4-(3h-pyrazolo [4, 3-f] quinolin-7-yl)-n-(2-(dimethylamino) ethyl) benzamide or hydroxamic acid compounds, compositions, and methods of use

Abstract

Compounds comprising N-(2-(dimethylamino) ethyl) benzamide or hydroxamic acid derivatives for inhibition of certain kinases are useful for the treatment of disease states mediated by certain kinases. Pharmaceutical compositions, conjugates, and methods of treating those kinase mediated disease states are also provided.

Description

priority

[0001] This application is related to and claims priority to U.S. Provisional Patent Application No. 63 / 348,034, filed on June 2, 2022. The contents of the foregoing application are hereby incorporated by reference into the present disclosure in their entirety. Technical Field

[0002] The present invention generally relates to compounds and pharmaceutical compositions comprising 4-(3H-pyrazolo[4,3-f]quinolin-7-yl)-N-(2-(dimethylamino)ethyl)benzamide or hydroxamic acid, which are potential anticancer or anti-inflammatory agents for treating diseases mediated by kinases, such as inflammation and cancer. Methods of treating those diseases are also provided. Background Art

[0003] This section introduces aspects that may be helpful in better understanding the present disclosure. Therefore, these statements should be read in this light and not as admissions of what is or is not prior art.

[0004] Cells contain more than 500 kinases that regulate processes such as the cell cycle, growth, migration, and immune responses. Several dysregulated kinases (i.e., kinases that have acquired gain-of-function mutations or are overexpressed) can drive cancer proliferation. Fabbro et al., Ten things you should know about protein kinases: IUPHAR Review 14, British J Pharmacology 172(11): 2675-2700 (2015). Small molecule inhibitors of cancer-driving kinases (e.g., BCR-ABL1 fusion protein, FMS-like tyrosine kinase 3 (FLT3) internal tandem duplication (FLT3-ITD), mutated or overexpressed anaplastic lymphoma kinase (ALK), epidermal growth factor receptor (EGFR), platelet-derived growth factor receptor (PDGFR), KIT, vascular endothelial growth factor (VEGFR), B-Raf, Bruton's tyrosine kinase (BTK), phosphatidyl-4,5-bisphosphate 3-kinase catalytic subunit delta (PI3Kδ), and Erb-B2) have achieved some clinical success. Fabbro, 25 years of small molecule weight kinase inhibitors: potentials and limitations, Molecular Pharmacology 87 (5): 766-775 (2015). Recent efforts have also focused on targeting other kinases to prevent cancer growth, such as cell cycle kinases (CDKs) and kinases that target histones, the cytoskeleton, or other processes that are important to the cell.

[0005] However, most kinase inhibitors to date have become ineffective due to the emergence of drug-resistant clones.

[0006] There are many mechanisms by which cancer cells develop resistance to kinase inhibitors, including copy number multiplication, additional kinase mutations (e.g., secondary mutations occurring in the tyrosine kinase domain of the FLT3-ITD kinase), or activation of alternative kinase pathways and / or downstream targets that can bypass inhibition of specific kinase targets. Lindblad et al., Aberrant activation of the PI3K / mTOR pathway promotes resistance to sorafenib in AML, Oncogene 35(39):5119-5131(2016).

[0007] For example, acute myeloid leukemia (AML) is a devastating disease that affects more than 20,000 people in the United States each year. About 30% of AML patients carry FLT3 kinase mutations, and for these patients, the prognosis is generally worse than for patients without FLT3 mutations. Lin & Chen, Advances in the drug therapies of acute myeloid leukemia (except acute promyelocytic leukemia), Drug Design, Development & Therapy 12: 1009-1017 (2018). Many FLT3 inhibitors have been developed and are being tested clinically, some of which, including midostaurin, quizartinib and gilteritinib, have been approved in the United States or Japan. Although these FLT3 inhibitors modestly improve the survival of AML patients carrying FLT3, resistance to approved FLT3 inhibitors has been observed in the clinic. Specifically, FLT3-F691I is resistant to midostaurin, while FLT3-F691L is resistant to gilteritinib and quizartinib. Scholl et al., Molecular mechanisms of resistance to FLT3 inhibitors in acute myeloid leukemia: ongoing challenges and future treatments, Cells 9(11): 2493(2020). In addition to FLT3 resistance mutations, FLT3-independent resistance pathways may also emerge after long-term FLT3i treatment, such as NRAS activation or mutations in other kinases. Alotaibi et al., Patterns of resistance differ inpatients with acute myeloid leukemia treated with type I versus type II FLT3inhibitors, Blood Cancer Discovery 2(2):125-134(2021); Kasi et al., Clonalevolution of AML on novel FMS-like tyrosine kinase-3(FLT3)inhibitor therapy with evolving actionable targets, Leukemia Research Reports 5:7-10(2016).In some cases, patients who were initially FLT3-positive became FLT3-negative during treatment; however, the cancer still progressed driven by alternative pathways. Schmalbrock et al., Clonal evolution of acute myeloid leukemia with FLT3-ITD mutation under treatment with midostinib, Blood 137(22):3093-3104(2021).

[0008] Kinase inhibitors that inhibit cancer driver kinases as well as downstream targets (kinases and non-kinase targets such as histone demethylases) and / or kinases that cooperate with the driver kinase can enhance efficacy and reduce the likelihood of developing resistance to the kinase inhibitor. However, a common challenge with such multidrug therapeutic approaches is promiscuous binding, which can lead to toxicity.

[0009] Therefore, there is an unmet need for drugs that can address FLT3 as well as other oncogenic pathways that may be engaged during treatment. Summary of the invention

[0010] In certain embodiments, a compound having the structure of Formula I, or a pharmaceutically acceptable salt thereof, is provided: in: X1 is each independently N, CH or C-halogen; X2 is each independently N, CH, C-halogen, C-CH n1 X m (where n1+m=3 and X is halogen), C-OCF3, C-OCHF2, C-OMe, C-Me, C-Et, C-tBu, C-iPr, CN-alkyl, CN-heteroalkyl, CO-alkyl or CO-heteroalkyl;

[0011] R2, R3 and R4 are each independently alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl; and Y is in: n is 1 to 5, R5 and R6 are each independently H, alkyl or heteroalkyl, and R6 may optionally form a ring structure, R7 are each independently H or alkyl, Het-Ar is a heteroaromatic ring, and W is NH, NMe, NEt, NCH2CH2OH, NCH2CH2OMe, O, SO or SO2, provided that Y is not NH2.

[0012] In certain embodiments, at least one of X1 and / or X2 is CF, CHF2, CF3 or CHF3. Wherein at least one of X1 and / or X2 is CF, CHF2, CF3 or CHF3. The compound may have a structure of Formula IA: or a pharmaceutically acceptable salt thereof, wherein: n2 is 1, 2 or 3; Q is each independently O, NH, substituted N, CH2, or substituted C, S, SO or SO2; and Each Q together forms a 6-membered, 7-membered or 8-membered ring or a bicyclic ring.

[0013] One or more of R5 and / or R6 may be Me. In certain embodiments, R7 is each independently H or Me. In certain embodiments, Y is and each R6 together forms morpholine, piperidine, piperazine or pyrrolidine. In certain embodiments, Y is and each R7 together forms a ring structure (e.g., wherein the ring structure includes cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl). In certain embodiments, Y is and each R7 together forms a ring structure (e.g., wherein the ring structure includes cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl). In certain embodiments, Y is and Het-Ar comprises imidazole, oxazole, pyrazole, triazole, isoxazole, isothiazole, tetrazole, oxadiazole, thiadiazole, pyrimidine or triazine. In certain embodiments, Y is And R5 is Me.

[0014] The compound may have the structure of Formula IB: Alternatively, the compound may be a pharmaceutically acceptable salt thereof, wherein: o is 0-2; and Each R8 is independently H, alkyl or heteroalkyl. Each R7 can form a ring structure together (for example, wherein the ring structure includes cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl). In certain embodiments of formula IB, one R8 is Me, Et or CF3.

[0015] In certain embodiments, the compound has the structure of Formula II: or a pharmaceutically acceptable salt thereof, wherein: X1 is each independently N, CH or C-halogen; X2 is each independently N, CH, C-halogen, C-CH n1 Xm (where n1+m=3 and X is halogen), C-OCF3, C-OCHF2, C-OMe, C-Me, C-Et, C-tBu, C-iPr, CN-alkyl, CN-heteroalkyl, CO-alkyl, or CO-heteroalkyl; Q is each independently O, NH, substituted N, CH2, or substituted C, S, SO or SO2, and each Q when combined together forms a 6-membered, 7-membered or 8-membered ring or a bicyclic ring; n3 is 1, 2, or 3; R4 is alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl; W is O, NH or NMe; and R9 is alkyl or heteroalkyl, provided that R9 is not H.

[0016] In certain embodiments, the compound or a pharmaceutically acceptable salt thereof has the following structure:

[0017] In certain embodiments, the compound has the structure or a pharmaceutically acceptable salt thereof.

[0018] The compound may have the following structure:

[0019] The compound may have the following structure:

[0020] The compound may have the following structure: Or it may be a pharmaceutically acceptable salt thereof.

[0021] The compound may have the following structure:

[0022] The compound may have the following structure:

[0023] The compound may have the following structure:

[0024] The compound may have the following structure:

[0025] The compound may have the following structure:

[0026] The compound may have the following structure:

[0027] A PROTAC conjugate is also provided. In certain embodiments, the PROTAC conjugate has a chemical structure of Formula X: A-L'-D (Formula X) or a pharmaceutically acceptable salt thereof, wherein: A is a group of any compound described herein; L' is a linker that binds A and D or is absent; and D is the ubiquitin pathway protein binding moiety.

[0028] Use of a compound herein or a conjugate herein or a pharmaceutically acceptable salt of a compound or conjugate herein in the manufacture of a medicament for treating a disease in a subject. In certain embodiments, the disease includes a disease in which modulation of a kinase can reduce the severity of the disease in the subject. For example, the disease can be cancer, diabetes, an inflammatory disease, or a neurological disease.

[0029] Also provided is a pharmaceutical composition. The pharmaceutical composition herein may include any one of the compounds herein, the conjugates herein, or pharmaceutically acceptable salts, N-oxides, hydrates, solvates, tautomers, or optical isomers of the compounds or conjugates herein; and a pharmaceutically acceptable carrier and / or diluent. The pharmaceutical composition may also include a pharmaceutically acceptable excipient.

[0030] Also provided is a method for treating a disease state (e.g., cancer) of a subject. In certain embodiments, the method includes administering to the subject a therapeutically effective amount of: a compound herein, a conjugate herein, or a pharmaceutically acceptable salt, N-oxide, hydrate, solvate, tautomer, or optical isomer of the compound or conjugate; or a pharmaceutical composition comprising one or more of a compound herein, a conjugate herein, or a pharmaceutically acceptable salt, N-oxide, hydrate, solvate, tautomer, or optical isomer of the compound or conjugate.

[0031] In certain embodiments of the method, the compound or conjugate includes a compound having the structure: or a pharmaceutically acceptable salt thereof.

[0032] In certain embodiments of the method, the compound or conjugate includes a compound having the structure: or a pharmaceutically acceptable salt thereof.

[0033] The method may also include administering a second therapy to the subject. The second therapy may include an effective amount of a chemotherapeutic agent, an immunotherapeutic agent, or a hormonal therapeutic agent; or radiation therapy.

[0034] The cancer may be acute myeloid leukemia (AML). In certain embodiments, the cancer is selected from AML, chronic myeloid leukemia, ovarian cancer, cervical cancer, pancreatic cancer, breast cancer, brain cancer, skin cancer, lung cancer, prostate cancer, lymphoma, leukemia, colon cancer, stomach cancer, head cancer, neck cancer, thyroid cancer, kidney cancer, liver cancer and stomach cancer.

[0035] In certain embodiments of the method, an effective amount of an IC of about 50 nM or less (e.g., 50 nM or less) is administered to a subject. 50 Inhibit CDK in a subject. In certain embodiments, an effective amount of about 30 nM or less (e.g., 30 nM or less) is administered to a subject. 50 Inhibiting CDK in a subject.

[0036] Administration of an effective amount to a subject can specifically inhibit FLT3 and haspin kinases in the subject, such as when a dual inhibitor compound is administered.

[0037] Compounds and / or conjugates described herein are also provided for use in treating a disease state regulated by one or more kinases. For example, the disease state may be cancer and the one or more kinases may be CDK, FLT3 and / or haspin kinases. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] In view of the following detailed description of various exemplary embodiments of the present disclosure, the embodiments disclosed herein and other features, advantages and aspects included therein and methods of achieving them will become apparent. These detailed descriptions will be better understood in conjunction with the accompanying drawings.

[0039] Figure 1 A schematic diagram showing the modification of compound HSD1217 to generate compounds HSH2177 and HSD1993, both of which are potent inhibitors of CDK2.

[0040] Figure 2A Fusion of indole to quinoline is shown to provide a novel hinge-binding 3H-pyrazolo[4,3-f]quinoline scaffold core.

[0041] Figure 2B Examples of C-7 substituted compounds with a 3H-pyrazolo[4,3-f]quinoline moiety that affect kinase targeting are shown.

[0042] Figure 3 Shown are graphical data relating to the percent viability of SNU-16 cell line (a human gastric cancer cell line) when treated with 1 μM of each identified compound.

[0043] Figures 4A-4D The SNU-16 cell line is shown relative to compound HSH2177 ( Figure 4A ), compound HSH3014( Figure 4B ), compound HSH3180( Figure 4C ) and compound HSH3194( Figure 4D ) related data of proliferation percentage.

[0044] Figure 5 Shown are graphical data relating to the percent viability of the KCL22-IR cell line (a chronic myeloid leukemia cell line) when treated with 1 μM of each of the identified compounds.

[0045] Figures 6A-6DThe KCL22-IR cell line is shown relative to compound HSH2177 ( Fig. 6A ), compound HSH3014( Figure 6B ), compound HSH3194( Figure 6C ) and compound HSH3180( Fig.6D ) related data of proliferation percentage.

[0046] Figure 7 Shown are graphical data relating to the percent viability of the Molm-14 (D835Y) cell line (a human AML cell line with a mutation at D835Y) when treated with 1 μM of each of the identified compounds.

[0047] Figures 8A-8D The Molm-14 (D835Y) cell line is shown relative to the compound HSH2177 ( Fig. 8A ), compound HSH3014( Figure 8B ), compound HSH3194( Figure 8C ) and compound HSH3180( Fig.8D ) related data of proliferation percentage.

[0048] Fig. 9 Shown are graphical data relating to the percent viability of the Molm-14 (F69IL) cell line (a human AML cell line with a mutation at F691L) when treated with 1 μM of each of the identified compounds.

[0049] Figures 10A-10C The Molm-14 (F69IL) cell line is shown relative to the compound HSH2177 ( Fig. 10A ), compound HSH3194( Fig. 10B ) and compound HSH3180( Fig. 10C ) related data of proliferation percentage.

[0050] Fig.11A Schematic representation of compound HSD1217 and ATP binding residues is shown.

[0051] Figures 11B-11D The haspin (PDB: 2WB8) and three initial compounds (compound 4 ( Fig. 11B ), compound 5 (Figure 2C) and compound 6 ( Fig.11D )) with hydrogen bonds represented as HYD, π-cation interactions as X, and π-π stacking as Y. Solvent exposure is represented as gray rings around atoms. Docking was performed and visualized by PyMol and Maestro.

[0052] Figures 12A-12C Enzyme binding and inhibition data for compound HSK205 are shown, where Fig. 12A The K of haspin kinase obtained from Eurofins is shown. d Data (each data point represents the mean, and error bars represent the SEM of three replicates), Fig. 12B The compound HSK205 showed that the IC 50 Data for FLT3-ITD enzyme assay in duplicate mode with 3-fold serial dilutions starting at 10 μM and Fig. 12C The compound HSK205 showed that the IC 50 Data for FLT3(D835Y) enzyme assay in duplicate mode with 3-fold serial dilutions starting from 10 μM (all data were fitted to a nonlinear regression equation using GraphPad Prism 9.0 software).

[0053] Fig.13A and 13B The results of Western blot analysis are shown, where Fig.13A Molm-14 cells treated with compound HSK205 (5 nM, 20 nM or 100 nM) or control for 24 hours (100 nM of gilteritinib was used as a positive control), and Fig. 13B Figure 2 shows the relative to actin loading control. Fig.13A The data were quantified for the bands in (values ​​are reported as the mean of three replicates, and the error bars represent the standard deviation (NS: p>0.05, *: p<0.05, **: p<0.01, ***p<0.001, ****p<0.0001).

[0054] Fig.14A and 14B The results of Western blot analysis are shown, where Fig.14A Molm-14 cells treated with compound HSK205 (5 nM, 20 nM or 100 nM) or control for 24 hours (5 nM of gilteritinib was used as a positive control), and Fig. 14B Figure 2 shows the relative to actin loading control. Fig.14A The data were quantified for the bands in (values ​​are reported as the mean of three replicates, and the error bars represent the standard deviation (NS: p>0.05, *: p<0.05, **: p<0.01, ***p<0.001, ****p<0.0001).

[0055] Fig.15The dose response curve of FLT3-driven AML cells treated with different concentrations of compound HSK205, compound HSK2150 or gilteritinib (a known FLT3 inhibitor) for 72 hours, wherein the data were fitted to a nonlinear regression equation using GraphPad Prism 9.0 software (each data point represents the mean, and the error bars represent the SEM of three replicates).

[0056] Fig.16 It is the data graph of DNA intercalation test. Mitoxantrone was used as positive control and DMSO was used as negative control. Compounds HSK205, DMSO and mitoxantrone were tested in the dose range of 1 mM to 10 nM. Data were fitted to a nonlinear regression equation using GraphPad Prism 9.0 software (each data point represents the average of three replicates).

[0057] While the present disclosure is susceptible to various modifications and alternative forms, exemplary embodiments thereof are shown by way of example in the drawings and are herein described in detail. DETAILED DESCRIPTION

[0058] Although the concepts of the present disclosure have been elaborated and described in detail in the specification herein, the results of the specification should be regarded as illustrative rather than restrictive; it should be understood that only illustrative embodiments have been shown and described and that all changes and modifications that come within the spirit of the disclosure are intended to be protected.

[0059] Sintim et al., U.S. Pat. No. 11,040,973, demonstrated that HSD1217, a 4-(3H-pyrazolo[4,3-f]quinolin-7-yl)benzamide-containing compound synthesized via a Doebner-Povarov multicomponent reaction, is a potent inhibitor of kinases, such as FLT3 kinase.

[0060] HSD1217 has the following structure:

[0061] However, the compound HSD1217 has only moderate inhibitory effects on CDKs such as CDK2 and CDK12, which are important kinase targets that have emerged as potential anticancer targets. For example, the IC of HSD1217 for CDK2 inhibition is 50 It is 185nM.

[0062] In certain embodiments, the present disclosure provides compounds comprising a 2-(dimethylamino)ethyl moiety or a hydroxamic acid in place of the benzamide moiety and can achieve unexpected and significantly improved kinase targeting compared to at least compound HSD1217 and other conventional CDK inhibitors.

[0063] In certain embodiments, the compounds herein are analogs of compound HSH2177 and / or compound HSD1993. Such compounds have the ability to inhibit CDK function and can be used to treat disease states where targeting CDK can lead to restoration of normal cell function, such as cancer, inflammatory diseases, and certain neurological diseases.

[0064] In addition, it has been recently reported that FMS-like tyrosine kinase-3 (FLT3) inhibitors have activity against other cancer-related kinases, such as dual FLT3 / MERTK, FLT3 / MNK, FLT3 / CDK, FLT3 / TOPK, FLT3 / PIM, or FLT3 / AURKA inhibitors, and simultaneous inhibition of FLT3 and other kinase pathways may bring some advantages in overcoming drug resistance. Minson et al., MRX2843, a novel dual MerTK-FLT3 inhibitor with activity against resistance-conferring FLT3 mutations in acute myeloid leukemia, Blood 124:3757(2014); Yen et al., Identification of dual FLT3 and MNK2 inhibitor for acute myeloid leukemia treatment using a structure-based virtual screening approach, Bioorganic Chemistry121:105675(2022); Goh et al., SB1317, a potent andorally active FLT3-CDK inhibitor with high anti-tumor efficacy in models of hematological malignancies, Blood 110(11):1593(2007); Dayal et al., Dual FLT3 / TOPK inhibitor with activity against FLT3-ITD secondary mutations potentlyinhibits acute acute mutations myeloid leukemia cell lines,Future Medicinal Chemistry 10(7):823-835(2018); Czardybon et al., A novel, dual pan-PIM / FLT3 inhibitor SEL24 exhibits broad therapeutic potential in acute myeloid leukemia, Oncotarget 9(24):16917-16931(2018); Taussig et al.,EP0042,a dual FLT3 and Aurora kinase inhibitor:preliminary results of an ongoing phase I / IIa first in human studyin patients with relapsed / refractory acute myeloid leukemia,Blood 140(Supplement 1):6222-6223(2022). For example, CCT245718, a dual FLT3 / Aurora A inhibitor may overcome FLT3-D835Y-mediated resistance. Tariq et al.,CCT245718,adual FLT3 / AuroraAinhibitor overcomes D835Y-mediated resistance to FLT3 inhibitors in acutemyeloid leukaemia cells,British J Cancer 125:966-974(2021). In another interesting report, it was shown that the frequency of resistance to a dual FLT3 / CDK4 inhibitor (AMG 925) was lower than that of other FLT3 inhibitors that do not inhibit CDK4 (e.g., quizartinib). Li et al., Discovery of AMG 925, a FLT3 and CDK4 dual kinase inhibitor with preferential affinity for the activated state of FLT3, J Medicinal Chemistry 57(8):3430-3449 (2014).

[0065] Despite the existence of these dual inhibitors, no dual inhibitors of FLT3 and haspin have been discovered to date. Haspin is a novel target in cancer and is the primary kinase that phosphorylates histone H3 at T3. Amoussou et al., Haspin: a promising target for the design of inhibitors as potent anticancerdrugs, Drug Discovery Today 23(2):409-415(2018). Recent preliminary reports suggest that reducing haspin by genetic means or by compound inhibition reduces leukemic proliferation. Taking advantage of the recent discovery that haspin is overexpressed and plays a key proliferative role in many cancers, in certain embodiments, optimized 3H-pyrazolo[4,3-f]quinolyl compounds are provided that exhibit improved activity against both FLT3 and haspin (i.e., 3H-pyrazolo[4,3-f]quinolyl dual FLT3 / haspin inhibitors).

[0066] In certain embodiments, compounds having the structure of Formula I are provided: or a pharmaceutically acceptable salt thereof, wherein: X1 is each independently N, CH or C-halogen; X2 is each independently N, CH, C-halogen, C-CH n1 X m (where n1+m=3 and X is halogen), C-OCF3, C-OCHF2, C-OMe, C-Me, C-Et, C-tBu, C-iPr, CN-alkyl, CN-heteroalkyl, CO-alkyl or CO-heteroalkyl; R2, R3 and R4 are each independently alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl; and Y is in: n is 1 to 5, R5 and R6 are each independently H, alkyl or heteroalkyl, and R6 may optionally form a ring structure, R7 are each independently H or alkyl, Het-Ar is a heteroaromatic ring, and W is NH, NMe, NEt, NCH2CH2OH, NCH2CH2OMe, O, SO or SO2. In certain embodiments, Y is not NH2 (ie, a compound of Formula I with the proviso that Y is not NH2).

[0067] At least one of X1 and / or X2 may be CF, CHF2, CF3 or CHF3. In certain embodiments, each X2 is CH. In certain embodiments, at least one X2 is CH. In certain embodiments, each X1 is CH. In certain embodiments, at least one X1 is CH.

[0068] One or more of R5 and / or R6 in Y may be Me. R7 in Y may each independently be H or Me. In certain embodiments, when Y is When Y is, each R6 together can form morpholine. In certain embodiments, when Y is When Y is, each R6 together can form piperidine. In certain embodiments, when Y is When Y is, each R6 together can form piperazine. In certain embodiments, when Y is When R6 is taken together, it can form pyrrolidine.

[0069] In certain embodiments, when Y is When, each R7 together may form a ring structure. The ring structure may include, for example, a cyclopropyl group. The ring structure may include a cyclobutyl group. The ring structure may include a cyclopentyl group. The ring structure may include a cyclohexyl group.

[0070] In certain embodiments, when Y is When, each R7 together may form a ring structure. The ring structure formed by each R7 may include, for example, a cyclopropyl group. The ring structure may include a cyclobutyl group. The ring structure may include a cyclopentyl group. The ring structure may include a cyclohexyl group.

[0071] In certain embodiments, Y is And Het-Ar includes imidazole, oxazole, pyrazole, triazole, isoxazole, isothiazole, tetrazole, oxadiazole, thiadiazole, pyrimidine or triazine.

[0072] In certain embodiments, Y is And R5 is Me.

[0073] In certain embodiments, the compound has the structure of Formula IA: or a pharmaceutically acceptable salt thereof, wherein: X1 is each independently N, CH or C-halogen; X2 is each independently N, CH, C-halogen, C-CH n1 X m (where n1+m=3 and X is halogen), C-OCF3, C-OCHF2, C-OMe, C-Me, C-Et, C-tBu, C-iPr, CN-alkyl, CN-heteroalkyl, CO-alkyl or CO-heteroalkyl; R4 is alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl; Y is in: n is 1 to 5, R5 and R6 are each independently H, alkyl or heteroalkyl, and R6 may optionally form a ring structure, R7 are each independently H or alkyl, Het-Ar is a heteroaromatic ring, and W is NH, NMe, NEt, NCH2CH2OH, NCH2CH2OMe, O, SO or SO2, Provided that Y is not NH2. n2 is 1, 2 or 3; Q is each independently O, NH, substituted N, CH2, or substituted C, S, SO or SO2; and Each Q together forms a 6-, 7- or 8-membered ring or a bicyclic ring.

[0074] The compound may have the structure of Formula IB: or a pharmaceutically acceptable salt thereof, wherein: X1 is each independently N, CH or C-halogen; X2 is each independently N, CH, C-halogen, C-CH n1 X m (where n1+m=3 and X is halogen), C-OCF3, C-OCHF2, C-OMe, C-Me, C-Et, C-tBu, C-iPr, CN-alkyl, CN-heteroalkyl, CO-alkyl or CO-heteroalkyl; R4 is alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl; R5 is H, alkyl or heteroalkyl; R7 are each independently H or alkyl; o is 0-2; and Each R8 is independently H, alkyl or heteroalkyl.

[0075] In certain embodiments, when the compound has the structure of formula IB, each R7 together forms a ring structure. The ring structure may include cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In certain embodiments, at least one R8 is Me, Et or CF3.

[0076] In certain embodiments, the compound has the following structure:

[0077] The compound may have one of the following structures, or a pharmaceutically acceptable salt of one of the following:

[0078] In certain embodiments, the compound has one of the following structures, or a pharmaceutically acceptable salt of one of the following:

[0079] In certain embodiments, the compound has one of the following structures, or a pharmaceutically acceptable salt of one of the following:

[0080] In certain embodiments, the compound has one of the following structures, or a pharmaceutically acceptable salt of one of the following:

[0081] In certain embodiments, the compound has one of the following structures, or a pharmaceutically acceptable salt of one of the following:

[0082] In certain embodiments, the compound has one of the following structures, or a pharmaceutically acceptable salt of one of the following:

[0083] In certain embodiments, the compound has one of the following structures, or a pharmaceutically acceptable salt of one of the following:

[0084] In certain embodiments, the compound may have the structure of Formula II: or a pharmaceutically acceptable salt thereof, wherein: Each X1 is independently N, CH or C-halogen; Each X2 is independently N, CH, C-halogen, C-CH n1 X m (where n1+m=3 and X is halogen), C-OCF3, C-OCHF2, C-OMe, C-Me, C-Et, C-tBu, C-iPr, CN-alkyl, CN-heteroalkyl, CO-alkyl or CO-heteroalkyl; Each Q is independently O, NH, substituted N, CH2, or substituted C, S, SO or SO2, and each Q when combined together forms a 6-membered, 7-membered or 8-membered ring or a bicyclic ring; n3 is 1, 2, or 3; R4 is alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl; W is O, NH or NMe; and R9 is alkyl or heteroalkyl, provided that R9 is not H.

[0085] The compound may have the following structure: Or a pharmaceutically acceptable salt of any of the above structures.

[0086] In certain embodiments, the compound has one of the following structures, or a pharmaceutically acceptable salt of one of the following:

[0087] In certain embodiments, the compound has one of the following structures, or a pharmaceutically acceptable salt of one of the following:

[0088] As mentioned above, the compound can be a CDK specific inhibitor. In certain embodiments, the compound is a dual haspin and FLT3 specific inhibitor. The compound can specifically bind to the receptor of CDK receptor or haspin and FLT3, thereby reducing off-target interactions, such as interactions with growth factor receptors. When it comes to small molecules / receptors or other binding pairs, "specific binding", "high affinity binding" or "specific" or "selective" binding means that under specified conditions, a specific compound or its recognition region is bound to a specific receptor or site (such as a CDK inhibition site, such as the ATP binding catalytic site of a tyrosine kinase domain), and will not be combined with other receptors present (such as off-target growth factor receptors) in large quantities. Specific binding or high affinity binding can also mean, for example, the affinity of a binding compound, a ligand or a binding composition to its target (for example, CDK inhibition or dual FLT3 / haspin inhibition) is usually higher than the affinity of the compound, a ligand or a binding composition to another receptor (for example, growth factor receptor) by at least 20%, at least 25%, at least 50% or at least 100% (2 times).

[0089] In typical embodiments, a molecule that specifically binds to a target will have at least about 10 6 Liter / mol (K O =10 ～ 6 The binding specificity of a compound to a CDK receptor or to a haspin and FLT3 receptor is determined using standard techniques known in the art and described herein.

[0090] In certain embodiments, the N-(2-(dimethylamino)ethyl)benzamide or hydroxamic acid derivatives herein can significantly enhance the activity against kinases, including CDKs. Figure 1 The modification of the previously known compound HSD1217 to the compounds HSH2177 and HSD1993 herein is shown. HSD1217 inhibits the IC of CDK2 50 The value is 185nM, and the IC of compound HSH2177 inhibits CDK2 50 The value can be 7nM, and the IC of compound HSD1993 inhibits CDK2 50 The value can be 4 nM. In addition, the IC values ​​of compound HSD2177 and compound HSD1993 for inhibiting CDK12 / CyclinK 50 The values ​​were 27 nM and 9 nM, respectively. Under similar experimental conditions, THZ531 and dinaciclib inhibited CDK12 with IC 50 The values ​​were 60 nM and 4 nM, respectively.

[0091] The present inventors have previously shown that the "fusion" of two known kinase hinge binders, indazole and quinoline (which are commonly found in U.S. Food and Drug Administration ("FDA") approved drugs such as axitinib and erlotinib), can produce 3H-pyrazolo[4,3-f]quinoline ( Figure 2A ), the 3H-pyrazolo[4,3-f]quinoline can bind to the hinge region of the kinase. Dayal et al., 3H-pyrazolo[4,3-f]quinoline-based kinase inhibitors inhibit the proliferation of acute myeloid leukemia cells in vivo, J Medicinal Chemistry 64(15):10981-10996(2021). It is worth noting that the substituents at the 7 position of the 3H-pyrazolo[4,3-f]quinoline moiety can determine which kinases are effectively inhibited. For example, pyrazole substitution at the 7 position can produce a kinase that binds to mutant FLT3 and K d Dayal et al., Dual FLT3 / TOPK inhibitor with activity against FLT3-ITD secondary mutations potently inhibits acute myeloid leukemia cell lines, Future Medicinal Chemistry 10(7):823-835(2018); Dayal et al., Potently inhibiting cancer cell migration with novel 3H-pyrazolo[4,3-f]quinoline boronic acid ROCK inhibitors, European J Medicinal Chemistry 180:449-456(2019)( Figure 2B ).

[0092] Compounds substituted with benzo[d]thiazolyl groups (such as SHD992) are potent CDK inhibitors, while compounds containing phenols (such as HSD972) (see Figure 2B ) may be an excellent haspin inhibitor (HSD972 haspin IC 50=14nM). Opoku-Temeng et al., Tetrahydro-3H-pyrazolo[4,3-a]phenanthridine-based CDKinhibitor, Chemical Communications 54(36):4521-4524(2018); Opoku-Temeng et al., 3H-pyrazolo[4,3-f]quinoline haspin kinase inhibitors and anticancer properties, Bioorganic Chemistry 78:418-426(2018). In addition to the 3H-pyrazolo[4,3-f]quinoline core, some other similar systems, such as the pyrazolo[3,4-h]quinoline scaffold, may also have anticancer activity. Nguyen et al., Induction of paraptotic cell death in breast cancer cells by a novel pyrazolo[3,4-h]quinoline derivative through ROS production and endoplasmic reticulumstress, Antioxidants 11(1):117(2022).

[0093] Although HDS972 and HSD1169 (see Figure 2B ) contain a 3H-pyrazolo[4,3-f]quinoline moiety, but they inhibit either haspin or FLT3, respectively; rather than inhibiting both haspin and FLT3. In certain embodiments, the compounds herein comprise a phenol moiety at position -7 replaced with a benzamide (compound HSD1217, see Figure 3 A), which inhibits haspin better than HDS972. In certain embodiments, the compounds herein are derivatives of compound HSD1217 that have been optimized for dual FLT3 / haspin binding.

[0094] The compounds described herein may contain one or more chiral centers, or may otherwise exist as multiple stereoisomers. It should be understood that, in one embodiment, the invention described herein is not limited to any particular stereochemical requirements, and the compounds and compositions, methods, uses, and medicaments comprising them may be optically pure, or may be any of a variety of stereoisomer mixtures, including racemates and other enantiomeric mixtures, other diastereomeric mixtures, etc. It should also be understood that such stereoisomer mixtures may include a single stereochemical configuration at one or more chiral centers, and a mixture of stereochemical configurations at one or more other chiral centers.

[0095] Similarly, the compounds described herein may include geometric centers, such as cis, trans, E and Z double bonds. It should be understood that, in another embodiment, the invention described herein is not limited to any particular geometric isomer requirements, and the compounds and compositions, methods, uses and medicaments including them may be pure, or may be any of a variety of geometric isomer mixtures. It should also be understood that such geometric isomer mixtures may include a single configuration at one or more double bonds, and a mixture of geometric configurations at one or more other double bonds.

[0096] It is expected that the compounds herein can be provided in the form of prodrugs. The term "prodrug" refers to an inactive derivative of a parent compound / drug that can be hydrolyzed, oxidized, or otherwise reacted under biological conditions (in vitro or in vivo) to provide an active compound, particularly a compound herein. Prodrugs can be created to overcome one or more obstacles to the effective use of potential active compounds, such as obstacles to the instability and / or possible toxicity of active compounds. Examples of prodrugs include, but are not limited to, derivatives and metabolites of compounds, including bio-hydrolyzable parts, such as bio-hydrolyzable amides, bio-hydrolyzable esters, bio-hydrolyzable carbamates, bio-hydrolyzable carbonates, bio-hydrolyzable ureides, and bio-hydrolyzable phosphate analogs. Specific prodrugs of compounds with carboxyl functional groups are lower alkyl esters of carboxylic acids. Carboxylic acid esters can be conveniently formed by any carboxylic acid moiety present on the esterified molecule. In certain embodiments, the prodrug form of the compounds herein may include a protecting group, which, in addition to protecting the compound from oxidation and the like, also allows targeting of specific sites (e.g., tumor microenvironment) in the subject's body. Prodrugs can generally be prepared using well-known methods, such as those described in Burger's Medicinal Chemistry and Drug Discovery 6th ed. (Donald J. Abraham ed., 2001, Wiley) and Design and Application of Prodrugs (H. Bundgaard ed., 1985, Harwood Academic Publishers GmbH). PROTAC Conjugates

[0097] Also provided are PROTAC conjugates comprising the compounds herein. The PROTAC conjugate may have a chemical structure of formula (X): A-L'-D(X) wherein A is a compound described herein (eg, a kinase inhibitor described herein); L' is a linker or is absent; and D is a ubiquitin pathway protein binding moiety.

