, 20 h O' *CF3PHRT-010066-4 PHRT-010066-5 PHRT-010066-6O, Ph,'R' OP" S-N. H 6 Bcc 010001-55 TFA. PCM, rt, 1 h NaH, DMF rt, 1 h PHRT-010066-6 PHRT-010066-9 PHRT-010066-7PHRT-010066 Step 1: N-(3-amino-4-cvclopropoxvphenvl)-N-r2-(diethvlamino)ethvl]-2.2.2-trifluoroacetamide (800.0 mg, 1.62 mmol) and 2,5-dichloro-N-methoxy-N-methylpyrido[4,3-d]pyrimidine-7- carboxamide (644.5 mg, 3.24 mmol) was dissolved in / -PrOH (20 mL), and the mixture was stirred at 120 °C overnight. After being cooled to room temperature, the reaction mixture was concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: CTECE / MeOH = 10 / 1, v / v) to yield N-(3-((7-chloropyrido[4,3-d]pyrimidin-2-yl)amino)-4- (trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-((lR,5S)-8-(2,2,2-trifluoroacetyl)-8 azabicyclo[3.2.1]octan-3-yl)acetamide as yellow solids (450.0 mg, 44% yield), m / z (ESI) calcd. for C27H31CIF3N7O4: 656, found: 657, [M+H]+.Sten2^N-(3-((7-chloropyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)-2,2,2- trifluoro-N-((lR,5S)-8-(2,2,2-trifluoroacetyl)-8-azabicyclo[3.2.1]octan-3-yl)acetamide (480.0 mg, 0.73 mmol), Pd(dppf)Cl2 (57.1 mg, 0.07 mmol) and K3PO4 (114.5 mg, 2.19 mmol) were placed into a single-neck round-bottom flask, followed by addition of 1,4-di oxane (20 mL) and H2O (4 mL). A septum capped the flask, and the mixture was bubbled with N2 flow through a needle for 10 minutes, after which 2-ethenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (283.4 mg, 1.84 mmol) was injected using a syringe. After stirring at 90 °C for 2 h, the reaction mixture was extracted from water with ethyl acetate (100 mL x 3 times). The combined organic layer was dried over anhydrous MgSCU and concentrated under reduced pressure. The residue was elutedwith from a silica Cl 8 column to yield 2,2,2-trifluoro-N-((lR,5S)-8-(2,2,2-trifluoroacetyl)-8-azabicyclo[3.2. l]octan-3 -yl)-N-(4-(tri fluorometh oxy)-3-((7-vinylpyrido[4, 3 -d]pyrimidin-2-y l)amino)phenylacetamide as yellow solids (380.0 mg, 78.0% yield), m / z (ESI), calcd. for C₂₇H₂₁F₉N₆O₃:648, found:649, [M+H]+.Step 3: 2.2.2-trifluoro-N-((lR.5S)-8-(2.2.2-trifluoroacetvl)-8-azabicvclor3.2.11octan-3-vl)-N-(4-(trifluoromethoxy)-3-((7-vinylpyrido[4,3-d]pyrimidin-2-yl)amino)phenyl)acetamide (380.0 mg, 0.73 mmol) were dissolved in THF (tetrahydrofuran; 20 mL), then OsC>4 (17.8 mg, 0.07 mmol) solution in THF (0.1 mL) was added to it, followed by addition of water (20 mL) and NaIO₄ (614.7 mg, 3.0 mmol). The mixture was stirred at room for 1 hour and extracted with ethyl acetate (50 mL x 3 times). The combined organic layer was dried over anhydrous MgSCU and concentrated under reduced pressure to yield 2,2,2-trifluoro-N-(3-((7-formylpyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)-N-((lR,5S)-8-(2,2,2-tr ifluoroacetyl)-8-azabicyclo[3.2.1]octan-3-yl)acetamide as yellow solids (380 mg, 98.0% yield), which was used in the next step without further purification, m / z (ESI), calcd. for C₂₆H₁₉F₉N₆O₄:650, found:651,[M+H]+.Step 4: To a solution of N-(diphenylphosphate) methylsulfonamide tert butyl ester (PHRT-010001-55; 50.8 mg, 1.44 mmol) in DMF (20 mL) was added NaH (60wt%; 240.4 mg, 5.76 mmol), and the mixture was stirred at room temperature for 30 minutes. Then 2,2,2-trifluoro-N-(3-((7-formylpyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)-N-((lR,5S)-8-(2,2,2-trifluoroacetyl)-8-azabicyclo[3.2.1]octan-3-yl)acetamide (380.0 mg, 0.59 mmol) was introduced before additional 1-hour stirring. Afterward, the reaction mixture was quenched by slowly adding water (0.5 mL), then extracted with ethyl acetate (200 mL x 3 times). The combined organic layer was dried over anhydrous MgSO₄ and concentrated under reduced pressure to yield tert-butyl(((E)-2-(2-((5-(((lR,5S)-8-azabicyclo[3.2.1]octan-3-yl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl) carbamate (300.0 mg, 75.0% yield), which was used in the next step without further purification, m / z (ESI), calcd. for C28H32F3N7O5S:635, found:636, [M+H]+.Step 5: Tert-butyl(((E)-2-(2-((5-(((lR,5S)-8-azabicyclo[3.2.1]octan-3-yl)amino)-2- (trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (160.0mg, 0.23 mmol) was dissolved in DCM (10 mL), then formaldehyde( 20.7 mg, 0.69 mmol) and NaBH(AcO)3 (38.2 mg, 0.69 mmol) was added to it. The reaction mixture was stirred at room temperature for 2 h, followed by concentration under reduced pressure. The residue was eluted with DCM / MeOH (v / v = 10 / 1) from a silica-gel column to afford tert-butyl (((E)-2-(2-((5-(((lR,5S)-8-methyl-8-azabicyclo[3.2.1]octan-3-yl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate as yellow solids (100.1 mg, 63% yield) m / z(ESI), calcd. for C29H34F3N7O5S: 649, found:650[M+H]+.Step 6: tert-butyl (((E)-2-(2-((5-(((lR,5S)-8-methyl-8-azabicyclo[3.2.1]octan-3-yl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (100.0 mg, 0.14 mmol) was dissolved in DCM (10 mL), then trifluoroacetic acid (2 mL) was added to that. The reaction mixture was stirred at room temperature for 0.5 h, followed by concentration under reduced pressure; the residue was purified by with reversed-phase flash chromatography ( Column: XBridge Prep OBD C18 Column, 30*150 mm, 5pm; mobile phase A: 10 mM aqueous solution of NH4HCO3, mobile phase B: ACN; flow rate: 60 mL / min; gradient: 27% B to 43% B in 9 min; wave length: 254 nm; RT: 7.8 min) to yield (E)-2-(2-((5-(((lR,5S)-8-methyl-8-azabicyclo[3.2.1]octan-3-yl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)ethene-l-sulfonamide as yellow solids (13.2 mg, 13.5%yield). m / z (ESI), calcd. for C26H30F3N7O4S: 549, found:550, [M+H]+. 1HNMR (300 MHz, DMSO-d6) 89.68 (s, 1H), 9.42 (s, 1H), 9.17 (s, 1H), 7.67 (s, 1H), 7.60 - 7.45 (m, 2H), 7.26 (s, 2H), 7.12 (d, J = 8.9 Hz, 1H), 6.92 (s, 1H), 6.48 (s, 1H), 5.81 (s, 1H), 3.09 (dd, J = 6.6, 4.2 Hz, 2H), 2.21 (s, 3H), 2.13 - 1.88 (m, 6H), 1.69 (d, J = 15.1 Hz, 2H), 1.24 (s, 1H).Following the teachings of the General Reaction Schemes and the synthesis procedure for Example 56 were prepared as shown in Table VII:TABLE VIIExample # Structure Resera ID?' Externa! 3D:> NMR L*. LCMS 'H NMR (400 MHz, DMSO-d6) 6947 (s, 1H), A, > XZ z M. > X7, 9.26 (s, 1H), 7.55 -7.46 (m, 2H), 7.37 (d, J =14.9 Hz, 1H), 7.22 (s, 2H), 6.88 (s, 1H), 6.78 xr (d, J = 8.4 Hz, 1H), 6.33 (dd, J =8.4, 2.2 Hz,° 'x 1H), 5 38 (d, J = 8.0 Hz, 1H), 3.45 (dt, J =8.1,57 RT-010201 PHRT-010095 4.1 Hz, 1H), 2.84 (s, 3H), 2.58 (ddd, J = 12.4, 508.3, [M+H]+ “Y^£rrfb7.0, 2.7 Hz, 1H), 2.44 (ddd, J = 19.8, 9.9, 7.1Hz, 3H), 2.23 (S, 3H), 1.96 - 1.88 (m, 2H),1.80 (td, J = 8.3, 4.2 Hz, 1H), 1.72 - 1.53 (m,4H), 069 (dq, J = 8.2, 3.9, 2 7 Hz, 2 H), 0.43(d, J = 3.9 Hz, 2H).'H NMR (400 MHz, DMSO-d6) 9.38 (s, 1H),9.14 (s, 1H), 7.68 (s, 1H), 7.44 (q, J = 149Hz, 2H), 7.29 (s, 2H), 7.09 (d, J = 8.7 Hz, 1H),6.38 (dd, J =8.8, 27 Hz, 1H), 5.16 (d, J = 8.058 RT-010194 PHRT-010107 544, [M+H]+Hz, 1H), 3.77 (tt, J = 65, 3.0 Hz, 1H), 2.61(ddd, J = 13.1, 7.3, 2.9 Hz, 1H), 2.50 - 2.39(m, 4H), 2.25 (s, 3H), 2.01 - 1.87 (m, 2H),1.76 - 150 (m, 4H), 0.70 - 0.52 (m, 4H).'H NMR (400 MHz, DMSO-d6) 69 72 (s, 1H),9.38 (s, 1H), 7.66 (s, 1H), 7.43 (q, J = 149Hz, 2H), 7.26 (s, 2H), 6.78 (d, J = 8.4 Hz, 1H),6.70 (s, 1H), 6.37 (dd, J = 8.4, 2.0 Hz, 1H),5.41 (d, J = 7.9 Hz, 1H), 3.48 - 3.41 (m, 1H),59 RT-010204 PHRT-O1O11O 528.2, [M+H]+2.61 - 2.54 (m, 1H), 2.47 - 2.35 (m, 2H),2.23 (s, 3H), 1.91 (q, J = 11.4, 9.3 Hz, 2H),1 78 (dq, J =84, 5 6, 43 Hz, 1H), 1 71 -1 50(m, 4H), 071 -0 60 (m, 2H), 043 (q, J =5 4Hz, 2H).'H NMR (400 MHz, DMSO-d6) 610.01 (s,1H), 943 (s, 1H), 7.69 (s, 1H), 7.55 -7.37(m, 2H), 7.28 (s, 2H), 7.11 (d, J = 8.8 Hz, 1H),6.89 (s, 1H), 6.51 (dd, J = 8.9, 2.7 Hz, 1H),5.85 (d, J = 7.7 Hz, 1H), 3.15 (d, J = 7.6 Hz,60 RT-010212 PHRT-010112 558.1, [M+H]+ •V6. O' “ 1H), 2 76 (d, J = 11.2 Hz, 2H), 2.18 (s, 3H),2.09 - 1.99 (m, 2H), 1.92 (d, J = 11.5 Hz, 2H),1.42 (q, J = 11.2, 98 Hz, 2H), 1.23 (s, 2H).19F NMR (376 MHz, DMSO-d6) 6 -56.93, - 73.42.'H NMR (400 MHz, DMSO-d6) 69 65 (s, 1H),9.35 (s, 1H), 7.58 (s, 1H), 7.39 (s, 2H), 7.25(s, 2H), 686- 6.61 (m, 2H), 6.37 (dd, J = 8.5,2.4 Hz, 1H), 5.39 (d, J = 8.0 Hz, 1H), 3.45 (dq,6 A X / kz-s'? J = 9.0, 4.9 Hz, 1H), 2.59 (ddd, J = 13.2, 7.3,1 C y- RT-010214 PHRT-010116 2.9 Hz, 1H), 2.42 (dd, J = 119, 8.7 Hz, 1H), 512.2, [M+H]+2.00 - 1.85 (m, 2H), 1.78 (td, J = 8.4, 4.3 Hz,1H), 1 65 (dddd, J = 23.1, 11.0, 8.3, 5.0 Hz,2H), 154 (t, J = 9.0 Hz, 2H), 0.67 (dt, J = 8.7,3.0 Hz, 2H), 0.48-0.37 (m, 2H). 