[0098] While protein inhibitors can block or reduce protein activity in cells, protein degradation in cells can also reduce activity or completely remove target proteins. Therefore, exploiting the protein degradation pathways of the cell can provide an additional way to reduce or remove protein activity. One of the main degradation pathways of the cell is called the ubiquitin-proteasome system. In this system, proteins are marked for degradation by the proteasome through ubiquitination. Ubiquitination of proteins can be achieved by ubiquitin pathway protein binding moieties, which bind to proteins and add ubiquitin molecules to the protein.

[0099] In order to take advantage of this degradation pathway, PROTAC has been developed. The term "PROTAC" refers to a proteolytic targeting chimeric molecule comprising a ubiquitin pathway protein binding portion, an optional linker and a targeting portion. PROTAC combines a ubiquitin pathway protein binding portion (e.g., E3 ubiquitin ligase) with a protein to be targeted for degradation. In order to promote protein degradation by proteasomes, PROTAC consists of a group bound to a ubiquitin ligase and a group bound to a protein targeted for degradation (optionally, these groups are connected to a linker). This molecular structure can bring the ubiquitin pathway protein binding portion to the vicinity of the protein, thereby making it ubiquitinated and marked for degradation.

[0100] The ubiquitin pathway protein binding portion is any suitable structure that can recognize and bind to ubiquitin pathway proteins. In general, ubiquitin pathway proteins are any entities or complexes that can catalyze or cause catalysis of ubiquitin or ubiquitin-like modified polypeptides (e.g., Nedd8, APG12 or ISG15 / UCRP) transferred to another protein. In certain embodiments, the ubiquitin pathway protein is a ubiquitin protein ligase or E3 protein. At least 100 different E3 proteins are encoded by the human genome. Winston et al., A Family of Mammalian F-Box proteins, Current Biology 9(20): 1180-1182(1999).

[0101] In certain embodiments, the ubiquitin pathway protein is a protein that participates in the ubiquitin-like pathway or its components, which transfers ubiquitin-like modified polypeptides (e.g., SUMO, Nedd8, APG12, or ISG15 / UCRP). The components of the ubiquitin-like modification pathway are generally homologs of the ubiquitin pathway. For example, the ubiquitin-like pathway of SUMO includes homologs of ubiquitin protein activating enzymes or E1 proteins, ubiquitin protein conjugating enzymes or E2 proteins, and ubiquitin ligases or E3 proteins.

[0102] In certain embodiments, the ubiquitin pathway protein binding portion is any suitable ligand of a ubiquitin pathway protein, such as a ubiquitin protein ligase or an E3 protein or a homologue thereof. In certain embodiments, the ubiquitin pathway protein binds to a peptide, a domain or region of a ubiquitin pathway protein ligand. In certain embodiments, the ubiquitin pathway protein binding portion recognizes and binds to a ubiquitin pathway protein in a regulated manner.

[0103] A component is a compound herein that binds to a target protein for degradation (eg, a targeted kinase inhibitor such as CDK, FLT3 or haspin kinase). The term "protein" includes oligopeptides and sufficiently long polypeptide sequences that can bind to the A component.

[0104] Particular A groups are any of the compounds described herein.

[0105] The compound and the ubiquitin pathway protein binding portion can be optionally connected to a linker. The linker can be any suitable linker. The linker can include atoms selected from C, N, O, S, Si and P; selected from C, N, O, S and P; or selected from C, N, O and S. The linker can have a main chain of varying lengths, so that there can be as few as two atoms in the linker main chain to up to 100 or more continuous atoms in the linker main chain. The "main chain" of the linker is the shortest continuous atomic chain that forms a covalent bond connection between A and D. In some embodiments, the multivalent linker has a branched main chain, and each branch is used as a part of the main chain linker until the end is reached.

[0106] For example, the linker can have a chain length of at least about 7 atoms. In some embodiments, the length of the linker is at least about 10 atoms. In some embodiments, the length of the linker is at least about 14 atoms. In some embodiments, the length of the linker is about 7 to about 31 atoms (e.g., about 7 to 31, 7 to about 31, or 7 to 31 atoms), about 7 to about 24 atoms (e.g., about 7 to 24, 7 to about 24, or 7 to 24 atoms) or about 7 to about 20 atoms (e.g., about 7 to 20, 7 to about 20, or 7 to 20 atoms). In some embodiments, the linker is about 14 to about 31 atoms (e.g., about 14 to 31, 14 to about 31, or 14 to 31 atoms), about 14 to about 24 atoms (e.g., about 14 to 24, 14 to about 24, or 14 to 24 atoms), or about 14 to about 20 atoms (e.g., about 14 to 20, 14 to about 20, or 14 to 20 atoms). In some embodiments, the chain length of the linker is at least 7 atoms, at least 14 atoms, at least 20 atoms, at least 25 atoms, at least 30 atoms, or at least 40 atoms; or 1 to 15 atoms, 1 to 5 atoms, 5 to 10 atoms, 5 to 20 atoms, 10 to 40 atoms, or 25 to 100 atoms.

[0107] The linker may comprise at least one carbon-carbon bond and / or at least one amide bond. The linker may comprise one or more L- or D- configurations, natural or non-natural amino acids, PEG monomers, PEG oligomers, PEG polymers, or any combination thereof. For linkers comprising one or more PEG units, unless otherwise indicated, all carbon atoms and oxygen atoms of the PEG units are part of the backbone.

[0108] In certain embodiments, a linker is a group comprising one or more covalently linked building blocks.

[0109] In certain embodiments, the linker group is an optionally substituted (poly) ethylene glycol, which has 1 to about 100 ethylene glycol units, about 1 to about 50 ethylene glycol units, 1 to about 25 ethylene glycol units, about 1 to 10 ethylene glycol units, 1 to about 8 ethylene glycol units and 1 to 6 ethylene glycol units, 2 to 4 ethylene glycol units, and / or an optionally substituted alkyl, wherein there is dispersed with optionally substituted O, N, S, P or Si atom. In certain embodiments, the linker group is substituted by aryl, phenyl, benzyl, alkyl, alkylene or heterocyclic group. In certain embodiments, the linker group is asymmetric. In certain embodiments, the linker group is symmetrical.

[0110] Alternatively, or in addition to chain length, in some embodiments, the linker may have suitable substituents that affect hydrophobicity or hydrophilicity. Thus, for example, the linker may have hydrophobic side chain groups, such as alkyl, cycloalkyl, aryl, aralkyl or similar groups, each of which is optionally substituted. If the linker includes one or more amino acids, the linker may contain hydrophobic amino acid side chains, such as one or more amino acid side chains from Phe and TyR, including substituted variants thereof, and analogs and derivatives of such side chains.

[0111] The linker may include a spacer (e.g., conjugated to and / or including a spacer). The spacer may be any suitable spacer. The spacer of the linker may include hydrophilic, hydrophobic, amphiphilic, non-peptide, peptide and / or aromatic monomers. The length of the spacer may be 1 to 30 (e.g., 1 to 30 carbon atoms, PEG with 1-30 units, etc.). Examples of hydrophilic spacers include, but are not limited to, polyethylene glycol polymers and derivatives thereof. Examples of hydrophobic spacers include, but are not limited to, pure or mixed branched hydrocarbons, fluorocarbons, alkanes, alkenes and / or alkyne polymers. Examples of amphiphilic spacers include, but are not limited to, pure or mixed phospholipids and / or derivatives thereof. Examples of peptide spacers include, but are not limited to, pure and mixed single, branched, L- or D- configurations, essential, non-essential, natural and non-natural amino acids and derivatives thereof. Examples of aromatic spacers include, but are not limited to, pure and mixed repeating quinones.

[0112] In some embodiments, the linker is formed by a click chemistry / click chemistry-derived synthesis method. Those skilled in the art understand that the terms "click chemistry" and "click chemistry-derived" generally refer to a class of small molecule reactions commonly used for conjugation, allowing a selected substrate to be connected to a specific molecule. Click chemistry is not a single specific reaction, but describes a way to generate products following examples from nature, which also generates substances by connecting small modular units. In many applications, click reactions connect biomolecules and reporter molecules. Click chemistry is not limited to biological conditions; the concept of "click" reactions has been used in pharmacology and various biomimetic applications. However, they are very useful in the detection, localization and identification of biomolecules.

[0113] Click reactions can be performed in a single reaction vessel, are typically undisturbed by water, produce minimal byproducts, and are “spring-loaded”—characterized by a high thermodynamic driving force that rapidly and irreversibly drives them to produce a single reaction product in high yield, with high reaction specificity (and, in some cases, both regio- and stereo-specificity). These properties make click reactions suitable for problems of isolating and targeting molecules in complex biological environments, where the products correspondingly need to be physiologically stable and any byproducts need to be nontoxic (e.g., to in vivo systems).

[0114] Salt

[0115] Compounds and conjugates may be present in the form of pharmaceutically acceptable salts. The term "pharmaceutically acceptable salts" refers to those salts whose counterions can be used in medicines. In various embodiments, such salts include, but are not limited to: 1) acid addition salts, which can be obtained by reacting the free base of the parent compound with an inorganic acid (e.g., hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, sulfuric acid, and perchloric acid, etc.) or with an organic acid (e.g., acetic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid, succinic acid, or malonic acid, etc.); or 2) salts formed when the acidic protons present in the parent compound are replaced by metal ions (e.g., alkali metal ions, alkaline earth ions, or aluminum ions); or salts formed when coordinated with organic bases (e.g., ethanolamine, diethanolamine, triethanolamine, trimethylamine, N-methylglucamine, etc.). Pharmaceutically acceptable salts are well known to those skilled in the art, and any such pharmaceutically acceptable salts are considered to be relevant to the embodiments described herein.

[0116] Pharmaceutically acceptable salts can be synthesized from parent compounds containing basic or acidic moieties by conventional chemical methods. In some cases, such salts can be prepared by reacting these compounds in free acid or base form with a stoichiometric amount of an appropriate base or acid in water or an organic solvent, or in a mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred. A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, the disclosure of which is incorporated herein by reference.

[0117] In various embodiments, suitable acid addition salts are formed from acids that form nontoxic salts. Illustrative examples include acetate, aspartate, benzoate, benzenesulfonate, bicarbonate / carbonate, bisulfate / sulfate, borate, camphorsulfonate, citrate, edisylate, ethanesulfonate, formates, fumarates, glucoheptonates, gluconates, glucuronates, hexafluorophosphates, hydroxybenzoates, hydrochlorides / chlorides, hydrobromides / bromides, hydroiodides / iodides, isethionates, lactates, malates, maleates, malonates, mesylates, methylsulfates, naphthoates, 2-naphthalenesulfonates, nicotinates, nitrates, orotates, oxalates, palmitates, pamoates, phosphates / hydrogenphosphates / dihydrogenphosphates, saccharates, stearates, succinates, tartrates, mesylates, and trifluoroacetates.

[0118] In various embodiments, suitable base salts are formed from bases that form non-toxic salts. Illustrative examples include arginine, benzathine, calcium, choline, diethylamine, diethanolamine, glycine, lysine, magnesium, meglumine, ethanolamine, potassium, sodium, tromethamine, and zinc salts. Hemisalts of acids and bases can also be formed, such as hemisulphate and hemicalcium salts.

[0119] In each embodiment of the present specification, it is understood that the formula includes and represents not only all pharmaceutically acceptable salts of the compound and conjugate, but also any and all hydrates and / or solvates of the compound formula or its salt. The term "solvate" refers to a compound or its salt, which also includes a stoichiometric or non-stoichiometric solvent bound by non-covalent intermolecular forces. When the solvent is water, the solvate is a hydrate.

[0120] It should be understood that some functional groups, such as hydroxyl, amino and the like, form complexes and / or coordination compounds with water and / or various solvents, having the physical forms of various compounds. Therefore, it should be understood that these molecular formulas include and represent those various hydrates and / or solvates.

[0121] Composition, route of administration and dosage

[0122] A composition is also provided, which includes the above-mentioned compound (e.g., a compound of Formula I, Formula IA, Formula IB or Formula II) or a conjugate (e.g., a conjugate of Formula X) and a pharmaceutically acceptable carrier or excipient. The term "composition" generally refers to any product containing more than one ingredient (including a compound or conjugate). The composition can be prepared from an independent compound or conjugate or from salts, solutions, hydrates, solvates and other forms of the compound and / or conjugate.

[0123] The term "pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers, diluents or encapsulating materials that are suitable for administration to humans or other vertebrates. The term "carrier" refers to an organic or inorganic natural or synthetic ingredient to which the active ingredient is combined to facilitate administration. The carrier can be an excipient. The choice of carrier can depend on factors such as the specific mode of administration, the effect of the carrier on solubility and stability, and the properties of the dosage form. Pharmaceutical compositions suitable for delivering the compounds described herein and methods for preparing the same can be found in, for example, Remington: The Science & Practice of Pharmacy, 21st edition (Lippincott Williams & Wilkins, 2005).

[0124] The components of the compositions may also be mixed with the compounds, or with each other, in such a manner that there is no interaction that would significantly impair the desired pharmaceutical efficacy.

[0125] For example, the composition can include Cremophor, polysorbate, nanoparticles, polymers or hydrogels. In certain embodiments, the pharmaceutical composition includes a variety of compounds and a pharmaceutically acceptable carrier. In certain embodiments, the pharmaceutical composition also includes at least one additional pharmaceutically active agent. At least one additional pharmaceutically active agent can be an agent that can be used to treat cancer.

[0126] Pharmaceutical compositions can be prepared by combining one or more compounds or conjugates with a pharmaceutically acceptable carrier and optionally one or more additional ingredients (e.g., pharmaceutically active ingredients). The formulations can be administered in pharmaceutically acceptable solutions, which typically contain pharmaceutically acceptable concentrations of salts, buffers, preservatives, compatible carriers, adjuvants, and optionally other therapeutic ingredients.

[0127] Pharmaceutically acceptable carriers may include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents and absorption delay agents, etc., and combinations thereof that are physiologically compatible. The carrier may be suitable for parenteral administration. Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the temporary preparation of sterile injection solutions or dispersions. Examples of such carriers (or excipients) include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers (e.g., polyethylene glycol). One or more other active agents may also be incorporated into the pharmaceutical composition.

[0128] The composition can be formulated as a liquid, such as a suspension or solution. Liquid preparations can include water, ethanol, polyethylene glycol, propylene glycol, methylcellulose or a suitable oil, and one or more emulsifiers and / or suspending agents. Liquid preparations can be prepared by solid reconstitution.

[0129] Pharmaceutical preparations (e.g., for parenteral administration) include aqueous solutions of active compounds in water-soluble form. In addition, suspensions of active compounds can be prepared as suitable oily injection suspensions. Aqueous suspensions can contain compounds mixed with suitable excipients alone or in combination with one or more other active agents. Excipients include suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and gum arabic; dispersants or wetting agents, such as naturally occurring phospholipids, such as lecithin; condensation products of alkylene oxides with fatty acids, such as polyoxyethylene stearate; condensation products of ethylene oxide with long-chain aliphatic alcohols, such as heptadecaethyleneoxyhexadecanoate; condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol, such as polyoxyethylene sorbitol monooleate; or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as polyoxyethylene sorbitan monooleate. Aqueous suspensions can also include one or more preservatives, such as ascorbic acid or ethyl, n-propyl or p-hydroxybenzoate, and one or more coloring agents. In certain embodiments, aqueous suspensions can also include suitable lipophilic solvents or carriers, including fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglyceride, or liposomes. Optionally, suspensions can also include suitable stabilizers or agents that increase compound solubility, to allow preparation of highly concentrated solutions.

[0130] Alternatively, the pharmaceutical composition can be in powder form and formulated with a suitable carrier (e.g., sterile pyrogen-free water) before use. Dispersible powders and granules suitable for preparing aqueous suspensions by adding water can provide the active ingredient blended with a suspending agent, a dispersant or a wetting agent and one or more preservatives. Other excipients, such as coloring agents, may also be present.

[0131] Suitable emulsifiers include naturally occurring gums, such as gum arabic or tragacanth; naturally occurring phospholipids, such as soybean lecithin; and esters, including partial esters derived from fatty acids and hexitol anhydrides, such as sorbitol monooleate, and condensation products of partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. Isotonic agents, such as sugars, polyols (such as mannitol, sorbitol or sodium chloride) may be included in the composition. Extended absorption of the injection composition may be achieved by including one or more agents (such as monostearate and gelatin) that delay absorption in the composition.

[0132] For medical or therapeutic use, an effective amount of the compound or composition can be administered to a subject by any method of delivering the compound as needed. Administration of the composition can be accomplished by any method known to those skilled in the art. Routes of administration include, but are not limited to, intravenous, intramuscular, intraperitoneal, intravesical (bladder), oral, subcutaneous, direct injection, mucosal (e.g., topical for eyes), inhalation, and topical.

[0133] Colorants and / or flavoring agents may be included. For example, the compound may be formulated (eg, by liposome or microsphere encapsulation) and then further included in an edible product, such as a refrigerated beverage containing colorants and flavoring agents.

[0134] Exemplary forms for oral administration include, but are not limited to, tablets, capsules, elixirs, syrups, and the like.

[0135] In certain embodiments, the compound can be directly administered into the bloodstream, muscle or internal organs. Suitable routes for such parenteral administration include intravenous, intraarterial, intraperitoneal, intrathecal, epidural, intraventricular, intraurethral, ​​intrasternal, intracranial, intratumoral, intramuscular, intranasal and subcutaneous. Suitable methods for parenteral administration include needle (including microneedle) syringe, needle-free syringe and infusion technology. When systemic delivery of compound and / or composition is required, compound and / or composition can be formulated for parenteral administration by injection, such as by push or continuous infusion. Injection preparations can exist in unit dosage form, such as in ampoules or more dosage containers, and are added with preservatives. Compositions can be in the form of suspensions, solutions or emulsions in oily or aqueous carriers, and can contain preparatons, such as suspending agents, stabilizers and / or dispersants.

[0136] Parenteral formulations are typically aqueous solutions that may contain carriers or excipients such as salts, carbohydrates and buffers (preferably pH 3-9), but for some applications they may be more suitably formulated as sterile non-aqueous solutions or in dry form to be used with a suitable carrier such as sterile pyrogen-free water.

[0137] Liquid preparations may be suitable for parenteral administration of the compound. Parenteral preparations are prepared under sterile conditions, for example, by lyophilization under sterile conditions, which can be easily accomplished using standard pharmaceutical techniques well known to those skilled in the art. The solubility of the compound can be increased by using appropriate formulation techniques (e.g., incorporation of a solubilizing agent).

[0138] Preparations for parenteral administration can be formulated to release immediately and / or slow release. The compound can be administered in the form of a slow release formulation, for example, in the form of a composition comprising a slow release polymer. The compound can be prepared with a carrier that protects it from rapid release, such as a controlled release formulation, including implants and microencapsulation delivery systems. Biodegradable biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, polylactic acid and polylactic acid, polyglycolic acid copolymers (PGLA). The method for preparing such preparations is generally known to those skilled in the art.

[0139] Sterile injection solutions can be prepared by adding the compound alone or in combination with one or more other active agents, in the desired amount, to an appropriate solvent, adding one or more combinations of the above ingredients as needed, and then filtering and sterilizing. Typically, dispersions are prepared by adding the compound to a sterile carrier containing a dispersion medium and any additional ingredients described above. For sterile powders for the preparation of sterile injection solutions, preferred preparation methods are vacuum drying and freeze drying, which obtain a powder of the active ingredient plus any additional desired ingredients from a previously sterile filtered solution, or these ingredients can be sterile filtered together.

[0140] Pharmaceutical compositions can be formulated into solutions, microemulsions, liposomes or other ordered structures suitable for high drug concentrations. The carrier can be a solvent or dispersion medium, containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.) and suitable mixtures thereof. Appropriate fluidity can be maintained, for example, by using a coating (e.g., lecithin), by maintaining the desired particle size in the case of a dispersion, and by using a surfactant.

[0141] Where appropriate, the compound or pharmaceutical composition containing the compound may be administered continuously.

[0142] Treatments and uses

[0143] Methods for treating kinase-mediated diseases are provided. In certain embodiments, the kinase-mediated disease is cancer. The method may include administering to a subject (e.g., a subject who is experiencing or is at risk of experiencing a kinase-mediated disease (e.g., cancer)) an effective amount of the above-mentioned compound, the above-mentioned conjugate, a pharmaceutically acceptable salt, N-oxide, hydrate, solvate, tautomer or optical isomer of the compound or conjugate, or any pharmaceutical composition herein. In certain embodiments, the compound has a structure of Formula I: or a pharmaceutically acceptable salt thereof, wherein: X1 is each independently N, CH or C-halogen; X2 is each independently N, CH, C-halogen, C-CH n1 X m (where n1+m=3 and X is halogen), C-OCF3, C-OCHF2, C-OMe, C-Me, C-Et, C-tBu, C-iPr, CN-alkyl, CN-heteroalkyl, CO-alkyl or CO-heteroalkyl; R2, R3 and R4 are each independently alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl; and Y is in: n is 1 to 5, R5 and R6 are each independently H, alkyl or heteroalkyl, and R6 may optionally form a ring structure, R7 are each independently H or alkyl, Het-Ar is a heteroaromatic ring, and W is NH, NMe, NEt, NCH2CH2OH, NCH2CH2OMe, O, SO or SO2, Provided that Y is not NH2.

[0144] In certain embodiments, the compound has the structure of Formula IA: or a pharmaceutically acceptable salt thereof, wherein: X1 is each independently N, CH or C-halogen; X2 is each independently N, CH, C-halogen, C-CH n1 X m (where n1+m=3 and X is halogen), C-OCF3, C-OCHF2, C-OMe, C-Me, C-Et, C-tBu, C-iPr, CN-alkyl, CN-heteroalkyl, CO-alkyl or CO-heteroalkyl; R4 is alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl; Y is in: n is 1 to 5, R5 and R6 are each independently H, alkyl or heteroalkyl, and R6 may optionally form a ring structure, R7 are each independently H or alkyl, Het-Ar is a heteroaromatic ring, and W is NH, NMe, NEt, NCH2CH2OH, NCH2CH2OMe, O, SO or SO2, Provided that Y is not NH2. n2 is 1, 2, or 3; and Q is each independently O, NH, substituted N, CH2, or substituted C, S, SO or SO2; and Each Q together forms a 6-, 7- or 8-membered ring or a bicyclic ring.

[0145] In certain embodiments, the compound has the structure of Formula IB: or a pharmaceutically acceptable salt thereof, wherein: X1 is each independently N, CH or C-halogen; X2 is each independently N, CH, C-halogen, C-CH n1 X m (where n1+m=3 and X is halogen), C-OCF3, C-OCHF2, C-OMe, C-Me, C-Et, C-tBu, C-iPr, CN-alkyl, CN-heteroalkyl, CO-alkyl or CO-heteroalkyl; R4 is alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl; R5 is H, alkyl or heteroalkyl; R7 are each independently H or alkyl, o is 0-2; and Each R8 is independently H, alkyl or heteroalkyl.

[0146] In certain embodiments, the compound has the structure of Formula II: or a pharmaceutically acceptable salt thereof, wherein: X1 is each independently N, CH or C-halogen; X2 is each independently N, CH, C-halogen, C-CH n1 X m(where n1+m=3 and X is halogen), C-OCF3, C-OCHF2, C-OMe, C-Me, C-Et, C-tBu, C-iPr, CN-alkyl, CN-heteroalkyl, CO-alkyl or CO-heteroalkyl; Q is each independently O, NH, substituted N, CH2, or substituted C, S, SO or SO2, and each Q when combined together forms a 6-membered, 7-membered or 8-membered ring or a bicyclic ring; n3 is 1, 2, or 3; R4 is alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl; W is O, NH or NMe; and R9 is alkyl or heteroalkyl, provided that R9 is not H.

[0147] The kinase-mediated disease may be selected from the group consisting of cancer, inflammatory diseases, diabetes, and neurological diseases. In certain embodiments, the kinase-mediated disease is cancer. The cancer may be acute myeloid leukemia (AML). The cancer may be chronic myeloid leukemia. The cancer may be gastric cancer. The cancer may be a drug-resistant cancer. The cancer may be selected from the group consisting of AML, chronic myeloid leukemia, ovarian cancer, cervical cancer, pancreatic cancer, breast cancer, brain cancer, skin cancer, lung cancer, prostate cancer, lymphoma, leukemia, colon cancer, gastric cancer, head cancer, neck cancer, thyroid cancer, kidney cancer, liver cancer, and stomach cancer.

[0148] In certain embodiments, the compound or conjugate administered to the subject includes the compound HSH2177. In certain embodiments, the compound or conjugate administered to the subject includes the compound HSH2177, and an effective amount of a drug that inhibits CDK, IC 50 About 50 nM or less (e.g., 50 nM or less). In certain embodiments, the compound or conjugate administered to a subject includes Compound 205. In certain embodiments, the compound or conjugate administered to a subject includes Compound 205, and an effective amount is administered to the subject to specifically inhibit FLT3 and haspin kinases in the subject.

[0149] In certain embodiments of the method, an effective amount of an IC of about 50 nM or less (e.g., 50 nM or less) is administered to a subject. 50 Inhibits CDK in a subject (e.g., with specificity). In certain embodiments of the method, the compound or pharmaceutical composition is administered at an IC of about 30 nM or less (e.g., 30 nM or less, 29 nM or less, 28 nM or less, 27 nM or less, 25 nM or less, ... 10 nM or less, or 5 nM or less). 50Inhibit CDK (e.g., with specificity). Administration of an effective amount to a subject can inhibit both FLT3 and haspin (e.g., with dual specificity). In certain embodiments, administration of an effective amount of Compound 205 or an analog thereof, e.g., with an IC of about 50 nM or less 50 (e.g. IC50nM or lower 50 ) dually inhibits both FLT3 and haspin kinases.

[0150] The terms "treat", "treating", "treated", "treatment" (for a disease or condition) are an approach to obtaining beneficial or desired results, including and preferably clinical results, and include, but are not limited to, one or more of the following: improving symptoms associated with the disease, curing the disease, reducing the severity of the disease or otherwise inhibiting the disease, delaying the progression of the disease, alleviating one or more symptoms associated with the disease, reducing the severity of the disease, inhibiting the onset of the disease, inhibiting the progression of the disease, improving the quality of life of patients with the disease, prolonging survival, and / or prophylactic or preventative treatment.

[0151] As used herein, "effective amount" refers to any amount of a compound or conjugate used in treatment, and refers to the amount of the compound or conjugate in a formulation that, when administered as part of a desired dosage regimen (to a subject, e.g., a human), is sufficient to achieve the desired biological effect (e.g., relief of symptoms, amelioration of symptoms, or slowing of the onset of a disease condition, according to clinically acceptable standards for the disease or condition being treated or for cosmetic purposes, e.g., at a reasonable benefit / risk ratio applicable to any medical treatment).

[0152] In certain embodiments, the method may further include administering to the subject a second therapy comprising an effective amount of a chemotherapeutic agent (e.g., when the disease state is cancer), an immunotherapeutic agent (e.g., when the disease state is an inflammatory disease), or a hormonal therapy agent. In certain embodiments, the second therapy may include radiation therapy.

[0153] It should be understood that in the methods described herein, the individual components of co-administration or combination can be administered simultaneously, continuously, separately or in the form of a single pharmaceutical formulation by any suitable means. When the co-administered compound or composition is administered in a separate dosage form, the number of doses administered per day for each compound can be the same or different. The compound, conjugate and / or composition can be administered by the same or different routes of administration and / or with a second therapy. The compound, conjugate or composition can be administered simultaneously in separate or single dosage forms at the same or different times during the treatment according to a simultaneous or alternating regimen.

[0154] In conjunction with the teachings provided herein, by selecting among various active compounds and weighing factors such as efficacy, relative bioavailability, patient weight, severity of adverse side effects and mode of administration, an effective prevention or treatment regimen can be formulated that does not cause significant undesirable toxicity but is effective for treating a particular subject. The effective amount of any particular application may vary depending on factors such as the disease or condition being treated, the specific compound being administered, the size of the subject, or the severity of the disease or condition. One of ordinary skill in the art can empirically determine the effective amount of a particular compound and / or other therapeutic agent without having to perform excessive experiments.

[0155] For example, for any compound, the effective amount can be first determined from an animal model. For compounds tested in humans and compounds known to exhibit similar pharmacological activity (e.g., other related active agents), the effective dose can also be determined from human data. Higher doses may be required for parenteral administration. The applied dose can be adjusted according to the relative bioavailability and efficacy of the administered compound. It is entirely within the capabilities of ordinary technicians to adjust the dose to achieve maximum efficacy according to the above methods and other methods well known in the art.

[0156] A maximum dose, i.e., the highest safe dose based on some medical judgment, may be used. Multiple doses may be used daily to achieve appropriate systemic levels of the compound. Appropriate systemic levels may be determined, for example, by measuring peak or sustained plasma levels of the drug in the patient.

[0157] "Dose" and "amount" are used interchangeably herein.

[0158] In general, for human subjects, the daily oral dose of the compound is about 0.01 mg / kg / day to 1,000 mg / kg / day. Oral doses of 0.5 mg / kg to 50 mg / kg administered once or more daily can produce therapeutic effects. The dose can be appropriately adjusted according to the mode of administration to achieve the desired local or systemic drug level. For example, the daily dose for intravenous administration can be one order of magnitude lower to several orders of magnitude. If the subject does not respond adequately at such a dose, a higher dose (or an effective higher dose through a different, more localized delivery route) can be used within the patient's tolerance range. Consider multiple daily dosing to achieve appropriate systemic levels of the compound.

[0159] For clinical use, any compound or conjugate can be administered in an amount equal to or equivalent to 0.2-2,000 milligrams (mg) of compound per kilogram of subject body weight per day. The compound can be administered in an amount equal to or equivalent to 2-2,000 mg of compound per kilogram of subject body weight per day. The compound can be administered in an amount equal to or equivalent to 20-2,000 mg of compound per kilogram of subject body weight per day. The compound can be administered in an amount equal to or equivalent to 50-2,000 mg of compound per kilogram of subject body weight per day. The compound can be administered in an amount equal to or equivalent to 100-2,000 mg of compound per kilogram of subject body weight per day. The compound can be administered in an amount equal to or equivalent to 200-2,000 mg of compound per kilogram of subject body weight per day. When a precursor or prodrug of a compound is to be administered, it is administered in an amount equal to (i.e., sufficient to deliver) the above-mentioned compound.

[0160] The preparation of the compound or its pharmaceutically acceptable salt can be applied to human subjects in an effective amount. Typical dosage ranges can be about 0.01 micrograms / kg to about 2 mg / kg body weight per day. The dosage of the drug to be administered may depend on variables such as the type and extent of the disease, the overall health of the particular subject, the particular compound administered, the excipient used to prepare the compound and its route of administration. Routine experiments can be used to optimize the dosage and frequency of administration of any particular compound or its pharmaceutically acceptable salt.

[0161] The compound or a pharmaceutically acceptable salt thereof can be administered at a concentration ranging from about 0.001 μg / kg to greater than about 500 mg / kg. For example, the concentration can be 0.001 μg / kg, 0.01 μg / kg, 0.05 μg / kg, 0.1 μg / kg, 0.5 μg / kg, 1.0 μg / kg, 10.0 μg / kg, 50.0 μg / kg, 100.0 μg / kg, 500 μg / kg, 1.0 mg / kg, 5.0 mg / kg, 10.0 mg / kg, 15.0 mg / kg, 20.0 mg / kg, 25.0 mg / kg, 30.0 mg / kg, 35.0 mg / kg, In some embodiments, the present invention relates to an aqueous solution of at least about 100 mg / kg, about 200 mg / kg, about 400 mg / kg, about 500 mg / kg, about 600 mg / kg, about 700 mg / kg, about 800 mg / kg, about 900 mg / kg, about 1000 mg / kg, about 1500 mg / kg, about 2000 mg / kg, about 2500 mg / kg, about 3000 mg / kg, about 3500 mg / kg, about 4000 mg / kg, about 4500 mg / kg, to greater than about 500.0 mg / kg or any incremental value thereof. It should be understood that all values ​​and ranges between these values ​​and ranges should be covered.

[0162] The compound or a pharmaceutically acceptable salt thereof can be administered in a dosage range of about 0.2 mg / kg / day to greater than about 100 mg / kg / day. For example, the dosage can be 0.2 mg / kg / day to 100 mg / kg / day, 0.2 mg / kg / day to 50 mg / kg / day, 0.2 mg / kg / day to 25 mg / kg / day, 0.2 mg / kg / day to 10 mg / kg / day, 0.2 mg / kg / day to 7.5 mg / kg / day, 0.2 mg / kg / day to 5 mg / kg / day, 0.25 mg / kg / day to 100 mg / kg / day, 0.25 mg / kg / day to 50 mg / kg / day, 0.25 mg / kg / day to 25 mg / kg / day, 0.25 mg / kg / day to 100 mg / kg / day, 0.25 mg / kg / day to 50 mg / kg / day, 0.25 mg / kg / day to 25 mg / kg / day, 0.25 mg / kg / day to 100 mg / kg / day. / day to 10mg / kg / day, 0.25mg / kg / day to 7.5mg / kg / day, 0.25mg / kg / day to 5mg / kg / day, 0.5mg / kg / day to 50mg / kg / day, 0.5mg / kg / day to 25mg / kg / day, 0.5mg / kg / day to 20mg / kg / day, 0.5mg / kg / day to 15mg / kg / day, 0.5mg / kg / day to 10mg / kg / day, 0.5mg / kg / day to 7.5mg / kg / day, 0.5mg / kg / day to 5mg / kg / day, 0.75mg / kg / day to 50mg / kg / day, 0.75mg / kg / day to 25mg / kg / day, 0.75mg / kg / day to 20mg / kg / day, 0.75mg / kg / day to 15mg / kg / day, 0.75mg / kg / day to 10mg / kg / day, 0.75mg / kg / day to 7.5mg / kg / day, 0.75mg / kg / day to 5mg / kg / day, 1.0mg / kg / day to 50mg / kg / day, 1.0mg / kg / day to 25mg / kg / day, 1.0mg / kg / day to 20mg / kg / day, 1.0mg / kg / day to 15mg / kg / day, 1.0mg / kg / day to 10mg / kg / day, 1.0mg / kg / day to 7.5mg / kg / day, 1.0mg / kg / day to 5mg / kg / day, 2mg / kg / day to 50mg / kg / day, 2mg / kg / day to 25mg / kg / day, 2mg / kg / day to 20mg / kg / day, 2mg / kg / day to 15mg / kg / day, 2mg / kg / day to 10mg / kg / day, 2mg / kg / day to 7.5mg / kg / day, or 2mg / kg / day to 5mg / kg / day.

[0163] The compound can be administered at a dosage ranging from about 0.25 mg / kg / day to about 25 mg / kg / day. For example, the dosage can be 0.25 mg / kg / day, 0.5 mg / kg / day, 0.75 mg / kg / day, 1.0 mg / kg / day, 1.25 mg / kg / day, 1.5 mg / kg / day, 1.75 mg / kg / day, 2.0 mg / kg / day, 2.25 mg / kg / day, 2.5 mg / kg / day, 2.75 mg / kg / day, 3.0 mg / kg / day, 3.25 mg / kg / day, 3.5 mg / kg / day, 3.75 mg / kg / day, 4.0 mg / kg / day, 4.25mg / kg / day, 4.5mg / kg / day, 4.75mg / kg / day, 5mg / kg / day, 5.5mg / kg / day, 6.0mg / kg / day, 6.5mg / kg / day, 7.0mg / kg / day, 7.5mg / kg / day, 8.0mg / kg / day, 8.5mg / kg / day, 9.0mg / kg / day, 9.5mg / kg / day, 10mg / kg / day, 11mg / kg / day, 12mg / kg / day, 13mg / kg g / day, 14mg / kg / day, 15mg / kg / day, 16mg / kg / day, 17mg / kg / day, 18mg / kg / day, 19mg / kg / day, 20mg / kg / day, 21mg / kg / day, 22mg / kg / day, 23mg / kg / day, 24mg / kg / day, 25mg / kg / day, 26mg / kg / day, 27mg / kg / day, 28mg / kg / day, 29mg / kg / day, 30mg / kg / day, 31mg / kg / day, 32mg / kg / day 50 mg / kg / day, 51 mg / kg / day, 52 mg / kg / day, 53 mg / kg / day, 54 mg / kg / day, 55 mg / kg / day, 56 mg / kg / day, 57 mg / kg / day, 58 mg / kg / day, 59 mg / kg / day, 60 mg / kg / day, 61 mg / kg / day, 62 mg / kg / day, 63 mg / kg / day, 64 mg / kg / day, 65 mg / kg / day, 66 mg / kg / day, 67 mg / kg / day, 68 mg / kg / day, 69 mg / kg / day or 70 mg / kg / day.

[0164] The compound, its pharmaceutically acceptable salt or its precursor can be administered at a concentration ranging from 0.01 micromolar to greater than or equal to 500 micromolar. For example, the dosage can be 0.01 micromolar, 0.02 micromolar, 0.05 micromolar, 0.1 micromolar, 0.15 micromolar, 0.2 micromolar, 0.5 micromolar, 0.7 micromolar, 1.0 micromolar, 3.0 micromolar, 5.0 micromolar, 7.0 micromolar, 10.0 micromolar, 15.0 micromolar, 20.0 micromolar, 25.0 micromolar, 30.0 micromolar, 35.0 micromolar, 40.0 micromolar, 45.0 micromolar, 46.0 micromolar, 47.0 micromolar, 48.0 micromolar, 49.0 micromolar, 50.0 micromolar, 51.0 micromolar, 52.0 micromolar, 53.0 micromolar, 54.0 micromolar, 56.0 micromolar, 57.0 micromolar, 58.0 micromolar, 59.0 micromolar, 60.0 micromolar, 61.0 micromolar, 62.0 micromolar, 63.0 micromolar, 64.0 micromolar, 65.0 micromolar, 66.0 micromolar, 67.0 micromolar, 68.0 micromolar, 69.0 micromolar, 70.0 micromolar, 71.0 micromolar, 72.0 micromolar, 73.0 micromolar, 74.0 micromolar, 75.0 micromolar, 76.0 micromolar, 77.0 micromolar, 78.0 micromolar, 79 In some embodiments, the present invention relates to an aqueous solution of at least about 500 micromolar, about 200 micromolar, about 400 micromolar, about 500 micromolar, about 600 micromolar, about 700 micromolar, about 800 micromolar, about 900 micromolar, about 1000 micromolar, about 1500 micromolar, about 2000 micromolar, about 2500 micromolar, about 3000 micromolar, about 3500 micromolar, about 4000 micromolar, about 4500 micromolar, to greater than about 5000 micromolar or any incremental value thereof. It should be understood that all values ​​and ranges between these values ​​and ranges should be covered.

[0165] The compound or a pharmaceutically acceptable salt thereof or a precursor thereof may be administered at a concentration ranging from 0.10 micrograms / mL to 500.0 micrograms / mL. For example, the concentration can be from 0.10 micrograms / mL, 0.50 micrograms / mL, 1 microgram / mL, 2.0 micrograms / mL, 5.0 micrograms / mL, 10.0 micrograms / mL, 20 micrograms / mL, 25 micrograms / mL, 30 micrograms / mL, 35 micrograms / mL, 40 micrograms / mL, 45 micrograms / mL, 50 micrograms / mL, 60.0 micrograms / mL, 70.0 micrograms / mL, 80.0 micrograms / mL, 90.0 micrograms / mL, 100.0 micrograms / mL, 150.0 micrograms / mL, 200.0 micrograms / mL, 250.0 micrograms / mL, 250.0 micrograms / mL, 300.0 micrograms / mL, 350.0 micrograms / mL, 400.0 micrograms / mL, 450.0 micrograms / mL, to greater than about 500.0 micrograms / mL or any incremental value thereof. It should be understood that all values ​​and ranges between these values ​​and ranges should be included.

[0166] Preparations can be used in the form of pharmaceutically acceptable solutions, which solutions can generally contain pharmaceutically acceptable concentrations of salt, buffers, preservatives, compatible carriers, adjuvants and optional other therapeutic ingredients. For therapeutic use, the compound of an effective amount can be administered to a subject by any means by which the compound is delivered to the desired surface. Pharmaceutical compositions can be administered by any means known to those skilled in the art. Routes of administration include, but are not limited to, intravenous, intramuscular, intraperitoneal, intravesical (bladder), oral, subcutaneous, direct injection (e.g., injection into a tumor or abscess), mucosal (e.g., local for eyes), suction and topical.

[0167] For intravenous and other parenteral administration routes, the compound can be formulated as a lyophilized preparation, a lyophilized preparation of the active compound inserted or encapsulated by liposomes, a lipid complex or a salt complex in an aqueous suspension. The lyophilized preparation is usually reconstituted in a suitable aqueous solution (e.g., sterile water or saline) shortly before administration.

[0168] For oral administration, the compound can be easily formulated by combining the active compound with a pharmaceutically acceptable carrier known in the art. Such a carrier enables the compound to be formulated into tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral ingestion by the subject to be treated. Pharmaceutical preparations for oral use can be obtained as solid excipients, optionally grinding the resulting mixture, and processing the granular mixture after adding suitable adjuvants (if necessary) to obtain tablets or dragee cores. Suitable excipients are particularly fillers, such as sugars, including lactose, sucrose, mannitol or sorbitol; cellulose preparations, such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidone (PVP). If necessary, a disintegrant can be added, such as cross-linked PVP, agar or alginic acid or a salt thereof, such as sodium alginate. Optionally, oral formulations may also be formulated in saline or a buffer (eg, ethylenediaminetetraacetic acid (EDTA)) for neutralizing internal acidic conditions, or may be administered without any carrier.

[0169] Oral dosage forms of the compound are also contemplated. The compound may be chemically modified so that oral delivery of the derivative is effective. Typically, the chemical modification contemplated is to attach at least one moiety to the compound itself, wherein the moiety allows (a) inhibition of acid hydrolysis; and (b) absorption from the stomach or intestine into the bloodstream. It is also desirable to improve the overall stability of the compound and increase circulation time in the body. Examples of such moieties include polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethyl cellulose, dextran, polyvinyl alcohol, PVP, and polyproline. Abuchowski and Davis, "Soluble Polymer-Enzyme Adducts," In: Enzymes as Drugs, Hocenberg and Roberts, eds., Wiley-Interscience, New York, NY, pp. 367-383 (1981); Newmark et al., J Appl Biochem 4: 185-189 (1982). Other polymers that may be used are poly-1,3-dioxolane and poly-1,3,6-thioxoctane.For pharmaceutical uses, as described above, polyethylene glycol moieties are suitable.

[0170] The release site of the compound herein or its pharmaceutically acceptable salt may be the stomach, small intestine (e.g., duodenum, jejunum, or ileum), or large intestine. Those skilled in the art have available formulations that will not dissolve in the stomach but will release the substance in the duodenum or elsewhere in the intestinal tract. Release may avoid the harmful effects of the gastric environment by protecting the compound or by releasing the compound or its pharmaceutically acceptable salt outside the gastric environment, for example in the intestinal tract.

[0171] To ensure complete gastric tolerance, a coating that is impermeable at least at pH 5.0 may be essential. Examples of more common inert ingredients used as enteric coatings are cellulose acetate trimellitate (CAT), hydroxypropyl methylcellulose phthalate (HPMCP), HPMCP 50, HPMCP 55, polyvinyl acetate phthalate (PVAP), Eudragit L30D, Aquateric, cellulose acetate phthalate (CAP), Eudragit L, Eudragit S and shellac. These coatings can be applied as mixed films.

[0172] Tablets may also be coated with coatings or coating mixtures that are not intended for gastric protection. This may include sugar coatings or coatings that make the tablet easier to swallow. Capsules may consist of a hard shell such as gelatin for delivery of dry therapeutic agents (e.g., powders); for liquid forms, a soft gelatin shell may be used. The outer shell material for cachets may be thick starch or other edible paper. For pills, lozenges, molded tablets, or tablet triturates, moist massing techniques may be used.

[0173] As mentioned above, the compound or its pharmaceutically acceptable salt may be included in the formulation as fine multi-particulates in the form of granules or pellets having a particle size of about 1 mm. The formulation of the material for capsule administration may also be a powder, lightly compressed plugs or even tablets.

[0174] Pharmaceutical formulations that can be used orally include push-in capsules made of gelatin, and soft sealed capsules made of gelatin and a plasticizer (e.g., glycerol or sorbitol). Push-in capsules may contain the active ingredient blended with a filler (e.g., lactose), a binder (e.g., starch), and / or a lubricant (e.g., talc or magnesium stearate) and an optional stabilizer. In soft capsules, the active compound may be dissolved or suspended in a suitable liquid, such as a fatty oil, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers may be added. Microspheres formulated for oral administration may also be used. Such microspheres are well defined in the art. All formulations for oral administration should have a dosage suitable for such administration.

[0175] For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

[0176] For topical administration, the compounds can be formulated into solutions, gels, ointments, creams, suspensions, etc., as is well known in the art. Systemic formulations include those designed for administration by injection, such as subcutaneous, intravenous, intramuscular, intrathecal, or intraperitoneal injection, as well as those designed for transdermal, transmucosal oral, or pulmonary administration.

[0177] For administration by inhalation, the compound is conveniently delivered in the form of an aerosol spray from a pressurized pack or nebulizer, using a suitable propellant, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges, for example, of gelatin, for use in an inhaler or insufflator may be formulated containing a powder mix of the compound and a suitable powder base, for example lactose or starch.

[0178] Pulmonary delivery of the compound (or its salt) is also contemplated. The compound is delivered to the lungs of a mammal upon inhalation and crosses the lung epithelial lining into the bloodstream. Other reports of inhaled molecules include Adjei et al., Pharm Res 7:565-569 (1990); Adjei et al., Int J Pharmaceutics 63:135-144 (1990) (leuprolide acetate); Braquet et al., J Cardiovasc Pharmacol 13(suppl. 5):143-146 (1989) (endothelin-1); Hubbard et al., Annal Int Med 3:206-212 (1989) (a1-antitrypsin); Smith et al., 1989, J Clin Invest 84:1145-1146 (a-1-proteinase); Oswein et al., 1990, "Aerosolization of Proteins," Proceedings of Symposium on Respiratory Drug Delivery II, Keystone, Colorado, March, (recombinant hepatocyte growth hormone); Debs et al., 1988, J Immunol 140:3482-3488 (interferon-γ and tumor necrosis factor α) and Platz et al., U.S. Pat. No. 5,284,656 (granulocyte colony stimulating factor; incorporated herein by reference). U.S. Pat. No. 5,451,569, issued Sept. 19, 1995 to Wong et al. (the disclosure of which is specifically incorporated herein by reference) describes a method and composition for pulmonary delivery of drugs for systemic effect.

[0179] A variety of mechanical devices designed for pulmonary delivery of therapeutic products are contemplated for use, including, but not limited to, nebulizers, metered dose inhalers, and powder inhalers, all of which are familiar to those skilled in the art.

[0180] Nasal delivery of pharmaceutical compositions is also contemplated. Nasal delivery allows the pharmaceutical composition to enter the bloodstream directly after the therapeutic product is applied to the nasal cavity without depositing the product in the lungs. Formulations for nasal delivery include formulations with dextran or cyclodextran.

[0181] When systemic delivery is required, the compound can be formulated for parenteral administration by injection, for example by bolus injection or continuous infusion. Formulations for injection can be in unit dose form, for example in ampoules or multidose containers, with the addition of preservatives. The composition can be in the form of a suspension, solution or emulsion in an oily or aqueous vehicle, and can contain preparatants, such as suspending agents, stabilizers and / or dispersants.

[0182] Pharmaceutical preparations for parenteral administration include aqueous solutions of the active compound in water-soluble form. In addition, the suspension of the active compound can be prepared into suitable oily injection suspensions. Suitable lipophilic solvents or carriers include fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions can contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol or dextran. Optionally, the suspension can also contain suitable stabilizers or agents that increase the solubility of the compound to allow the preparation of high concentration solutions.

[0183] Alternatively, the active compound may be in powder form for constitution with a suitable vehicle (eg, sterile pyrogen-free water) before use.

[0184] The compounds may also be formulated in rectal or vaginal compositions such as suppositories or retention enemas, eg, containing conventional suppository bases such as cocoa butter or other glycerides.

[0185] In addition to the above-mentioned preparations, the compound can also be formulated into a long-acting preparation. Such a long-acting preparation can be formulated with a suitable polymer or hydrophobic material (e.g., formulated as an emulsion in an acceptable oil) or an ion exchange resin, or formulated as a poorly soluble derivative, e.g., a poorly soluble salt.

[0186] The pharmaceutical composition may also contain suitable solid or gel phase carriers or excipients. Examples of these carriers or excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols.

[0187] Suitable liquid or solid pharmaceutical preparation forms are, for example, aqueous solutions or saline solutions for suction, microcapsules, endocytosis agents, dosage forms coated on microscopic gold particles, dosage forms contained in liposomes, atomizers, aerosols, pills for implanting skin, or dosage forms dried on sharp objects to scrape into the skin. Pharmaceutical compositions also include granules, powders, tablets, coated tablets, (micro) capsules, suppositories, syrups, emulsions, suspensions, creams, drops or preparations with sustained release of active compounds, in which excipients and additives and / or adjuvants as described above are generally used, such as disintegrants, adhesives, coating agents, swelling agents, lubricants, flavoring agents, sweeteners or solubilizing agents. Pharmaceutical compositions are applicable to various drug delivery systems. For a brief review of drug delivery methods, see Langer R, Science 249:1527-1533 (1990).

[0188] Disintegrants can be included in the solid dosage form formulation of the therapeutic agent. Materials used as disintegrants include, but are not limited to, starch, including the commercial disintegrant Explotab based on starch. Sodium starch glycolate, Amberlite, sodium carboxymethylcellulose, super branched starch, sodium alginate, gelatin, orange peel, acid carboxymethylcellulose, natural sponge and bentonite can all be used. Another form of disintegrant is an insoluble cation exchange resin. Powdered gums can be used as disintegrants and binders, and these can include powdered gums, such as agar, carrageenan or tragacanth. Alginic acid and its sodium salt can also be used as disintegrants.

[0189] Binders can be used to hold the compound together to form a hard tablet, including materials from natural products, such as gum arabic, gum tragacanth, starch and gelatin. Other binders include methylcellulose (MC), ethylcellulose (EC) and carboxymethylcellulose (CMC). PVP and hydroxypropylmethylcellulose (HPMC) can both be used in alcohol solutions to form particles of the therapeutic agent.

[0190] Friction reducers may be added to the formulation of the therapeutic agent to prevent sticking during formulation. Lubricants may be used as a layer between the therapeutic agent and the mold wall and may include, but are not limited to, stearic acid (including its magnesium and calcium salts), polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils, and waxes. Soluble lubricants may also be used, such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycols of various molecular weights, Carbowax 4000 and 6000.

[0191] Glidants may be added to improve the flow properties of the drug during formulation and to aid rearrangement during compression. Glidants may include starch, talc, fumed silica, and hydrated aluminosilicates.

[0192] In order to help the therapeutic agent dissolve in the water environment, a surfactant can be added as a wetting agent. The surfactant can include anionic detergents, such as sodium lauryl sulfate, dioctyl sodium sulfosuccinate and dioctyl sodium sulfonate. Cationic detergents that can be used include benzalkonium chloride and benzethonium chloride. Possible nonionic detergents that can be included in the preparation as a surfactant include lauryl alcohol 400, polyethylene glycol 40 stearate, polyoxyethylene hydrogenated castor oil 10, 50 and 60, glyceryl monostearate, polysorbate 40, 60, 65 and 80, sucrose fatty acid esters, methylcellulose and carboxymethylcellulose. These surfactants can be present in the preparation of the compound or its derivatives alone or in a mixture of different proportions.

[0193] The compound can be administered as such (pure) or in the form of a pharmaceutically acceptable salt. When used in medicine, the salt should be pharmaceutically acceptable, but non-pharmaceutically acceptable salts can be conveniently used to prepare pharmaceutically acceptable salts thereof. Such salts include, but are not limited to, salts prepared from the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, maleic acid, acetic acid, salicylic acid, p-toluenesulfonic acid, tartaric acid, citric acid, methanesulfonic acid, formic acid, malonic acid, succinic acid, naphthalene-2-sulfonic acid and benzenesulfonic acid. In addition, such salts can be prepared as alkali metal or alkaline earth metal salts, such as sodium salts, potassium salts or calcium salts of carboxylic acid groups.

[0194] Inert substances may be used to dilute or increase the volume of the compound. These diluents may include carbohydrates, particularly mannitol, α-lactose, anhydrous lactose, cellulose, sucrose, modified dextran, and starch. Certain inorganic salts may also be used as fillers, including tricalcium phosphate, magnesium carbonate, and sodium chloride. Some commercially available diluents are Fast-Flo, Emdex, STA-Rx1500, Emcompress, and Avicell.

[0195] Suitable buffers include acetic acid and its salts (1-2% w / v); citric acid and its salts (1-3% w / v); boric acid and its salts (0.5-2.5% w / v); phosphoric acid and its salts (0.8-2% w / v). Suitable preservatives include benzalkonium chloride (0.003-0.03% w / v); chlorobutanol (0.3-0.9% w / v); parabens (0.01-0.25% w / v) and thimerosal (0.004-0.02% w / v).

[0196] The pharmaceutical composition contains an effective amount of the compound described herein and optionally one or more other therapeutic agents, contained in a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers, diluents or encapsulating substances, which are suitable for administration to humans or other vertebrates. The term "carrier" refers to a natural or synthetic organic or inorganic ingredient with which the active ingredient is combined to facilitate application. The components of the pharmaceutical composition can also be mixed with the compound, and with each other, in a manner such that there is no interaction that would significantly impair the desired drug efficacy.

[0197] Therapeutic agents, specifically including but not limited to the compounds described herein, can be provided in the form of particles. "Particles" as used herein refer to nanoparticles or microparticles (or larger particles in some cases), which can be composed of compounds described herein or other therapeutic agents in whole or in part. The particles can contain therapeutic agents in a core surrounded by a coating, including but not limited to an enteric coating. The therapeutic agent can also be dispersed throughout the particles. The therapeutic agent can also be adsorbed into the particles. The particles can have release kinetics in any order, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, and any combination thereof. In addition to therapeutic agents, the particles can also include any materials conventionally used in the pharmaceutical and medical fields, including but not limited to erodible, non-erodible, biodegradable or non-biodegradable materials or combinations thereof. The particles can be microcapsules containing compounds in a solution or semi-solid state. The particles can have almost any shape.

[0198] Non-biodegradable and biodegradable polymer materials can be used to manufacture particles for delivering therapeutic agents. Such polymers can be natural or synthetic polymers. Polymers are selected based on the time period of required release. Particularly interesting bioadhesive polymers are included in Sawhney et al., Macromolecules 26: The bioerodible hydrogel described in 581-587 (1993), the teaching of which is hereby incorporated by reference into this paper. These include polyhyaluronic acid, casein, gelatin, gelatin, polyanhydride, polyacrylic acid, alginate, chitosan, poly (methyl methacrylate), poly (ethyl methacrylate), poly (butyl methacrylate), poly (isobutyl methacrylate), poly (hexyl methacrylate), poly (isodecyl methacrylate), poly (dodecyl methacrylate), poly (phenyl methacrylate), poly (methyl acrylate), poly (isopropyl acrylate), poly (isobutyl acrylate) and poly (octadecyl acrylate).

[0199] Compounds can be included in controlled release systems. The term "controlled release" is intended to refer to any drug-containing formulation, wherein the mode and profile of drug release from the formulation is controlled. This refers to immediate release formulations and non-immediate release formulations, which include but are not limited to sustained release and delayed release formulations. The term "slow release" (also referred to as "extended release") refers to a pharmaceutical formulation that gradually releases the drug over a long period of time in its conventional sense, and can cause the blood level of the drug to be substantially constant over a long period of time. The term "delayed release" refers to a pharmaceutical formulation in which there is a time delay between the administration of the formulation and the release of the drug from the formulation in its conventional sense. "Delayed release" may or may not involve gradually releasing the drug over a long period of time, and therefore may or may not be "slow release".

[0200] Long-term sustained-release implants are particularly suitable for the treatment of chronic diseases. "Long-term" release as used herein refers to that the implant is constructed and arranged to deliver the active ingredient at the therapeutic level for at least 7 days, and up to 30-60 days. Long-term sustained-release implants are well known to those of ordinary skill in the art, including some of the above-mentioned release systems.

[0201] Depending on the route of administration, various permissible doses are contemplated herein, including doses falling within about 10 6 Up to 10 11 The dosage can be a single dose or divided dose, and can be administered according to various schedules, including qd (once a day), bid (twice a day), tid (three times a day), or even once every other day, once a week, once a month, once a quarter, etc. In each case, it should be understood that the effective amount described herein corresponds to the administration situation, or corresponds to the total daily, weekly, monthly or quarterly dose determined by the dosing schedule.

[0202] In addition to the exemplary dosages and dosing regimens described herein, the attending diagnostician or physician can also determine the effective amount of any one of the compounds or mixtures of compounds described herein by using known techniques and / or by observing the results obtained under similar circumstances. In determining the effective amount or dosage, the attending diagnostician or physician will consider many factors, including but not limited to the species of mammal (including humans), its size, age and general health, the specific disease or condition involved, the degree of involvement or severity of the disease or condition, the response of the individual patient, the specific compound administered, the mode of administration, the bioavailability characteristics of the administered formulation, the selected dosage regimen, the use of concurrent medications, and other relevant circumstances.

[0203] In certain embodiments, there is provided a compound, a conjugate, or a pharmaceutically acceptable salt of the compound or conjugate for use in the manufacture of a medicament for treating a disease in a subject. The compound or conjugate may be any compound or conjugate herein. The subject's disease may be a disease in which modulation of a kinase reduces severity or otherwise treats such a disease in the subject. The disease may be any disease state described herein. In certain embodiments, the disease is cancer, diabetes, an inflammatory disease, or a neurological disease.

[0204] Any of the compounds and / or conjugates herein may be used to treat a disease state regulated by one or more kinases. The disease state may be cancer, and the one or more kinases may be CKD, FLT3 and / or haspin kinases.

[0205] Those skilled in the art will recognize that various modifications may be made to the above specific embodiments. The embodiments should not be limited to the specific limitations described. Other embodiments are also possible.

[0206] While compounds, conjugates, and pharmaceutical compositions have been illustrated and described in detail in the foregoing description, they should be considered illustrative rather than restrictive, and it should be understood that only certain embodiments have been shown and described and that all changes and modifications that fall within the spirit of the invention are desired to be protected.

[0207] The scope of the compounds, compositions and methods of the present invention is intended to be defined by the following claims. However, the disclosure may be implemented in a manner different from that specifically explained and illustrated without departing from its spirit or scope. It will be appreciated by those skilled in the art that various alternatives to the embodiments described herein may be adopted when implementing the claims without departing from the spirit and scope defined by the following claims.

[0208] The use of any section headings is intended to aid in reading the document and should not be construed as limiting. In addition, the information associated with a section heading may appear within or outside of that particular section.

[0209] All patents, patent application publications, journal articles, textbooks, and other publications mentioned in this document are indicative of the level of skill of those skilled in the art to which the present disclosure pertains. All such publications are incorporated herein by reference to the same extent as if each publication were specifically and individually indicated to be incorporated by reference. If there is an inconsistency in usage between this document and those documents incorporated by reference, the usage in the incorporated references shall be deemed supplementary to that of this document; for irreconcilable inconsistencies, the usage in this document shall prevail.

[0210] Various techniques and mechanisms of the present disclosure sometimes describe a connection or link between two components. Unless otherwise specified or the context clearly indicates a difference, words such as attached, linked, coupled, connected, bound, and similar terms and their inflectional morphemes are used interchangeably. These words and expressions do not necessarily mean direct connection, but include connections through intermediate components. It should be noted that a connection between two components does not necessarily mean a direct, unimpeded connection, as various other components may exist between the two components. Therefore, unless otherwise specified, connection does not necessarily mean a direct, unimpeded connection.

[0211] Some definitions

[0212] As used herein, the following terms and phrases shall have the following meanings. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0213] The term "about" can allow for a certain degree of variation in values ​​or ranges, for example, within 10%, within 5%, or within 1% of a stated value or a stated range limit. The term "substantially" can allow for a certain degree of variation in values ​​or ranges, for example, within 90%, within 95%, within 99%, within 99.5%, within 99.9%, within 99.99%, or within at least about 99.999% or more of a stated value or a stated range limit.

[0214] Unless the context clearly dictates otherwise, the terms "a", "an" or "the" are used to include one or more. Unless otherwise specified, the term "or" is used to refer to a non-exclusive "or". In addition, it should be understood that the expressions or terms used herein (not otherwise defined) are used for descriptive purposes only and not for limiting purposes.

[0215] Unless otherwise specified, the term "or" is used to mean a non-exclusive "or." Further, it is to be understood that the phraseology or terminology employed herein (not otherwise defined) is for the purpose of description only and not for the purpose of limitation.

[0216] The term "substituted" as used herein refers to the replacement of one or more hydrogen atoms contained in a functional group by one or more non-hydrogen atoms. The term "functional group" or "substituent" as used herein refers to a group that can be substituted or has been substituted on a molecule. Examples of substituents or functional groups include, but are not limited to, halogens (e.g., F, Cl, Br, and I); oxygen atoms in groups such as hydroxyl, alkoxy, aryloxy, aralkyloxy, oxo (carbonyl), carboxyl (including carboxylic acid, carboxylate, and carboxylate); sulfur atoms in groups such as thiol, alkyl and aryl sulfide, sulfoxide, sulfone, sulfonyl, and sulfonamide; nitrogen atoms in groups such as amine, azide, hydroxylamine, cyano, nitro, N-oxide, hydrazide, and enamine; and other heteroatoms in various other groups.

[0217] The term "alkyl" as used herein refers to a group having 1 to about 20 carbon atoms (C1-C 20 ), 1 to 12 carbon atoms (C1-C 12 ), substituted or unsubstituted straight and branched chain alkyl and cycloalkyl groups having 1 to 8 carbon atoms (C1-C8), or in certain embodiments 1 to 6 carbon atoms (C1-C6). Examples of straight chain alkyl groups include alkyl groups having 1 to 8 carbon atoms, such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl. Examples of branched chain alkyl groups include, but are not limited to, isopropyl, isobutyl, sec-butyl, tert-butyl, neopentyl, isopentyl, and 2,2-dimethylpropyl. The term "alkyl" as used herein includes n-alkyl, isoalkyl, and anteisoalkyl groups as well as other branched forms of alkyl groups. Representative substituted alkyl groups may be substituted one or more times by any of the groups listed herein, such as amino, hydroxyl, cyano, carboxyl, nitro, sulfenyl, alkoxy, and halogen groups.

[0218] The term "alkenyl" as used herein refers to substituted or unsubstituted straight and branched divalent alkenyl and cycloalkenyl groups having 2 to 20 carbon atoms (C2-C 20 ), 2 to 12 carbon atoms (C2-C 12 ), 2 to 8 carbon atoms (C2-C8), or in certain embodiments, 2 to 4 carbon atoms (C2-C4) and at least one carbon-carbon double bond. Examples of straight-chain alkenyl groups include alkenyl groups having 2 to 8 carbon atoms, such as -CH=CH-, -CH=CHCH2-, etc. Examples of branched alkenyl groups include, but are not limited to, -CH=C(CH3)-, etc.

[0219] An alkynyl group is a fragment containing an open point of attachment on a carbon atom that would form when a hydrogen atom bonded to a triple-bonded carbon is removed from an alkyne molecule. The term "hydroxyalkyl" as used herein refers to an alkyl group as defined herein substituted with at least one hydroxy (-OH) group.

[0220] The term "cycloalkyl" as used herein refers to substituted or unsubstituted cyclic alkyl groups, such as, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. In some embodiments, the cycloalkyl group may have 3 to about 8-12 ring members, while in other embodiments, the number of ring carbon atoms ranges from 3 to 4, 5, 6, or 7. In some embodiments, the cycloalkyl group may have 3 to 6 carbon atoms (C3-C6). Cycloalkyl groups also include polycyclic cycloalkyl groups, such as, but not limited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, and carbyl, as well as fused rings, such as, but not limited to, decahydronaphthyl, etc.

[0221] The term "acyl" as used herein refers to a group containing a carbonyl moiety, wherein the group is bonded through a carbonyl carbon atom. The carbonyl carbon atom is also bonded to another carbon atom, which may be part of a substituted or unsubstituted alkyl, aryl, aralkylcycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl, etc. In the special case where the carbonyl carbon atom is bonded to hydrogen, the group is a "formyl", i.e., an acyl group as defined herein. The acyl group may include 0 to about 12-40, 6-10, 1-5, or 2-5 additional carbon atoms bonded to the carbonyl group. Acryloyl is an example of an acyl group. Acyl groups may also include heteroatoms as defined herein. Nicotinoyl (pyridyl-3-carbonyl) is an example of an acyl group as defined herein. Other examples include acetyl, benzoyl, phenylacetyl, pyridylacetyl, cinnamoyl, and acryloyl, etc. When the group of carbon atoms bonded to the carbonyl carbon atom contains a halogen, the group is referred to as a "haloacyl". An example is trifluoroacetyl.

[0222] The term "aryl" as used herein refers to a substituted or unsubstituted cyclic aromatic hydrocarbon containing no heteroatoms in the ring. Thus, aryl includes, but is not limited to, phenyl, azulenyl, heptenyl, biphenyl, indoletenyl, fluorenyl, phenanthrenyl, triphenyl, pyrenyl, naphthenyl, In some embodiments, the aryl group contains about 6 to about 14 carbon atoms (C6-C 14 ) or 6 to 10 carbon atoms (C6-C 10 ). Aryl can be unsubstituted or substituted, as defined herein. Representative substituted aryl groups can be monosubstituted or multiply substituted, such as, but not limited to, 2-, 3-, 4-, 5- or 6-substituted phenyl or 2-8 substituted naphthyl, which can be substituted with carbon or non-carbon groups (such as those listed herein).

[0223] As used herein, the terms "aralkyl" and "arylalkyl" refer to an alkyl group as defined herein, wherein a hydrogen bond or carbon bond of the alkyl group is replaced by an aryl bond as defined herein. Representative aralkyl groups include benzyl and phenethyl groups, as well as fused (cycloalkylaryl)alkyl groups, such as 4-ethyl-indanyl. Aralkenyl groups are alkenyl groups as defined herein, wherein a hydrogen bond or carbon bond of the alkyl group is replaced by an aryl bond as defined herein.

[0224] The term "heterocyclyl" as used herein refers to substituted or unsubstituted aromatic and non-aromatic ring compounds containing 3 or more ring members, one or more of which are heteroatoms, such as but not limited to B, N, O and S. Therefore, the heterocyclyl can be a cycloheteroalkyl or heteroaryl, or if it is polycyclic, any combination thereof. In some embodiments, the heterocyclyl includes 3 to about 20 ring members, while other such groups have 3 to about 15 ring members. In some embodiments, the heterocyclyl includes a heterocyclyl containing 3 to 8 carbon atoms (C3-C8), 3 to 6 carbon atoms (C3-C6) or 6 to 8 carbon atoms (C6-C8).

[0225] Heteroaromatic ring is an embodiment of heterocyclic radical. The phrase "heterocyclic radical" includes fused ring species, including those ring species comprising fused aromatic and non-aromatic groups. Representative heterocyclic radicals include but are not limited to pyrrolidinyl, azetidinyl, piperidinyl, piperazinyl, morpholinyl, chromanyl, indolinone, isoindolinone, furanyl, pyrrolidinyl, pyridyl, pyrazinyl, pyrimidinyl, triazine, thienyl, tetrahydrofuranyl, pyrrolyl, oxazolyl, oxadiazolyl, imidazolyl, triazolyl, tetrazolyl, benzoxazoline, benzothiazoline and benzimidazoline.

[0226] The term "heterocycloalkyl" as used herein refers to an alkyl group in which a hydrogen bond or a carbon bond in an alkyl group as defined herein is replaced by a heterocyclyl bond as defined herein. Representative heterocycloalkyl groups include, but are not limited to, furan-2-ylmethyl, furan-3-ylmethyl, pyridin-3-ylmethyl, tetrahydrofuran-2-ylmethyl and indol-2-ylpropyl.

[0227] The term "heteroaralkyl" as used herein refers to an alkyl group as defined herein wherein a hydrogen or carbon bond of the alkyl group is replaced by a bond to a heteroaryl group as defined herein.

[0228] The term "alkoxy" as used herein refers to an oxygen atom connected to an alkyl group (including a cycloalkyl group), as defined herein. Examples of straight chain alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, etc. Examples of branched chain alkoxy groups include, but are not limited to, isopropoxy, sec-butoxy, tert-butoxy, isopentyloxy, isohexyloxy, etc. Examples of cyclic alkoxy groups include, but are not limited to, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, etc. Alkoxy groups may also include double or triple bonds and may also include heteroatoms. For example, allyloxy is an alkoxy group defined herein. Methoxyethoxy is also an alkoxy group defined herein, and in the case where two adjacent atoms of the structure are substituted by it, methylenedioxy is also an alkoxy group defined herein.

[0229] The term "amine" as used herein refers to primary, secondary and tertiary amines having, for example, the formula N(group)3, wherein each group can independently be H or non-H, such as alkyl, aryl, etc. Amines include, but are not limited to, R-NH2, such as alkylamines, arylamines, alkylarylamines; R2NH, wherein each R is independently selected, such as dialkylamines, diarylamines, arylalkylamines, heterocyclic amines, etc.; and R3N, wherein each R is independently selected, such as trialkylamines, dialkylarylamines, alkyldiarylamines, triarylamines, etc. The term "amine" as used herein also includes ammonium ions.

[0230] The term "amino" as used herein refers to -NH2, -NHR, -NR2, -NR3 + Substituents of the form wherein each R is independently selected, and the protonated form of each, but -NR3 + Except for, it cannot be protonated. Therefore, any compound substituted by amino group can be regarded as amine. "Amino group" referred to herein can be primary, secondary, tertiary or quaternary amino group. "Alkylamino" includes monoalkylamino, dialkylamino and trialkylamino.

[0231]

[0046] The terms "halo," "halogen," or "halide" group, as used herein, alone or as part of another substituent, refer, unless otherwise stated, to a fluorine, chlorine, bromine, or iodine atom.

[0232] The term "haloalkyl" as used herein includes monohaloalkyl, polyhaloalkyl (wherein all halogen atoms may be the same or different) and perhaloalkyl (wherein all hydrogen atoms are substituted by halogen atoms (e.g., fluorine). Examples of haloalkyl include trifluoromethyl, 1,1-dichloroethyl, 1,2-dichloroethyl, 1,3-dibromo-3,3-difluoropropyl, perfluorobutyl, -CF(CH3)2, and the like.

[0233] As used herein, the term "optionally substituted" or "optionally substituted" means that the group in question is unsubstituted or substituted with one or more specified substituents. When the group in question is substituted with multiple substituents, the substituents may be the same or different. When the terms "independently," "independently are," and "independently selected from" are used, it means that the group in question may be the same or different. Certain terms defined herein may appear multiple times in a structure, and when they appear, each term should be defined independently of the other terms.