19F NMR(376 MHz, DMSO-d6) 6 -7049.Example 62Example 62 Step 1: Tert-butyl (E)-4-(3-((7-(2-(N-(tert-butoxycarbonyl)sulfamoyl)vinyl)-5-chloropyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)piperazine-l-carboxylate (60.0 mg, 0.37 mmol) was dissolved in DMF (6 mL), then CsF (75.0 mg, 0.49 mmol) was added. The mixture was stirred at 80 °C for 3 hours, then extracted with ethyl acetate (50 mL x 3 times). The combined organic layer was dried over tetrahydrous MgSO₄ and concentrated under reduced pressure; the residue was purified with a C18 column (eluent: ACN / H2O, 0% to 100% in 30 min) to yield crude tert-butyl (E)-4-(3-((7-(2-(N-(tert-butoxycarbonyl)sulfamoyl)vinyl)-5-fluoropyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)piperazine-l -carboxylate as yellow solids (38.0 mg, 64% yield), which was used in the next step without further purification, m / z (ESI) calcd. for C30H35F4N7O7S: 713.2, found: 714.2, [M+H]+.Step 2: (E)-4-(3-((7-(2-(N-(tert-butoxvcarbonvl3sulfamovl)vinvl)-5-fluoropyridor4.3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)piperazine-l-carboxylate (38.0 mg, 0.05 mmol) was dissolved in the mixture of DCM (5mL) and trifluoroacetic acid, and the solution was stirred for 0.5 hours. Afterward, the reaction mixture was concentrated under the reduced pressure to afford trifluoroacetate, (E)-2-(5-fluoro-2-((5-(piperazin-l-yl)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)ethene-l -sulfonamide as yellow solid (50.2 mg), which was used in the next step without further purification, m / z (ESI) calcd. for C20H19F4N7O3S: 513.1, found: 514.1, [M+H]+.Step 3; The above obtained trifluoroacetate, (E)-2-(5-fluoro-2-((5-(piperazin-l-yl)-2-(trifluoromethoxy )phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)ethene-l -sulfonamide (50.2 mg, 0.05 mmol), was dissolved in DCM (3 mL), then formaldehyde (60% aqueous solution; 8.7 mg, 0.20 mmol) was P O added to it. The mixture was stirred for 30 minutes before adding sodium triacetoxyborohydride (12.3 mg, 0.05 mmol). After being stirred for another 1 hour, the reaction mixture was diluted with water and extracted with MeOH / DCM (1 / 10, v / v). The combined organic layer was dried over anhydrous MgSO₄ and concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography (column: XBridge Prep OBD Cl 8 Column, 30*150 mm, 5pm; mobile phase A: 10 mmol / L NH4HCO3 aqueous solution, mobile phase B: ACN; flow rate: 60 mL / min; gradient: 32% B to 53%B in 9 min; wavelength: 254nm / 220nm; RT: 8.03 min) to yield (E)-2-(5-fluoro-2-((5-(4-methylpiperazin-l-yl)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)ethene-l -sulfonamide (2.0 mg, 7.1 % overall yield of last two steps ) as yellow solid, m / z (ESI) calcd. for C21H21F4N7O3S:527.1, found: 528.1, [M+H]+.Following the teachings of the General Reaction Schemes and the synthesis procedure for Example 62 were prepared as shown in Table VIII:TABLE VIIIExample Structure Resero iD • External ID NMR; - 'H NMR (400 MHz, DMSO-d6) 9.77 (s, 1H), 9.37 (s, 1H), 7.63 (s, 1H), 7.40 (d, J = 6.6 Hz, 2H), 7.26 (s, 2H), 7.12 (s, 1H), 6.90 (d, J = 8.6 Hz, 1H), 6.77 (dd, 63 RT-O1O195 PHRT-010114 J = 8.6, 2.4 Hz, 1H), 3.15 - 3.02 (m, 4H), 2.47 - 2.40 (m, 4H), 2.21 (s, 3H), 1.90 — 1.77 (m, 1H), 0.77 — 0.67 (m, 2H), 0.50 (q, J = 5.6 Hz, 2H).19F NMR (376 MHz, DMSO-d6) 6 -70.06.1H NMR (400 MHz, DMSO-d6) 69.68 (s, 1H), 9.35 (s, 1H), 7.57 (s, 1H), 7.39 (s, 2H), 7.28 (s, 2H), 6.78 (d, J = 8.4 Hz, 2H), 6.43 (dd, J = 8.4, 2.4 Hz, 1H), 5.38 (d, J = 7.9 Hz, 1H), 3.20 - 3.03 (m, 1H), 2.73 64 RT-O1O2O5 PHRT-O1O115 (dt, J = 11.9, 3.8 Hz, 2H), 2.16 (s, 3H), 1.99 (t, J =10.8 Hz, 2H), 1.90 (d, J = 11.2 Hz, 2H), 1.82 - 1.75 (m, 1H), 1.39 (q, J = 11.5, 9.7 Hz, 2H), 0.73 -0.59 (m, 2H), 0.52 -0.33 (m, 2H). 19F NMR (376 MHz, DMSO-d6) 5 -70.46.1H NMR (400 MHz, DMSO-d6) 59.96 (s, 1H), 9.40 (s, 1H), 7.62 (s, 1H), 7.49 - 7.36 (m, 2H), 7.27 (bs, 1H), 7.11 (d, J = 8.9 Hz, 1H), 6.87 (s, 1H), 6.50 (dd, 65 RT-O1O215 PHRT-010118 J = 9.0, 2.8 Hz, 1H), 5.84 (d, J = 7.7 Hz, 1H), 3.20 - 3.07 (m, 1H), 2.73 (d, J = 11.0 Hz, 2H), 2.16 (s, 3H), 2.06 - 1.84 (m, 4H), 1.45- 1.36 (m, 2H). 19F NMR (376 MHz, DMSO-d6) 5 -56.98, -69.85.Example Structure;x. Resero 8D=:. T. Eixternai ID tri NMR1H NMR (400 MHz, DMSO-d6) 6 9.96 (s, 1H), 9.40 (s, 1H), 7.64 (s, 1H), 7.41 (d, J = 3.6 Hz, 2H), 7.33 (d, J = 34 8 Hz, 2H), 7.12 (d, J = F 8.8 Hz, 1H), 6.82 (s, 1H), 6.45 (dd, J = 9.0,2.8 Hz, 1H), 5.89 (d, J = 7 8 Hz, 1H), 3.56 - 66 RT-010216 PH RT-010119Z"*A o''b 3.46 (m, 1H), 2 69 - 2.56 (m, 2H), 2.48 - 2.42 (m, 2H), 2 26 (s, 3H), 1.99 - 1.90 (m, 2H), 1.75 -1.69 (m, 2H), 1.61 -1.52 (m, 2H); 19F NMR (376 MHz, DMSO-d6) 6 -56.99, - 69.83..’H NMR (400 MHz, DMSO-d6) 9.37 (s, 1H), 9.16 (s, 1H), 7.68 (s, 1H), 7.57 (s, 1H), 7.47 (d, J = 14.9 Hz, 1H), 7.41 (s, 1H), 7.26 (s, 2H), 7.23 (s, 1H), 6.79 (dd, J = 8.9, 2.9 Hz, 1H), 67 RT-010196 PH RT-010120 3.84 (dq, J = 6.0, 2.8 Hz, 1H), 3.11 — 2.98(m, 4H), 2 47 (d, J = 4 9 Hz, 4H), 2 23 (s, 3H), 0.70 (q, J = 6 7, 5.6 Hz, 2H), 0.62 (d, J = 6.3 Hz, 2H) 19F NMR (376 MHz, DMSO-d6) 6 - 70.20.1H NMR (400 MHz, DMSO-d6) 6 9.36 (s, 1H), 9.16 (s, 1H), 7.60 (s, 1H), 7.47 - 7.36 F (m, 2H), 7 29 (s, 2H), 7.17 (s, 1H), 7.10 (d, J = 8.8 Hz, 1H), 6.46 (dd, J = 8.8, 2.8 Hz, 1H), 5.25 (s, 1H), 3.78 (tt, J = 6.1, 2.9 Hz, 1H), 68 A A RT-010206 PH RT-O1O1213.27 (s, 1H), 3.07 (d, J = 11.4 Hz, 3H), 2.56 Ala ' ° (d, J = 11 0 Hz, 2H), 2 46 (s, 3H), 2 01 (d, J =12 1 Hz, 2H), 1 51 (q, J = 12 7, 11.8 Hz, 2H), 0.68 - 0.63 (m, 2H), 0.61 - 0.57 (m, 2H). 19F NMR (376 MHz, DMSO-d6) 6 -70.13, -73.45.1H NMR (400 MHz, DMSO-d6) 6 9.35 (s, c 1H), 9.09 (s, 1H), 7.61 (s, 1H), 7.40 (s, 2H),7.29 (s, 2H), 7.09 (d, J = 8.8 Hz, 1H), 6.42 - ~rAH,i- 6.32 (m, 1H), 5 15 (s, 1H), 3.78 (dt, J = 8 8, 69 A A A^ AX ^,9, O ONK= RT-010197 PH RT-010122 4.2 Hz, 1H), 2.62 (dq, J = 9.7, 3.5, 2 4 Hz, p r X1- 1 'S< c 1H), 2.50 - 2.39 (m, 4H), 2.26 (s, 3H), 1.94 A\ o H (td, J = 13 2, 11 6, 5 5 Hz, 2H), 1 76 - 1 52 > (m, 4H), 0 69 -0.56 (m, 4H). 19F NMR (376MHz, DMSO-d6) 6 -70.15.Example 70Cl PHRT-010001-24 HCHO STAB Pd(dppf)CI2, K3PO4.DCM, rt, 2 h 1 4-dioxane, 90 °C, 6 Step 1 Step 2 Step 3O, Ph V OPh" S-NH 6 Boe 010001-55 7TA, DCM, NaH DMF, rt. 2 h rt 05 h Step 5 Step 670-5Example 70 Step 1: The mixture of tert-butyl 4-(N-(3-amino-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoroacetamido)azepane-l -carboxylate (1.0 g, 2.11 mmol), 2,7-dichloropyrido [4, 3-d] pyridine (843.2 mg, 4.23 mmol) and i-PrOH (40 mL) was stirred at 120 °C for overnight. After being cooled to room temperature, the mixture was concentrated under reduced pressure, and the residue was purified with a silica gel column (eluent: CFLCh / MeOH = 10 / 1, v / v) to yield N-(azepan-4-yl)-N-(3-((7-chloropyrido[4, 3-d]pyrimidin-2-yl)amino)-4-(tri fluoromethoxy )phenyl)-2,2,2-trifluoroacetamide (501.0 mg, 43% yield), m / z (ESI) calcd. for C₂₇H₂₇ClF₆N₆O₄: 548.17, found: 549.17, [M+H]+.Step 2: N-(azepan-4-yl)-N-(3-((7-chloropyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoroacetamide (500.0 mg, 0.91 mmol) was dissolved in DCM (50 mL), followed by addition of formaldehyde (40% aqueous solution; 174.1 mg, 5.81 mmol). After being stirred for 10 minutes, NaBH(AcO)₃ (488.2 mg, 2.31 mmol) was added, and the mixture was stirred for about 0.5 h. Afterward, the reaction mixture was diluted with water and extracted with DCM / MeOH (10 / 1, v / v). The combined organic phases were dried over anhydrous MgSO₄ and concentrated under reduced pressure to yield N-(3-((7-chloropyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-(l-methylazepan-4-yl)acetamide as yellow solid (430.0 mg, 99% yield), which was used in the next step withoutfurther purification, m / z (ESI) calcd. for C₂₃H₂₁ClF₆N₆O₂: 562.13, found: 563.13, [M+H]⁺.Step 3: N-(3-((7-chloropyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-(1-methylazepan-4-yl)acetamide (430.1 mg, 0.76 mmol), Pd(dppf)Cl₂ (62.4 mg, 0.07 mmol) and K3PO4 (325.2 mg, 2.28 mmol) were loaded into a single-neck round-bottom flask, followed by addition of solvent mixture of 1,4-dioxane and H₂O (20 mL, 5 / 1, v / v). A septum capped the flask, and the mixture was bubbled with N2 flow through a needle for 10 minutes, after which 4,4,5,5-tetramethyl-2-vinyl-l,3,2-dioxaborolane (353.2 mg, 2.29 mmol) was injected using a syringe. After stirring at 90 °C for 3 h, the reaction mixture was diluted with water and extracted with ethyl acetate (100 mL x 3 times). The combined organic layer was dried over anhydrous MgSO₄ and concentrated under reduced pressure. The residue was purified with a silica-gel column (eluent: DCM / MeOH = 10 / 1, v / v) to yield N1-(l-methylazepan-4-yl)-4-(trifluoromethoxy)-N3-(7-vinylpyrido[4,3-d]pyrimidin-2-yl)benzene-l,3-diamine as yellow solid (286.3 mg, 67%). m / z (ESI) calcd. for C₂₅H₂₄F₆N₆O₂: 554.19, found: 555.19, [M+H]⁺.Step 4: N¹-(1-methylazepan-4-yl)-4-(trifluoromethoxy)-N³-(7-vinylpyrido[4,3-d]pyrimidin-2-yl)benzene-1,3-diamine (286.0 mg, 0.51 mmol) was dissolved in THF (tetrahydrofuran; 4 mL), then OsO₄ (25.9 mg, 0.10 mmol) solution in THF (1 mL) was added there, followed by addition of water (5 mL) and NaIO₄ (439.0 mg, 2.06 mmol). The mixture was stirred for 1 hour and extracted with ethyl acetate (50 mL x 3 times). The combined organic layer was dried over anhydrous MgSO4and concentrated under reduced pressure. The residue was purified with a Cl 8 column to yield 2-((5-((l-methylazepan-4-yl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidine-7-carbaldehyde as yellow solid (116.0 mg, 40% yield), which was used in the next step without further purification, m / z (ESI) calcd. for C₂₄H₂₂F₆N₆O₃: 556.17, found: 557.17, [M+H]+.Step 5: Tert-butyl (((diphenylphosphoryl)methyl)sulfonyl)carbamate (164.3 mg, 0.41 mmol) was dissolved in DMF (10 mL), and NaH (60wt%; 66.9 mg, 2.78 mmol) was then added. The mixture was stirred at room temperature for 30 minutes, after which 2-((5-((l-methylazepan-4-yl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidine-7-carbaldehyde (116.0 mg, 0.21 mmol) was introduced to it, followed by additional 1-hour stirring at room temperature. Afterward, the reaction mixture was quenched by slowly adding water (0.5 mL), then extractedwith ethyl acetate (50 mL x 3 times). The combined organic phases were dried over anhydrous MgSO4and concentrated under reduced pressure, and the residue was purified with a silica-gel column (DCM / MeOH = 10 / 1, v / v)) to yield tert-butyl (E)-((2-(2-((5-((l-methylazepan-4-yl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (60.2 mg, 45% yield) as yellow solid.m / z (ESI) calcd. for C₂₈H₃₄F₃N₇O₅S: 637.23, found: 638.23, [M+H]+.Step 6: Tert-butyl (E)-((2-(2-((5-((l-methylazepan-4-yl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (55.2 mg, 0.08 mmol) was dissolved in DCM (dichloromethane; 5.0 mL), then trifluoroacetic acid (1.0 mL) was added to it. The reaction mixture was stirred at room temperature for 0.5 h and then concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography (column: X Bridge Prep OBD C18 Column, 30*150 mm, 30*150 mm, 5pm; mobile phase A: 10 mmol / L NH4HCO3 aqueous solution + 0.05% ammonium aqueous solution, mobile phase B: ACN; flow rate: 60 mL / min; gradient: 25% to 50% in 9 minutes; wavelength: 254nm / 220nm; RT: 7.38 min) to obtain (E)-2-(2-((5-((l-methylazepan-4-yl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)ethene-l -sulfonamide (2.4 mg, 5.2% yield) as yellow solids, m / z (ESI) calcd. for C₂₃H₂₆F₃N₇O₃S: 537.18, found: 538.18, [M+H]+.JH NMR (400 MHz, DMSO-d6) 5: 9.42 (s, 1H), 9.17 (s, 1H), 7.68 (s, 1H), 7.57 (d, J = 15.0 Hz, 1H), 7.46 (d, J = 15.0 Hz, 1H), 7.11 (dd, J = 8.9, 1.5 Hz, 1H), 6.98 (s, 1H), 6.42 (dd, J = 8.9, 2.8 Hz, 1H), 3.52 - 3.47 (m, 1H), 2.62 - 2.54 (m, 2H), 2.50 - 2.40 (m, 2H), 2.24 (s, 3H), 2.00 - 1.91 (m, 2H), 1.75 - 1.52 (m, 2H);19F NMR (376 MHz, DMSO-d₆) δ: -57.00.Following the teachings of the General Reaction Schemes and the synthesis procedure for Example 70 were prepared as shown in Table IX:TABLE IXExample # Structure ReseroJD ExternalJD NMR: ■’H NMR (300 MHz, DMSO-dB) 9.37 (s, 1H), 9.14 (s, 1H), 8.72 (s, 1H), 7.66 (s, 1H), 7.54 (d, J = 14.9 Hz, 1H), 7.43 (d, J = 14.9 Hz, 1H), 7.36 (s, 1H), 7.26 (s, 2H), 7.07 (d, J = 8.7 71 RT-010185 PHRT-010101Hz, 1H), 6.32 (dd, J = 8.8, 2.7 Hz, 1H), 5.15 (s, 1H), 3.77 uv (tt, J = 6.1, 3.1 Hz, 1H), 2.71 — 2.52 (m, 3H), 2.03 — 1.84(m, 3H), 1.83 — 1.39 (m, 5H), 0.72 — 0.54 (m, 4H)Example: *. Structure i.7. Resero ID.*. Eixtemai ID NMR1H NMR (400 MHz, DMSO-d6) 6 9.38 (s, 1H), 9.35 (s, 1H), 9.12 (s, 1H), 7.62 (s, 1H), 7.55 (d, J = 14.9 Hz, 1H), 7.43 (d, J = 14.9 Hz, 1H), 7.26 (s, 2H), 6.91 (s, 1H), 6.78 (d, J = 8.4 Hz, 1H), 6.40 (dd, J = 8.4, 2.2 Hz, 1H), 5.35 (d, J = 7.7 Hz, 1H), 3.12 (d, J = 7.1 72 RT-010187 PHRT-010103Hz, 1H), 2.74 (d, J = 11.6 Hz, 2H), 2.16 (s, 3H), 1.99 (t, J = 11.2 Hz, 2H), 1.94 - 1.87 (m, 2H), 1.84 - 1.77 (m, 1H), 1.39 (dt, J = 13.9, 7.0 Hz, 2H), 0.74 - 0.64 (m, 2H), 0.44 (q, J = 4.9, 4.4 Hz, 2H). 19F NMR (376 MHz, DMSO-d6) δ -73.45.1H NMR (400 MHz, DMSO-d6) 6 9.38 (s, 1H), 9.34 (s, 1H), 9.12 (s, 1H), 7.63 (s, 1H), 7.55 (d, J = 15.0A S. NH2Hz, 1H), 7 44 (d, J = 14 9 Hz, 1H), 7.26 (s, 2H), 6.88(d, J = 4.4 Hz, 1H), 6 78 (d, J = 8.4 Hz, 1H), 6.35 (dd, 73 RT-010188 PHRT-010104X-c^'" J = 8 4, 2 4 Hz, 1H), 5 39 (s, 1H), 2 64 - 2 56 (m,2H), 2.47 - 2.41 (m, 2H), 1.99 - 1.88 (m, 2H), 1.85 - 1.77(m, 1H), 1.77 - 1.47 (m, 5H), 0.76 - 0.65 (m, 2H), 0.46 - 0.42 (m, 2H).1H NMR (400 MHz, DMSO-d6) 9.39 (s, 1H), 9.15 (s, 1H), 8.74 (s, 1H), 7.65 (s, 1H), 7.56 (d, J = 14.9 Hz, HN^> A^^S'NHS1H), 7.49 — 7.35 (m, 2H), 7.27 (s, 2H), 7.08 (d, J =8.7 Hz, 1H), 6.39 (dd, J = 8.8, 2.7 Hz, 1H), 5.10 (d, J 73 RT-010193 PHRT-010100H = 7.9 Hz, 1H), 3.79 (dt, J = 8.8, 4.4 Hz, 1H), 3.16 —3.07 (m, 1H), 2.80 — 2.71 (m, 2H), 2.18 (s, 3H), 2.05 — 1.90 ( m, 4H), 1.46 — 1.35 (m, 2H), 0.71 — 0.57 (m, 4H).1H NMR (300 MHz, DMSO-d6) 6 9.72 (s, 1H), 9.38 Cl (s, 1H), 7.64 (s, 1H), 7.46 (d, J = 14.9 Hz, 1H), 7.38(d, J = 15.0 Hz, 1H), 7.27 (s, 2H ), 6.76 (t, J = 7.0 Hz, 2H), 6.42 (dd, J = 8.4, 2.4 Hz, 1H), 5.37 (d, J = 7.9 74 RT-010203 PHRT-010109Hz, 1H), 3.14 — 3.03 (m, 1H), 2.72 (d, J = 10.7 Hz, 2H), 2.14 (s, 3H), 2.01 -1.80 (m, 4H), 1.78 (ddd, J = 13.3, 8.2, 5.3 Hz, 1H), 1.37 (q, J = 10.9 Hz, 2H), 0.73 - 0.58 (m, 2H), 0.45 - 0.40 (m, 2H).1H NMR (400 MHz, DMSO-d6) 8 9.31 (s, 1H), 9.08 (s, 1H), 7.59 (s, 1H), 7.41 (d, J = 14.9 Hz, 1H), 7.33 (d, J = 15.0 Hz, 1H), 7.20 (s, 1H), 7.11 (s, 1H), 7.02 (d, J = 8.7 Hz, 1H), 6.37 (dd, J = 8.8, 2.8 Hz, 1H), 75 RT-010209 PHRT-010125 5.05 (d, J = 7.9 Hz, 1H), 3 70 (tt, J = 6.2, 3.0 Hz, 1H),3.05 (s, 1H), 2.84 - 2.72 (m, 2H), 2.25 (q, J = 7.1 Hz, 2H), 2.00 - 1.79 (m, 4H), 1.37 - 1.22 (m, 2H), 0.93 (t, J = 7.2 Hz, 3H), 0.61 - 0.56 (m, 2H), 0.54 - 0.49 (m, 2H)1H NMR (400 MHz, DMSO-d6) 6 9.38 (s, 1H), 9.16 C! (s, 1H), 7.67 (s, 1H), 7.48 (d, J = 15.0 Hz, 1H), 7.41(d, J = 14.9 Hz, 1H), 7.29 (s, 2H), 7.18 (s, 1H), 7.09 (d, J = 8 8 Hz, 1H), 6 44 (dd, J = 8 8, 2 8 Hz, 1H), 76 RT-010202 PHRT-010106 5.14 (s, 1H), 3.77 (ddd, J = 8.6, 5.8, 2.8 Hz, 1H), -W H )' “ 3.12 (d, J = 14.2 Hz, 1H), 2.80 (d, J = 11.0 Hz, 2H),2.22 (s, 3H), 2.10 (t, J = 11.2 Hz, 2H), 1.94 (d, J = 12.4 Hz, 2H), 1.52 - 1.35 (m, 2H), 0.75 - 0.63 (m,2H), 0.61 - 0.56 (m, 2H).Example 78HCI. MeOH, 1 4-dioxane, STAB.40 °C 20 h DOM, rt 2 h. Step 1 Step 278-1 78-2Step 1: Tert-butyl (E)-((2-(2-((5-(N-(4-(4-acetylpiperazin-1-yl)cyclohexyl)-2,2,2-trifluoroacetamido)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (100.0 mg, 0.41 mmol) and HCl (4 M in 1,4-dioxane; 2.5 mL) were loaded into a thick-wall glass vial, followed by addition of MeOH (2.5 mL). The mixture was stirred at 40 °C for around 20 hours and then concentrated