[0234] The terms "patient" and "subject" are used interchangeably and include human patients, laboratory animals (e.g., rodents (e.g., mice, rats, or hamsters), rabbits, monkeys, chimpanzees), livestock (e.g., dogs, cats, or rabbits), agricultural animals (e.g., cattle, horses, pigs, sheep, goats, livestock animals raised for food production), or captive wild animals (e.g., bears, pandas, lions, tigers, leopards, elephants, zebras, giraffes, gorillas, dolphins, or whales). The patient to be treated is preferably a mammal, particularly a human.

[0235] The terms "protein," "polypeptide," and "peptide" refer to compounds comprising amino acids linked by peptide bonds and are used interchangeably. Example

[0236] The following examples serve to illustrate the present disclosure. The examples are not intended to limit the scope of the claims in any way. Example 1 Synthesis of CDK inhibitor compounds

[0237] Unless otherwise stated, all reagents and solvents were purchased from commercial sources and used as received. Unless otherwise stated, all reactions were performed in sealed vials with screw caps. 1 H and 13 C NMR spectra were obtained in (CD3)2SO as solvent using a 500 MHz or 800 MHz spectrometer with Me4Si as internal standard. Chemical shifts are reported in parts per million (δ) and are calibrated using residual undeuterated solvent as internal reference.

[0238] 1 H NMR spectral data are reported as follows: Chemical shift (δ ppm) (multiplicity, coupling constant (Hz), integration). Multiplicity is reported as follows: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, or a combination thereof.

[0239] General Method A

[0240] In a 250 mL round bottom flask equipped with a stirring bar, 5-aminoindazole (10 mmol) and the appropriate aldehyde (4-formylbenzoic acid or 4-formylbenzonitrile) (10 mmol) were dissolved in 150 mL of 200 degree ethanol. The mixture was refluxed at 120°C for 2 hours, then the appropriate ketone (20 mmol) and a catalytic amount of hydrochloric acid were added. It was stirred at reflux for another 12 hours. The reaction was concentrated and purified under 40% ethyl acetate and 60% hexane.

[0241] General Method B

[0242] In a 50 mL round bottom flask equipped with a stirring bar, 4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzonitrile (850 mg, 2.53 mmol) and NaOH (532 mg, 13 mmol) were dissolved in ethanol (4 mL) and water (8 mL). This was added to the round bottom flask and refluxed for 12 hours. After about an hour, the mixture was completely dissolved. After the reaction was complete, the solvent was evaporated under reduced pressure, diluted with water, and acidified to pH 5 with HCl. Then stirred for 30 minutes to ensure that the pH was stable. The product was then collected by vacuum filtration.

[0243] General Method C

[0244] In a 20 mL vial equipped with a stirring bar, the acid (0.5 mmol) was dissolved in N,N-dimethylformamide (DMF) (2 mL) and N,N-diisopropylethylamine (DIPEA) (12.5 equiv.) Bis(2-oxo-3-oxazolidinyl)phosphonyl chloride (2.7 equiv.) was added to the solution and stirred at 50°C for 30 minutes, then the corresponding amine (0.5 mmol) was added and stirred at 50°C overnight. DMF was concentrated under reduced pressure and then purified with 15% methanol and 85% ethyl acetate.

[0245] General Method D

[0246] In a 20 mL vial equipped with a stirring bar, the acid (0.5 mmol) and 4-dimethylaminopyridine (12 mg), N-methylmorpholine (1 mmol) and (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine 3-oxide hexafluorophosphate, azabenzotriazole tetramethyluronium hexafluorophosphate) were dissolved in DMF (2 mL) and stirred at room temperature. After 30 minutes, 1-methylpiperazine (2 mmol) was added and stirred at room temperature overnight. DMF was concentrated under reduced pressure and the reaction was purified with 25% methanol and 75% ethyl acetate.

[0247] General Method E

[0248] In a microwave vial, the acid (0.5 mmol), fluoro-N,N,N',N'-bis(tetramethylene)formamidine hexafluorophosphate (0.6 mmol), DIPEA (1.75 mmol) and dichloromethane (2 mL) were stirred at room temperature for 30 minutes. Then the amine (0.38 mmol) was added and stirred in a sand bath at 80°C overnight. The reaction mixture was concentrated and purified with 10% methanol and 90% ethyl acetate.

[0249] General Method F

[0250] The acid intermediate (0.5 mmol) and hexafluorophosphate azabenzotriazole tetramethyl urea) were sealed in a 20 mL vial equipped with a stirring bar with a septum, evacuated and a N2 balloon was added. Anhydrous DMF (5 mL) and DIPEA (1.2 mL) were added to this reaction vial. The reaction was heated at 100 ° C for 10 minutes, then the amine (1 mmol) was added to the reaction and stirred overnight at 100 ° C. The reaction mixture was concentrated and purified by silica gel chromatography.

[0251] General Method G (Synthesis of Compound HSH3165)

[0252] In a 25 mL round bottom flask equipped with a stir bar, N-(2-aminoethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide was dissolved in trifluoroacetic acid (2 mL) and dichloromethane (1 mL) and stirred overnight at room temperature. The reaction was concentrated to give the pure product compound HSH3165.

[0253] General Methods

[0254] To a solution of compound HSH3107 (0.5 mmol) in anhydrous tetrahydrofuran (THF) (720 μL) at 0°C, lithium aluminum hydride (1.5 mmol, 1 M THF solution) was added. The mixture was heated to 50°C for 2 hours. It was cooled to 0°C, treated with water, and then stirred at room temperature for 3 minutes.

[0255] The solid was filtered through a celite bed, washed with THF, dried over sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography with [hexane:ethyl acetate (30:70 to 0:100)].

[0256] Intermediates:

[0257] Synthesis of 4-(8,9,10,11-tetrahydro-3H-8,11-methylenepyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid:

[0258] In a 250 mL round bottom flask equipped with a stirring bar, 5-aminoindazole (2 mmol) and 4-formylbenzoic acid (2 mmol) were dissolved in 10 mL of 200 proof ethanol. The mixture was refluxed at 120°C for 2 hours, then bicyclo[2.2.1]heptan-2-one (4 mmol) and a catalytic amount of hydrochloric acid were added. It was stirred at reflux for another 12 hours. The precipitate was washed with ethanol and used in the next step without further purification.

[0259] 1H NMR(800MHz, (CD3)2SO) δ8.75(s,1H),8.12(dd,J=20.4,7.8Hz,2H),7.97(d,J=7.9Hz,2H),7.86(q,J=9.3Hz,2H),2.95(s,1H),2.18 (dd,J=27.2,13.6Hz,2H),1.83(d,J=8.9Hz,1H),1.67(d,J=9.0Hz,1H),1.60(s,1H),1.35(d,J=10.6Hz,1H),1.22(t,J=9.9Hz,1H). 13 C NMR (201 MHz, (CD3)2SO) δ 167.64, 152.79, 147.96, 145.12, 144.02, 139.74, 130.72, 130.16, 129.86, 129.12, 128.07, 127.20, 121.04, 116.74, 115.88, 49.88, 43.24, 42.39, 26.88, 25.31. HRMS (ESI) m / z calculated value C 22 H 18 N3O2[M+H] + 356.1393, measured value 356.1392.

[0260] 4-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzonitrile

[0261] Synthesized using Method A: off-white solid (1.5 g, 10 mmol, 46.3%). 1 H NMR(500MHz,(CD3)2SO)δ8.55(s,1H),7.94–7.90(m,2H),7.88–7.80(m,2H),7.78–7. 74(m,2H),3.33(s,2H),2.75(t,J=6.1Hz,2H),2.04–1.97(m,2H),1.79–1.72(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 155.09, 146.02, 143.76, 142.80, 138.90, 136.43, 132.38, 130.59, 129.59, 129.18, 122.28, 119.23, 116.43, 114.66, 111.07, 29.64, 28.64, 22.51, 22.43. HRMS (ESI) m / z calculated value C 21 H 17 N4[M+H]+ 325.1444, measured value 325.1448.

[0262] 4-(9-Methyl-3H-pyrazolo[4,3-f]quinolin-7-yl)benzonitrile

[0263] Synthesized using Method A: off-white solid (0.840 g, 10 mmol, 30%). 1 H NMR (500MHz, (CD3)2SO) δ8.61–8.53(m,1H),8.44–8.36(m,2H),8.13(s,1H),7.97–7.88(m,4H),2.93(s,3H). 13 C NMR (126 MHz, (CD3)2SO) δ 151.30, 146.36, 144.74, 143.57, 138.94, 136.03, 133.06, 129.73, 127.94, 122.68, 120.92, 119.24, 116.71, 115.79, 111.79, 22.71. HRMS (ESI) m / z calculated value C 18 H 13 N4[M+H] + 285.1135, measured value 285.1136.

[0264] 4-(9-Cyclopropyl-3H-pyrazolo[4,3-f]quinolin-7-yl)benzonitrile

[0265] Synthesized using Method A: off-white solid (0.485 g, 10 mmol, 14.7%). 1 H NMR (500MHz, (CD3)2SO) δ 8.80 (s, 1H), 8.40 (d, J = 8.3Hz, 2H), 8.00–7.86 (m, 5H), 2.62 (m, 1H), 1.39–1.24 (m, 2H), 1.00 (q, J = 5.3, 4.8Hz, 2H). 13 C NMR (126 MHz, (CD3)2SO) δ 151.73, 149.26, 146.24, 143.69, 138.93, 136.66, 133.04, 129.80, 128.16, 123.61, 119.30, 117.24, 116.48, 116.06, 111.73, 40.40, 40.24, 40.06, 39.90, 39.73, 39.57, 39.40, 16.56, 7.80. HRMS (ESI) m / z calculated value C20 H 15 N4[M+H] + 311.1291, measured value 311.1293.

[0266] 4-(1-Methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzonitrile

[0267] Synthesized using Method A: off-white solid (402 mg, 2 mmol, 59.4%). 1 H NMR (500MHz, (CD3)2SO) δ7.88(d,J=8.1Hz,2H),7.71(d,J=7.9Hz,4H),3.37(s,2H),2.83(s,3H),2.74–2.65(m,2H),1.75(m,4H). 13 C NMR (126 MHz, (CD3)2SO) δ 154.5, 146.0, 142.7, 133.1, 132.4, 130.7, 130.5, 129.8, 128.2, 127.9, 124.2, 124.1, 119.2, 114.9, 111.0, 49.1, 31.5, 27.8, 22.4, 22.1. HRMS (ESI) m / z calculated value C 18 H 14 N3O2[M+H] + 339.1604, measured value 339.1605.

[0268] 3-Fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzonitrile

[0269] Synthesized using Method A: off-white solid (1.42 g, 10 mmol, 41.5%). 1 H NMR(500MHz,(CD3)2SO)δ8.56(s,1H),7.94(dd,J=9.5,1.7Hz,1H),7.88(d,J=9.1Hz,1H),7.85–7.78(m,2H) ,7.68(t,J=7.5Hz,1H),3.36(t,J=6.6Hz,2H),2.60(t,J=6.2Hz,2H),2.06–1.92(m,2H),1.85–1.71(m,2H). 13 C NMR(126MHz,(CD3)2SO)δ160.31(d, 1J=248Hz),150.62,143.80,142.73,138.94,136.47,134.29(d, 2 J=16Hz),133.27,129.92,129.44,129.26,122.57,120.29( 2 J=26Hz),118.00,116.40,114.82,113.18(d, 3 J = 10 Hz), 29.47, 27.26, 22.51, 22.09. HRMS (ESI) m / z calculated value C 21 H 16 N4[M+H] + 343.1353, measured value 343.1352.

[0270] 2-Fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzonitrile

[0271] Synthesized using Method A: off-white solid (0.220 g, 5 mmol, 12.9%). 1 H NMR(500MHz,(CD3)2SO)δ8.59(s,1H),8.00(t,J=7.4Hz,1H),7.91–7.77(m,2H),7.73(d,J=10.4Hz,1 H),7.63(d,J=8.0Hz,1H),3.35(s,2H),2.80(t,J=6.2Hz,2H),2.09–1.97(m,2H),1.82–1.73(m,2H). 13 CNMR(126MHz,(CD3)2SO)δ163.68(d, 1 J=255Hz),153.84,149.06,149.00,143.76,143.02,136.44,133.99,129 .56,129.22,126.85,122.55,117.69,117.53,114.91,114.46,99.94(d, 2 J = 15 Hz), 29.66, 28.46, 22.48, 22.40. HRMS (ESI) m / z calculated value C 21 H 16 N4[M+H] + 343.1353, measured value 343.1352.

[0272] 3-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzonitrile

[0273] Synthesized using Method A: off-white solid (0.665 g, 10 mmol, 20.2%). 1 H NMR(500MHz,(CD3)2SO)δ8.54(s,1H),7.98(s,1H),7.91–7.82(m,3H),7.80(d,J=9.1 Hz,1H),7.67(t,J=7.8Hz,1H),2.74(t,J=6.2Hz,2H),2.01–1.91(m,2H),1.73(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 154.67, 143.86, 142.83, 142.51, 134.35, 132.95, 131.92, 129.75, 129.55, 129.22, 127.67, 122.31, 119.13, 116.16, 115.89, 115.09, 111.75, 29.62, 28.62, 22.47, 22.42. HRMS (ESI) m / z calculated value C 21 H 17 N4[M+H] + 325.1448, measured value 325.1449.

[0274] 3-Methoxy-4-(8,9,10,11-tetrahydro-3H-naphtho[1,2-e]indazol-7-yl)benzonitrile

[0275] Synthesized using Method A: off-white solid (0.255 g, 10 mmol, 7.21%). 1 H NMR(500MHz, (CD3)2SO) δ8.55(s,1H),7.85(d,J=9.0Hz,1H),7.80(s,1H),7.59(s,1H),7.51(dd,J=7.6,1.4Hz,1H),7.41(d,J=7.7 Hz,1H),3.78(s,3H),3.34(d,J=16.9Hz,2H),2.59(s,1H),2.42(d,J=16.5Hz,1H),1.98(dd,J=13.7,6.5Hz,2H),1.84–1.68(m,2H). 13C NMR (126 MHz, (CD3)2SO) δ 157.37, 153.80, 143.74, 143.68, 141.82, 136.31, 135.78, 131.79, 130.19, 129.49, 125.20, 119.17, 116.47, 115.27, 114.35, 114.31, 112.47, 56.56, 29.41, 26.94, 22.62, 22.13. HRMS (ESI) m / z calculated value C 22 H 19 N4O[M+H] + 355.1553, measured value 355.1553.

[0276] 3-Methoxy-4-(8,9,10,11-tetrahydro-3H-naphtho[1,2-e]indazol-7-yl)benzonitrile

[0277] Synthesized using Method A: off-white solid (0.719 g, 6.14 mmol, 33.1%). 1 H NMR(500MHz, (CD3)2SO) δ8.51(s,1H),7.84(q,J=8.9Hz,2H),7.77(d,J=7.8Hz,1H),7.38(s,1H),7.23(d,J= 7.9Hz,1H),3.96(s,3H),3.28(t,J=6.7Hz,2H),2.74(t,J=6.3Hz,2H),2.05–1.90(m,2H),1.81–1.67(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 161.15, 155.30, 148.08, 143.56, 142.68, 138.88, 136.39, 133.64, 129.57, 129.14, 122.31, 116.82, 116.44, 114.57, 113.51, 100.26, 56.99, 29.59, 28.50, 22.47, 22.41. HRMS (ESI) m / z calculated value C 22 H 19 N4O[M+H] + 355.1553, measured value 355.1552.

[0278] 4-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid

[0279] Synthesized using method B: off-white solid (0.812 g, 2 mmol 93.6%). 1 H NMR(500MHz,(CD3)2SO)δ8.75(s,1H),8.26(d,J=9.2Hz,1H),8.19(d,J=9.2Hz,1H),8.15–8.11(m,2H),7.85 –7.80(m,2H),3.46(t,J=6.5Hz,2H),2.76(t,J=6.1Hz,2H),2.03(dd,J=6.0,2.9Hz,2H),1.83–1.76(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 167.17, 151.31, 151.03, 140.41, 138.09, 137.05, 134.78, 132.69, 131.32, 130.42, 129.69, 123.45, 122.02, 120.11, 115.06, 30.67, 28.05, 21.85, 21.72. HRMS (ESI) m / z calculated value C 21 H 18 N3O2[M+H] + 344.1393, measured value 344.1392.

[0280] 4-(9-Methyl-3H-pyrazolo[4,3-f]quinolin-7-yl)benzoic acid

[0281] Synthesized using Method B: off-white solid (0.462 g, 3 mmol, 50.8%). 1 H NMR (500MHz, (CD3)2SO) δ 8.64 (d, J = 6.1 Hz, 1H), 8.33 (m, 2H), 8.22 (d, J = 6.0 Hz, 1H), 8.15 (dd, J = 9.2, 6.3 Hz, 1H), 8.09 (m, 3H), 3.01 (d, J = 6.2 Hz, 3H). 13 C NMR (126 MHz, (CD3)2SO) δ 167.34, 151.05, 148.04, 143.57, 140.70, 139.95, 134.55, 132.31, 130.16, 128.18, 126.62, 122.86, 121.93, 118.55, 116.28, 23.05. HRMS (ESI) m / z calculated value C 18 H 14 N3O2[M+H] + 304.1081, measured value 304.1081.

[0282] 4-(9-Cyclopropyl-3H-pyrazolo[4,3-f]quinolin-7-yl)benzoic acid

[0283] Synthesized using Method B: off-white solid (0.464 g, 2.2 mmol, 64.1%). 1 H NMR(500MHz, (CD3)2SO)δ8.90(s,1H),8.36(d,J=8.2Hz,2H),8.21(d,J=9.2Hz,1H),8.13–8.07(m, 3H), 7.98 (s, 1H), 2.74 (d, J = 5.4Hz, 1H), 1.39 (dd, J = 8.5, 2.4Hz, 2H), 1.13 (dd, J = 5.5, 2.2Hz, 2H). 13 CNMR (126 MHz, (CD3)2SO) δ 167.35, 152.74, 151.36, 143.33, 140.68, 140.08, 135.01, 132.36, 130.10, 128.48, 126.47, 123.83, 118.75, 118.02, 115.98, 17.03, 8.58. HRMS (ESI) m / z calculated value C 20 H 16 N3O2[M+H] + 330.1237, measured value 330.1238.

[0284] 4-(3,8,9,10,11,12-Hexahydrocyclohepta[c]pyrazolo[4,3-f]quinolin-7-yl)benzoic acid

[0285] Synthesized using Method A: off-white solid (0.522 g, 4 mmol, 36.5%). 1 H NMR(500MHz, (CD3)2SO) δ8.65(s,1H),8.04(d,J=7.8Hz,2H),7.84(d,J=9.2Hz,1H),7.77(d,J=9.1Hz,1H),7.5 8(d,J=7.8Hz,2H), 3.50(d,J=7.2Hz,2H), 2.93(d,J=8.4Hz,2H), 1.84(d,J=9.5Hz,4H), 1.59(d,J=7.8Hz,2H). 13C NMR (126 MHz, (CD3)2SO) δ 167.57, 155.09, 148.75, 146.07, 144.73, 140.15, 135.20, 134.37, 134.16, 130.47, 129.81, 129.76, 129.46, 121.59, 116.02, 31.16, 31.10, 30.12, 27.28, 24.99. HRMS (ESI) m / z calculated value C 22 H 20 N3O2[M+H] + 358.1550, measured value 358.1550.

[0286] 4-(3,8,10,11-Tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzoic acid

[0287] Synthesized using Method A: off-white solid (0.220 g, 0.76 mmol, 21.2%). 1 H NMR (500MHz, (CD3)2SO) δ 8.68 (s, 1H), 8.13–7.99 (m, 4H), 7.76 (d, J = 8.4Hz, 2H), 4.77 (s, 2H), 4.17 (t, J = 5.8Hz, 2H), 3.49 (d, J = 5.9Hz, 2H). 13 C NMR (126 MHz, (CD3)2SO) δ 167.32, 150.67, 143.75, 141.15, 140.39, 140.21, 134.52, 132.11, 129.85, 129.79, 128.41, 125.87, 122.36, 118.50, 115.52, 66.34, 64.17, 29.11. HRMS (ESI) m / z calculated value C 20 H 16 N3O3[M+H] + 346.1186, measured value 346.1187.

[0288] 4-(9,9-dioxido-3,8,10,11-tetrahydropyrazolo[4,3-f]thiopyrano[3,4-c]quinolin-7-yl)benzonitrile

[0289] Synthesized using Method A: off-white solid (1.68 g, 4.75 mmol, 89.7%). 1H NMR(800MHz, (CD3)2SO) δ8.74(s,1H),8.12(d,J=8.1Hz,2H),8.05(d,J=9.0Hz,1H),7.93(d,J =9.0Hz, 1H), 7.73 (d, J = 8.1Hz, 2H), 4.52 (s, 2H), 4.04 (d, J = 6.7Hz, 2H), 3.72 (d, J = 6.5Hz, 2H). 13 C NMR (201 MHz, (CD3)2SO) δ 167.37, 154.71, 143.89, 142.68, 140.28, 140.13, 135.10, 131.40, 129.94, 129.79, 128.36, 122.56, 121.45, 117.96, 115.45, 51.54, 45.94, 30.62. HRMS (ESI) m / z calculated value C 20 H 16 N3O4S[M+H] + 394.0856, measured value 394.0856.

[0290] 4-(3,8,9,10-Tetrahydrocyclopenta[c]pyrazolo[4,3-f]quinolin-7-yl)benzoic acid

[0291] Synthesized using Method A: off-white solid (518 mg, 4 mmol, 39.3%). 1 H NMR (500MHz, (CD3)2SO) δ 8.48 (s, 1H), 8.12–8.08 (m, 2H), 8.01–7.93 (m, 4H), 3.43 (t, J = 7.6Hz, 2H), 3.21 (t, J = 7.5Hz, 2H), 2.29–2.18 (m, 2H). 13 C NMR (126 MHz, (CD3)2SO) δ 167.39, 153.03, 149.45, 142.46, 141.75, 139.61, 137.07, 133.84, 131.71, 129.75, 129.51, 126.17, 119.93, 117.67, 116.07, 34.02, 32.73, 24.73. HRMS (ESI) m / z calculated value C 20 H 16 N3O2[M+H] + 330.1237, measured value 330.1236.

[0292] 4-(8,9-Dihydro-3H-cyclobuta[c]pyrazolo[4,3-f]quinolin-7-yl)benzoic acid

[0293] Synthesized using Method A: off-white solid (0.030 g, 3 mmol, 3.2%). 1 H NMR(500MHz,(CD3)2SO)δ8.37(d,J=0.9Hz,1H),8.29–8.25(m,2H),8.11–8.0 6(m,2H),7.91–7.83(m,2H),3.77(dd,J=5.3,2.6Hz,2H),3.70–3.64(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 167.53, 150.32, 146.59, 145.26, 141.88, 137.99, 131.57, 130.26, 130.16, 127.32, 117.98, 115.63, 115.59, 115.48, 115.01, 32.33, 30.41. HRMS (ESI) m / z calculated value C 19 H 14 N3O2[M+H] + 316.1080, measured value 316.1080.

[0294] 4-(5-Fluoro-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid

[0295] Synthesized using Method A: off-white solid (0.145 g, 1.38 mmol, 29.1%). 1 H NMR (800MHz, (CD3)2SO) δ8.62 (s, 1H), 8.07 (d, J = 7.7Hz, 2H), 7.71 (dd, J = 23.9, 8.8H z,3H),3.34–3.30(m,2H),2.82–2.73(m,2H),2.06–1.96(m,2H),1.82–1.70(m,2H). 13 C NMR(201MHz,(CD3)2SO)δ167.51,157.66( 1 J=250Hz),155.81,144.92,143.35,134.62,134.55,134.49,130.72,130.68,129.72,129.43,123.01,112.66,99.67( 2 J = 24 Hz), 29.63, 28.69, 22.27, 22.15. HRMS (ESI) m / z calculated value C21 H 17 FN3O2[M+H] + 362.1299, measured value 362.1300.

[0296] 3-Fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid

[0297] Synthesized using Method B: off-white solid (1.58 g, 4.44 mmol, 98.5%). 1 H NMR(500MHz, (CD3)2SO) δ8.64(s,1H),8.06(d,J=22.2Hz,3H),7.72(d,J=11.2Hz,1H),7.63(d,J=8 .1Hz,1H),3.31(t,J=6.6Hz,2H),2.76(t,J=6.2Hz,2H),2.04–1.91(m,2H),1.76(q,J=6.0Hz,2H). 13 C NMR(126MHz,(CD3)2SO)δ167.55,165.09,162.00,159.95,157.23,150.10,132 .37,131.56,130.83,126.27,123.27,123.14,121.70,120.78,120.70,118.99( 2 J = 22 Hz), 115.01, 30.42, 28.01, 21.88, 21.75. HRMS (ESI) m / z calculated value C 21 H 17 FN3O2[M+H] + 362.1299, measured value 362.1300.

[0298] 5-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)thiophene-2-carboxylic acid

[0299] Synthesized using Method A: off-white solid (0.545 g, 4 mmol, 39.0%). 1H NMR(500MHz, (CD3)2SO) δ8.58(s,1H),7.94(dd,J=9.1,0.9Hz,1H),7.87(d,J=9.2Hz,1H),7.76(d,J=3.8Hz,1 H),7.62(d,J=4.0Hz,1H),3.30(t,J=6.5Hz,2H),3.02(t,J=6.2Hz,2H),2.01–1.92(m,2H),1.86–1.78(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 163.22, 147.97, 146.86, 145.85, 141.63, 140.23, 136.54, 134.86, 133.59, 129.90, 129.82, 127.01, 122.73, 117.50, 115.69, 30.18, 28.68, 22.22, 22.03. HRMS (ESI) m / z calculated value C 19 H 16 N3O2S[M+H] + 350.0958, measured value 350.0960.

[0300] 3-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid

[0301] Synthesized using Method B: off-white solid (0.453 g, 2 mmol, 65.9%). 1 H NMR(500MHz,(CD3)2SO)δ8.72(s,1H),8.20(t,J=1.8Hz,1H),8.13–8.02(m,3H),7.91(m,1H),7.7 0(t,J=7.7Hz,1H),3.45(t,J=6.5Hz,2H),2.77(t,J=6.2Hz,2H),2.09–1.99(m,2H),1.79(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 167.28, 152.64, 140.35, 139.35, 136.86, 134.90, 134.08, 131.63, 130.72, 130.61, 130.47, 129.20, 128.96, 124.55, 123.02, 118.61, 115.51, 30.36, 28.37, 22.09, 22.01. HRMS (ESI) m / z calculated value C 21 H 18 N3O2[M+H]+ 344.1393, measured value 344.1394.

[0302] 3-Methoxy-4-(8,9,10,11-tetrahydro-3H-naphtho[1,2-e]indazol-7-yl)benzoic acid

[0303] Synthesized using Method B: off-white solid (0.193 g, 0.75 mmol, 68.2%). 1 H NMR(500MHz,(CD3)2SO)δ8.79(s,1H),8.23(d,J=9.2Hz,1H),8.14(d,J=9.2Hz,1H),7.78–7.72(m,2H),7 .61(d,J=7.7Hz,1H),3.50(d,J=6.8Hz,2H),2.62(t,J=6.3Hz,2H),2.07–1.98(m,2H),1.85–1.76(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 167.10, 157.26, 151.83, 148.67, 140.49, 136.49, 134.97, 134.81, 132.47, 131.50, 126.57, 123.61, 122.16, 121.28, 120.55, 115.09, 112.62, 56.61, 30.58, 26.84, 21.83, 21.38. HRMS (ESI) m / z calculated value C 22 H 21 N3O3[M+H] + 374.1499, measured value 374.1497.

[0304] 2-Methoxy-4-(8,9,10,11-tetrahydro-3H-naphtho[1,2-e]indazol-7-yl)benzoic acid

[0305] Synthesized using Method B: off-white solid (814 mg, 2.31 mmol, 94.3%). 1H NMR(800MHz, (CD3)2SO) δ8.52(d,J=17.7Hz,1H),7.86(d,J=9.0Hz,1H),7.84–7.80(m,1H),7.78(d,J=7.7Hz,1H),7.39(d,J =1.5Hz,1H),7.23(dd,J=7.7,1.6Hz,1H),3.96(s,3H),3.23(m,2H),2.72(t,J=6.2Hz,2H),1.95(m,2H),1.77–1.68(m,2H). 13 C NMR (201 MHz, (CD3)2SO) δ 161.02, 155.15, 147.91, 143.41, 142.64, 138.75, 136.30, 133.59, 129.43, 129.03, 122.32, 122.20, 116.82, 116.25, 114.53, 113.32, 100.02, 56.86, 29.47, 28.41, 22.32, 22.26. HRMS (ESI) m / z calculated value C 22 H 21 N3O3[M+H] + 374.1499, measured value 374.1497.

[0306] tert-Butyl 4-(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoyl)piperazine-1-carboxylate

[0307] Synthesized using Method C: off-white solid (100 mg, 0.5 mmol, 39.1%). 1 H NMR(800MHz, (CD3)2SO) δ8.59(s,1H),7.93–7.78(m,2H),7.64(dd,J=8.7,2.9Hz,2H),7.52(dd,J=8.1,2.6H z,2H),3.70–3.54(m,2H),3.54–3.38(m,5H),2.80(t,J=6.4Hz,2H),2.01(m,2H),1.76(m,2H),1.42(s,9H). 13C NMR (201 MHz, (CD3)2SO) δ 169.48, 161.41, 156.00, 154.23, 143.69, 142.45, 142.42, 138.47, 136.29, 135.38, 129.51, 129.19, 127.09, 121.98, 116.21, 114.53, 79.59, 47.31, 44.95, 43.73, 41.90, 29.58, 28.81, 28.43, 22.50, 22.41. HRMS (ESI) m / z calculated value C 30 H 34 N5O3[M+H] + 512.2656, measured value 512.2655.

[0308] N-(2-(Dimethylamino)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH2177)

[0309] Synthesized using method C: off-white solid (30 mg, 0.5 mmol, 14.65%). 1 H NMR(500MHz,(CD3)2SO)δ8.58(t,J=5.8Hz,2H),7.96–7.90(m,2H),7.88–7.79(m,2H),7.67–7.62(m,2H),3.40–3.27(m, 7H),2.77(t,J=6.1Hz,2H),2.39–2.25(m,7H),2.13(s,3H),2.00(m,4.4,2.8Hz,2H),1.74(m,2H),1.68(q,J=7.0Hz,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.33, 156.14, 143.83, 142.54, 134.34, 129.64, 129.44, 129.27, 127.29, 122.07, 56.27, 55.23, 53.17, 46.20, 38.50, 29.67, 28.88, 26.69, 22.58, 22.49. HRMS (ESI) m / z calculated value C 25 H 28 N5O[M+H] + 414.2288, measured value 414.2285.

[0310] N-(3-(4-methylpiperazin-1-yl)propyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH2195)

[0311] Synthesized using Method C: off-white solid (10 mg, 0.5 mmol, 4.1%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.58 (t, J = 5.8 Hz, 2H), 7.96–7.90 (m, 2H), 7.88–7.79 (m, 2H), 7.67–7.62 (m, 2H), 3.31–3.28 (m, 3H), 2.77 (t, J = 6.1 Hz, 2H), 2.42–2.23 (m, 8H), 2.13 (s, 3H), 2.00 (m, 2H), 1.74 (m, 2H), 1.68 (q, J = 7.0 Hz, 2H). HRMS (ESI) m / z calculated value C 29 H 35 N6O[M+H] + 483.2867, measured value 483.2868.

[0312] N-(Cyclopropylmethyl)-4-(8,9,10,11-tetrahydro-3H-naphtho[1,2-e]indazol-7-yl)benzamide (Compound HSH3001)

[0313] Synthesized using Method C: off-white solid (20 mg, 0.5 mmol, 10.1%). 1 H NMR(500MHz, (CD3)2SO) δ8.64(t,J=5.7Hz,1H),8.57(s,1H),8.04–7.91(m,2H),7.84(q,J=12.4,10.6Hz,2H),7.69–7.57(m,2H), 3.18(t,J=6.2Hz,2H),2.76(t,J=6.3Hz,2H),2.03–1.95(m,2H),1.73(m,2H),1.06(m,1H),0.51–0.36(m,2H),0.31–0.17(m,2H). 13C NMR (126 MHz, (CD3)2SO) δ 166.33, 156.12, 143.73, 143.54, 142.68, 138.73, 136.42, 134.33, 129.53, 129.44, 129.34, 127.37, 122.03, 116.42, 114.57, 44.02, 29.68, 28.85, 22.55, 22.47, 11.53, 4.20, 3.80. HRMS (ESI) m / z calculated value C 25 H 25 N4O[M+H] + 397.2023, measured value 397.2021.

[0314] N-(3-(Piperidin-1-yl)propyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3004)

[0315] Synthesized using Method C: off-white solid (23 mg, 0.5 mmol, 9.8%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.59 (s, 2H), 7.93 (s, 2H), 7.83 (d, J = 19.7 Hz, 2H), 7.63 (s, 2H), 2.76 (s, 2H), 2.32 (s, 6H), 2.00 (s, 2H), 1.72 (d, J = 19.4 Hz, 4H), 1.48 (s, 4H), 1.36 (s, 2H). HRMS (ESI) m / z calculated value C 29 H 34 N5O[M+H] + 468.2758, measured value 468.2759.

[0316] N-(2-(Dimethylamino)ethyl)-4-(8,9,10,11-tetrahydro-3H-8,11-methylenepyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3014)

[0317] Synthesized using Method F: off-white solid (10 mg, 0.5 mmol, 4.7%). 1H NMR(500MHz,(CD3)2SO)δ8.69(m,1H),8.47(d,J=4.4Hz,1H),8.28(d,J=8.3Hz,1H), 8.02(d,J=7.9Hz,2H),7.91(d,J=8.0Hz,2H),7.84(m,1H),7.29(m,1H),4.36(s,1H), 3.75(d,J=3.2Hz,1H),2.87(t,J=6.5Hz,2H),2.53(s,6H),2.21–2.14(m,2H),1.85( d, J=8.9Hz, 1H), 1.71 (d, J=8.8Hz, 1H), 1.36 (t, J=9.5Hz, 1H), 1.23 (t, J=9.4Hz, 1H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.68, 152.84, 148.81, 148.28, 142.68, 139.85, 139.78, 134.95, 134.15, 132.47, 129.57, 128.93, 128.20, 127.87, 119.88, 57.59, 49.98, 44.38, 43.32, 42.47, 36.55, 26.96, 25.42. HRMS (ESI) m / z calculated value C 26 H 28 N5O[M+H] + 426.2289, measured value 426.2287.

[0318] N-(2-methoxyethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3016)

[0319] Synthesized using Method C: off-white solid (53 mg, 0.5 mmol, 26.25%). 1 H NMR(500MHz, (CD3)2SO) δ8.61(t,J=5.1Hz,1H),8.56(s,1H),7.98–7.94(m,2H),7.84(d,J=4.9Hz,2H),7.68–7.6 1(m,2H),3.51–3.43(m,4H),3.32(d,J=4.0Hz,2H),3.28(s,3H),2.76(t,J=6.1Hz,2H),1.99(m,2H),1.73(m,2H). 13C NMR (126 MHz, (CD3)2SO) δ 166.50, 156.18, 143.93, 143.62, 142.54, 138.73, 136.42, 134.06, 129.65, 129.45, 129.33, 127.37, 121.99, 116.44, 114.52, 70.96, 58.42, 39.53, 29.68, 28.86, 22.57, 22.48. HRMS (ESI) m / z calculated value C 24 H 25 N4O2[M+H] + 401.1972, measured value 401.1970.