under reduced pressure. The residue was eluted with CH2Cl2 / MeOH (v / v = 10 / 1) from a silica-gel column to afford (E)-2,2,2-trifluoro-N-(4-(piperazin-l-yl)cyclohexyl)-N-(3-((7-(2-sulfamoylvinyl)pyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)acetamide as yellow solids (46.0 mg, 67.0%yield). m / z(ESI) calcd. for C₂₆H₃₁F₃N₈O₃S: 592.2, found: 593.2, [M+H]+.Step 2: (E)-2,2,2-trifluoro-N-(4-(piperazin-l-yl)cyclohexyl)-N-(3-((7-(2-sulfamoylvinyl)pyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)acetamide (36.0 mg, 0.06 mmol) and cyclopropanecaboxaldehyde (12.6 mg, 0.18 mmol) was dissolved in DCM (3mL), and the mixture was stirred at room temperature for 10 min prior to addition of sodium triacetoxyborohydride (38.2 mg, 0.18 mmol). The reaction mixture was stirred at room temperature for 2 h, followed by concentration under reduced pressure; the residue was purified by high-pressure preparation HPLC (Column: XBridge Prep C18 OBD Column, 30*150 mm, 5pm; mobile Phase A: 10 mM aqueous solution of NH4HCO3 + 0.05% NH3H2O, mobile Phase B: ACN; flow rate: 60 mL / min; Gradient: 14% B to 40% B in 9 min; Wave Length: 254 nm / 220 nm, RT: 8.18 min) to yield(E)-2-(2-((5-(((lr,4r)-4-(4-(cyclopropylmethyl)piperazin-l-yl)cyclohexyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3 -d]pyrimidin-7-yl)ethene- 1 -sulfonamide as yellow solids (7.5 mg, 20.1% yield). MS (ESI) calcd. For C₃₀H₃₇F₃N₈O₃S: 646.2 m / z, found LCMS: 647.2 [M+H]+. ¹H NMR (300 MHz, Methanol-d4) δ 9.35 (s, 1H), 9.13 (s, 1H), 7.78 (s, 1H), 7.68 - 7.48 (m, 3H), 7.11 - 7.04 (m, 1H), 6.46 (ddd, J = 19.1, 8.8, 2.7 Hz, 1H), 4.68 - 4.52 (m, 3H), 2.79 - 2.68 (m, 4H), 2.66 (s, 7H), 2.41 - 2.26 (m, 5H), 2.14 - 2.03 (m, 2H), 1.79 - 1.65 (m, 2H), 1.55 - 1.41 (m, 2H), 1.35 - 1.18 (m, 3H), 0.96 - 0.85 (m, 1H), 0.56 (d, J =7.7 Hz, 2H), 0.20 - 0.12 (m, 2H).19F NMR (282 MHz, Methanol-d4) 8 -59.98.Following the teachings of the General Reaction Schemes and the synthesis procedure for Example 78 were prepared as shown in Table X:TABLE XExample # Structure Name NMR LCMS (E)-2-(2-((5-(((lS,4s)-4-(4- 1H NMR (300 MHz, Methanol-d4) 6 methylpiperazin-1- 9.36 (d, J = 0.6 Hz, 1H), 9.13 (s, 1H), yl)cyclohexyl)amino)-2- 7.71 -7.60 (m, 3H), 7.54 (d, J = 15.0 Hz, (trifluoromethoxy)phenyl)amino)py 1H), 7.11 (dd, J = 8.9, 1.4 Hz, 1H), 6.5179 xcw rido[4,3-d]pyrimidin-7-yl)ethene-l- (dd, J = 9.0, 2.8 Hz, 1H), 3.63 (d, J =6.3 607.2 [M+H]+NH;sulfonamide Hz, 1H), 2.94-2.35 (m, 9H), 2.31 (s,3H), 2.12 - 1.99 (m, 2H), 1.73 (q, J =10.9, 9.0 Hz, 6H). 19F NMR (282 MHz,Methanol-d4) 8-59.97.(E)-2-(2-((2-cyclopropoxy-5-(4- 1H NMR (500 MHz, DMSO-d6) 89.40 methylpiperazin-1- (s, 1H), 9.15 (s, 1H), 8.80 (s, 1H), 7.75 yl)phenyl)amino)pyrido[4,3- (s, 2H), 7.57 (d, J = 14.9 Hz, 1H), 7.51d]pyrimidin-7-yl)ethene-l- (d, J = 15.0 Hz, 1H), 7.20 (d, J = 8.9 Hz,80V / S. A-. o'b sulfonamide 3H), 6.74 (dd, J =8.9, 2.9 Hz, 1H), 3.86 482.2, [M+H] +(tt, J = 6.1, 3.0 Hz, 1H), 3.08 (t, J = 4.9Hz, 4H), 2.23 (s, 3H), 1.23 (s, 1H), 0.75-0.67 (m, 2H), 0.64 (q, J =4.8, 4.0 Hz,2H).(E)-2-(2-((2-cyclopropyl-5-(4- 1H NMR (300 MHz, DMSO-d6) 89.45 methylpiperazin-1- (s, 1H), 9.37 (s, 1H), 9.11 (s, 1H), 7.67 yl)phenyl)amino)pyrido[4,3- (s, 1H), 7.54 (d, J = 15.0 Hz, 1H), 7.45d]pyrimidin-7-yl)ethene-l- (d, J = 14.9 Hz, 1H), 7.23 (s, 3H), 6.8881 sulfonamide (d, J =8.5 Hz, 1H), 6.73 (dd, J = 8.6, 2.6 466.2, [M+H] +Hz, 1H), 3.09 (t, J = 5.0 Hz, 4H), 2.44 (d,J = 5.0 Hz, 4H), 2.20 (s, 3H), 1.89 - 1.81(m, 1H), 0.79 -0.67 (m, 2H), 0.54-0.43 (m, 2H).Example 8282-1 82-2 Example 82 Step 1: Tert-butyl (E)-((2-(5-chloro-2-((2-cyclopropoxy-5-(4-methylpiperazin-l-yl)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (35.0 mg, 0.05 mmol), Pd(dppf)Cl₂ (4.6 mg, 0.005 mmol) and K3PO4 (36.2 mg, 0.15 mmol) were loaded into a singleneck round-bottom flask, followed by addition of solvent mixture of 1,4-di oxane (3 mL). The flask was capped by a septum, and the mixture was bubbled with N2 flow through a needle for 10 minutes, after which Methylboronic acid (6.7 mg, 0.10 mmol) was injected using a syringe. After being stirred at 90 °C for 4 h, the reaction mixture was diluted with water and extracted with ethyl acetate (100 mL x 3 times). The combined organic layer was dried over anhydrous MgSO₄ and concentrated under reduced pressure; and the residue was eluted with a silica-gel column (eluent: MeOH / CH2C12= 1 / 10, v / v) to yield tert-butyl (E)-((2-(2-((2-cyclopropoxy-5-(4-methylpiperazin-l-yl)phenyl)amino)-5-methylpyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate as yellow solids (14.0 mg, 51.0% yield), m / z (ESI) calcd. for C₂₉H₃₇N₇O₅S: 595.3, found: 596.20, [M+H]+.Step 2; Tert-butyl (E)-((2-(2-((2-cyclopropoxy-5-(4-methylpiperazin-l-yl)phenyl)amino)-5-methylpyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (14.0 mg, 0.02 mmol) was dissolved in 1,4-di oxane (1 mL), then HCl-dioxane (4 M, 1 mL) was added thereto. The reaction mixture was stirred at room temperature for 0.5 h, followed by concentration under reduced pressure. The residue was purified by reversed-phase flash chromatography (column: Xselect CSH C18 OBD, 30*150 mm, 5pm; mobile phase A: 0.1% FA aqueous solution, mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: 3% B to 33% B in 7 min; wave length: 254 nm; RT: 7.55 min) to obtain (E)-2-(2-((2-cyclopropoxy-5-(4-methylpiperazin-l-yl)phenyl)amino)-5-methylpyrido[4,3-d]pyrimidin-7-yl)ethene-l-sulfonamide (3.5 mg, 30.0% yield) as yellow solids, m / z (ESI), calcd. for C₂₄H₂₉N₇O₃S: 495.2, found: 496.2, [M+H]⁺. ¹H NMR (300 MHz, DMSO-d₆) δ 9.51 (s, 1H), 8.70 (s, 1H), 7.83 (s, 1H), 7.60- 7.39 (m, 3H), 7.21 (d, J= 10.5 Hz, 3H), 6.73(dd, J= 8.9, 2.9 Hz, 1H), 3.86 (ddt, J= 8.9, 5.9, 3.1 Hz, 1H), 3.15 -3.05 (m, 4H), 2.86 (s, 3H), 2.54 (d, J= 4.6 Hz, 2H), 2.27 (s, 3H), 1.23 (s, 2H), 0.75 - 0.62 (m, 4H).Following the teachings of the General Reaction Schemes and the synthesis procedure for Example 82 were prepared as shown in Table XI:TABLE XIExample # Structure i - ReseroJD; * ExternaMD NMR; *1H NMR (400 MHz, DMSO-d6) δ 9.59 (s, 1H), 9.50 (s, 1H), 8.71 (s, 1H), 7.56 (d, J = 14.8 Hz, 1H), 7.53 (s, 1H), 7.46 (s, 1H), 7.40 (d, J = 14.9 Hz, 1H), 7.25 (s, 2H), 7.11 (d, J = 8.4 Hz, 1H), 6.44 (d, J = 8.3 Hz, 83 RT-010191 PHRT-010091 1H), 5.43 (s, 1H), 3.80 (s, 1H), 3.50 (d, J = 11.9 Hz,2H), 3.09 (q, J = 11.5, 10.6 Hz, 2H), 2.86 (s, 3H), 2.79 (s, 3H), 2.18 (d, J = 13.6 Hz, 2H), 1.98 (s, 1H), 1.57 (q, J = 12.5, 11.7 Hz, 2H), 0.68 (d, J = 5.9 Hz, 2H), 0.64 (d, J = 5.0 Hz, 2H).’H NMR (400 MHz, DMSO-d6) 9.49 (s, 1H), 8.61 (s, 1H), 7.60 - 7.50 (m, 1H), 7.45 (s, 1H), 7.39 (d, J = 14.8 Hz, 1H), 7.25 (s, 1H), 7.08 (d, J = 8.7 Hz, 1H), 6.32 (dd, J = 8.8, 2.7 Hz, 1H), 5.13 (s, 1H), 3.79 (tt, 84 x iv RT-010192 PHRT-010092„A, L Z~\ O’ J = 6.1, 3.1 Hz, 1H), 2.86 (s, 3H), 2.70 - 2.59 (m,1H), 2.55 (dd, J = 7.9, 3.3 Hz, 3H), 2.49 - 2.38 (m, 1H), 2.28 (s, 2H), 1.95 (ddd, J = 22.0, 12.4, 5.2 Hz, 2H), 1.82 - 1.26 (m, 4H), 0.96 - 0.27 (m, 4H)1H NMR (400 MHz, DMSO-d6) δ 9.47 (s, 1H), 9.27 (s, 1H), 7.52 (d, J = 14.9 Hz, 1H), 7.45 (s, 1H), 7.36 (d, J = 14.8 Hz, 1H), 7.22 (s, 2H), 6.91 (s, 1H), 6.77 (d, J = 8.4 Hz, 1H), 6.38 (dd, J = 8.5, 2.4 Hz, 1H), 85 RT-010200 PHRT-010094 5.34 (d, J = 7.9 Hz, 1H), 3.17 - 3.04 (m, 1H), 2.83(s, 3H), 2.72 (dt, J = 12.0, 3.7 Hz, 2H), 2.15 (s, 3H), 2.04 - 1.87 (m, 4H), 1.81 (ddd, J = 13.6, 8.4, 5.3 Hz, 1H), 1.38 (qd, J = 11.2, 3.7 Hz, 2H), 0.74 - 0.62 (m, 2H), 0.46 - 0.35 (m, 2H).1H NMR (300 MHz, DMSO-d6) δ 9.69 (s, 1H), 9.52 (s, 1H), 7.55 (d, J = 4.4 Hz, 1H), 7.51 (s, 1H), 7.45 - 7.38 (m, 2H), 7.23 (d, J = 10.0 Hz, 3H), 6.84 (dd, J = 86 HNA / ^^XNH' RT-010179 PHRT-0100969.0, 3.0 Hz, 1H), 3.30 - 3.23 (m, 4H), 3.19 - 3.12 (m, 4H), 2.84 (s, 3H), 2.21 (s, 3H), 1.22 (s, 2H). 19F NMR (282 MHz, DMSO-d6) δ -56.80.Example # Structure Resero ID y External ID y NMR y 1H NMR (400 MHz, DMSO-d6) δ 9.55 (d, J = 25.2 Hz, 2H), 7.57 - 7.48 (m, 2H), 7.39 (d, J = 14.9 Hz, 1H), 7.23 (s, 2H), 7.09 (d, J = 8.6 Hz, 1H), 7.02 (s, 1H), 6.46 (d, J = 8.9 Hz, 1H), 87 J RT-010210 PHRT-010097 5.81 (d, J = 7.6 Hz, 1H), 3.13 (s, 1H), 2.86 (s, 3H), 2.74 (d, J = 11.2 Hz, 2H), 2.16 (s, 3H),1.97 (dd, J = 33.5, 11.4 Hz, 4H), 1.42 (q, J = 10.1 Hz, 2H). 