[0320] 4-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-(thiazol-2-yl)benzamide (Compound HSH3017)

[0321] Synthesized using method C: off-white (16 mg, 0.5 mmol, 7.5%). 1 H NMR(500MHz,(CD3)2SO)δ8.59(s,1H),8.20(d,J=8.0Hz,2H),7.92–7.83(m,2H),7.73(d,J=8.0Hz,2H),7 .57(d,J=3.6Hz,1H),7.29(d,J=3.7Hz,1H),2.80(t,J=6.0Hz,2H),2.07–1.94(m,2H),1.85–1.65(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 165.35, 159.34, 155.86, 145.24, 143.70, 142.66, 138.72, 136.47, 136.42, 131.86, 129.77, 129.64, 129.34, 128.37, 122.12, 116.43, 114.65, 114.38, 29.69, 28.84, 22.57, 22.47. HRMS (ESI) m / z calculated value C 24 H 20 N5OS[M+H] + 426.1383, measured value 426.1379.

[0322] N-Cyclopropyl-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3019)

[0323] Synthesized using Method C: off-white solid (25.2 mg, 0.5 mmol, 13.2%). 1 H NMR(500MHz, (CD3)2SO) δ8.56(s,1H),8.52(d,J=4.3Hz,1H),7.92(d,J=7.9Hz,2H),7.84(t,J=9.7Hz,2H),7.62(d,J=7.9 Hz,2H),3.34(s,2H),2.88(m,1H),2.74(t,J=6.1Hz,2H),2.03–1.94(m,2H),1.79–1.68(m,2H),0.70(m,2H),0.60(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 167.72, 156.13, 143.87, 143.63, 142.53, 138.74, 136.41, 134.09, 129.62, 129.39, 129.25, 127.33, 122.02, 116.41, 114.52, 29.65, 28.85, 23.59, 22.55, 22.47, 6.27. HRMS (ESI) m / z calculated value C 24 H 23 N4O[M+H] + 383.1866, measured value 383.1865.

[0324] N-(3-methoxypropyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3088)

[0325] Synthesized using Method F: off-white solid (13 mg, 0.5 mmol, 6.3%). 1 H NMR(500MHz, (CD3)2SO) δ8.55(t,J=9.3Hz,1H),7.95–7.91(m,2H),7.88–7.82(m,2H),7.65–7.60(m,2H),7. 52(t,J=5.5Hz,1H),3.24(s,3H),3.21(s,2H),3.15(m,2H),2.76(t,J=6.1Hz,2H),2.00(m,2H),1.76(m,6H). 13C NMR (126 MHz, (CD3)2SO) δ 166.43, 161.39, 156.19, 143.84, 143.64, 142.57, 138.73, 136.43, 134.28, 129.64, 129.44, 127.31, 122.01, 116.45, 114.53, 70.24, 69.75, 58.40, 42.45, 37.09, 29.69, 28.86, 22.56. HRMS (ESI) m / z calculated value C 25 H 27 N4O2[M+H] + 415.2129, measured value 415.2129.

[0326] (4-Methylpiperazin-1-yl)(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)methanone (Compound HSH3107)

[0327] Synthesized using Method D: off-white solid (510 mg, 4 mmol, 30%). 1 H NMR(500MHz,(CD3)2SO)δ8.57(s,1H),7.86(d,J=9.0Hz,1H),7.82(s,1H),7.61(d,J=7.9Hz,2H),7.47(d,J=8.0Hz,2H),3 .53(s,4H),3.34(t,J=6.8Hz,2H),2.79(t,J=6.3Hz,2H),2.39–2.32(m,4H),2.22(s,3H),2.07–1.98(m,2H),1.77(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 169.38, 156.15, 143.80, 142.53, 142.44, 135.81, 129.62, 129.53, 129.25, 128.15, 127.02, 122.09, 121.02, 116.34, 114.47, 55.02, 46.01, 29.66, 28.85, 22.61, 22.53. HRMS (ESI) m / z calculated value C 26 H 28 N5O[M+H] + 426.2289, measured value 426.2290.

[0328] N-(2-(Diethylamino)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3124)

[0329] Synthesized using Method F: off-white solid (66 mg, 0.5 mmol, 30%). 1 H NMR(800MHz, (CD3)2SO) δ8.59(s,1H),8.39(t,J=5.7Hz,1H),7.99–7.91(m,2H),7.87(d,J=9.0Hz,1H),7.83(d,J=9.0Hz,1H),7.68–7.63(m,2H), 3.40–3.32(m,4H),2.78(t,J=6.2Hz,2H),2.60(t,J=7.2Hz,2H),2.53(q ,J=7.2Hz,4H),2.04–2.00(m,2H),1.78–1.73(m,2H),1.01–0.97(m,6H). 13 C NMR (201 MHz, (CD3)2SO) δ 166.28, 156.08, 143.81, 142.44, 136.28, 134.30, 129.56, 129.36, 129.16, 127.16, 122.01, 116.24, 116.22, 116.18, 114.63, 51.99, 47.25, 38.12, 29.58, 28.76, 22.50, 22.42, 12.43. HRMS (ESI) m / z calculated value C 27 H 32 N5O[M+H] + 442.2601, measured value 442.2600.

[0330] N-(2-(Pyridin-2-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3130)

[0331] Synthesized using Method F: off-white solid (32 mg, 0.5 mmol, 14.3%). 1H NMR(500MHz,(CD3)2SO)δ8.67(m,1H),8.58(s,1H),8.51–8.43(m,2H),7.88(m,4H),7.64(d,J=8.2Hz,2H),7.32–7.26(m,2 H),3.57(q,J=6.7,6.7,6.6Hz,2H),2.90(t,J=7.1,7.1Hz,2H),2.76(t,J=6.1,6.1Hz,2H),2.05–1.96(m,2H),1.73(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.51, 156.13, 149.92, 148.98, 143.95, 143.68, 142.56, 134.11, 129.62, 129.48, 129.30, 129.11, 127.29, 124.76, 122.05, 120.96, 114.65, 114.47, 34.68, 29.67, 28.86, 22.57, 22.48. HRMS (ESI) m / z calculated value C 28 H 26 N5O[M+H] + 448.2132, measured value 448.2130.

[0332] N-(3-morpholinopropyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3146)

[0333] Synthesized using Method F: off-white solid (92.9 mg, 0.5 mmol, 39.5%). 1 H NMR(500MHz,(CD3)2SO)δ8.57(t,J=5.4Hz,2H),7.96–7.90(m,2H),7.85(q,J=9.0Hz,2H),7.67–7.61(m,2H),3.5 6(t,J=4.6Hz,5H),3.30(d,J=6.5Hz,3H),2.77(t,J=6.1Hz,2H),2.34(d,J=6.7Hz,6H),2.00(m,2H),1.72(m,4H). 13C NMR (126 MHz, (CD3)2SO) δ 166.37, 156.20, 143.83, 143.74, 143.59, 142.57, 136.42, 134.32, 129.64, 129.45, 129.34, 127.29, 122.00, 116.42, 114.52, 66.69, 56.59, 53.84, 38.29, 29.68, 28.86, 26.46, 22.57, 22.49. HRMS (ESI) m / z calculated value C 28 H 32 N5O2[M+H] + 470.2550, measured value 470.2548.

[0334] N-(2-(Piperidin-1-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3117)

[0335] Synthesized using Method F: off-white solid (10 mg, 0.5 mmol, 4.3%). 1 H NMR (800MHz, DMSO-d6) δ8.59 (s, 1H), 8.40 (t, J = 5.7Hz, 1H), 7.95 (d, J = 8.1Hz, 2H), 7.87 (d,J=9.0Hz,1H),7.82(d,J=9.2Hz,1H),7.65(d,J=8.1Hz,2H),3.42(q,J=6.4Hz,2H),3. 33(t,J=6.8Hz,2H),2.78(t,J=6.3Hz,2H),2.47(t,J=7.1Hz,2H),2.44–2.34(m,4H),2. 01(p,J=6.4Hz,2H),1.75(p,J=6.1Hz,2H),1.50(p,J=5.6Hz,4H),1.38(q,J=6.2Hz,2H). 13 C NMR (201 MHz, DMSO-d6) δ 166.29, 156.06, 143.89, 143.83, 143.74, 142.43, 136.09, 134.28, 129.55, 129.35, 129.15, 127.20, 122.00, 116.26, 114.66, 58.09, 54.49, 37.47, 29.58, 28.76, 26.03, 24.45, 22.50, 22.42. HRMS (ESI) m / z calculated value C 28 H 32N5O[M+H] + 454.2601, measured value 454.2600.

[0336] N-(2-(Dimethylamino)ethyl)-4-(3,8,9,10-tetrahydrocyclopenta[c]pyrazolo[4,3-f]quinolin-7-yl)benzamide (Compound HSH3159)

[0337] Synthesized using Method F: off-white solid (10.8 mg, 0.25 mmol, 10.8%). 1 H NMR(500MHz, (CD3)2SO) δ8.95(t,J=5.6Hz,1H),8.51(s,1H),8.07(d,J=8.1Hz,2H),7.99(d,J=8.1Hz ,2H),7.90(q,J=9.2Hz,2H),3.67(q,J=5.8Hz,2H),3.50(t,J=7.6Hz,2H),2.83(s,6H),2.28(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.89, 151.26, 149.87, 143.36, 142.48, 141.78, 136.49, 136.34, 133.79, 133.45, 129.25, 128.86, 127.87, 119.54, 116.52, 56.49, 42.90, 35.12, 33.57, 33.10, 25.01. HRMS (ESI) m / z calculated value C 24 H 26 N5O[M+H] + 400.2132, measured value 400.2131.

[0338] N-(Cyanomethyl)-4-(3,8,9,10-tetrahydrocyclopenta[c]pyrazolo[4,3-f]quinolin-7-yl)benzamide (Compound HSH3160)

[0339] Synthesized using Method F: off-white solid (12.5 mg, 0.25 mmol, 13.6%). 1H NMR(500MHz, (CD3)2SO) δ9.30(t,J=5.5Hz,1H),8.47(s,1H),8.01(d,J=1.2Hz,4H),7.93(d,J=10.0Hz,1H ),7.87(d,J=9.0Hz,1H),4.35(d,J=5.3Hz,2H),3.49(t,J=7.6Hz,2H),3.29(t,J=7.5Hz,2H),2.27(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.88, 151.15, 149.88, 144.93, 143.81, 138.32, 136.42, 135.28, 132.76, 129.27, 129.09, 127.85, 119.50, 118.17, 116.69, 114.96, 33.56, 33.08, 28.27, 24.98. HRMS (ESI) m / z calculated value C 22 H 18 N5O[M+H] + 368.1506, measured value 368.1504.

[0340] Tert-butyl (2-(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide)ethyl)carbamate (Compound HSH3150)

[0341] Synthesized using Method F: off-white solid (170 mg, 0.5 mmol, 70%). 1 H NMR(500MHz, (CD3)2SO) δ8.56(s,1H),8.41(s,1H),7.95(d,J=8.0Hz,2H),7.84(d,J=10.1Hz,2H),7.63(d,J=7.9Hz,2H),6.78(s,1H),3.34(d t, J=13.1, 6.4Hz, 4H), 3.15 (q, J=5.7Hz, 2H), 2.76 (q, J=10.7, 8.4Hz, 2H), 2.00 (h, J=9.1, 7.3Hz, 2H), 1.75 (h, J=9.6, 7.7Hz, 2H), 1.38 (s, 9H). 13CNMR (126 MHz, (CD3)2SO) δ 166.74, 156.26, 156.20, 144.00, 143.78, 143.74, 142.53, 136.36, 136.31, 134.32, 129.66, 129.35, 129.24, 127.35, 122.08, 114.51, 78.22, 40.55, 29.65, 28.81, 28.80, 28.74, 22.58, 22.52. HRMS (ESI) m / z calculated value C 28 H 32 N5O3[M+H] + 486.2499, measured value 486.2494.

[0342] N-(2-aminoethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (compound HSH3165)

[0343] Synthesized using Method G: off-white solid (85 mg, 0.245 mmol, 89%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.63 (s, 1H), 8.58 (s, 1H), 7.99 (d, J = 8.1 Hz, 2H), 7.92 (s, 2H), 7.88–7.79 (m, 2H), 7.66 (d, J = 8.1 Hz, 2H), 3.57 (q, J = 6.0 Hz, 2H), 3.35 (d, J = 6.5 Hz, 2H), 3.06 (t, J = 6.2 Hz, 2H), 2.77 (t, J = 6.2 Hz, 2H), 2.05–1.98 (m, 2H), 1.82–1.69 (m, 2H). HRMS (ESI) m / z calculated value C 23 H 24 N5O[M+H] + 386.1975, measured value 386.1973.

[0344] N-(2-(Methylsulfonyl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3175)

[0345] Synthesized using Method E: off-white solid (6.7 mg, 0.5 mmol, 3%). 1H NMR(500MHz, (CD3)2SO) δ8.81(t,J=5.6Hz,1H),8.57(s,1H),7.99–7.88(m,2H),7.85(q,J=9.2Hz,2H),7.70–7.6 0(m,2H),3.76–3.63(m,2H),3.41(t,J=6.9Hz,2H),3.05(s,3H),2.76(t,J=6.1Hz,2H),2.00(m,2H),1.74(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.71, 156.13, 144.19, 143.62, 142.60, 138.74, 136.43, 133.68, 129.64, 129.56, 129.34, 127.37, 122.02, 116.44, 114.56, 53.38, 41.26, 33.90, 29.68, 28.85, 22.56, 22.47. HRMS (ESI) m / z calculated value C 24 H 25 N4O3S[M+H] + 449.1642, measured value 449.1642.

[0346] N-(2-sulfamoylethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3184)

[0347] Synthesized using Method F: off-white solid (10 mg, 0.5 mmol, 2.23%). 1 H NMR(500MHz,(CD3)2SO)δ8.58(s,2H),7.97–7.93(m,2H),7.89–7.79(m,2H),7.67–7.62(m,2H),3.77–3.67(m, 2H), 3.34 (t, J=6.5Hz, 2H), 3.29 (dd, J=8.1, 6.2Hz, 2H), 2.77 (t, J=6.2Hz, 2H), 2.07–1.98 (m, 2H), 1.76 (m, 2H). 13C NMR (126 MHz, (CD3)2SO) δ 166.75, 156.12, 144.22, 143.88, 142.57, 133.96, 129.77, 129.64, 129.48, 129.23, 128.11, 127.34, 127.11, 122.14, 116.28, 54.24, 35.33, 29.66, 28.80, 22.58, 22.52. HRMS (ESI) m / z calculated value C 23 H 24 N5O3S[M+H] + 450.1594, measured value 450.1591.

[0348] (4-Isopropylpiperazin-1-yl)(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)methanone (Compound HSH3182)

[0349] Synthesized using Method F: off-white solid (56 mg, 0.5 mmol, 24.6%). 1 H NMR(500MHz,(CD3)2SO)δ8.56(s,1H),7.91–7.77(m,2H),7.68–7.57(m,2H),7.47(d,J=8.0Hz,2H),3.51(s,4H), 2.78(t,J=6.1Hz,2H),2.67(m,1H),2.46(d,J=5.3Hz,3H),2.01(m,2H),1.81–1.71(m,2H),0.97(d,J=6.5Hz,6H). 13 C NMR (126 MHz, (CD3)2SO) δ 169.32, 156.19, 143.83, 143.77, 142.59, 142.37, 136.23, 135.81, 129.63, 129.50, 129.29, 127.06, 122.09, 116.35, 114.55, 54.23, 48.71, 29.66, 28.84, 22.58, 22.51, 18.56. HRMS (ESI) m / z calculated value C 28 H 32 N5O[M+H] + 454.2601, measured value 454.2598.

[0350] N-(Prop-2-yn-1-yl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3199)

[0351] Synthesized using Method F: Off-white solid (14 mg, 0.2 mmol, 18.4%). 1 H NMR (500 MHz, (CD3)2SO) δ 9.02 (t, J = 5.7 Hz, 1H), 8.59 (s, 1H), 7.96 (d, J = 8.0 Hz, 2H), 7.85 (t, J = 11.3 Hz, 2H), 7.65 (d, J = 8.0 Hz, 2H), 4.09 (d, J = 6.5 Hz, 2H), 3.23 (s, 1H), 3.13 (s, 1H), 2.76 (t, J = 6.3 Hz, 2H), 2.06–1.94 (m, 2H), 1.74 (d, J = 5.7 Hz, 2H). HRMS (ESI) m / z calculated value C 24 H 21 N4O[M+H] + 381.1710, measured value 381.1709.

[0352] N-(1-((Dimethylamino)methyl)cyclopropyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3192)

[0353] Synthesized using Method F: off-white solid (18.2 mg, 0.5 mmol, 8.3%). 1 H NMR(500MHz, (CD3)2SO) δ8.59(d,J=9.7Hz,2H),7.98–7.90(m,2H),7.84(d,J=11.5Hz,2H),7.65–7.55(m,2H),2.7 5(t,J=6.1Hz,2H),2.53(s,2H),2.26(s,6H),2.00(m,2H),1.73(m,2H),0.85–0.76(m,2H),0.68(q,J=4.8Hz,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.90, 156.21, 143.84, 143.68, 142.57, 138.78, 138.70, 136.40, 134.40, 129.64, 129.31, 127.48, 122.05, 116.40, 114.54, 63.91, 49.07, 45.90, 31.57, 29.67, 28.88, 22.57, 22.49, 12.82. HRMS (ESI) m / z calculated value C 27H 30 N5O[M+H] + 440.2444, measured value 440.2443.

[0354] N-(2-(Dimethylamino)ethyl)-4-(3,8,9,10,11,12-hexahydrocyclohepta[c]pyrazolo[4,3-f]quinolin-7-yl)benzamide (Compound HSH3194)

[0355] Synthesized using Method F: off-white solid (13.5 mg, 0.5 mmol, 6.3%). 1 H NMR(500MHz, (CD3)2SO) δ8.62(s,1H),8.32(q,J=5.7,4.9Hz,1H),7.96–7.90(m,2H),7.84(dd,J=9.1,0.9Hz,1H),7.78(d,J=9.1Hz,1H) ,7.60–7.51(m,2H),3.54(t,J=4.9Hz,2H),3.41(td,J=6.7,5.4Hz,2H),3.00–2.96(m,2H),2.22(s,6H),1.93–1.82(m,4H),1.63(m,2H). 13 CNMR (126 MHz, (CD3)2SO) δ 166.51, 155.33, 148.69, 146.38, 144.72, 144.41, 139.87, 135.25, 134.27, 129.78, 129.49, 127.32, 121.51, 118.84, 116.05, 58.62, 45.59, 37.93, 31.20, 31.10, 30.14, 27.30, 25.03. HRMS (ESI) m / z calculated value C 26 H 30 N5O[M+H] + 428.2444, measured value 428.2442.

[0356] N-(2-(Dimethylamino)ethyl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide (Compound HSH3195)

[0357] Synthesized using Method F: off-white solid (57 mg, 0.5 mmol, 27.4%). 1H NMR(500MHz,(CD3)2SO)δ8.59(s,1H),8.34(t,J=5.6Hz,1H),7.98–7.83(m,4H),7.67–7.64(m,2H), 4.78(d,J=1.5Hz,2H),4.16(t,J=5.9Hz,2H),3.43–3.37(m,4H),2.46(d,J=6.8Hz,2H),2.21(s,6H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.36, 161.45, 153.03, 144.49, 142.38, 139.50, 134.89, 129.53, 129.16, 127.53, 127.50, 127.05, 121.54, 118.63, 116.09, 66.84, 64.42, 58.66, 45.65, 37.99, 28.74. HRMS (ESI) m / z calculated value C 24 H 26 N5O2[M+H] + 416.2081, measured value 416.2080.

[0358] 4-(8,9-Dihydro-3H-cyclobuta[c]pyrazolo[4,3-f]quinolin-7-yl)-N-(2-(dimethylamino)ethyl)benzamide (Compound HSH3197)

[0359] Synthesized using Method F: off-white solid (9 mg, 0.095 mmol, 24.5%). 1 H NMR(500MHz,(CD3)2SO)δ8.39–8.32(m,2H),8.26–8.20(m,2H),8.02–7.96(m,2H),7.93–7.81(m,2H), 3.79–3.74(m,2H),3.70–3.64(m,2H),3.42(td,J=6.7,5.5Hz,2H),2.52(t,J=6.8Hz,2H),2.26(s,6H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.42, 161.50, 150.26, 146.88, 145.25, 140.35, 137.83, 135.37, 130.23, 130.15, 128.15, 127.23, 127.10, 117.87, 115.04, 58.54, 45.48, 37.81, 32.32, 30.39. HRMS (ESI) m / z calculated value C 23 H24 N5O[M+H] + 386.1975, measured value 386.1974.

[0360] N-(2-(Pyrrolidin-1-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3180)

[0361] Synthesized using Method F: off-white solid (9 mg, 0.5 mmol, 4.1%). 1 H NMR(500MHz,(CD3)2SO)δ8.56(s,1H),8.34(t,J=5.7Hz,1H),7.98–7.93(m,2H),7.87–7.78(m,2H),7.66–7.61(m,2H),3.43(d,J=5.8Hz,2H),3 .32(t,J=6.6Hz,2H),2.76(t,J=6.1Hz,2H),2.62(t,J=6.9Hz,2H),2.53 –2.47(m,4H),2.04–1.95(m,2H),1.79–1.72(m,2H),1.72–1.65(m,4H). 13 CNMR (126 MHz, (CD3)2SO) δ 166.44, 161.41, 156.16, 143.96, 143.90, 142.49, 140.03, 134.43, 129.65, 129.39, 129.19, 127.30, 122.11, 116.29, 115.28, 55.38, 54.10, 39.25, 29.64, 28.82, 23.75, 22.59, 22.52. HRMS (ESI) m / z calculated value C 27 H 30 N5O[M+H] + 440.2444, measured value 440.2443.

[0362] (4-(3,8,9,10,11,12-Hexahydrocyclohepta[c]pyrazolo[4,3-f]quinolin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3181)

[0363] Synthesized using Method F: off-white solid (74 mg, 0.4 mmol, 42.1%). 1H NMR(800MHz, (CD3)2SO) δ8.63(s,1H),7.85(d,J=9.0Hz,1H),7.79(s,1H),7.54(d,J=8.0Hz,2H),7.48(d,J=8.1Hz, 2H),3.61(s,2H),3.57–3.50(m,3H),3.02–2.96(m,2H),2.40–2.26(m,4H),2.20(s,3H),1.87(m,4H),1.63(m,2H). 13 C NMR (201 MHz, (CD3)2SO) δ 169.22, 161.12, 155.16, 148.67, 144.49, 142.75, 138.93, 135.49, 135.25, 129.65, 129.57, 127.02, 121.36, 116.07, 114.83, 55.12, 54.94, 54.28, 45.96, 45.05, 31.26, 31.00, 30.06, 27.28, 24.89. HRMS (ESI) m / z calculated value C 27 H 30 N5O[M+H] + 440.2445, measured value 440.2444.

[0364] (4-(5-Fluoro-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3183)

[0365] Synthesized using Method F: off-white solid (37 mg, 0.38 mmol, 22%). 1 H NMR(800MHz, (CD3)2SO) δ8.55(s,1H),7.71(d,J=10.0Hz,1H),7.64(d,J=7.6Hz,2H),7.50(d,J=7.0Hz,2H),3.65(s,2H),3.30( s, 2H), 2.79 (d, J = 6.7Hz, 2H), 2.34 (d, J = 68.6Hz, 4H), 2.20 (d, J = 6.3Hz, 3H), 1.99 (q, J = 6.6, 6.1Hz, 2H), 1.74 (q, J = 6.2Hz, 2H). 13 C NMR(201MHz,(CD3)2SO)δ169.18,157.78( 1J = 250 Hz), 156.11, 142.96, 141.87, 136.43, 135.83, 134.28, 130.75, 130.71, 129.53, 127.01, 122.72, 112.86, 98.40, 55.14, 54.64, 47.52, 45.98, 41.85, 29.59, 28.83, 22.31, 22.19. HRMS (ESI) m / z calculated value C 26 H 27 FN5O[M+H] + 444.2194, measured value 444.2196.

[0366] Piperidin-1-yl(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)methanone (Compound HSH3185)

[0367] Synthesized using Method F: off-white solid (14 mg, 0.5 mmol, 6.8%). 1 H NMR(800MHz, (CD3)2SO) δ8.57(s,1H),7.91–7.83(m,2H),7.63(d,J=8.1Hz,2H),7.47(d,J=8.0Hz,2H),3.61(s,2H),3.37 (t,J=6.7Hz,4H),2.81(t,J=6.2Hz,2H),2.02(td,J=6.1,3.3Hz,2H),1.81–1.72(m,2H),1.67–1.62(m,2H),1.55(m,4H). 13 C NMR (201 MHz, (CD3)2SO) δ 169.15, 156.17, 143.61, 142.44, 142.08, 138.69, 136.30, 136.27, 129.55, 129.44, 129.26, 126.68, 121.90, 116.40, 114.40, 40.50, 29.62, 28.82, 25.73, 24.47, 22.52, 22.44. HRMS (ESI) m / z calculated value C 26 H 27 N4O[M+H] + 411.2179, measured value 411.2178.

[0368] N-cyclohexyl-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (compound HSH3186)

[0369] Synthesized using Method F: off-white solid (13 mg, 0.5 mmol, 6.5%). 1 H NMR(800MHz,(CD3)2SO)δ8.59(s,1H),8.25–8.20(m,1H),8.00–7.94(m,2H),7. 90–7.82(m,2H),7.64(td,J=4.7,2.4Hz,2H),3.82(tt,J=7.5,4.0Hz,1H),3.27( d,J=5.7Hz,2H),2.85–2.73(m,2H),2.03(q,J=6.2Hz,2H),1.87(d,J=11.4Hz,2 H),1.80–1.73(m,4H),1.66–1.60(m,1H),1.42–1.29(m,4H),1.20–1.13(m,1H). 13 C NMR (126 MHz, (CD3)2SO) δ 165.56, 156.21, 143.73, 143.64, 142.54, 138.71, 136.39, 134.62, 134.55, 129.65, 129.32, 127.45, 122.03, 116.42, 114.54, 48.86, 32.92, 29.67, 28.87, 25.77, 25.45, 22.58, 22.50. HRMS (ESI) m / z calculated value C 27 H 29 N4O[M+H] + 425.2336, measured value 425.2336.

[0370] 1-(4-(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoyl)piperazin-1-yl)ethan-1-one (Compound HSH3187)

[0371] Synthesized using Method F: off-white solid (13.8 mg, 0.5 mmol, 5.7%). 1H NMR(500MHz,,(CD3)2SO)δ8.56(s,1H),7.86(d,J=9.1Hz,1H),7.81(d,J=9.8Hz,1H),7.65–7.61(m,2H),7.54–7.49 (m,2H),3.57–3.50(m,6H),3.34(q,J=9.8,7.4Hz,4H),2.79(t,J=6.1Hz,2H),2.05–1.97(m,5H),1.79–1.73(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 169.72, 169.08, 156.13, 143.82, 142.60, 142.57, 135.50, 129.62, 129.57, 129.27, 127.15, 122.10, 118.03, 116.51, 116.36, 114.50, 46.07, 41.38, 29.65, 28.84, 22.58, 22.52, 21.57. HRMS (ESI) m / z calculated value C 27 H 28 N5O2[M+H] + 454.2237, measured value 454.2241.

[0372] (4-Methylpiperazin-1-yl)(5-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)thiophen-2-yl)methanone (Compound HSH3200)

[0373] Synthesized using Method F: off-white solid (17 mg, 0.5 mmol, 7.9%). 1 H NMR(800MHz, (CD3)2SO) δ8.56(s,1H),7.88(d,J=9.0Hz,1H),7.83–7.76(m,1H),7.57(d,J=3.9Hz,1H),7.40(d,J=3.8Hz,1H),3.6 8(t,J=5.2Hz,4H),3.34(s,2H),3.11(t,J=6.2Hz,2H),2.39(t,J=5.2Hz,4H),2.23(s,3H),2.05–2.00(m,2H),1.89–1.85(m,2H). 13C NMR (201 MHz, (CD3)2SO) δ 162.76, 148.32, 147.82, 143.33, 143.23, 138.76, 138.42, 136.38, 129.71, 129.21, 128.95, 127.50, 121.98, 116.25, 114.98, 54.98, 45.93, 29.89, 28.86, 22.43, 22.19. HRMS (ESI) m / z calculated value C 24 H 26 N5OS[M+H] + 432.1852, measured value 432.1853.

[0374] (4-Methylpiperazin-1-yl)(3-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)methanone (Compound HSH3201)

[0375] Synthesized using Method F: off-white solid (20 mg, 0.5 mmol, 9.4%). 1 H NMR(800MHz, (CD3)2SO) δ8.57(s,1H),7.91–7.80(m,2H),7.65(d,J=7.8Hz,1H),7.55(d,J=7.2Hz,2H),7.44(d,J=7.8H z,1H),3.78–3.55(m,4H),2.79(t,J=6.3Hz,2H),2.43–2.30(m,5H),2.19(s,3H),2.04–1.96(m,2H),1.81–1.71(m,2H). 13 C NMR (201 MHz, (CD3)2SO) δ 169.21, 155.93, 143.73, 142.49, 141.33, 138.93, 136.36, 136.12, 130.51, 129.53, 129.16, 128.53, 127.78, 126.60, 121.99, 116.23, 114.49, 54.76, 47.68, 45.88, 29.58, 28.80, 22.47, 22.44. HRMS (ESI) m / z calculated value C 26 H 28 N5O[M+H] + 426.2289, measured value 426.2290.

[0376] N-(Cyanomethyl)-4-(9-methyl-3H-pyrazolo[4,3-f]quinolin-7-yl)benzamide (Compound HSH3202)

[0377] Synthesized using Method F: off-white solid (23 mg, 0.5 mmol, 13.5%). 1 H NMR (800MHz, (CD3)2SO) δ9.26 (m, 1H), 8.60 (s, 1H), 8.41 (d, J = 8.7Hz, 2H), 8.19 (d, J = 9.0Hz, 1H), 8.05 (d, J = 8.7Hz, 2H), 8.01–7.91 (m, 2H), 4.36 (q, J = 5.6Hz, 2H), 2.97 (d, J = 8.4Hz, 3H). 13 CNMR (201 MHz, (CD3)2SO) δ 166.77, 152.12, 146.15, 144.54, 142.37, 136.05, 133.37, 129.68, 128.27, 127.19, 122.31, 120.83, 118.02, 116.80, 115.64, 28.19, 22.71. HRMS (ESI) m / z calculated value C 20 H 16 N5O[M+H] + 342.1349, measured value 342.1353.

[0378] N-(Cyanomethyl)-3-fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3203)

[0379] Synthesized using Method F: off-white solid (70 mg, 0.5 mmol, 35%). 1 H NMR(500MHz, (CD3)2SO) δ8.59(s,1H),7.88(d,J=9.1Hz,1H),7.81(d,J=9.1Hz,1H),7.51(t,J=7.6Hz,1H),7.35( m,2H),3.35(t,J=6.5Hz,2H),2.64(t,J=6.3Hz,2H),2.52(s,2H),2.04–1.96(m,2H),1.79(dd,J=7.5,4.2Hz,2H). 13 C NMR(126MHz,(CD3)2SO)δ169.03,158.32( 1 J=245.7),151.69,143.97,142.42,140.00,139.02( 3J=7.56Hz),134.97,131.90,130.08,129.76( 2 J=16.4Hz),129.44,123.46,122.44,116.21,115.64,114.58( 2 J=23.9Hz),34.73,29.47,27.38,22.59,22.19.

[0380] (2-Fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3204)

[0381] Synthesized using Method F: yellow solid (18 mg, 0.25 mmol, 8%). 1 H NMR(500MHz, (CD3)2SO) δ8.57(s,1H),7.86(d,J=9.0Hz,1H),7.81(s,1H),7.47(t,J=5.1Hz,3H),3.68( s,2H),2.81(t,J=6.4Hz,2H),2.42–2.27(m,5H),2.22(d,J=7.7Hz,4H),2.01(m,2H),1.82–1.71(m,2H). 13 CNMR(126MHz,(CD3)2SO)δ164.36,161.15,158.69(d, 1 J=247Hz),154.77,144.56(d, 3 J=7.56Hz),143.85,142.72,142.45,129.56,129.22,128.81,126.08,123.87(d, 2 J=17.6Hz),122.30,121.84,116.75(d, 2 J = 21 Hz), 116.22, 55.25, 46.10, 45.98, 29.66, 28.68, 22.54, 22.48. HRMS (ESI) m / z calculated value C 26 H 27 FN5O[M+H] + 444.2194, measured value 444.2194.

[0382] (3-Fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3205)

[0383] Synthesized using Method F: off-white solid (56 mg, 0.5 mmol, 25.3%). 1 H NMR(500MHz, (CD3)2SO) δ8.58(s,1H),7.88(d,J=9.1Hz,1H),7.82(s,1H),7.52(t,J=7.6Hz,1H),7.38–7.29(m,2H),3.56(s,4H),3.41–3 .30(m,2H),3.27–3.11(m,2H),2.65(t,J=6.2Hz,2H),2.46(s,2H),2.28(s,3H),2.00(p,J=6.5Hz,2H),1.80(tq,J=9.2,6.2,4.4Hz,2H). 13 C NMR(126MHz,(CD3)2SO)δ167.89,160.36(d, 1 J=245.7Hz),158.41,151.64,143.87,142.44,138.89,138.35(d, 3 J=7.56Hz),136.34,132.08,130.11,129.98,129.44,123.42,122.42,116.44,114.61(d, 2 J = 23 Hz), 54.65, 54.55, 45.62, 29.47, 27.40, 22.59, 22.19. HRMS (ESI) m / z calculated value C 26 H 27 FN5O[M+H] + 444.2194, measured value 444.2193.

[0384] (2-Methoxy-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3206)

[0385] Synthesized using Method F: off-white solid (44 mg, 0.5 mmol, 19.31%). 1H NMR(800MHz,(CD3)2SO)δ8.61–8.53(m,1H),7.87(dd,J=9.0,4.2Hz,1H),7.84(s,1H),7.26(dd,J =7.6,4.2Hz,1H),7.22(d,J=4.1Hz,1H),7.15(dd,J=7.9,3.8Hz,1H),3.84(d,J=4.4Hz,3H),3.72 –3.64(m,1H),3.61(d,J=16.7Hz,1H),3.33–3.28(m,2H),3.25–3.17(m,2H),2.80(q,J=6.1,5.6H z,2H),2.34(m,3H),2.26–2.22(m,1H),2.21(d,J=4.3Hz,3H),2.01(m,2H),1.76(m,J=6.1Hz,2H). 13 C NMR (201 MHz, (CD3)2SO) δ 166.75, 156.28, 155.04, 143.49, 143.38, 142.43, 138.71, 136.29, 129.52, 129.26, 127.66, 125.47, 121.98, 121.76, 116.47, 114.47, 112.59, 56.11, 55.13, 54.68, 46.61, 45.98, 41.34, 29.57, 28.71, 22.47, 22.42. HRMS (ESI) m / z calculated value C 27 H 30 N5O2[M+H] + 456.2394, measured value 456.2394.

[0386] N-(1-(Dimethylamino)propan-2-yl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 3207)

[0387] Synthesized using Method F: off-white solid (31 mg, 0.5 mmol, 14.5%). 1H NMR(800MHz, (CD3)2SO) δ8.57(d,J=8.7Hz,1H),8.08(d,J=8.3Hz,1H),7.94(d,J=7.9 Hz,2H),7.88–7.80(m,2H),7.62(d,J=8.0Hz,2H),4.22–4.15(m,1H),2.76(t,J=6.2H z,2H),2.51–2.47(m,2H),2.47(dd,J=12.2,7.4Hz,1H),2.28(dd,J=12.2,6.7Hz,1H) ,2.20(d,J=5.4Hz,6H),2.03–1.99(m,2H),1.79–1.73(m,2H),1.18(d,J=6.7Hz,3H). 13 C NMR (126 MHz, (CD3)2SO) δ 165.91, 156.21, 143.89, 142.49, 139.11, 136.26, 134.66, 129.66, 129.31, 129.22, 127.35, 127.09, 122.10, 116.31, 114.63, 64.90, 45.93, 43.77, 29.64, 28.82, 22.59, 22.53, 19.49. HRMS (ESI) m / z calculated value C 26 H 30 N5O[M+H] + 428.2444, measured value 428.2445.

[0388] (4-(2-Hydroxyethyl)piperazin-1-yl)(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)methanone (Compound HSH3208)

[0389] Synthesized using Method F: off-white solid (27 mg, 0.75 mmol, 7.9%). 1 H NMR(500MHz, (CD3)2SO) δ8.56(s,1H),7.87–7.75(m,2H),7.61(d,J=8.1Hz,2H),7.47(d,J=8.1Hz,2H),4.28(t,J=5.3Hz,1H),3.63–3.42(m ,6H),3.34(dq,J=10.4,5.0Hz,2H),3.20(d,J=14.4Hz,2H),2.79(t,J=6.2Hz,2H),2.48–2.42(m,4H),2.06–1.95(m,2H),1.80–1.71(m,2H).13 C NMR(126MHz,(CD3)2SO)δ169.30,156.16,143.81,142.52,142.44,138.86,136.38,135.82,129.62, 129.53,129.27,127.03,122.06,116.50,114.36,60.52,59.02,53.58,29.66,28.85,22.61,22.53.

[0390] (4-Methylpiperazin-1-yl)(4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)phenyl)methanone (Compound HSH3209)

[0391] Synthesized using Method F: off-white solid (5 mg, 0.5 mmol, 4.7%). 1 H NMR(800MHz, (CD3)2SO) δ8.60(s,1H),7.92(d,J=9.0Hz,1H),7.86(d,J=9.3Hz,1H),7.69–7.62(m,2H),7.53–7.4 8(m,2H),4.81(s,2H),4.16(t,J=5.9Hz,2H),3.65(s,2H),3.40(t,J=6.1Hz,4H),2.47–2.28(m,4H),2.24(s,3H). 13 C NMR (201 MHz, (CD3)2SO) δ 169.13, 152.89, 144.33, 140.77, 139.40, 138.84, 136.15, 135.67, 129.41, 129.25, 127.42, 127.20, 121.39, 116.04, 115.07, 66.80, 64.35, 54.79, 47.28, 45.81, 41.78, 40.49, 28.62. HRMS (ESI) m / z calculated value C 25 H 26 N5O2[M+H] + 428.2081, measured value 428.2080.

[0392] N-(1-methylpiperidin-4-yl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3210)

[0393] Synthesized using Method F: off-white solid (83 mg, 0.75 mmol, 25%). 1 H NMR(500MHz, (CD3)2SO) δ8.56(s,1H),8.20(d,J=7.6Hz,1H),7.95(d,J=8.1Hz,2H),7.88–7.75(m,2H),7.61(d,J=8.1H z,2H),3.87–3.70(m,1H),3.32(t,J=6.6Hz,2H),2.85–2.69(m,4H),2.19(s,3H),2.08–1.95(m,4H),1.84–1.59(m,6H).

[0394] (4-(9,9-dioxido-3,8,10,11-tetrahydropyrazolo[4,3-f]thiopyrano[3,4-c]quinolin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3211)

[0395] Synthesized using Method F: off-white solid (53 mg, 0.75 mmol, 14.9%). 1 H NMR(800MHz, (CD3)2SO) δ8.70(s,1H),8.02(d,J=9.0Hz,1H),7.91–7.84(m,1H),7.68(d,J=8.0Hz,2H),7.63(d ,J=8.1Hz,2H),4.58(s,2H),4.02(d,J=6.8Hz,2H),3.72(t,J=6.7Hz,2H),3.38(d,J=35.2Hz,8H),2.78(s,3H). 13 C NMR (201 MHz, (CD3)2SO) δ 169.34, 155.40, 151.42, 144.71, 140.93, 138.97, 136.64, 135.23, 129.78, 129.29, 129.16, 127.56, 122.30, 121.13, 115.67, 52.47, 51.65, 45.95, 42.57, 40.40, 30.51. HRMS (ESI) m / z calculated value C 25 H 26 N5O3S[M+H] + 476.1751, measured value 476.1750.

[0396] N-(2-(Dimethylamino)ethyl)-4-(9,9-dioxido-3,8,10,11-tetrahydropyrazolo[4,3-f]thiopyrano[3,4-c]quinolin-7-yl)benzamide (Compound HSH3212)

[0397] Synthesized using Method F: off-white solid (68 mg, 0.75 mmol, 19.6%). 1 H NMR(800MHz, (CD3)2SO) δ8.71(s,1H),8.50(t,J=4.8Hz,1H),8.01(dt,J=9.3,4.2Hz,3H),7.92–7.84(m,1H),7.71–7.61(m,2H ), 4.51 (s, 2H), 4.01 (d, J = 9.4Hz, 2H), 3.71 (d, J = 9.0Hz, 2H), 3.42 (p, J = 6.9Hz, 2H), 2.48–2.40 (m, 2H), 2.21 (d, J = 3.9Hz, 6H). 13 C NMR (201 MHz, (CD3)2SO) δ 166.13, 155.51, 144.76, 141.95, 139.88, 138.96, 135.59, 134.85, 129.54, 129.37, 127.64, 122.25, 121.13, 117.19, 115.61, 58.58, 51.72, 45.93, 45.66, 37.90, 30.48. HRMS (ESI) m / z calculated value C 24 H 26 N5O3S[M+H] + 464.1750, measured value 464.1751.

[0398] N-(2-(Pyrrolidin-1-yl)ethyl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide (Compound HSH3214)

[0399] Synthesized using Method F: off-white solid (99 mg, 0.75 mmol, 29.9%). 1H NMR(500MHz,,(CD3)2SO)δ8.57(s,1H),8.38(t,J=5.7Hz,1H),8.03–7.93(m,2H),7.90(d,J=9.2Hz,1H),7.84(d,J=9.1Hz,1H),7.71–7.61(m,2H),4. 78(s,2H),4.14(t,J=5.9Hz,2H),3.44(t,J=6.3Hz,2H),3.39(q,J=6.9,6. 0Hz, 2H), 2.63 (t, J = 6.9Hz, 2H), 2.58–2.48 (m, 4H), 1.68 (h, J = 3.2Hz, 4H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.35, 153.00, 144.49, 142.39, 139.93, 139.45, 134.92, 134.66, 134.53, 129.53, 129.17, 127.54, 127.47, 121.52, 116.09, 66.86, 64.42, 55.36, 54.10, 39.23, 28.73, 23.74. HRMS (ESI) m / z calculated for C 26 H 28 N5O2[M+H] + 442.2237, measured value 442.2238.

[0400] Piperazine-1-yl(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)methanone (Compound HSH3215)

[0401] Synthesized using Method G: off-white solid (37 mg, 0.26 mmol, 34.6%). 1 H NMR (800 MHz, (CD3)2SO) δ8.59 (s, 1H), 7.87 (d, J = 9.0 Hz, 1H), 7.82 (d, J = 9.0 Hz, 1H), 7.63 (d, J = 8.0 Hz, 2H), 7.50 (d, J = 7.9 Hz, 2H), 3.77–3.51 (m, 4H), 3.33 (d, J = 6.7 Hz, 2H), 2.86 (m, 4H), 2.79 (t, J = 6.2 Hz, 2H), 2.01 (m, 2H), 1.76 (m, 2H). HRMS (ESI) m / z calculated value C 25 H 26 N5O[M+H] + 412.2132, measured value 412.2133.

[0402] N-(2-(Dimethylamino)ethyl)-5-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)thiophene-2-carboxamide (Compound HSH3216)

[0403] Synthesized using Method F: off-white solid (35 mg, 0.5 mmol, 16.7%). 1 H NMR (500MHz, (CD3)2SO) δ8.54(s,1H),8.28(t,J=5.8Hz,1H),7.85(d,J=9.1Hz,1H),7.77(d,J=9.1Hz,1H),7.72(d,J=4.1Hz,1H),7.57(d,J =4.1Hz,1H),3.36(q,J=6.8Hz,2H),3.30(s,2H),3.08(t,J=6.1Hz,2H),2.43(t,J=6.8Hz,2H),2.19(s,6H),2.00(m,2H),1.90–1.81(m,2H). 13 C NMR (201 MHz, (CD3)2SO) δ 161.63, 148.93, 148.44, 143.45, 143.24, 140.75, 138.98, 136.37, 129.23, 128.96, 128.80, 128.30, 122.04, 116.06, 115.02, 58.44, 45.41, 37.53, 29.89, 28.90, 22.43, 22.17. HRMS (ESI) m / z calculated value C 23 H 26 N5OS[M+H] + 420.1852, measured value 420.1850.

[0404] N-(2-(Dimethylamino)ethyl)-3-fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3217)

[0405] Synthesized using Method F: off-white solid (53 mg, 0.75 mmol, 16.4%). 1H NMR(800MHz, (CD3)2SO) δ8.61(s,1H),8.53(t,J=5.6Hz,1H),7.89(d,J=9.0Hz,1H),7.83(dd,J=7.8,1.8Hz,2H),7.79(m,1H),7.57(t,J=7.6Hz,1H),3 .44(q,J=6.7Hz,3H),3.36(t,J=6.8Hz,2H),2.63(t,J=6.4Hz,2H),2.54(t ,J=6.8Hz,2H),2.28(s,6H),2.02(q,J=6.2,5.0Hz,2H),1.84–1.77(m,2H). 13 C NMR(201MHz,(CD3)2SO)δ165.08,160.01(d, 1 J=245Hz),151.56,143.82,142.39,137.00,136.96,131.99,131.78,131 .76,131.48,131.39,129.93,129.40,123.66,122.38,116.31,114.76(d, 2 J = 24 Hz), 58.33, 45.38, 37.76, 29.39, 27.28, 22.48, 22.08. HRMS (ESI) m / z calculated value C 25 H 27 FN5O[M+H] + 432.2194, measured value 432.2190.

[0406] 3-Fluoro-N-(2-(pyrrolidin-1-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3218)

[0407] Synthesized using Method F: off-white solid (146 mg, 0.75 mmol, 42.5%). 1H NMR(500MHz, (CD3)2SO) δ8.59(s,1H),8.46(t,J=5.7Hz,1H),7.87(d,J=9.1Hz,1H),7.81(m,2H),7.77(dd,J=10.7,1.8Hz,1H),7.54(t,J=7.6Hz,1H ),3.46–3.38(m,2H),3.34(t,J=6.5Hz,2H),2.62(dd,J=8.3,5.6Hz,4H), 2.51(m,4H),2.05–1.94(m,2H),1.84–1.73(m,2H),1.68(h,J=3.2Hz,4H). 13 C NMR(126MHz,(CD3)2SO)δ165.12,160.48(d, 1 J=245Hz),151.65,143.96,142.42,137.22,137.17,131.84,131.80,131 .57,131.43,129.98,129.48,123.70,122.47,116.23,115.52,114.86(d, 2 J = 24 Hz), 55.28, 54.08, 39.36, 29.46, 27.36, 23.74, 22.57, 22.17. HRMS (ESI) m / z calculated value C 27 H 29 FN5O[M+H] + 458.2350, measured value 458.2350.

[0408] (2-Methoxy-4-(8,9,10,11-tetrahydro-3H-naphtho[1,2-e]indazol-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3222)

[0409] Synthesized using Method F: off-white solid (77 mg, 0.75 mmol, 22.6%). 1H NMR(800MHz,(CD3)2SO)δ8.61–8.53(m,1H),7.87(dd,J=9.0,4.2Hz,1H),7.84(s,1H),7.26(dd,J =7.6,4.2Hz,1H),7.22(d,J=4.1Hz,1H),7.15(dd,J=7.9,3.8Hz,1H),3.84(d,J=4.4Hz,3H),3.72 –3.64(m,1H),3.61(d,J=16.7Hz,1H),3.33–3.28(m,2H),3.25–3.17(m,2H),2.80(q,J=6.1,5.6H z,2H),2.34(m,3H),2.26–2.22(m,1H),2.21(d,J=4.3Hz,3H),2.01(m,2H),1.76(m,J=6.1Hz,2H). 13 C NMR (201 MHz, (CD3)2SO) δ 166.75, 156.28, 155.04, 143.49, 143.38, 142.43, 138.71, 136.29, 129.52, 129.26, 127.66, 125.47, 121.98, 121.76, 116.47, 114.47, 112.59, 56.11, 55.13, 54.68, 46.61, 45.98, 41.34, 29.57, 28.71, 22.47, 22.42. HRMS (ESI) m / z calculated value C 27 H 30 N5O2[M+H] + 456.2394, measured value 456.2394.

[0410] (4-(9-cyclopropyl-3H-benzo[e]indazol-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3223)

[0411] Synthesized using Method F: off-white solid (69 mg, 0.75 mmol, 22.4%). 1 H NMR(800MHz, (CD3)2SO) δ8.84(s,1H),8.32(d,J=8.1Hz,2H),7.98–7.91(m,3H),7.52(d,J=8.2Hz,2H) ,3.63(s,2H),2.65(s,1H),2.45–2.23(m,4H),2.20(s,3H),1.35–1.28(m,2H),1.04(t,J=5.3Hz,2H). 13C NMR (201 MHz, (CD3)2SO) δ 169.21, 152.76, 148.76, 146.13, 140.35, 138.60, 136.59, 136.43, 129.70, 127.72, 127.34, 123.08, 116.89, 116.52, 115.63, 54.95, 47.48, 45.90, 41.98, 40.43, 16.55, 7.65. HRMS (ESI) m / z calculated value C 25 H 26 N5O[M+H] + 412.2132, measured value 412.2132.

[0412] 7-(4-((4-methylpiperazin-1-yl)methyl)phenyl)-8,9,10,11-tetrahydro-3H-naphtho[1,2-e]indazole (Compound HSH3225)

[0413] Synthesized using Method H: off-white solid (30 mg, 0.5 mmol, 14.6%). 1 H NMR(500MHz,(CD3)2SO)δ8.56(s,1H),7.86–7.76(m,2H),7.48(d,J=8.0Hz,2H),7.37(d,J=8.0Hz,2H),3.52(s,2H),3.33(t, J=6.6Hz,2H),2.78(t,J=6.2Hz,2H),2.42(t,J=4.9Hz,4H),2.38–2.27(m,4H),2.16(s,3H),2.01(m,2H),1.79–1.70(m,2H). 13 C NMR (201 MHz, (CD3)2SO) δ 169.39, 158.54, 158.39, 156.06, 143.74, 142.51, 142.32, 135.48, 129.52, 129.48, 129.20, 127.01, 121.99, 116.94, 116.15, 47.82, 45.28, 42.13, 40.37, 29.57, 28.78, 22.48, 22.40. HRMS (ESI) m / z calculated value C 26 H 30 N5[M+H] + 412.2495, measured value 412.2496.

[0414] (4-(Methylsulfonyl)piperazin-1-yl)(4-(3,8,10,11-tetrahydroisochromeno[5,6-e]indazol-7-yl)phenyl)methanone (Compound HSH3126)

[0415] Synthesized using Method F: off-white solid (127 mg, 1 mmol, 25.9%). 1 H NMR(500MHz, (CD3)2SO) δ8.52(s,1H),7.88(t,J=10.6Hz,2H),7.65(d,J=8.1Hz,2H),7.54(d,J=8.1Hz,2H ),4.78(s,2H),4.13(t,J=5.9Hz,2H),3.65(m,4H),3.36(t,J=5.9Hz,2H),3.25–3.17(m,4H),2.91(s,3H). 13 C NMR(126MHz,(CD3)2SO)δ169.50,152.93,144.29,141.17,139.43,138.80,135.91,135.80,1 29.50,129.38,127.51,127.39,121.39,116.31,115.07,66.87,64.42,45.85,35.09,28.69.

[0416] N-(2-(Dimethylamino)ethyl)-4-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3127)

[0417] Synthesized using Method F: off-white solid (38.4 mg, 0.5 mmol, 17.9%). 1 H NMR(800MHz, (CD3)2SO) δ8.28(t,J=5.7Hz,1H),7.97–7.94(m,2H),7.74(d,J=8.9Hz,2H),7.63–7.60(m,2H),3. 45–3.39(m,4H),2.87(s,3H),2.80–2.76(m,2H),2.47(t,J=6.9Hz,2H),2.22(s,6H),1.80(h,J=4.7,3.3Hz,4H). 13C NMR (201 MHz, (CD3)2SO) δ 166.38, 155.59, 144.25, 143.86, 142.87, 142.43, 141.24, 134.32, 129.78, 129.25, 128.25, 127.35, 127.23, 123.83, 114.51, 58.64, 45.61, 40.64, 37.97, 31.47, 27.87, 22.41, 22.08. HRMS (ESI) m / z calculated value C 26 H 30 N5O[M+H] + 428.2444, measured value 428.2445.

[0418] 4-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-(2-(pyrrolidin-1-yl)ethyl)benzamide (Compound HSH3128)

[0419] Synthesized using Method F: off-white solid (40.7 mg, 0.3 mmol, 29.92%). 1 H NMR(800MHz, (CD3)2SO) δ8.33(t,J=5.6Hz,1H),7.98–7.94(m,2H),7.76–7.67(m,2H),7.64–7.59(m,2H),3.48–3.39(m,4H),2.87 (s,3H),2.78(t,J=6.0Hz,2H),2.64(t,J=7.0Hz,2H),2.57–2.51(m,J=2.8,2.0Hz,4H),1.84–1.77(m,4H),1.70(h,J=3.1Hz,4H). 13 C NMR (201 MHz, (CD3)2SO) δ 166.36, 155.59, 144.35, 144.28, 143.85, 142.43, 134.34, 129.77, 129.24, 128.25, 127.35, 127.24, 127.08, 123.83, 114.27, 55.30, 54.01, 40.65, 39.18, 31.47, 27.87, 23.68, 22.41, 22.08. HRMS (ESI) m / z calculated value C 28 H 32 N5O[M+H] + 454.2601, measured value 454.2603.

[0420] (4-(9-methyl-3H-pyrazolo[4,3-f]quinolin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone

[0421] Synthesized using Method D: off-white solid (53 mg, 0.5 mmol, 27.5%). 1 H NMR(800MHz, (CD3)2SO) δ8.60(d,J=6.8Hz,1H),8.34(m,2H),8.16(d,J=6.9Hz,1H),7.96(dt,J=31.9,8.0Hz,2H),7.54 (d,J=7.8Hz,2H),3.56–3.41(m,3H),2.97(d,J=6.8Hz,4H),2.35(s,4H),2.22(d,J=6.9Hz,3H),1.67(d,J=6.2Hz,1H). 13 C NMR (201 MHz, (CD3)2SO) δ 175.1, 169.2, 152.5, 146.3, 144.5, 140.2, 136.7, 136.1, 129.6, 128.0, 127.8, 127.2, 122.2, 120.6, 116.7, 54.9, 46.0, 22.7. HRMS (ESI) m / z calculated value C 23 H 25 N5O[M+H] + 386.1975, measured value 386.1974.

[0422] (4-Methylpiperazin-1-yl)(4-(3,8,9,10-tetrahydrocyclopenta[c]pyrazolo[4,3-f]quinolin-7-yl)phenyl)methanone

[0423] Synthesized using Method D: off-white solid (71 mg, 0.5 mmol, 34.5%). 1 H NMR(800MHz, (CD3)2SO) δ8.48(s,1H),7.97(d,J=7.8Hz,2H),7.93(s,1H),7.88(d,J=8.9Hz,1H),7.5 3(d,J=7.7Hz,2H),3.52(s,2H),3.31(d,J=7.4Hz,2H),2.33–2.26(m,3H),2.23(s,4H),1.66(s,2H). 13C NMR (201 MHz, (CD3)2SO) δ 169.2, 151.3, 149.7, 148.9, 148.5, 141.5, 136.0, 135.2, 129.2, 128.9, 128.1, 127.3, 121.0, 119.4, 116.7, 114.8, 54.9, 46.0, 42.5, 33.5, 33.1, 24.9. HRMS (ESI) m / z calculated value C 25 H 27 N5O[M+H] + 412.2132, measured value 412.2133.

[0424] (4-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone

[0425] Synthesized using Method D: off-white solid (43 mg, 0.5 mmol, 19.5%). 1 H NMR(500MHz, (CD3)2SO) δ7.75–7.64(m,2H),7.58(d,J=8.1Hz,2H),7.46(d,J=8.1Hz,2H),3.38(dt,J=1 5.1, 5.2Hz, 3H), 2.84 (s, 3H), 2.77 (t, J = 6.3Hz, 2H), 2.41–2.31 (m, 5H), 2.21 (s, 5H), 1.82–1.73 (m, 5H). 13 C NMR (126 MHz, (CD3)2SO) δ 171.8, 169.4, 161.2, 155.6, 142.5, 136.8, 135.7, 131.9, 129.8, 129.5, 128.3, 127.9, 127.2, 127.1, 123.9, 55.0, 54.4, 46.0, 45.1, 31.6, 28.0, 22.5, 22.2. HRMS (ESI) m / z calculated value C 27 H 30 N5O[M+H] + 440.2444, measured value 440.2445.

[0426] N-((1H-imidazol-4-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0427] Synthesized using Method D: off-white solid (111 mg, 0.5 mmol, 52.6%). 1 H NMR(800MHz, (CD3)2SO) δ8.99(t,J=7.1Hz,1H),8.05–8.01(m,2H),7.89(d,J=8.9Hz,1H),7.83(d,J=6.0Hz,2H),7.65(d ,J=7.9Hz,2H),7.05(s,1H),4.47(d,J=5.5Hz,2H),2.78(t,J=6.3Hz,2H),2.02(p,J=6.3Hz,2H),1.76(p,J=5.8Hz,2H). 13 C NMR (201 MHz, (CD3)2SO) δ 166.3, 156.1, 150.6, 143.9, 142.5, 140.1, 135.0, 134.0, 129.5, 129.3, 128.8, 127.5, 122.0, 120.6, 117.1, 36.2, 29.6, 28.8, 22.5, 22.4. HRMS (ESI) m / z calculated value C 25 H 23 N6O[M+H] + 423.1927, measured value 423.1927.

[0428] N-(2-(1H-imidazol-1-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0429] Synthesized using Method D: off-white solid (84 mg, 0.5 mmol, 38.5%). 1 H NMR (800MHz, (CD3)2SO) δ8.74(t,J=5.5Hz,1H),8.57(s,1H),7.93(d,J=7.8Hz,2H),7.89(d,J=8.9Hz,1H),7.86–7.80(m,1H),7.65(d,J=7.8Hz,2H) ,7.63(s,1H),7.18(s,1H),6.89(s,1H),4.22(t,J=6.2Hz,2H),3.63(q,J =5.9Hz,2H),2.78(t,J=6.2Hz,2H),2.06–1.99(m,2H),1.80–1.73(m,2H). 13C NMR (201 MHz, (CD3)2SO) δ 166.8, 156.1, 145.0, 144.1, 142.5, 138.8, 137.8, 136.3, 133.9, 129.5, 129.4, 128.7, 127.3, 126.9, 120.0, 118.4, 116.4, 114.5, 45.6, 40.9, 29.6, 28.8, 22.5, 22.4. HRMS (ESI) m / z calculated value C 26 H 25 N6O[M+H] + 437.2084, measured value 437.2081.

[0430] N-((1H-imidazol-2-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0431] Synthesized using Method D: off-white solid (65 mg, 0.5 mmol, 30.8%). 1 H NMR(800MHz, (CD3)2SO) δ9.17(t,J=5.9Hz,1H),8.57(s,1H),8.07(d,J=7.8Hz,2H),7.89(d,J=9.0Hz,1H),7.82(d,J=9.3Hz,1H),7 .65(d,J=7.8Hz,2H),6.93(s,2H),4.56(d,J=5.6Hz,2H),2.77(t,J=6.3Hz,2H),2.00(q,J=6.8,6.3Hz,2H),1.75(p,J=5.6Hz,2H). 13 C NMR (201 MHz, (CD3)2SO) δ 166.5, 156.0, 146.7, 145.5, 144.1, 143.7, 142.5, 138.8, 136.2, 133.8, 129.5, 129.3, 129.2, 127.6, 127.5, 122.0, 119.1, 116.2, 114.6, 37.5, 29.6, 28.8, 22.5, 22.4. HRMS (ESI) m / z calculated value C 25 H 23 N6O[M+H] + 423.1927, measured value 423.1926.

[0432] N-((1-methyl-1H-imidazol-2-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0433] Synthesized using Method D: off-white solid (68 mg, 0.5 mmol, 31.2%). 1 H NMR(500MHz, (CD3)2SO) δ8.56(s,1H),8.37(d,J=4.3Hz,3H),8.16(d,J=8.3Hz,3H),7.04(d,J=13.1Hz,1H),6.78(d,J=18.9Hz ,1H),4.56(d,J=5.3Hz,2H),3.68(s,5H),2.77(t,J=6.2Hz,3H),1.88(dp,J=127.7,5.7Hz,6H),1.23(dd,J=14.7,7.0Hz,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.5, 156.1, 147.2, 144.2, 142.5, 139.9, 134.9, 133.8, 129.5, 129.4, 129.2, 127.7, 127.6, 126.9, 125.7, 122.2, 120.4, 119.2, 36.0, 32.9, 29.7, 28.8, 22.6, 22.5. HRMS (ESI) m / z calculated value C 26 H 26 N6O[M+H] + 437.2084, measured value 437.2081.

[0434] N-((4H-1,2,4-triazol-3-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0435] Synthesized using Method D: off-white solid (79 mg, 0.5 mmol, 46.0%). 1 H NMR(500MHz,(CD3)2SO)δ8.58(s,1H),8.43–8.06(m,2H),8.01(d,J=7.8Hz,2H),7.84(q,J=9.1Hz,2H),7.65(d,J =7.8Hz, 2H), 4.61 (d, J = 5.6Hz, 2H), 3.34 (d, J = 13.1Hz, 2H), 2.78 (t, J = 6.1Hz, 2H), 1.89 (dp, J = 125.7, 6.1Hz, 4H). 13C NMR (126 MHz, (CD3)2SO) δ 166.7, 161.5, 156.2, 148.6, 144.2, 143.9, 142.6, 133.9, 129.7, 129.4, 129.2, 127.6, 125.4, 122.1, 120.3, 119.8, 116.3, 36.8, 29.7, 28.8, 22.6, 22.5. HRMS (ESI) m / z calculated value C 24 H 22 N7O[M+H] + 424.1880, measured value 424.1877.

[0436] N-((1H-imidazol-2-yl)methyl)-3-fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0437] Synthesized using Method D: off-white solid (109 mg, 1.0 mmol, 49.5%). 1 H NMR(500MHz,(CD3)2SO)δ8.59(s,1H),7.93–7.78(m,4H),7.57(t,J=7.6Hz,1H),7.16(s,2 H), 4.65 (d, J = 5.5Hz, 2H), 3.35 (t, J = 6.5Hz, 2H), 2.62 (t, J = 6.2Hz, 2H), 2.04–1.74 (m, 4H). 13 CNMR(126MHz,(CD3)2SO)δ165.6,159.5(d, 1 J=252Hz),151.6,151.1,145.3,144.0,142.5,136.5,136.1,131.9,130.0,129.5,126.2,125.4,124.1,122.5,121.2,120.4,116.2,115.3(d, 2 J = 25 Hz), 36.9, 29.5, 27.4, 22.6, 22.2. HRMS (ESI) m / z calculated value C 25 H 22 FN6O[M+H] + 441.1833, measured value 441.1830.

[0438] N-((1H-imidazol-2-yl)methyl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide

[0439] Synthesized using Method D: off-white solid (117 mg, 0.5 mmol, 55.2%). 1 H NMR(500MHz,(CD3)2SO)δ8.59(s,1H),8.04(d,J=7.9Hz,2H),7.94–7.83(m,2H),7.67(d,J=7.9Hz,2 H), 6.95 (s, 2H), 4.79 (s, 2H), 4.55 (d, J = 5.6Hz, 2H), 4.16 (t, J = 5.8Hz, 2H), 3.41 (d, J = 11.8Hz, 2H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.6, 156.2, 153.0, 147.0, 145.5, 142.6, 139.6, 139.5, 134.4, 129.5, 129.2, 129.2, 127.8, 127.5, 122.0, 121.6, 118.8, 116.2, 71.2, 66.9, 64.4, 37.6, 28.7, 25.7. HRMS (ESI) m / z calculated value C 24 H 21 N6O2[M+H] + 425.1720, measured value 425.1716.

[0440] N-((1H-imidazol-2-yl)methyl)-4-(9,9-dioxido-3,8,10,11-tetrahydropyrazolo[4,3-f]thiopyrano[3,4-c]quinolin-7-yl)benzamide

[0441] Synthesized using Method D: off-white solid (98 mg, 0.5 mmol, 41.5%). 1 H NMR (500MHz, (CD3)2SO) δ8.69 (s, 1H), 8.05 (dd, J = 47.7, 8.4Hz, 3H), 7.88 (d, J = 9.1Hz, 1H), 7.68 (d, J = 7. 8Hz, 2H), 7.19 (s, 2H), 4.67 (d, J = 5.4Hz, 2H), 4.47 (s, 2H), 4.02 (t, J = 6.7Hz, 2H), 3.69 (t, J = 6.8Hz, 2H). 13C NMR (126 MHz, (CD3)2SO) δ 166.8, 155.5, 145.5, 145.0, 142.5, 139.2, 136.0, 134.3, 129.6, 129.4, 128.1, 126.7, 125.4, 122.3, 121.3, 121.0, 120.1, 115.7, 52.0, 46.3, 36.8, 30.4. HRMS (ESI) m / z calculated value C 24 H 21 N6O3S[M+H] + 473.1390, measured value 473.1386.

[0442] N-((1H-tetrazolyl-5-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0443] The final compound was synthesized via a "click" reaction:

[0444] To a solution of alkyne (1 eq.) in DMF (2 mL) and water (0.5 mL) was added TMS-azide (2 eq.). A catalytic amount of copper sulfide pentahydrate and sodium ascorbate were added. The reaction mixture was stirred at 100 °C overnight. The reaction mixture was rotary evaporated, diluted in ethyl acetate and washed with water, dried over sodium sulfate. It was then purified by column chromatography using 9% methanol, 1% aqueous ammonium hydroxide and 90% ethyl acetate.

[0445] Off-white solid. (36 mg, 1.0 mmol, 34.4%). 1 H NMR(500MHz, (CD3)2SO) δ8.79(d,J=3.9Hz,1H),8.26–8.08(m,4H),7.83(dd,J=8.1,3.8Hz,2H),4.59(dd,J=236.2 ,5.5Hz,2H),3.50(t,J=6.2Hz,2H),2.78(t,J=6.2Hz,2H),2.11–2.00(m,2H),1.82(ddt,J=11.9,9.4,4.1Hz,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.6, 155.3, 151.5, 140.5, 137.4, 135.4, 131.3, 130.3, 130.1, 128.1, 128.0, 123.4, 122.5, 120.2, 117.9, 115.2, 33.9, 30.7, 28.3, 28.1, 21.9. HRMS (ESI) m / z calculated value C23 H 21 N8O[M+H] + 425.1833, measured value 425.1831.

[0446] N-((4-methyl-1H-imidazol-2-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0447] Method D: Off-white solid (79 mg, 0.5 mmol, 36.2%). 1 H NMR(500MHz,(CD3)2SO)δ8.58(s,1H),8.09–7.97(m,2H),7.90–7.77(m,2H ),7.68–7.61(m,2H),6.69–6.64(m,1H),4.50(d,J=5.6Hz,2H),2.78(t,J= 6.2Hz,2H),2.12(d,J=1.1Hz,4H),2.05(d,J=16.2Hz,1H),2.01(td,J=9.0 ,7.4,4.7Hz,2H),1.77(tq,J=8.5,6.0,4.3Hz,2H),1.24(d,J=6.8Hz,1H). 13 CNMR(126MHz,(CD3)2SO)δ166.7,156.2,150.0,144.6,144.3,143.9,142.5,133.9,131.2,1 29.6,129.4,129.2,127.6,122.1,120.3,118.0,116.3,37.5,29.7,28.8,22.6,22.5,11.9.

[0448] (S)-N-((Tetrahydrofuran-2-yl)methyl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide

[0449] Synthesized using Method D: off-white solid (98 mg, 0.5 mmol, 45.7%). 1H NMR(500MHz,(CD3)2SO)δ8.62(s,1H),8.55(t,J=5.8Hz,1H),8.00–7.86(m,4H),7.6 9–7.65(m,2H),4.79(d,J=1.8Hz,2H),4.16(t,J=5.9Hz,2H),3.79–3.69(m,2H),3.6 3(td,J=8.0,6.7Hz,2H),3.51(dd,J=8.5,5.4Hz,2H),3.43(d,J=11.9Hz,3H),2.57– 2.49(m,1H),1.97(dtd,J=12.2,8.0,5.6Hz,1H),1.64(ddt,J=12.5,7.7,6.1Hz,1H). 13 C NMR(126MHz,(CD3)2SO)δ166.6,152.5,143.7,141.6,140.6,135.2,131.3,129.7,129.6, 129.3,128.7,127.8,127.7,121.8,116.0,71.1,67.3,66.7,64.4,42.6,31.0,30.1,28.8.

[0450] (R)-N-((Tetrahydrofuran-2-yl)methyl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide

[0451] Synthesized using Method D: off-white solid (121 mg, 0.5 mmol, 56.5%). 1 H NMR(500MHz,(CD3)2SO)δ8.60(s,1H),8.05(d,J=7.9Hz,2H),7.97–7.90(m,2H),7.86(d,J=9.2H z, 1H), 7.69 (dd, J = 17.1, 7.9Hz, 2H), 4.79 (s, 2H), 4.15 (t, J = 5.9Hz, 2H), 3.41 (d, J = 6.0Hz, 3H). 13 C NMR(126MHz,(CD3)2SO)δ167.5,152.8,144.5,144.0,139.6,131.2,129.7,129.5,1 29.4,129.2,127.6,127.5,121.6,116.1,71.1,67.3,66.8,64.4,42.6,30.11,28.7.

[0452] N-(Tetrahydro-2H-pyran-4-yl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide

[0453] Synthesized using Method D: off-white solid (96 mg, 0.5 mmol, 44.8%). 1 H NMR(500MHz, (CD3)2SO) δ8.27(d,J=7.6Hz,1H),7.94(dd,J=31.0,8.4Hz,4H),7.65(d,J=8.2Hz,2H),4.78(s,2H),4.16(t,J=5.9Hz,2H),4.04(t dd,J=13.4,9.3,5.1Hz,1H),3.97–3.83(m,2H),3.41(td,J=11.5,2.3Hz,4H),1.81(ddd,J=12.4,4.7,2.1Hz,2H),1.63(qd,J=11.8,4.4Hz,2H). 13 CNMR(126MHz,(CD3)2SO)δ165.9,156.9,153.1,148.7,144.4,142.4,139.5,139.1,136. 0,135.1,129.6,129.1,127.7,121.5,116.3,115.2,66.9,66.7,64.4,46.4,32.9,28.7.

[0454] N-((3-methyloxetan-3-yl)methyl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide

[0455] Synthesized using Method D: off-white solid (87 mg, 0.5 mmol, 40.6%). 1 H NMR(500MHz,(CD3)2SO)δ8.58(t,J=6.1Hz,1H),8.06–7.79(m,2H),7.77–7.58(m,1H),4.80(s,1H) ,4.52(d,J=5.7Hz,1H),4.28–4.08(m,2H),3.47(dd,J=50.8,5.9Hz,2H),3.18(s,3H),1.29(s,1H). 13CNMR(126MHz,(CD3)2SO)δ167.3,158.7,153.0,144.4,142.5,139.5,139.4,134.9, 129.6,129.2,129.0,127.7,121.5,116.3,80.0,79.8,66.9,64.4,46.2,28.7,22.5.