19F NMR (376 MHz, DMSO-d6) δ -56.96.1H NMR (400 MHz, DMSO-d6) δ 9.57 (s, 1H), 9.52 (s, 1H), 7.54 (d, J = 14.9 Hz, 1H), 7.51 (s, 1H), 7.39 (d, J = 14.9 Hz, 1H), 7.23 (s, 2H), 7.09 (d, J = 8.1 Hz, 1H), 6.97 (s, 1H), 6.41 (dd, J = 8.9, 2.8 Hz, 1H), 5.84 (d, J = 7.7 88 RT-010211 PHRT-010098Hz, 1H), 3.47 (dt, J = 8.6, 4.4 Hz, 1H), 2.86 (s, 3H), 2.62 -2.53 (m, 2H), 2.48 - 2.40 (m, 2H), 2.24 (s, 3H), 2.04 - 1.83 (m, 2H), 1.72 - 1.63 (m, 2H), 1.61 - 1.55 (m, 2H). 19F NMR (376 MHz, DMSO-d6) δ -56.98.Example 89Step 1: The mixture of N-(3-amino-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-(4-(4-(2,2,2-trifluoroacetyl)piperazin-1-yl)phenyl)acetamide (1.4 g, 2.51 mmol), 2,7-dichloropyridine [4, 3-d] pyrimidine (1.0 g, 5.11 mmol) and z-PrOH (60 mL) was stirred at 120 °C for overnight. After being cooled to room temperature, the mixture was concentrated under reduced pressure, and the residue was purified with a silica gel column (eluent: CH2CI2 / EA = 2 / 1, v / v) to yield N-(3-((7-chloropyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-(4-(4-(2,2,2-trifluoroacetyl)piperazin-l-yl)phenyl)acetamide as yellow solid (587.1 mg, 33% yield), m / z (ESI) calcd. for C₂₈H₁₉ClF₉N₇O₃: 707.11, found: 708.11, [M+H]+.Step 2: N-(3-((7-chloropyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)-2,2,2-trifluoro-N-(4-(4-(2,2,2-trifluoroacetyl)piperazin-1-yl)phenyl)acetamide (587.1 mg, 0.83 mmol), Pd(dppf)Cl₂ (109.2 mg, 0.13 mmol) and K₃PO₄ (235.8 mg, 1.63 mmol) were loaded into a singleneck round-bottom flask, followed by addition of solvent mixture of 1,4-dioxane and H₂O (20 mL, v / v = 5 / 1). The flask was capped by a septum, and the mixture was bubbled with N2 flow through a needle for 10 minutes, after which 4,4,5,5-tetramethyl-2-vinyl-l,3,2-dioxaborolane (383.3 mg, 2.48 mmol) was injected using a syringe. After being stirred at 90 °C for 3 h, the reaction mixture was diluted with water and extracted with ethyl acetate (100 mL x 3 times). Thecombined organic layer was dried over anhydrous MgSO₄ and concentrated under reduced pressure; and the residue was purified with a silica-gel column (eluent: DCM / MeOH = 10 / 1, v / v) to yield 2,2,2-trifluoro-N-(4-(4-(2,2,2-trifluoroacetyl)piperazin-l-yl)phenyl)-N-(4-(trifluoromethoxy)-3-((7-vinylpyrido[4,3-d]pyrimidin-2-yl)amino)phenyl)acetamide as yellow solid (302.3 mg, 52% yield), m / z (ESI) calcd. for C30H22F9N7O3: 699.16, found: 700.16, [M+H]+.Step 3; 2.2.2-trifluoro-N-(4-(4-(2.2.2-trifluoroacetyl)piperazin-l-yl)phenyl)-N-(4-(trifluoromethoxy)-3-((7-vinylpyrido[4,3-d]pyrimidin-2-yl)amino)phenyl)acetamide (142.2 mg, 0.21 mmol) was dissolved in THF (tetrahydrofuran; 4 mL), then OsO4(2.6 mg, 0.05 mmol) solution in THF (1 mL) was added thereto, followed by addition of water (5 mL) and sodium periodate (233.1 mg, 2.67 mmol). The mixture was stirred at room for 1 hour and subsequently extracted with ethyl acetate (100 mLx 3 times). The combined organic layer was dried over anhydrous MgSO4and concentrated under reduced pressure to yield 2,2,2-trifluoro-N-(3-((7-formylpyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)-N-(4-(4-(2,2,2-trifluoroacetyl)piperazin-l-yl)phenyl)acetamide as yellow solid (140.3 mg, 90% yield), which was used in the next step without further purification, m / z (ESI) calcd. for C29H20F9N7O4: 701.14, found: 702.14, [M+H]+.Step 4: tert-butyl (((diphenylphosphoryl)methyl)sulfonyl)carbamate (73.2 mg,0.18 mmol) was dissolved in DMF (5 mL), and NaH (60wt%; 17.8 mg, 0.74 mmol) was then added. The mixture was stirred at room temperature for 30 minutes, after which 2,2,2-trifluoro-N-(3-((7-formylpyrido[4,3-d]pyrimidin-2-yl)amino)-4-(trifluoromethoxy)phenyl)-N-(4-(4-(2,2,2-trifluoroacetyl)piperazin-l-yl)phenyl)acetamide (130.1 mg, 0.18 mmol) was introduced thereto, followed by additional 1-hour stirring at room temperature. Afterward, the reaction mixture was quenched by slow addition of water (0.5 mL), then extracted with ethyl acetate (50 mL * 3 times). The combined organic layer was dried over anhydrous MgSO4and concentrated under reduced pressure, and the residue was purified with a silica-gel column (eluent: DCM / MeOH = 10 / 1, v / v) to yield tert-butyl (E)-((2-(2-((5-((4-(piperazin-l-yl)phenyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate as yellow solid (40.2 mg, 68% yield), m / z (ESI) calcd. for C31H33F3N8O5S: 686.22, found: 687.22, [M+H]+.IllStep 5; Tert-butyl (E)-((2-(2-((5-((4-(piperazin-l -yl)phenyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (40.1 mg, 0.42 mmol) was dissolved in DMF (4 mL), followed by addition of formaldehyde (40% aqueous solution; 1.7 mg, 0.05 mmol). After being stirred for 10 minutes, NaBH(AcO)3(36.9 mg, 0.17 mmol) was added thereto. The mixture was stirred at room temperature for about 0.5 h then diluted with water and extracted with DCM / MeOH (10 / 1, v / v). The combined organic phase was dried over anhydrous MgSO₄ and concentrated under reduced pressure, the residue was purified with a silica-gel column (eluent: DCM / MeOH = 4 / 1, v / v) to yield tert-butyl (E)-((2-(2-((5-((4-(4-methylpiperazin-l-yl)phenyl)amino)-2-(tri fluoromethoxy )phenyl)amino)pyrido[4, 3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (30.1 mg, 75% yield) as yellow solid, m / z (ESI) calcd. for C32H35F3N8O5S: 700.24, found: 701.24, [M+H]+.Step 6; Tert-butyl (E)-((2-(2-((5-((4-(4-methylpiperazin-l-yl)phenyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)vinyl)sulfonyl)carbamate (30.1 mg, 0.04 mmol) was dissolved in DCM (dichloromethane; 2.5 mL), then trifluoroacetic acid (0.5 mL) was added thereto. The reaction mixture was stirred at room temperature for 0.5 h, then concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography (column: Xselect CSH C18 OBD column, 30*150 mm; mobile phase A: 0.05% TFA aqueous solution, mobile phase B: ACN; flow rate: 60 mL / min; gradient: 19% B to 49% B in 7 min; wave length: 254nm; RT: 6.7 min) to obtain trifluoroacetate, (E)-2-(2-((5-((4-(4-methylpiperazin-l-yl)phenyl)amino)-2-(trifluoromethoxy)phenyl)amino)pyrido[4,3-d]pyrimidin-7-yl)ethene-l-sulfonamide as yellow solids (2.7 mg, 8.7% yield), m / z (ESI) calcd. for C27H27F3N8O3S: 600.19, found: 601.19, [M+H]+. 'H NMR (400 MHz, Acetonitrile-ds) 59.24 (s, 1H), 9.04 (s, 1H), 8.37 (s, 1H), 7.55 (d, J = 14.9 Hz, 1H), 7.44 (s, 1H), 7.39 (d, J = 7.6 Hz, 1H), 7.15 (t, J = 9.8 Hz, 3H), 7.00 (d, J = 8.4 Hz, 2H), 6.60 (d, J = 8.9 Hz, 1H), 3.63 (d, J = 13.3 Hz, 2H), 3.50 (s, 4H), 3.18 (t, J = 12.3 Hz, 2H), 2.81 (s, 3H).19F NMR (376 MHz, Acetonitrile-d3) 8 -58.90, -75.66.Following the teachings of the General Reaction Schemes and the synthesis procedure for Example 89 were prepared as shown in Table XII:TABLE XllExample # Structure Name NMR LCMS (S, E)-2-(2-((5-((l-ethylpyrrolidin-3- 1H NMR (400 MHz, Methanol-d4) 8yl)amino)-2- 9.39 -9.36 (m, 1H), 9.14 (s, 1H), 7.75 (trifluoromethoxy)phenyl)amino)py (s, 1H), 7.69 -7.63 (m, 2H), 7.53 (d, J - rido[4,3-d]pyrimidin-7-yl)ethene-l- 14.9 Hz, 1H), 7.21 - 7.16 (m, 1H), 6.51XXX - sulfonamide (dd, J = 8.9, 2.8 Hz, 1H), 3.35 (s, 5H), 90 524.25, [M+H]+2.99 (s, 1H), 2.86 (s, 1H), 2.23 - 2.16H °"b(m, 2H), 2.03 (s, 3H). 19F NMR (376MHz, Methanol-d4) 6 -59.79 --60.23(m), -76.78 --77. 19F NMR (376 MHz,Methanol-d4) 6 -60.01, -77.02.(S, E)-2-(2-((5-((l- 1H NMR (500 MHz, Methanol-d4) 8 isopropylpyrrolidin-3-yl)amino)-2- 9.37 (s, 1H), 9.15 (s, 1H), 7.78 (s, 1H), (trifluoromethoxy)phenyl)amino)py 7.72 - 7.63 (m, 2H), 7.56 (d, J = 14.9 Hz,rido[4,3-d]pyrimidin-7-yl)ethene-l- 1H), 7.13 (dd, J =9.0, 1.5 Hz, 1H), 6.47'Ll sulfonamide (dd, J = 8 9, 2 8 Hz, 1H), 4 07 (p, J = 5 4Hz, 1H), 3.21 (dd, J = 10.1, 7.1 Hz, 1H),91 538.2, [M+H]+2.81 (p, J = 8.9, 8.0 Hz, 2H), 2.60 (dd, J =H 10.0, 5.1 Hz, 1H), 2.54 (p, J = 6.6 Hz,1H), 2.40 (dq, J = 14.7, 7.6 Hz, 1H), 1.81(dq, J = 13.1, 6.8 Hz, 1H), 1.31 (s, 1H),1.19 (dd, J = 6.4, 4.0 Hz, 6H). 19F NMR(471 MHz, Methanol-d4) 6 -59.98.EXAMPLE A

[0111] This example illustrates that exemplary compounds of the present invention inhibit PLK1 enzymatic activity.

[0112] Compounds were solubilized in DMSO to produce 10 mM stock solutions and diluted to 100 pM working solutions in DMSO. A series of 10, three-fold dilutions of each compound working solution was dispensed (Tecan D300e digital dispenser) on a 384-well white ProxiPlate (PerkinElmer, #6008280) in two technical replicates, yielding dilution series with final concentrations in 10 pL total reaction volume ranging from 0.05 to 1000 nM. Volasertib (MedChemExpress, #HY-12137) & GSK461364 (MedChemExpress, #HY-50877) were used as positive controls (1.0 pM final concentration, 5 technical replicates each). The volume was made up to 4 pL using a kinase buffer (PerkinElmer, HTRF® KinEASE-STK SI kit # 62ST1PEB), supplemented with 1.0 mM DTT & 5.0 mM MgCl2, and that same assay buffer without compounds was used as a negative control. DMSO was normalized to 1% final concentration across all wells.

[0113] All subsequent assay components were diluted using the kinase buffer supplemented with 1 mM DTT & 5 mM MgCl2. A 25 nM PLK1 protein (Carna Biosciences, #05-157) working solution was prepared, and 2 pL was dispensed into each well, resulting in a 5 nMfinal concentration. A 10 pM STK substrate 1 -biotin solution (PerkinElmer, HTRF® KinEASE-STK SI kit #62ST1PEB) was prepared according to manufacturer’s instructions, and 2 pL was dispensed into each well, resulting in a 2 pM final concentration. Finally, a 100 pM ATP solution (Sigma- Aldrich, A7699) was prepared, and 2 pL was added to each well, resulting in a 20 pM final concentration, initiating the kinase reaction. The plate was sealed, shaken briefly to mix the reaction contents well, centrifuged, and incubated for 90 min at 37 °C.