[0456] N-(2-Hydroxy-2-methylpropyl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide

[0457] Synthesized using Method D: off-white solid (72 mg, 0.5 mmol, 34.6%). 1 H NMR(500MHz,(CD3)2SO)δ8.19(t,J=6.1Hz,1H),8.06–7.78(m,4H),7.74–7.56(m,2H),4 .80 (s, 2H), 4.16 (t, J = 5.9Hz, 2H), 3.36 (dd, J = 43.4, 6.0Hz, 4H), 1.15 (d, J = 1.8Hz, 6H). 13 C NMR(126MHz,(CD3)2SO)δ168.3,154.0,144.4,141.6,141.0,139.7,134.9,131.7,128.7,1 27.5,121.4,126.3,121.3,120.4,117.0,113.6,71.5,67.8,64.4,50.4,28.7,26.5,26.3.

[0458] General synthetic scheme for hydroxamic acid analogs: Solution 1

[0459] General procedure for amide coupling:

[0460] In a 20mL capped vial, the acid intermediate (0.5mmol), O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (0.7mmol) and hexafluorophosphate azabenzotriazole tetramethylurea (HATU) (1.3 equivalents) were dissolved in anhydrous DMF (3mL). DIPEA (5 equivalents) was then added. The reaction was stirred overnight at 80°C. After completion, the reaction was concentrated and purified by silica gel column chromatography using ethyl acetate and hexane (30:70 to 70:30).

[0461] General procedure for deprotection of tetrahydropyranyl ethers:

[0462] In a 5 mL vial, the tetrahydropyranyl protected intermediate (0.25 mmol) was dissolved in dichloromethane (1.5 mL). Then trifluoroacetic acid or hydrochloric acid (2 equivalents) was added. The reaction was refluxed in a capped vial for 2 hours to give the desired deprotected product.

[0463] N-Hydroxy-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSD1993): Solution 2

[0464] According to Scheme 2, solution A was prepared by suspending the substrate ester (0.25 mmol) in methanol (2 mL), and a separate solution B was prepared by dissolving crushed potassium hydroxide (KOH) (10 equivalents) and hydroxylamine hydrochloride (5 equivalents) in methanol (4 mL) and stirring at ambient temperature for 30 minutes. The solution B filtrate was added to solution A and stirring was continued overnight. After completion, the solvent was removed under reduced pressure and the resulting solid was dissolved in water. The pH was adjusted to 7 with aqueous HCl (1N), the resulting precipitate was filtered and dried under high vacuum, and purified by washing and recrystallization from ethanol to obtain the desired product as an off-white solid (85 mg, 24%).

[0465] 1 H NMR(500MHz,DMSO-d6)δ11.31(s,1H),9.09(s,1H),8.57(s,1H),7.90–7.83(m,3H),7.81-7.80(m,1H), 7.63(d,J=8.3Hz,2H),3.34–3.27(m,2H),2.76(t,J=6.1Hz,2H),2.02–1.94(m,2H),1.78–1.67(m,2H); 13 C NMR(126MHz,DMSO-d6)δ164.5,162.7,156.0,143.9,143.7,142.5,138.7,136 .3,132.4,129.5,129.2,127.0,122.0,116.3,114.7,29.6,28.8,22.5,22.4.

[0466] 4-(1-Methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide (Compound HSD19931P)

[0467] Off-white solid (41%); 1 H NMR(500MHz,DMSO-d6)δ11.72(s,1H),7.87(d,J=8.0Hz,2H),7.77–7.67(m,2H),7.62(d,J=7.9Hz,2H),5.03(d,J=3.0Hz,1H), 4.07(t,J=9.7Hz,1H),3.57–3.52(m,1H),3.46–3.35(m,3H),2.83(s,3H),2.75(s,2H),1.82–1.69(m,7H),1.63–1.50(m,3H); 13 C NMR (126MHz, DMSO-d6) δ164.5,155.6,144.3,143.9,143.0,142.5,141.2,131.9,129.9,129.5, 128.4,127.4,123.8,114.8,114.6,101.4,61.8,31.7,28.3,28.0,25.2,22.5,22.1,20.4,18.7.

[0468] N-Hydroxy-4-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSD19931)

[0469] Off-white solid (58%); 1 H NMR (500MHz, DMSO-d6) δ11.40(s,1H),8.00(d,J=9.1Hz,1H),7.94(d,J=7.9Hz,2H),7.83(d,J=9.1Hz,1H),7.73(d, J=7.9Hz,2H),3.55(t,J=5.5Hz,2H),2.91(s,3H),2.76(t,J=5.8Hz,2H),2.48(t,J=2.0Hz,2H),1.89–1.73(m,4H).

[0470] 4-(9,9-dioxido-3,8,10,11-tetrahydropyrazolo[4,3-f]thiopyrano[3,4-c]quinolin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide

[0471] Off-white solid (36%); 1H NMR(500MHz,DMSO-d6)δ11.77(s,1H),8.68(s,1H),8.00(dd,J=9.1,0.9Hz,1H),7.96–7.89(m,3H),7.70–7.62(m,2H),5.08–4.9 8(m,1H),4.54–4.44(m,2H),4.12–3.96(m,3H),3.69(t,J=6.6Hz,2H),3.59–3.51(m,1H),1.80–1.67(m,3H),1.63–1.48(m,3H); 13 C NMR (126MHz, DMSO) δ164.3,155.4,144.6,142.5,139.1,136.6,132.7,129.7,129.5, 127.8,122.4,121.1,116.3,115.9,101.5,61.8,51.7,45.9,30.6,28.3,25.2,18.7.

[0472] 4-(9,9-dioxido-3,8,10,11-tetrahydropyrazolo[4,3-f]thiopyrano[3,4-c]quinolin-7-yl)-N-hydroxybenzamide (Compound 19932)

[0473] Off-white solid (61%); 1 H NMR (500MHz, DMSO-d6) δ11.34(s,1H),9.12(s,1H),8.70(d,J=12.2Hz,1H),8.00(d,J=9.0Hz,1H ), 7.91 (d, J = 8.0Hz, 2H), 7.64 (d, J = 8.0Hz, 2H), 4.50 (s, 2H), 4.01 (s, 2H), 3.69 (t, J = 6.7Hz, 2H).

[0474] 2-Fluoro-N-((tetrahydro-2H-pyran-2-yl)oxy)-5-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0475] Off-white solid (40%); 1H NMR(500MHz,DMSO-d6)δ11.49(s,1H),8.59(s,1H),7.92–7.81(m,1H),7.7 8–7.72(m,1H),7.69(dd,J=6.7,2.3Hz,1H),7.40(t,J=9.9,8.5Hz,1H),5.0 3(s,1H),4.06–3.90(m,1H),3.51(d,J=11.3Hz,1H),3.37–3.32(m,2H),2. 78(t,J=6.1Hz,2H),2.01(t,J=6.0Hz,2H),1.84–1.65(m,5H),1.53(s,3H).

[0476] 2-Fluoro-N-hydroxy-5-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSD19933)

[0477] Off-white solid (52%); 1 H NMR (500MHz, DMSO-d6) δ11.12(s,1H),8.73(s,1H),8.06(d,J=9.2Hz,1H),7.92(d,J=9.1Hz,1H),7.85–7.78(m, 2H),7.49(t,J=9.4Hz,1H),3.42(t,J=6.4Hz,2H),2.78(t,J=6.1Hz,2H),2.12–1.99(m,2H),1.87–1.73(m,2H).

[0478] 4-(9-methyl-3H-pyrazolo[4,3-f]quinolin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide

[0479] Off-white solid (39%); 1 H NMR(500MHz,DMSO-d6)δ11.74(s,1H),8.58(s,1H),8.37(d,J=8.1Hz,2H),8.21(s,1H),8.00–7.88(m,4H),5.11 –4.92(m,1H),4.09(d,J=11.5Hz,1H),3.54(dd,J=9.7,5.4Hz,1H),2.97(s,3H),1.73(s,3H),1.60–1.49(m,3H); 13C NMR (126MHz, DMSO) δ164.4,152.2,146.1,144.6,142.0,138.5,136.1,132.9,129. 8,128.2,127.1,122.2,120.9,116.9,115.7,101.5,61.8,28.4,25.2,22.8,18.8.

[0480] N-Hydroxy-4-(9-methyl-3H-pyrazolo[4,3-f]quinolin-7-yl)benzamide (Compound 19934)

[0481] Off-white solid (60%); 1 H NMR (500MHz, DMSO-d6) δ11.36(s,1H),8.64(s,1H),8.32(d,J=8.4Hz,2H),8.23(s,1H),8.04–7.96(m,2H),7.94(d,J=8.4Hz,2H),2.98(s,3H).

[0482] 4-(9-Cyclobutyl-3H-pyrazolo[4,3-f]quinolin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide (Compound HSD19935P)

[0483] Off-white solid (53%); 1 H NMR(500MHz,Methanol-d4)δ8.57(s,1H),8.25(d,J=8.1Hz,2H),8.07(s,1H),8.02(d,J =8.5Hz,1H),7.96(d,J=8.1Hz,2H),7.89(d,J=9.2Hz,1H),5.11(d,J=2.9Hz,1H),4.43(s ,1H),4.17(t,J=10.7Hz,1H),3.72–3.53(m,1H),2.80(s,2H),2.76–2.67(m,2H),2.40–2 .31(m,2H),2.29–2.20(m,1H),2.11–2.01(m,1H),1.96–1.78(m,2H),1.74–1.56(m,2H).

[0484] 4-(9-Cyclobutyl-3H-pyrazolo[4,3-f]quinolin-7-yl)-N-hydroxybenzamide (Compound HSD19935)

[0485] Off-white solid (55%); 1 H NMR(500MHz,DMSO-d6)δ11.35(s,1H),8.65(s,1H),8.39(d,J=8.4Hz,2H),8.19(s,1H),7.97–7.90(m,4 H), 4.44 (p, J = 8.1Hz, 1H), 2.71–2.57 (m, 2H), 2.41–2.29 (m, 2H), 2.27–2.08 (m, 1H), 2.03–1.85 (m, 1H).

[0486] N-Hydroxy-4-(1-methyl-9-morpholino-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSD19936)

[0487] Yellow solid (68%); 1 H NMR (500MHz, DMSO-d6) δ11.44(s,1H),11.04(s,1H),7.95(d,J=8.1Hz,3H),7.84(d,J=9.0Hz,1H),7.73(d,J=7.9Hz,2H),4.0 1–3.94(m,2H),3.82–3.72(m,5H),3.55–3.51(m,2H),3.32–3.17(m,4H),3.08–3.00(m,1H),2.90(s,3H),1.88–1.82(m,1H).

[0488] 4-(5-Fluoro-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-hydroxybenzamide (Compound HSD19937)

[0489] Off-white solid (59%); 1 H NMR (500MHz, DMSO-d6) δ11.33(s,1H),8.61(s,1H),7.87(d,J=8.3Hz,2H),7.71(d,J=10.0Hz,1H),7. 67–7.60(m,2H),3.30(t,J=6.3Hz,2H),2.78(t,J=6.1Hz,2H),2.03–1.94(m,2H),1.82–1.68(m,2H).

[0490] N-methoxy-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSD199310)

[0491] Off-white solid (65%); 1 H NMR(500MHz,DMSO-d6)δ11.83(s,1H),8.57(s,1H),7.97–7.78(m,4H),7.66(d,J=8.2Hz,2H), 3.74(s,3H),3.36–3.32(m,2H),2.76(t,J=6.1Hz,2H),2.08–1.90(m,2H),1.82–1.47(m,2H); 13 C NMR (126MHz, DMSO) δ164.3,156.0,144.4,143.6,142.6,138.7,136.4,131. 9,129.6,129.3,127.2,122.0,116.4,114.5,63.7,29.6,28.8,22.5,22.4.

[0492] 5-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)picolinamide

[0493] Off-white solid (57%); 1 H NMR (500MHz, DMSO-d6) δ11.90(s,1H),8.79(d,J=2.2Hz,1H),8.19(dd,J=8.0,2.1Hz,1H),8.09(d,J=8.0Hz,1H),7.76(s,1 H),5.11(d,J=2.7Hz,1H),4.24–4.11(m,1H),3.54–3.36(m,4H),2.95–2.75(m,6H),1.84–1.66(m,7H),1.62–1.52(m,3H); 13 C NMR (126MHz, DMSO) δ161.5,152.8,149.1,149.0,144.1,143.1,142.8,141.3,139.4,138.5,12 9.9,128.8,122.0,115.1,114.5,100.9,61.4,31.7,28.2,27.9,25.2,22.4,22.1,20.4,18.4.

[0494] N-Hydroxy-5-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)picolinamide (Compound HSD199312)

[0495] Off-white solid (70%); 1 H NMR (500MHz, DMSO-d6) δ11.58(s,1H),8.84(s,1H),8.26(dd,J=8.1,2.2Hz,1H),8.13(d,J=8.0Hz,1H),7.94( d,J=9.2Hz,1H),7.80(d,J=9.1Hz,1H),3.64–3.41(m,2H),2.90(s,3H),2.84–2.74(m,2H),1.99–1.72(m,4H).

[0496] 6-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)nicotinamide

[0497] Off-white solid (50%); 1 H NMR (500MHz, DMSO-d6) δ11.91(s,1H),9.07–8.84(m,1H),8.27(dd,J=8.2,2.3Hz,1H),7.91(d,J=8.2Hz,1H),7.80–7.55(m,1H),5.05(s, 1H),4.14–3.98(m,1H),3.56(dd,J=10.2,5.2Hz,1H),3.42(s,2H),3.05–2.94(m,2H),2.86(s,3H),1.86–1.66(m,7H),1.64–1.45(m,3H).

[0498] N-Hydroxy-6-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)nicotinamide (HSD199313)

[0499] Off-white solid (61%); 1H NMR (500MHz, DMSO-d6) δ11.52(s,1H),9.03(d,J=2.2Hz,1H),8.31(dd,J=8.1,2.3Hz,1H),7.96(d,J=8.1Hz,1H),7 .89(d,J=9.1Hz,1H),7.79(d,J=9.1Hz,1H),3.66–3.24(m,2H),2.99–2.94(m,2H),2.89(s,3H),1.97–1.48(m,4H).

[0500] N-Hydroxy-4-(3,8,10,11-tetrahydropyrazolo[4,3-f]thiopyrano[3,4-c]quinolin-7-yl)benzamide (Compound HSD199315)

[0501] Off-white solid (56%); 1 H NMR(500MHz,DMSO-d6)δ11.38(s,1H),8.73(s,1H),8.03(d,J=9.1Hz,1H),7.96–7.91(m,2H),7 .89(d,J=9.1Hz,1H),7.76–7.70(m,2H),3.88(s,2H),3.68–3.62(m,1H),3.17(t,J=6.1Hz,2H).

[0502] 4-(5-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide

[0503] Off-white solid (38%); 1 H NMR(500MHz,DMSO-d6)δ8.52(s,1H),7.90–7.81(m,2H),7.77–7.73(m,1H),7.72–7.69(m,2H),5.09–4.97(m,1H),4.15–4. 00(m,1H),3.60–3.49(m,1H),2.81(t,J=6.1Hz,2H),2.69(s,3H),2.08–1.96(m,2H),1.79–1.60(m,5H),1.60–1.49(m,3H).

[0504] N-Hydroxy-4-(5-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSD199316)

[0505] Off-white solid (62%); 1 H NMR(500MHz,DMSO-d6)δ11.30(s,1H),8.53(s,1H),7.90–7.83(m,2H),7.75(s,1H),7.73–7.6 6(m,2H),3.32(t,J=6.6Hz,2H),2.81(t,J=6.1Hz,2H),2.08–1.96(m,3H),1.81–1.68(m,3H).

[0506] 4-(5-Methoxy-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide

[0507] Off-white solid (51%); 1 H NMR (500MHz, DMSO-d6) δ11.73 (s, 1H), 8.46 (d, J = 7.2 Hz, 1H), 7.88 (d, J = 8. 3Hz,2H),7.65–7.59(m,2H),7.17(s,1H),5.04(d,J=3.0Hz,1H),4.12–4.04 (m,1H),3.96(s,3H),3.55(dd,J=9.4,5.9Hz,1H),3.38–3.29(m,2H),2.74 (t,J=6.0Hz,2H),2.03–1.94(m,2H),1.77–1.71(m,5H),1.60–1.52(m,3H).

[0508] N-Hydroxy-4-(5-methoxy-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSD199317)

[0509] Off-white solid (70%); 1 H NMR(500MHz,DMSO-d6)δ11.35(s,1H),8.52(s,1H),8.00–7.80(m,2H),7.73–7.56(m,2H),7.29(s,1H),3.9 9(d,J=1.4Hz,3H),3.32(t,J=6.4Hz,2H),2.72(t,J=6.1Hz,2H),2.19–1.94(m,2H),1.75(q,J=5.5Hz,2H). Example 2 Evaluation of CDK inhibitor compounds in cancer cells

[0510] Molm-14 (human acute myeloid leukemia (AML) cell line), Molm-14 D835Y (a human AML cell line with a mutation at D835Y), Molm-14 F691L (a human AML cell line with a mutation at F691L, which is a common cause of acquired FLT3 inhibitor resistance), SNU-16 (a human gastric cancer cell line) and KCL22-IR (a chronic myeloid leukemia cell line) were cultured in RPMI-1640 medium containing L-glutamine and sodium bicarbonate, supplemented with 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin. The cells were seeded in 96-well plates at 2.1E4 cells / mL, 190 μL per well. The plates were incubated in a 5% CO2 incubator for 24 hours. The cells were then treated with the desired concentration of the compound herein or dimethyl sulfoxide (DMSO) control and incubated in a 5% CO2 incubator for 72 hours.

[0511] Then, the cells were treated with CellTiter-Blue Viability Assay, incubated in a 5% CO2 incubator for 3 hours, and then the absorbance was measured at 570 nm using a Cytation 5 plate reader. The data were analyzed using GraphPad prism. Figure 3-10C The results are shown. Example 3 Structural analysis of the docking of compound HSD1217 with Haspin

[0512] To optimize compounds for dual FLT3 and haspin kinase inhibition, the docking scores of several dual FLT3 / haspin inhibitor compounds were evaluated.Determining the three-dimensional structure of the protein-receptor complex is an important component of structure-based drug design.

[0513] Briefly, the crystal structure of haspin (PDB: 2WB8) was downloaded from the Protein Data Bank and edited to remove any existing bound ligands within the active site of the protein. A two-dimensional ligand was constructed in ChemDraw 21.0.0 and transferred to a three-dimensional mol2 file using OpenBabel. Docking was performed using Glide software. Waters were removed prior to docking to haspin. Validation of the docking experiment with haspin kinase was based on the previously generated docking pose for HSD972. Visualization was performed using PyMol and Maestro.

[0514] The docking structure of compound HSD1217 and haspin was analyzed ( Fig.11A ) shows that the benzamide-NH2 part has a hydrogen bond interaction with the key residue D687 and is located in a region containing H651 and D649 residues, which are partially bound to the ATP phosphate. Therefore, polar groups (such as imidazole) with hydrogen bond donor ability attached to benzamide can reach H651 or D649 residues while maintaining hydrogen bonds with D687 residues to provide additional interactions and increase ligand affinity for proteins. Since H651, D649 or D687 are essential for the binding of ATP to haspin, it is speculated that inhibitors bound to these residues will have a higher chance of resisting kinase mutations that can eliminate inhibitor binding. Eswaran et al., Structure and functional characterization of the atypical human kinase haspin, Proceedings National Academy Science USA 106(48): 20198-20203(2009).

[0515] Computer simulation analysis of compounds 4, 5 and 6 (analogs of compound HSD1217) containing imidazole moieties, respectively, showed that all three compounds can form key interactions with at least one ATP binding residue ( Figures 11B-11D ). For docking analysis of 3H-pyrazolo[4,3-f]quinoline compounds with FLT3, see Dayal et al. (2021), supra. Example 4 Kinase assays and structure-activity relationship studies

[0516] Using the preliminary compounds identified in Example 3, structure-activity relationship (SAR) studies were performed to explore how modifications affect haspin and / or FLT3 inhibition, and new imidazole-containing pyrazole-based compounds (described herein) were identified that are dual FLT3 / haspin inhibitors that inhibit both kinases at ultra-low (subnanomolar) concentrations.

[0517] Multicomponent reactions (MCR) are often used to rapidly synthesize large numbers of highly functionalized analogs in a single pot for the construction of bioscreening libraries. Here, compounds were synthesized using a Doebner-like MCR followed by amidation (Scheme 3). Solution 3 Analogs were synthesized by Doebner-like MCR followed by amidation (a) (i) EtOH, reflux, 2 h and (ii) ketone (2 eq), catalytic HCL, reflux, 12 h; (b) HATU, DIPEA, DMF, 60 °C, 12 h.

[0518] For the initial test library, amines containing imidazole or its bioisostere were the focus, as it is thought that such heterocyclic amines (present in other drugs such as nilotinib) may promote ligand binding to protein residues, see above. In a Doebner-like MCR process, amines and aldehydes react to form a Schiff base, which subsequently reacts with an added ketone in the presence of an acid catalyst (HCl).

[0519] After obtaining the MCR product A (see Scheme 3), the final amide product (5-18) was formed by amidation reaction using commercially available amines, HATU (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine 3-oxide hexafluorophosphate, azabenzotriazole tetramethyl urea hexafluorophosphate) and Hünig's base (DIPEA).

[0520] After establishing the library of amide kinase inhibitors, kinase inhibition and AML cell line (Molm-14, containing FLT3-ITD) growth inhibition were analyzed (Table 1). More specifically, Haspin and FLT3 inhibition were evaluated internally by ADP Glo kinase assay (Promega, Madison, WI). All compounds were screened for kinases and Molm-14 cells at a concentration of 50 nM, and HSD1169 and gilteritinib were used as positive controls. The reported values ​​are the average of three replicates.

[0521] Briefly, the ADP-Glo ​​Kinase Assay Kit was used to measure haspin inhibition. Kinase buffer (SignalChem Cat#K01-09; 25 mM MOPS, pH 7.2, 12.5 mM β-glycerophosphate, 25 mM MgCl2, 2 mM EDTA) was added with dithiothreitol (DTT) (0.2 mM) and bovine serum albumin (BSA) (40 μg / mL) immediately before use. The reaction volume was 5 μL, containing 9 ng / μL kinase, 10 μM ATP, 50 nM inhibitor and appropriate peptide substrate (20 μM), incubated for 3 hours, and then quenched with an equal volume (5 μL) of GLO reagent. After 40 minutes, the reaction was performed on a Cytation TM 10 μL of DET reagent was added and incubated for 1 hour before visualization on a 5-cell imaging multimode reader (BioTek). Table 1. Structure, kinase inhibition and cell growth inhibition data of the evaluated compounds.

[0522] SAR studies support the notion that the imidazole ring is critical for the inhibition of haspin and FLT3 activity, as even subtle modifications can significantly affect kinase inhibition. Compounds 4, 5, and 6 all contain an imidazole moiety, albeit in different substitution patterns; however, their inhibitory properties against the tested kinases are different.

[0523] Compound 6 (2-substituted imidazole) showed better inhibitory effects on haspin and FLT3 (77% and 76%, respectively) than compound 5 (1-substituted imidazole) (33% and 31%, respectively) and compound 4 (4-substituted imidazole) (54% and 49%, respectively).

[0524] Since compound 6 showed the best inhibition against the kinases tested, the substitution pattern on compound 6 was investigated to potentially improve potency. Replacing the -NH on compound 6 with -NMe (as in compound 7) reduced haspin and FLT3 inhibition as well as AML growth inhibition. For example, at 50 nM, compound 6 inhibited haspin by 77%, while the -NMe analog (compound 7) inhibited the same enzyme by 26%. For FLT3, the potency dropped from 76% inhibition for compound 6 at 50 nM to 14% inhibition for compound 7. Replacing the imidazole unit in compound 6 with other 5-membered bioisosteres (triazole (compound 8), tetrazole (compound 10), or thiazole (compound 13)) resulted in compounds that inhibited both kinases and AML cell proliferation, but the effects were generally less than compound 6. Extending the carbon chain length between the amide and imidazole by one unit to give compound 15 had no significant effect on the inhibition of either kinase.

[0525] The effect of fluorine substitution on lead compounds is routinely investigated, as it is well known that fluorine can be used to modulate drug properties (target binding as well as general drug properties such as metabolic stability). Therefore, the benzamide moiety of compound 6 was fluorinated to give compounds 9 and 11; however, FLT3 and haspin kinase inhibition were not significantly affected (comparing compounds 9 and 11 with compound 6, Table 1).

[0526] Some imidazole-containing drugs, such as nilotinib, have their imidazole units methylated at position 4. Therefore, the extent to which the methylation of the imidazole unit in compound 6 affects kinase inhibition was also investigated. Methylation of the imidazole unit significantly increased the potency against haspin, FLT3, and Molm-14 cell proliferation. Compound 14 (also known as compound HSK205) inhibited haspin and FLT3 by 97% and 94%, respectively, at only 50 nM, indicating that compound HSK205 is a super potent inhibitor of FLT3 and haspin (see below). In fact, at a concentration of only 50 nM, compound HSK205 almost completely inhibited (96%) the growth of Molm-14 (FLT3-ITD) cells.

[0527] To validate the laboratory results, compound HSK205 was sent to the contract research organizations Eurofins and Reaction Biology to obtain binding data for haspin (Eurofins) and enzyme inhibition data for FLT3 (Reaction Biology).

[0528] The K of compound HSK205 binding to haspin d =0.55 nM (K of the haspin inhibitor CHR-6494) d <0.17nM)( Fig. 12A ).

[0529] For FLT3 inhibition, at 10 μM ATP, compound HSK205 has an IC of 2.0 for both FLT3-ITD and FLT3-D835Y. 50 <0.5nM, while under similar conditions, gilteritinib has IC 50 =0.7nM and IC 50 <0.5nM( Figures 12B-12C ).

[0530] These SAR studies confirm previous reports that the 3H-pyrazolo[4,3-f]quinoline moiety can target FLT3 and that this moiety is highly tunable as substitutions at the C7 position can be used to target other kinases. For example, compound HSD1169, also a 3H-pyrazolo[4,3-f]quinoline-containing compound with a fluorinated pyrazole, is a strong inhibitor of FLT3 but a weak inhibitor of haspin, whereas changing the C7 moiety to a benzamide yields a dual FLT3 / haspin inhibitor. Example 5 Target binding and in vitro biological activity

[0531] We then assessed whether the compound HSK205 indeed inhibited haspin in AML cells, and performed immunoblotting analysis to confirm the inhibition of histone H3 (Thr3) phosphorylation by haspin ( Fig.13A ). Briefly, Molm-14 cells were treated with compound HSK205 (5 nM or 20 nM or 100 nM) or DMSO control or gilteritinib (negative control) at a higher concentration of 100 nM and harvested 24 hours after treatment. The samples were then analyzed for phosphorylation of histone H3 (Thr3).

[0532] More specifically, cells were cultured in a medium supplemented with 10% fetal bovine serum and 1× penicillin-streptomycin (Gibco TM ) in RPMI medium until confluence. Cells were plated at 5000 cells / well in a 96-well plate. After incubation at 37°C in a 5% CO2 incubator for 24 hours, compounds diluted in the same medium were added to the indicated final concentrations and incubated at 37°C in a 5% CO2 incubator for 72 hours. Cell viability was determined by cell viability assay (Promega, Madison, WI) and the cells were plated on a Cytator 3 h after addition. TM The absorbance was read at 570 nm on a 5-cell imaging multi-function reader (BioTek). Appropriate vehicle and no-cell controls were included. 50 The readings were confirmed by two scientists on two different days.

[0533] Acute myeloid leukemia cells (Molm-14) were incubated with the specified compounds at different concentrations. Cells were collected and lysed, and protein concentrations were quantified according to the BSA standard. After protein denaturation, proteins were separated by SDS-PAGE electrophoresis and transferred to NC membranes. After blocking, incubation with specific primary antibodies overnight, incubation with peroxidase-coupled secondary antibodies at room temperature for 2 hours, and peroxidase activity was detected using a CCD camera LAS-4000 (Fujifilm) with SuperSignal West Pico reagent (ThermoScientific). Bands were standardized using the housekeeping gene actin. The reported values ​​represent the mean of three replicates, and the error bars represent SD. All specific antibodies were purchased from Cell Signaling.

[0534] The results showed that compound HSK205 could effectively reduce the phosphorylation of histone H3 (Thr3), the effect of 20nM was more significant than 5nM, and the effect of 100nM was more significant than 20nM ( Fig. 13B ). However, gilteritinib did not show haspin inhibition in Molm-13 cells.

[0535] In addition, FLT3 target binding was assessed in Molm-14 cells to confirm the FLT3 inhibition observed in the preliminary kinase screen of Example 4 (Table 1). Cells were treated for 2 hours, harvested, and then analyzed. Administration of compound HSK205 to Molm-14 cells resulted in a decrease in phosphorylated FLT3 ( Fig.14A and 14B ).

[0536] Many FLT3 inhibitors have been tested in the clinic, and some, such as gilteritinib, midostaurin, and quizartinib, have been approved; however, targeted FLT mutations that reduce AML sensitivity to these drugs have been observed. Schollet al. (2020), supra. Therefore, the compound HSK205 was evaluated against the Molm-14 cell line FLT3-ITD-D835Y and F691L, which have resistance mutations commonly seen in the clinic ( Fig.15 ). Notably, compound HSK205 exhibited low nanomolar GI against Molm-14-ITD-F691L (2.2 nM) and Molm14-ITD-D835Y (4.3 nM). 50 value.

[0537] To advance further compound development, DNA intercalation assays were performed to exclude compounds with high affinity for DNA. Compound HSK205 did not intercalate into DNA, despite containing multiple rings that could enhance DNA intercalation and unwanted toxicity. Fig.16 Under similar experimental conditions, mitoxantrone (a known DNA intercalator) did bind to DNA ( Fig.16 ). Example 6 Chemical properties and synthesis of compound HSK205 and its analogs

[0538] Regarding Examples 3-5, unless otherwise noted, all reagents and solvents were purchased from commercial sources and used as received. 1 H and 13 C NMR spectra were obtained in DMSO-d6 as solvent using a 500 MHz spectrometer or an 800 MHz spectrometer with Me4Si as internal standard. Chemical shifts are reported in parts per million (δ) and are calibrated using residual undeuterated solvent as internal reference.

[0539] 1H NMR spectral data are reported as follows: chemical shift (δ ppm) (multiplicity, coupling constant (Hz), integration). Multiplicity is reported as follows: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, or combinations thereof. High resolution mass spectra (HRMS) were obtained using an electron spray ionization (ESI) technique and a TOF mass spectrometer. New compounds were identified by 1 HNMR, 13 CNMR and HRMS data were used for characterization, and HPLC was used to report the purity of the final compounds.

[0540] 4-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid (Compound 1)

[0541] In a round-bottom flask equipped with a stirring bar, the appropriate indazole (1 equivalent) and the appropriate aldehyde (1 equivalent) were dissolved in ethanol. The mixture was refluxed at 120°C for 2 hours, and then the appropriate ketone (2 equivalents) and catalytic amount of hydrochloric acid were added. It was stirred for another 12 hours under reflux. The reaction solvent was evaporated using a rotary evaporator, and purified by column chromatography using 10% methanol in ethyl acetate to obtain compound 1 as an off-white solid (2.38 grams, 69.4%).

[0542] 1 H NMR(500MHz,(CD3)2SO)δ8.75(s,1H),8.26(d,J=9.2Hz,1H),8.19(d,J=9.2Hz,1H),8.15–8.11(m,2H),7.85 –7.80(m,2H),3.46(t,J=6.5Hz,2H),2.76(t,J=6.1Hz,2H),2.03(dd,J=6.0,2.9Hz,2H),1.83–1.76(m,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 167.17, 151.31, 151.03, 140.41, 138.09, 137.05, 134.78, 132.69, 131.32, 130.42, 129.69, 123.45, 122.02, 120.11, 115.06, 30.67, 28.05, 21.85, 21.72. HRMS (ESI) m / z calculated value C 21 H 17 N3O2[M+H] + 344.1393, measured value 344.1392.

[0543] 3-Fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid (Compound 2)

[0544] Compound 2 was obtained according to the same synthetic procedure as compound 1 as an off-white solid (636 mg, 58.7%). 1 HNMR(500MHz, (CD3)2SO) δ8.64(s,1H),8.06(d,J=22.2Hz,3H),7.72(d,J=11.2Hz,1H),7.63(d,J= 8.1Hz, 1H), 3.31 (t, J = 6.6Hz, 2H), 2.76 (t, J = 6.2Hz, 2H), 2.04–1.91 (m, 2H), 1.76 (q, J = 6.0Hz, 2H). 13 C NMR(126MHz,(CD3)2SO)δ167.55,165.09,162.00,159.95,157.23,150.10,132 .37,131.56,130.83,126.27,123.27,123.14,121.70,120.78,120.70,118.99( 2 J = 22 Hz), 115.01, 30.42, 28.01, 21.88, 21.75. HRMS (ESI) m / z calculated value C 21 H 16 FN3O2[M+H] + 362.1299, measured value 362.1300.

[0545] 2-Fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid (Compound 3)

[0546] Compound 3 was obtained according to the same synthetic procedure as compound 1 as an off-white solid (556 mg, 51.3%). 1 HNMR(500MHz, (CD3)2SO) δ8.76(d,J=8.6Hz,1H),8.16(d,J=14.3Hz,2H),8.04(t,J=7.7Hz,1H),7.67(d,J=11.1Hz,1 H), 7.60 (d, J = 8.1Hz, 1H), 3.47 (t, J = 6.6Hz, 2H), 2.80 (t, J = 6.2Hz, 2H), 2.05 (p, J = 6.0Hz, 2H), 1.81 (p, J = 5.8Hz, 2H). 13C NMR(126MHz,(CD3)2SO)δ165.04,162.04,159.99,150.59,140.43,138.54, 134.91,132.33,130.91,126.13,123.73,123.41,120.87,119.44,118.75( 2 J = 20 Hz), 115.32, 109.16, 30.48, 28.04, 21.99, 21.86. HRMS (ESI) m / z calculated value C 21 H 16 FN3O2[M+H] + 362.1299, measured value 362.1301.

[0547] N-((1H-imidazol-4-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 4)

[0548] The acidic intermediate (1.3 eq.) and HATU (1.5 eq.) were sealed with a septum in a 20 mL screw cap vial equipped with a stirring bar, the air was evacuated by a vacuum pump, and a nitrogen balloon was added to the reaction vessel to fill the vessel with an inert gas. Anhydrous DMF (2 mL) and DIPEA (8 eq.) were added to the reaction vial. The mixture was heated at 60 ° C for 30 minutes, and then the amine (1 eq.) was added. The reaction was stirred at 60 ° C overnight.

[0549] The reaction solvent was evaporated using a rotary evaporator and purified by column chromatography using 1% ammonium hydroxide, 9% methanol and 90% ethyl acetate to give compound 4 as an off-white solid (111 mg, 52.6%).

[0550] 1 H NMR (500MHz, (CD3)2SO) δ8.81(t,J=5.6Hz,1H),8.57(s,1H),7.99(d,J=8.2Hz,2H),7.94–7.76(m,2H),7.67–7.63(m,2H),7.62(s,1H) ,6.98(s,1H),4.46(d,J=5.6Hz,2H),3.32(t,J=6.5Hz,2H),2.76(t,J=6.1Hz,2H),2.03–1.95(m,2H),1.74(dp,J=9.1,3.2,2.6Hz,2H). 13C NMR (126 MHz, (CD3)2SO) δ 166.39, 156.17, 144.01, 143.85, 142.53, 135.85, 135.21, 134.22, 129.65, 129.56, 129.42, 129.24, 127.47, 122.11, 117.09, 116.30, 36.81, 29.65, 28.83, 22.58, 22.50. HRMS (ESI) m / z calculated value C 25 H 22 N6O[M+H] + 423.1928, measured value 423.1911.