[0114] After the kinase reaction finished, the plate seal was removed, 0.5 pM Streptavidin-XL665 (PerkinElmer, HTRF® KinEASE-STK SI kit #62ST1PEB) working solution was prepared according to manufacturer’s instructions in the detection buffer (PerkinElmer, HTRF® KinEASE-STK SI kit 62ST1PEB) and 5 pL was dispensed into each well, resulting in 0.125 pM final concentration. Then, 5 pL of the Antibody-cryptate (PerkinElmer, HTRF® KinEASE-STK SI kit 62ST1PEB) solution was added to each well. The plate was sealed again, shaken briefly, centrifuged, and incubated for 60 minutes at room temperature (25 °C). The FRET signal was then read on a PHERAstar® FSX microplate reader (BMG LABTECH) using 620 nm / 665 nm wavelengths for excitation / emission, respectively. Signal values were normalized for the background signal, and the acceptor / donor signal ratio was calculated and plotted against compound concentrations using GraphPad Prism™ software to generate semi-log concentrationresponse curves (CRCs). ICso values were determined using nonlinear regression of a variable slope model.TABLE XIIIExample # PLKI HTRF CRC / IC50 (nM) Example if PLK1 HIRE CRC / IC50(nM) Example if PLK1 HIRE CRC / IC50 (nM) 1 587 32 > 1000 62 4.62 > 1000 33 355.2 63 2.6> 10003 34 625.2 64 25.4941.84 > 1000 35 514.6 655 118.1 36 > 1000 666 83.1 37 > 1000 67 5.594.57 143.2 38 6892.990.08 48.5 39 69 33.288.69 266.7 40 3.9 70 19.42.910 405.3 41 71 65.43.611 > 1000 42 72 106.5 12 56.3 43 73 76.35.5 265.6 13 > 1000 44 744.4 129.9 14 > 1000 45 8.3 75 515 > 1000 46 76 916 78.7 47 12 7717 955.6 48 26.2 78 > 1000 18 10.1 49 11.5 79 143.4402.1 24.019 50 > 1000 80292.9 10.120 13.5 51 136.6 81 36.811.321 62.7 52 378.6 827.052.822 53 173.1 83 1.748.4> 100023 54 858.1 84 2.1> 1000899.524 55 754.4 85 3.1919.725 > 1000 56 8.5 86 4.8824.726 57 1.9 87792.827 > 1000 58 2.5 8828 347.8 59 21.1 89 355.973.629 60 90 800.774.5571.030 61 91 290.8597.631 N / AEXAMPLE B

[0115] This example illustrates the reactivity of glutathione in the reduced state (GSH) at pH 7.5 with exemplary compounds of the present invention.ANALYSES OF COMPOUND REACTIVITY WITH GSH (USP, CATALOG # 1294820) WERE PERFORMED IN A TIME-DEPENDENT EXPERIMENT THROUGH THE USE OF LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY (LC-MS) DETECTION. THE PEAK AREA OF THE COMPOUND NORMALIZED WITH THE PEAK AREA OF THE INTERNAL STANDARD (IS), RHODAMINE B (SIGMA ALDRICH, CATALOG # R6626), WAS CALCULATED IN ORDER TO DETERMINE THE HALF-LIFE (T½).

[0116] Compounds were solubilized in DMSO to obtain 10 mM concentrations. Dilutions of each compound were made in acetonitrile (LC-MS grade) to obtain 100 pM concentrations. An assay buffer, 50 mM Tris pH 7.5 with 150 mM NaCl, was deoxidized prior to testing by keeping it under a nitrogen stream for approximately 1 hour. GSH and the IS were solubilized in the assay buffer to obtain 1.2 mM and 25 pM concentrations, respectively.

[0117] To 86 pl of 1.2 mM GSH solution, 4 pl of 25 pM Rhodamine B solution was added into a glass vial with an insert. To initiate the reaction, 10 pL of the 100 pM compound solution was added to the vial, and the reaction mixtures were immediately injected into an LC-MS system, followed by repeated injections at fixed time intervals up to 24 hours. An autosampler temperature was set to 25°C to maintain a steady incubation temperature.Compounds were separated on an ACQUITY UPLC® HSS T3 1.8 pm, 2.1x100 mm column (Waters) at temperature 40°C with a 10-95% gradient of 0.1% formic acid in acetonitrile for 5.5 min at a flow rate of 0.25 mL / min using the ultra-high-performance liquid chromatography Dionex UltiMate 3000 system (Thermo Scientific) and analyzed using a Q Exactive1MPlus mass spectrometer equipped with an electrospray ionization ion source (Thermo Scientific). The mass spectrometer was operated in a standard Full MS mode in a scan range of 200 to 600 m / z.TABLE XIVExample # GSH Reactivity / GSH (mi n) Exampie # GSH Reactivity / GSH V / i (min) Example # GSH feactivity / GSH tta (min) 1 330 32 257 62 422 33 231 633 239 34 383 644 408 35 408 655 582 36 257 666 506 37 239 677 290 38 365 688 217 39 347 699 363 40 36 70 21810 299 41 209 71 36511 330 42 65 72 23112 367 43 7313 333 44 67 74 28914 215 45 7515 365 46 7616 363 47 7717 340 48 78 32918 462 49 517 79 24719 277 50 270 80 28920 248 51 352 81 36521 277 52 265 82 21722 239 53 244 8323 277 54 226 8424 224 55 259 8525 263 56 195 86 19326 289 57 8727 222 58 8828 468 59 89 53729 365 60 90 25730 408 61 91 34731 224

[0118] From the obtained chromatograms, peak areas of the test compounds and the internal standard (IS) were integrated using the Chromeleon™ Chromatography Data System, andthe area ratios between the test compounds and IS and a natural logarithm (In) from the obtained area ratios were calculated. The f / 2 value for each compound was determined from the plot curve of In (area ratio) versus time and the equation t½ =lN2 / (-slope).

Claims

We claim:

1. A compound of Formula (I):R1Formula (I)wherein:R1is hydrogen, halogen, hydroxy, aniline, C1-C6 alkyl, cycloalkyl, and haloalkyl;R2is hydrogen, alkoxy, or -O-haloalkyl optionally substituted with one or more alkoxy groups;R3is hydrogen, heterocyclyl optionally substituted with one or more R4, -NH-L1- N(RARB), -NH-C(0)0CH3, -NH-L2-cycloalkyl or -NH-L2-heterocyclyl, wherein the cycloalkyl or the heterocyclyl is optionally substituted with one or more R5;R6is OCF3, SCH3, cyclopropane, O-cyclopropane, OCH3, CH2CH3, azetidine, or S(O)(O)CH3;R7is NH2, NHCH3, or CH3;each R4is independently C1-C4 alkyl, cyclopropyl, hydroxyalkyl, -i -N R^R®), halogen, O, O-CH3, NH2, -C(O)O-t-butyl; cycloalkyl optionally substituted with NH2, -L3-aryl; or -L4-heterocyclyl;each R5is independently C1-C4 alkyl, N(CH3)(CH3), L2-aryl, or -L2-heterocyclyl, wherein L2-heterocyclyl is optionally substituted with one or more C1-C4 alkyl, C1-C4 alkylcycloalkyl, C(0)CH3 or aryl;each L1is C1-C4 alkylene;each L2is a bond or C1-C4 alkylene;each L3is -CH2-O-CH2-;each L4is a bond or methylene;each RAis hydrogen or C1-C3 alkyl;each RBis hydrogen or C1-C3 alkyl;and pharmaceutically acceptable salts thereof.

2. The compound of claim 1, wherein R1is halogen.

3. The compound of claim 2, wherein the halogen is chlorine, fluorine, or bromine.

4. The compound of claim 1, wherein R1is hydroxy.

5. The compound of claim 1, wherein R1is C1-C6 alkyl.

6. The compound of claim 5, wherein the C1-C6 is alkyl, is methyl, ethyl, or isopropyl.

7. The compound of claim 1, wherein R1is cycloalkyl.

8. The compound of claim 7, wherein the cycloalkyl is cyclopropyl.

9. The compound of claim 1, wherein R1is haloalkyl.

10. The compound of claim 9, wherein the haloalkyl is fluoromethyl, difluoromethyl, or trifluoromethyl.

11. The compound of claim 1, wherein R1is aniline.

12. The compound of claim 11, wherein the aniline is NH2.

13. The compound of any of claims 1-12, wherein R2is alkoxy or -O-haloalkyl.

14. The compound of claim 13, wherein R2is alkoxy.

15. The compound of claim 14, wherein the alkoxy is methoxy or propoxy.

16. The compound of claim 13, wherein R2is -O-haloalkyl.

17. The compound of claim 16, wherein the haloalkyl is difluoromethyl.

18. The compound of any of claims 1-17, wherein R6is OCF3.

19. The compound of any of claims 1-18, wherein R7is NH2.

20. The compound of any of claims 1-19, wherein R3is heterocyclyl optionally substituted with one or more R4, -NH-L1-N(RARB), -NH-L2-cycloalkyl or -NH-L2-heterocyclyl, wherein the cycloalkyl or the heterocyclyl is optionally substituted with one or more R5.

21. The compound of claim 20, wherein R3is heterocyclyl optionally substituted with one or more R4.

22. The compound of claim 21, wherein the heterocyclyl is 1,4-diazapanyl, 1 -methyl- 1, 4- diazapanyl or 1, 4-dimethyl-l,4-di(ll-oxidaneyl)-114,414-piperazinyl.

23. The compound of claim 21, wherein the heterocyclyl is pyrrolidinyl, piperidinyl, or piperazinyl, each optionally substituted with one or more R4, wherein R4is C1-C4 alkyl, hydroxyalkyl, -L1-N(RARB), -C(O)O-t-butyl, -L3-aryl, or -L4-heterocyclyl.

24. The compound of claim 23, wherein the heterocyclyl is pyrrolidinyl substituted with one R4, and R4is L4-heterocyclyl, wherein L4is methylene, and the heterocyclyl is pyrrolidinyl.

25. The compound of claim 23, wherein the heterocyclyl is piperidinyl substituted with one R4, and R4is L4-heterocyclyl, wherein L4is a bond, and the heterocyclyl is pyrrolidinyl.

26. The compound of claim 23, wherein the heterocyclyl is piperazinyl, and R4is C1-C4 alkyl, hydroxyalkyl, -L1-N(RARB), -C(O)O-t-butyl; or -L3-aryl.

27. The compound of claim 26, wherein the piperazinyl is substituted with one R4, and R4is C1-C4 alkyl, wherein the C1-C4 alkyl is methyl or ethyl.

28. The compound of claim 26, wherein the piperazinyl is substituted with one R4, and R4is - L’-NCR^R13).

29. The compound of claim 26, wherein the piperazinyl is substituted with one R4, and R4is - C(O)O-t-butyl.

30. The compound of claim 26, wherein the piperazinyl is substituted with two R4groups, wherein one R4 group is C1-C4 alkyl and the second R4 group is hydroxyalkyl or -L3- aryl.

31. The compound of claim 20, wherein R3is -NH-L1-N(RARB).

32. The compound of claim 31, wherein L1is propylene and RAand RBare each methyl.

33. The compound of claim 31, wherein L1is propylene and RAand RBare each ethyl.

34. The compound of claim 20, wherein R3is -NH-L2-heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more R5.

35. The compound of claim 20, wherein R3is NH-L2-cycloalkyl, wherein the cycloalkyl is optionally substituted with one or more R536. The compound of claim 34, wherein L2is a bond, and the heterocyclyl is pyrrolidinyl optionally substituted with one or more R5.

37. The compound of claim 36, wherein the pyrrolidinyl is substituted with one R5, wherein R5is ethyl or isopropyl.

38. The compound of claim 34, wherein L2is a C1-C4 alkylene, and the heterocyclyl is pyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl, each optionally substituted with one or more R5.

39. The compound of claim 38, wherein L2is ethylene or propylene.

40. The compound of claim 39, wherein the heterocyclyl is piperazinyl optionally substituted with one or more R5, wherein R5is methyl.

41. The compound of claim 34, wherein L2is dimethylethylene, and the heterocyclyl is pyrrolidinyl, piperidinyl, or morpholinyl.

42. The compound of claim 38, wherein L2is dimethylethylene, and the heterocyclyl is piperazinyl optionally substituted with one or more R5, wherein R5is methyl.

43. A compound of Formula (I), wherein the compound is:Cl Clpharmaceutically acceptable salts thereof44. A pharmaceutical composition comprising a compound of Formula (I), a pharmaceutically acceptable salt, and at least one pharmaceutically acceptable excipient or elixir.

45. A method of treating a PLK1 -mediated disease or disorder in a subject in need thereof, comprising administering a therapeutically effective amount of a compound of claims 1- 43 or a pharmaceutical salt thereof, or a pharmaceutical composition of claim 44.

46. The method of claim 45, wherein the PLK1 -mediated disease or disorder is cancer.

47. The method of claim 46, wherein the PLKl-mediated cancer is melanoma, non-small cell lung cancer (NSCLC), carcinomas of the head and neck, esophageal, pharynx, breast, liver, endometrium, colorectum, ovary, pancreas or prostate.

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