[0551] N-(2-(1H-imidazol-1-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 5)

[0552] Compound 5 was obtained according to the same synthetic procedure as compound 4 as an off-white solid (84 mg, 38.5%). 1 HNMR(800MHz, (CD3)2SO) δ8.74(t,J=5.5Hz,1H),8.57(s,1H),7.93(d,J=7.8Hz,2H),7.89(d,J=8.9Hz,1H),7.86–7.80(m,1H),7.65(d,J=7.8Hz,2H ),7.63(s,1H),7.18(s,1H),6.89(s,1H),4.22(t,J=6.2Hz,2H),3.63(q,J =5.9Hz,2H),2.78(t,J=6.2Hz,2H),2.06–1.99(m,2H),1.80–1.73(m,2H). 13 C NMR (201 MHz, (CD3)2SO) δ 166.82, 156.07, 144.98, 144.06, 142.49, 138.75, 137.79, 136.28, 133.87, 129.51, 129.39, 128.73, 127.30, 126.85, 119.97, 118.38, 116.37, 114.52, 45.60, 40.91, 29.60, 28.76, 22.50, 22.41. HRMS (ESI) m / z calculated value C 26 H 24 N6O[M+H] + 437.2084, measured value 437.2081.

[0553] N-((1H-imidazol-2-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 6)

[0554] Compound 6 was obtained according to the same synthetic procedure as compound 4 as an off-white solid (65 mg, 30.8%). 1 HNMR(800MHz, (CD3)2SO) δ9.17(t,J=5.9Hz,1H),8.57(s,1H),8.07(d,J=7.8Hz,2H),7.89(d,J=9.0Hz,1H),7.82(d,J=9.3Hz,1H), 7.65(d,J=7.8Hz,2H), 6.93(s,2H), 4.56(d,J=5.6Hz,2H), 2.77(t,J=6.3Hz,2H), 2.00(q,J=6.8,6.3Hz,2H), 1.75(p,J=5.6Hz,2H). 13 CNMR (201 MHz, (CD3)2SO) δ 166.48, 156.04, 146.73, 145.52, 144.05, 143.70, 142.46, 138.80, 136.15, 133.75, 129.47, 129.33, 129.16, 127.57, 127.50, 122.02, 119.08, 116.22, 114.55, 37.50, 29.58, 28.75, 22.49, 22.41. HRMS (ESI) m / z calculated value C 25 H 22 N6O[M+H] + 423.1927, measured value 423.1926.

[0555] N-((1-methyl-1H-imidazol-2-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 7)

[0556] Compound 7 was obtained according to the same synthetic procedure as compound 4 as a light yellow solid (68 mg, 31.2%). 1HNMR(500MHz, (CD3)2SO) δ8.56(s,1H),8.37(d,J=4.3Hz,3H),8.16(d,J=8.3Hz,3H),7.04(d,J=13.1Hz,1H),6.78(d,J=18.9Hz,1H) ,4.56(d,J=5.3Hz,2H),3.68(s,3H),3.58(s,2H)2.77(t,J=6.2Hz,3H),1.88(dp,J=127.7,5.7Hz,6H),1.23(dd,J=14.7,7.0Hz,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.48, 156.10, 147.20, 144.21, 142.53, 139.90, 134.92, 133.82, 129.52, 129.41, 129.19, 127.72, 127.58, 126.87, 122.20, 119.24, 36.04, 32.94, 29.66, 28.80, 22.59, 22.52. HRMS (ESI) m / z calculated value C 26 H 25 N6O[M+H] + 437.2084, measured value 437.2081.

[0557] N-((4H-1,2,4-triazol-3-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 8)

[0558] Compound 8 was obtained according to the same synthetic procedure as compound 4 as an off-white solid (79 mg, 37.4%). 1 HNMR(500MHz,(CD3)2SO)δ8.58(s,1H),8.43–8.06(m,2H),8.01(d,J=7.8Hz,2H),7.84(q,J=9.1Hz,2H),7.65(d,J =7.8Hz, 2H), 4.61 (d, J = 5.6Hz, 2H), 3.34 (d, J = 13.1Hz, 2H), 2.78 (t, J = 6.1Hz, 2H), 1.89 (dp, J = 125.7, 6.1Hz, 4H). 13C NMR (126 MHz, (CD3)2SO) δ 166.66, 161.54, 156.17, 148.63, 144.20, 143.89, 142.55, 133.95, 129.66, 129.41, 129.24, 127.56, 122.14, 119.78, 116.29, 36.76, 29.66, 28.81, 22.59, 22.53. HRMS (ESI) m / z calculated value C 24 H 21 N7O[M+H] + 424.1880, measured value 424.1877.

[0559] N-((1H-imidazol-2-yl)methyl)-3-fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 9)

[0560] Compound 9 was obtained according to the same synthetic procedure as compound 4 as an off-white solid (109 mg, 49.5%). 1 HNMR(500MHz,(CD3)2SO)δ8.59(s,1H),7.93–7.78(m,4H),7.57(t,J=7.6Hz,1H),7.16(s,2 H), 4.65 (d, J = 5.5Hz, 2H), 3.35 (t, J = 6.5Hz, 2H), 2.62 (t, J = 6.2Hz, 2H), 2.04–1.74 (m, 4H). 13 C NMR(126MHz,(CD3)2SO)δ165.63,161.85,159.52( 1 J=246.4Hz),151.59,151.13,145.34,144.18,143.97,142.50,136.45,131.9 8,131.87,129.97,129.47,129.06,124.10,122.51,121.17,116.22,115.16( 2 J = 20 Hz), 36.92, 29.47, 27.37, 22.56, 22.17. HRMS (ESI) m / z calculated value C 25 H 21 FN6O[M+H] + 441.1833, measured value 441.1830.

[0561] N-((1H-tetrazolyl-5-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 10)

[0562] To a solution of alkyne (1 eq.) in DMF (2 mL) and water (0.5 mL) was added TMS-azide (2 eq.). Catalytic amounts of copper sulfide pentahydrate and sodium ascorbate were added. The reaction mixture was stirred at 100 °C overnight. The reaction solvent was evaporated off using a rotary evaporator, and the residue was diluted with ethyl acetate, washed with water, and dried over sodium sulfate. It was then purified by column chromatography using 90% ethyl acetate and 10% hexane to give an off-white solid (36 mg, 34%).

[0563] 1 H NMR(500MHz, (CD3)2SO) δ9.43(dt,J=33.1,5.5Hz,1H),8.79(d,J=3.9Hz,1H),8.33–8.08(m,4H),7.83(dd,J=8.1,3.9Hz,2H) ,4.82(d,J=5.6Hz,2H),3.50(t,J=6.2Hz,2H),2.78(t,J=6.2Hz,2H),2.15–1.98(m,2H),1.82(ddt,J=11.9,9.4,4.1Hz,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.55, 155.31, 151.46, 140.50, 137.36, 135.41, 131.33, 130.29, 130.13, 128.06, 128.01, 123.41, 122.54, 120.16, 117.93, 115.22, 33.87, 30.66, 28.29, 28.13, 21.92, 21.80. HRMS (ESI) m / z calculated value C 23 H 20 N8O[M+H] + 425.1833, measured value 425.1831.

[0564] N-((1H-imidazol-2-yl)methyl)-2-fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 11)

[0565] Compound 11 was obtained according to the same synthetic procedure as compound 4 as an off-white solid (87 mg, 39.5%). 1HNMR(500MHz, (CD3)2SO) δ8.82(q,J=4.9Hz,1H),8.55(s,1H),7.83(dq,J=22.1,9.0,7.4Hz,3H),7.50(t,J=10.4Hz,2H),6.97 (s, 2H), 4.55 (d, J = 5.6Hz, 2H), 3.25 (t, J = 6.7Hz, 2H), 2.76 (t, J = 6.1Hz, 2H), 1.96 (p, J = 6.5, 6.1Hz, 2H), 1.72 (p, J = 5.7Hz, 2H). 13 C NMR(126MHz,(CD3)2SO)δ164.00,161.89,160.52( 1 J=249.4Hz),154.65,149.62,145.80,145.73,145.06,143.72,142.71,1 38.50,130.58,129.58,129.18,125.69,122.93,122.82,122.25,117.29( 2 J = 23 Hz), 116.18, 37.66, 29.62, 28.70, 22.48, 22.40. HRMS (ESI) m / z calculated value C 25 H 21 FN6O[M+H] + 441.1873, measured value 441.1837.

[0566] N-((1H-pyrrol-3-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 12)

[0567] Compound 12 was obtained according to the same synthetic procedure as compound 4 as a light brown solid (77 mg, 36.5%). 1 HNMR(500MHz, (CD3)2SO) δ8.47(s,1H),8.01(d,J=8.0Hz,2H),7.79(q,J=9.1Hz,2H),7.60(d,J=7.9Hz,2H),6.74(t,J=2.1Hz,1H),6.67(q,J=2.4Hz,1 H),6.08(q,J=2.3Hz,1H),4.41(d,J=5.6Hz,2H),3.53(p,J=6.6Hz,2H),3. 17(t,J=6.4Hz,2H), 1.89(q,J=7.6,6.3Hz,2H), 1.63(p,J=6.1,5.7Hz,2H). 13C NMR (126 MHz, (CD3)2SO) δ 166.30, 165.17, 162.82, 156.02, 143.84, 143.75, 142.45, 134.41, 129.52, 129.36, 129.11, 127.37, 127.01, 121.99, 120.98, 118.03, 116.28, 116.22, 107.90, 54.09, 36.02, 22.38, 22.33, 12.65. HRMS (ESI) m / z calculated value C 26 H 23 N5O[M+H] + 422.1981, measured value 422.1971.

[0568] 4-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-(thiazol-2-ylmethyl)benzamide (Compound 13)

[0569] Compound 13 was obtained according to the same synthetic procedure as compound 4 as an off-white solid (65 mg, 29.8%). 1 HNMR(500MHz, (CD3)2SO)δ9.40(t,J=6.0Hz,1H),8.57(s,1H),8.07–7.98(m,2H),7.90–7.78(m,2H),7.73(d,J=3.3Hz,1H),7.72–7 .65(m,2H),7.61(d,J=3.3Hz,1H),4.80(d,J=5.9Hz,2H),3.33(s,2H),2.78(t,J=6.1Hz,2H),2.05–1.96(m,2H),1.84–1.71(m,2H). 13 CNMR (126 MHz, (CD3)2SO) δ 169.89, 166.81, 156.08, 144.47, 142.64, 142.58, 136.40, 133.52, 129.66, 129.63, 129.29, 127.52, 127.07, 122.13, 120.49, 114.52, 41.53, 29.67, 28.83, 22.58, 22.51. HRMS (ESI) m / z calculated value C 25 H 21 N5OS[M+H] + 440.1545, measured value 440.1547.

[0570] N-((4-methyl-1H-imidazol-2-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 14, also known as Compound HSK205)

[0571] Compound 14 was obtained according to the same synthetic procedure as compound 4 as an off-white solid (89 mg, 40.7%). 1 HNMR(500MHz,(CD3)2SO)δ9.01(t,J=5.7Hz,1H),8.57(s,1H),8.05–7.99(m,2H),7.89–7.80(m,2H),7.68–7.63(m,2H),6.63–6.58(m,1H),4.46( d,J=5.7Hz,2H),2.77(t,J=6.1Hz,2H),2.09(d,J=1.0Hz,3H),2.07(s,1H),2.01(dd,J=7.8,3.9Hz,2H),1.97(s,1H),1.75(dd,J=7.6,4.2Hz,2H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.47, 156.23, 149.06, 144.60, 144.08, 143.78, 142.59, 138.74, 136.44, 133.83, 132.46, 129.67, 129.44, 127.61, 122.02, 116.46, 114.56, 105.50, 60.23, 37.63, 29.71, 28.87, 22.58, 14.56. HRMS (ESI) m / z calculated value C 26 H 24 N6O[M+H] + 437.2079, measured value 437.2083.

[0572] N-(2-(1H-imidazol-2-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 15)

[0573] Compound 15 was obtained according to the same synthetic procedure as compound 4 as an off-white solid (73 mg, 33.4%). 1HNMR(500MHz,(CD3)2SO)δ8.63(t,J=5.6Hz,1H),8.57(s,1H),7.93–7.89(m,2H),7 .87–7.77(m,2H),7.64(d,J=8.3Hz,2H),7.16(d,J=21.5Hz,1H),6.90(d,J=10.4Hz, 1H),4.21(t,J=6.1Hz,2H),3.63(q,J=5.9Hz,2H),3.42(q,J=6.0Hz,2H),2.76(t,J =6.1Hz, 2H), 1.99 (ddd, J = 15.1, 7.5, 4.5Hz, 2H), 1.73 (ddd, J = 9.0, 7.2, 4.2Hz, 2H). 13 CNMR (126 MHz, (CD3)2SO) δ 166.94, 161.91, 156.11, 144.18, 143.82, 142.54, 137.83, 134.00, 129.65, 129.49, 129.24, 128.72, 128.67, 127.35, 122.11, 120.11, 119.92, 116.35, 45.82, 38.81, 29.65, 28.83, 22.57, 22.49. HRMS (ESI) m / z calculated value C 26 H 24 N6O[M+H] + 437.2084, measured value 437.2070.

[0574] N-((4-methyl-1H-imidazol-2-yl)methyl)-4-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0575] According to the same synthetic procedure as compound HSK205, a light yellow solid compound (119 mg, 52.9%) was obtained. 1 H NMR(500MHz, (CD3)2SO)δ8.01(d,J=7.9Hz,2H),7.79–7.65(m,2H),7.61(d,J=8.0Hz,2H),6.65(s,1H ), 4.48 (d, J = 4.9Hz, 2H), 3.40 (s, 2H), 2.85 (d, J = 5.8Hz, 3H), 2.75 (s, 2H), 2.10 (s, 3H), 1.77 (s, 4H). 13C NMR (201 MHz, (CD3)2SO) δ 165.37, 159.24, 155.88, 151.77, 145.26, 143.68, 142.67, 138.74, 136.48, 131.87, 129.78, 129.65, 129.36, 128.38, 122.14, 116.45, 114.66, 114.39, 41.00, 39.19, 31.16, 29.69, 28.85, 22.57, 22.47. HRMS (ESI) m / z calculated value C 27 H 26 N6O[M+H] + 451.2241, measured value 451.2230.

[0576] 3-Fluoro-4-(9-hydroxy-1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-(1-(4-methyl-1H-imidazol-2-yl)ethyl)benzamide

[0577] According to the same synthetic procedure as compound HSK205, an off-white solid compound (98 mg, 39.3%) was obtained. 1 HNMR(500MHz,(CD3)2SO)δ8.43(d,J=7.0Hz,2H),7.95–7.87(m,2H),7.81(d d,J=8.9,2.5Hz,1H),5.31(dq,J=7.0,4.8Hz,1H),4.09–4.00(m,2H),3.90(d ,J=4.9Hz,2H),3.48(dd,J=17.5,7.1Hz,1H),3.38(ddd,J=15.6,9.1,6.4Hz ,1H),3.30–3.18(m,2H),2.28(s,3H),2.05–1.97(m,3H),1.86–1.75(m,3H). 13 C NMR(201MHz,(CD3)2SO)δ165.89,160.87,159.26,150.77,147.34,144.09,141.82,137.27,131.54,130.91,129.13,128.54 ,128.31,128.19,127.76,126.99,126.58,117.67,117.01,115.13,56.76,51.42,36.68,30.21,26.56,21.85,19.44,14.02.

[0578] 3-Fluoro-4-(9-hydroxy-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-((4-methyl-1H-imidazol-2-yl)methyl)benzamide

[0579] According to the same synthetic procedure as compound HSK205, a yellow solid compound (87 mg, 37.1%) was obtained. 1 HNMR(500MHz,(CD3)2SO)δ8.74(s,1H),7.98(d,J=9.2Hz,2H),7.92–7.79(m,3H),7.12–7.07(m,1H),4.49(s,2H),4.01(tt d,J=7.9,6.0,4.2Hz,1H),3.90(d,J=6.1Hz,2H),3.39–3.25(m,2H),3.18(ddd,J=15.6,8.8,6.6Hz,2H),1.95–1.79(m,3H). 13 CNMR (201 MHz, (CD3)2SO) δ 165.69, 160.34, 158.28, 149.06, 147.30, 144.08, 142.58, 138.14, 133.23, 131.46, 129.87, 129.03, 127.91, 127.72, 127.55, 126.98, 126.74, 119.20, 115.16, 109.66, 60.19, 39.52, 29.71, 28.94, 23.67, 13.96. HRMS (ESI) m / z calculated value C 26 H 23 FN6O2[M+H] + 471.1967, measured value 471.1963.

[0580] 3-Fluoro-N-((4-methyl-1H-imidazol-2-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0581] According to the same synthetic procedure as compound HSK205, an off-white solid compound (69 mg, 30.4%) was obtained. 1HNMR(500MHz,(CD3)2SO)δ8.72(s,1H),7.94(d,J=8.9Hz,1H),7.90–7.81(m,2H),7.0 9–7.01(m,2H),4.56(s,2H),3.37–3.28(m,4H),1.93–1.85(m,4H),1.83–1.73(m,3H). 13 C NMR(201MHz,(CD3)2SO)δ166.57,166.51,161.57,159.58,153.97,153.91,15 0.38,144.34,142.00,141.52,135.30,135.27,135.21,132.37,132.31,131. HRMS (ESI) m / z calculated value C 26 H 23 FN6O[M+H] + 455.1927, measured value 455.1936.

[0582] N-((4-methyl-1H-imidazol-2-yl)methyl)-2-(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)acetamide

[0583] According to the same synthetic procedure as compound HSK205, a light yellow solid compound (70 mg, 30.6%) was obtained. 1 HNMR(500MHz,(CD3)2SO)δ8.68(s,1H),7.97(d,J=9.2Hz,2H),7.83–7.74(m,4H),6.45(dt,J=8.4,0.9Hz, 1H), 4.40 (s, 2H), 4.21 (s, 2H), 3.60 (t, J = 0.9Hz, 2H), 3.31–3.21 (m, 4H), 2.04–1.91 (m, 2H), 1.86 (s, 3H). 13C NMR (201 MHz, (CD3)2SO) δ 169.09, 153.62, 148.76, 143.62, 141.18, 141.07, 136.44, 135.77, 135.50, 132.39, 128.85, 128.70, 126.44, 126.02, 119.66, 116.66, 116.39, 113.46, 41.38, 36.96, 27.86, 27.39, 22.62, 21.75, 14.26. HRMS (ESI) m / z calculated value C 27 H 26 N6O[M+H] + 451.2253, measured value 451.2251.

[0584] 4-(1-methoxy-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-((4-methyl-1H-imidazol-2-yl)methyl)benzamide

[0585] According to the same synthetic procedure as compound HSK205, a white solid compound (109 mg, 46.7%) was obtained. 1 HNMR(500MHz,(CD3)2SO)δ8.64(s,1H),8.04–7.97(m,2H),7.88–7.78(m,2H),7.69–7.61(m,2H),6.61–6.56(m,1H),4.44(d,J=5.6Hz,2H),4 .07(s,3H),2.79(t,J=6.2Hz,2H),2.11(d,J=1.0Hz,3H),2.07(s,1H),2.02(dd,J=7.6,3.9Hz,2H),1.94(s,1H),1.71(dd,J=7.2,4.0Hz,2H).

[0586] N-(1-(4-methyl-1H-imidazol-2-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0587] According to the same synthetic procedure as compound HSK205, a light yellow solid compound (101 mg, 44.8%) was obtained. 1H NMR(500MHz,(CD3)2SO)δ8.75(s,1H),7.98–7.86(m,4H),7.81(d,J=9.1Hz,2H),6.82(dd,J=6.1,0.9Hz,1H), 5.04(dq,J=7.0,4.8Hz,1H),3.31–3.21(m,4H),1.93–1.84(m,4H),1.82(d,J=4.8Hz,3H),1.81–1.74(m,3H). 13 C NMR(201MHz,(CD3)2SO)δ165.83,153.97,151.95,143.62,141.16,141.17,139.49,136.75,135.11,132.38,128 .52,126.64,126.48,126.02,119.66,117.14,116.66,113.46,46.03,27.86,27.39,22.62,21.75,19.16,13.27.

[0588] N-(1-(4-methyl-1H-imidazol-2-yl)ethyl)-4-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0589] According to the same synthetic steps as compound HSK205, a light orange solid compound (23 mg, 9.9%) was obtained. 1 HNMR(500MHz, (CD3)2SO) δ7.96–7.87(m,4H),7.80(d,J=9.3Hz,2H),6.85(dd,J=6.1,0.6Hz,1H),5.01(dq,J=6.6,4.2Hz ,1H),3.34(d,J=13.5Hz,2H),3.27–3.21(m,4H),2.28–2.22(m,3H),2.09(s,3H),1.93–1.81(m,2H),1.85–1.77(m,3H). 13 C NMR(201MHz,(CD3)2SO)δ165.79,153.88,151.87,144.21,143.70,141.64,140.87,139.49,136.75,135.21,128.52 ,126.81,126.61,125.46,120.93,117.33,117.14,112.10,46.11,27.67,27.53,22.58,21.73,19.21,18.68,13.61.

[0590] N-(2-(4-methyl-1H-imidazol-2-yl)propan-2-yl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0591] According to the same synthetic procedure as compound HSK205, an off-white solid compound (84 mg, 36.2%) was obtained. 1 HNMR(500MHz,(CD3)2SO)δ8.61(s,1H),7.99–7.92(m,2H),7.92–7.85(m,2H),7.81(d,J=8.8Hz,1H ),6.46–6.41(m,1H),3.31(s,3H)3.12(s,3H),1.93–1.83(m,4H),1.83–1.73(m,4H),1.79(s,3H). 13 C NMR(201MHz,(CD3)2SO)δ166.23,153.96,151.23,143.62,141.18,141.07,139.41,136.56,136.13,132.39,128 .52,127.37,126.48,126.02,119.66,116.93,116.66,113.46,50.07,27.86,27.52,27.39,22.62,21.75,12.60.

[0592] N-(2-(4-methyl-1H-imidazol-2-yl)propan-2-yl)-4-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0593] According to the same synthetic procedure as compound HSK205, a white solid compound (109 mg, 45.6%) was obtained. 1 HNMR(500MHz,(CD3)2SO)δ7.98–7.88(m,2H),7.91–7.85(m,2H),7.46–7.41(m,2H),3.3 5(s,3H),3.21(s,3H),2.30(s,3H),1.92–1.84(m,4H),1.80–1.71(m,4H),1.72(s,3H). 13C NMR (201 MHz, (CD3)2SO) δ 166.19, 153.98, 151.27, 144.21, 143.70, 141.64, 140.87, 139.41, 136.56, 136.13, 128.52, 127.37, 126.81, 125.46, 120.93, 117.33, 116.93, 112.10, 50.07, 27.67, 27.53, 27.52, 22.62, 21.75, 18.68, 10.60. HRMS (ESI) m / z calculated value C 29 H 30 N6O[M+H] + 479.2411, measured value 479.2422.

[0594] N-((4-ethyl-1H-imidazol-2-yl)methyl)-4-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0595] According to the same synthetic procedure as compound HSK205, a yellow solid compound (43 mg, 18.5%) was obtained. 1 HNMR(500MHz,(CD3)2SO)δ8.07–7.96(m,4H),7.80(d,J=9.2Hz,2H),6.23(d,J=4.9Hz,1H),4.48(s,2H),3.39(d, J=13.1Hz,2H),3.29–3.20(m,4H),2.70(q,J=7.1Hz,2H),1.93–1.82(m,2H),1.75(s,3H),1.28(t,J=6.6Hz,3H). 13 C NMR (201 MHz, (CD3)2SO) δ 165.86, 153.98, 150.50, 144.23, 143.72, 141.64, 140.89, 139.56, 139.48, 134.48, 128.52, 126.95, 126.80, 125.44, 120.93, 117.36, 116.07, 112.10, 36.72, 27.64, 27.56, 22.59, 21.78, 20.65, 18.79, 13.33. HRMS (ESI) m / z calculated value C 28 H 28 N6O[M+H] + 465.2347, measured value 465.2340.

[0596] 4-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-((4-(trifluoromethyl)-1H-imidazol-2-yl)methyl)benzamide

[0597] According to the same synthetic steps as compound HSK205, an orange-yellow solid compound (217 mg, 88.4%) was obtained. 1 H NMR (500MHz, (CD3)2SO) δ8.66 (s, 1H), 7.99–7.87 (m, 4H), 7.81 (d, J = 9.1Hz, 2H), 6.72(dq,J=4.1,2.3Hz,1H),4.57(s,2H),3.32–3.21(m,4H),1.92–1.83(m,4H). 13 C NMR(201MHz,(CD3)2SO)δ165.89,153.96,150.15,150.11,150.08,150.04,143. 62,141.18,141.07,139.56,134.48,132.39,129.61,129.31,129.01,128.71,1 HRMS (ESI) m / z calculated value C 26 H 21 F3N6O[M+H] + 491.1781, measured value 491.1785.

[0598] 4-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-((4-(trifluoromethyl)-1H-imidazol-2-yl)methyl)benzamide

[0599] According to the same synthetic procedure as compound HSK205, an off-white solid compound (197 mg, 78.1%) was obtained. 1H NMR(500MHz,(CD3)2SO)δ7.97–7.87(m,4H),7.80(d,J=8.8Hz,2H),6.68(dq,J=4.0,1.8Hz,1 H), 4.49 (s, 2H), 3.18 (d, J = 10.9Hz, 2H), 3.27–3.20 (m, 4H), 1.85 (s, 3H), 1.83–1.76 (m, 2H). 13 C NMR(201MHz,(CD3)2SO)δ165.91,153.98,150.16,150.13,150.09,150.03,144.2 1,143.70,141.64,140.87,139.56,134.48,129.62,129.37,129.05,128.70,128. 54,126.99,126.81,125.46,124.40,122.21,120.93,120.04,117.85,117.33,116.37,116.35,116.29,116.27,112.10,36.79,27.67,27.53,22.59,21.67,18.68. HRMS (ESI) m / z calculated value C 27 H 23 F3N6O[M+H] + 505.1925, measured value 505.1918.

[0600] N-Methyl-N-((4-methyl-1H-imidazol-2-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0601] 1 H NMR(500MHz,(CD3)2SO)δ9.01(t,J=5.7Hz,1H),8.55(s,1H),8.07–7.99(m, 2H),7.88–7.80(m,2H),7.68–7.63(m,2H),6.63–6.58(m,1H),4.42(d,J=5. 7Hz,2H),2.76(t,J=6.3Hz,2H),2.61(s,3H),2.09(d,J=1.0Hz,3H),2.05(s ,1H),2.01(dd,J=7.8,3.9Hz,2H),1.94(s,1H),1.77(dd,J=7.5,4.1Hz,2H).

[0602] N-((4-methyl-1H-imidazol-2-yl)methyl)-4-(3H-pyrazolo[4,3-f]quinolin-7-yl)benzamide

[0603] 1 H NMR(500MHz,(CD3)2SO)δ8.99(t,J=5.5Hz,1H),8.52(s,1H),8.46-8.39(m,2H),8.11–8.01(m,2H),7 .89–7.81(m,2H),7.69–7.62(m,2H),6.61–6.55(m,1H),4.44(d,J=5.6Hz,2H),2.08(d,J=1.1Hz,3H).

[0604] N-((4-methyl-1H-imidazol-2-yl)methyl)-4-(3',8',10',11'-tetrahydrospiro[oxetane-3,9'-pyrazolo[4,3-a]phenanthridine]-7'-yl)benzamide

[0605] 1 H NMR(500MHz,(CD3)2SO)δ9.00(t,J=5.6Hz,1H),8.55(s,1H),8.08–7.96(m,2H),7.89– 7.80(m,2H),7.66–7.61(m,2H),6.66–6.59(m,1H),4.43(d,J=5.6Hz,2H),3.89(d,J=8. 1Hz,2H),3.65(d,J=8.2Hz,2H),2.77(t,J=6.1Hz,2H),2.61(s,3H),2.09(d,J=1.1Hz,3 H), 2.04 (s, 1H), 2.01 (dd, J = 7.7, 3.6Hz, 2H), 1.91 (s, 1H), 1.72 (dd, J = 7.7, 4.2Hz, 2H).

[0606] N-((4-methyl-1H-imidazol-2-yl)methyl)-4-(2,3,3',5,6,8',10',11'-octahydrospiro[pyran-4,9'-pyrazolo[4,3-a]phenanthridine]-7'-yl)benzamide

[0607] 1H NMR(500MHz,(CD3)2SO)δ8.59(s,1H),8.12–7.99(m,2H),7.86–7.79(m,2H),7.66–7.60(m,2H),6.69–6.58(m,1H),4.44(d,J=5.5Hz,2H),3.71(dt,J=10.8,2.8Hz,2H),3.62(dt,J=10.8,6.5Hz,2H),2.77(t,J=6.1Hz,2H),2.61(s,3H),2.09(d,J=1.1Hz,3H),2.04(s,1H),2.01(dd,J=7.7,3.6Hz,2H),1.97-1.88(m,5H),1.72(dd,J=7.7,4.2Hz,2H)。

Claims

1. A compound having a structure of Formula I: or a pharmaceutically acceptable salt thereof, wherein: X1 is each independently N, CH or C-halogen; X2 is each independently N, CH, C-halogen, C-CH n1 X m wherein n1+m=3 and X is halogen, C-OCF3, C-OCHF2, C-OMe, C-Me, C-Et, C-tBu, C-iPr, CN-alkyl, CN-heteroalkyl, CO-alkyl or CO-heteroalkyl; R2, R3 and R4 are each independently alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl; and Y is in: n is 1 to 5, R5 and R6 are each independently H, alkyl or heteroalkyl, and R6 may optionally form a ring structure, R7 are each independently H or alkyl, Het-Ar is a heteroaromatic ring, and W is NH, NMe, NEt, NCH2CH2OH, NCH2CH2OMe, O, SO or SO2, Provided that Y is not NH2.

2. The compound according to claim 1, wherein at least one of X1 and / or X2 is CF, CHF2, CF3 or CHF3.

3. The compound according to claim 1, which has the structure of Formula IA: or a pharmaceutically acceptable salt thereof, wherein: n2 is 1, 2 or 3; Q is each independently O, NH, substituted N, CH2, or substituted C, S, SO or SO2; and Each Q together forms a 6-membered, 7-membered or 8-membered ring or a bicyclic ring.

4. The compound according to claim 1, wherein one or more of R5 and / or R6 is Me.

5. The compound according to claim 1, wherein each R7 is independently H or Me.

6. The compound according to claim 1, wherein Y is And each R6 together forms morpholine, piperidine, piperazine or pyrrolidine.

7. The compound according to claim 1, wherein Y is And each R7 together forms a ring structure.

8. The compound of claim 7, wherein the ring structure comprises cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

9. The compound according to claim 1, wherein Y is And each R7 together forms a ring structure.

10. The compound of claim 9, wherein the ring structure comprises cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

11. The compound according to claim 1, wherein Y is And Het-Ar includes imidazole, oxazole, pyrazole, triazole, isoxazole, isothiazole, tetrazole, oxadiazole, thiadiazole, pyrimidine or triazine.

12. The compound according to claim 1, wherein Y is And R5 is Me.

13. The compound according to claim 3, which has the structure of Formula IB: or a pharmaceutically acceptable salt thereof, wherein: o is 0 to 2; and Each R8 is independently H, alkyl or heteroalkyl.

14. The compound of claim 13, wherein each R7 together forms a ring structure.

15. The compound of claim 14, wherein the ring structure comprises cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

16. A compound according to any one of claims 13-15, wherein at least one R8 is Me, Et or CF3.

17. A compound having the structure of formula II: or a pharmaceutically acceptable salt thereof, wherein: X1 is each independently N, CH or C-halogen; X2 is each independently N, CH, C-halogen, C-CH n1 X m wherein n1+m=3 and X is halogen, C-OCF3, C-OCHF2, C-OMe, C-Me, C-Et, C-tBu, C-iPr, CN-alkyl, CN-heteroalkyl, CO-alkyl, or CO-heteroalkyl; Q is each independently O, NH, substituted N, CH2, or substituted C, S, SO or SO2, and each Q when combined together forms a 6-membered, 7-membered or 8-membered ring or a bicyclic ring; n3 is 1, 2, or 3; R4 is alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl; W is O, NH or NMe; and R9 is alkyl or heteroalkyl, provided that R9 is not H.

18. The compound according to claim 1 or 17 or a pharmaceutically acceptable salt thereof, which has the following structure:

19. The compound according to claim 1 or 3, having the following structure: or a pharmaceutically acceptable salt thereof.

20. The compound according to claim 2 or 3, having the following structure:

21. The compound according to claim 1, having the following structure:

22. The compound according to claim 1, having the following structure: or a pharmaceutically acceptable salt thereof.

23. The compound according to claim 1, having the following structure:

24. The compound according to claim 1, having the following structure:

25. The compound according to claim 1, having the following structure:

26. A compound according to claim 1 or claim 3 having the following structure:

27. A compound according to claim 1 or claim 3 having the following structure:

28. A compound according to claim 1 or claim 3 having the following structure:

29. A PROTAC conjugate having a chemical structure of Formula X: A—L'—D (Formula X) or a pharmaceutically acceptable salt thereof, wherein: A is a group of a compound according to any one of claims 1 to 28; L' is a linker that binds A and D or is absent; and D is the ubiquitin pathway protein binding moiety.

30. Use of a compound according to any one of claims 1-28, a conjugate according to claim 29, or a pharmaceutically acceptable salt of said compound or conjugate in the manufacture of a medicament for treating a disease in a subject, wherein modulation of a kinase reduces the severity of the disease in said subject.

31. The use according to claim 30, wherein the disease is cancer, diabetes, an inflammatory disease or a neurological disease.

32. A pharmaceutical composition comprising: A compound according to any one of claims 1 to 28, a conjugate according to claim 29, or a pharmaceutically acceptable salt, N-oxide, hydrate, solvate, tautomer or optical isomer of the compound or conjugate; and Pharmaceutically acceptable carrier and / or diluent.

33. The pharmaceutical composition of claim 32, further comprising a pharmaceutically acceptable excipient.

34. A method of treating cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of: A compound according to any one of claims 1 to 28, a conjugate according to claim 29, or a pharmaceutically acceptable salt, N-oxide, hydrate, solvate, tautomer or optical isomer of the compound or conjugate; or A pharmaceutical composition comprising the compound according to any one of claims 1 to 28, the conjugate according to claim 29, or one or more of the pharmaceutically acceptable salts, N-oxides, hydrates, solvates, tautomers or optical isomers of the compound or conjugate.

35. The method of claim 34, wherein the compound or conjugate comprises a compound according to claim 22.

36. The method of claim 34, wherein the compound or conjugate comprises a compound of claim 19.

37. The method of claim 34, further comprising administering to the subject a second therapy comprising: an effective amount of a chemotherapeutic, immunotherapeutic, or hormonal therapeutic agent; or Radiation therapy.

38. The method of any one of claims 34-37, wherein the cancer is acute myeloid leukemia (AML).

39. The method of any one of claims 34-37, wherein the cancer is selected from AML, chronic myeloid leukemia, ovarian cancer, cervical cancer, pancreatic cancer, breast cancer, brain cancer, skin cancer, lung cancer, prostate cancer, lymphoma, leukemia, colon cancer, gastric cancer, head cancer, neck cancer, thyroid cancer, kidney cancer, liver cancer, and stomach cancer.

40. The method of claim 35, wherein an effective amount is administered to the subject at an IC of about 50 nM or less, such as 50 nM or less. 50 and inhibiting a CDK in the subject.

41. The method of claim 40, wherein the IC 50 About 30 nM or less, such as 30 nM or less.

42. The method of claim 36, wherein administering to the subject an effective amount specifically inhibits FLT3 and haspin kinases in the subject.

43. A compound according to any one of claims 1 to 28 or a conjugate according to claim 29 for use in treating a disease state modulated by one or more kinases.

44. The compound or conjugate of claim 43, wherein the disease state is cancer and the one or more kinases are CDK, FLT3 and / or haspin kinases.

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