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, pharmaceutical compositions, conjugates, and methods of treating these kinase-mediated disease states are also provided, which contain N-(2-(dimethylamino)ethyl)benzamide- or hydroxamic acid derivatives for use in the inhibition of certain kinases for use in certain kinase-mediated disease states.

Description

Technical Field

[0001] Cross - Reference to Related Applications This patent application claims the benefit of, and priority to, U.S. Provisional Patent Application No. 63 / 348,034, filed on June 2, 2022. The content of the aforementioned application is hereby incorporated by reference in its entirety into this disclosure.

[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 - potential anti - cancer or anti - inflammatory agents for the treatment of kinase - mediated diseases such as inflammation and cancer. Methods of treating these diseases are also provided.

Background Art

[0003] This chapter introduces aspects that may help facilitate a better understanding of the present disclosure. Accordingly, these descriptions should be construed in light of this and should not be understood as an admission as to what is prior art or what is not prior art.

[0004] Cells contain over 500 kinases, which control diverse processes such as the cell cycle, growth, migration, and immune response. Some deregulated kinases (i.e., kinases that have achieved 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 driver 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 receptor (VEGFR), B-Raf, Bruton's tyrosine kinase (BTK), phosphatidylinositol-4,5-bisphosphate 3-kinase enzyme subunit delta (PI3Kδ), and Erb-B2) have had 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 been made to target other kinases that halt cancer growth, such as cell cycle kinases (CDKs), and kinases that target other processes important to histones, the cytoskeleton, or the cell.

[0005] However, most of the kinase inhibitors to date have been rendered ineffective due to the emergence of resistant clones.

[0006] Multiple mechanisms are major factors in cancer cell resistance to kinase inhibitors, and such mechanisms include multiplication of copy number, further kinase mutations (such as secondary mutations occurring in the tyrosine kinase domain of 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 over 20,000 people each year in the United States. Approximately 30% of AML patients carry mutations in the FLT3 kinase internally, and for these patients, the prognosis is usually worse than those without FLT3 mutations. Lin & Chen, Advances in the drug therapies of acute myeloid leukemia (except acute wpromyelocytic leukemia), Drug Design, Development & Therapy 12: 1009-1017 (2018). Many FLT3 inhibitors have been developed and tested in the clinic, and several, including midostaurin, quizartinib, and gilteritinib, are approved in the United States or Japan. Although these FLT3 inhibitors have gradually improved the survival rate of AML patients with FLT3 internally, resistance to the 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, resistance pathways independent of FLT3, such as NRAS activation, or mutations in other kinases can also emerge after extended FLT3i treatment.Alotaibi et al., Patterns of resistance differ in patients with acute myeloid leukemia treated with type I versus type II FLT3 inhibitors, Blood Cancer Discovery 2(2): 125-134 (2021); Kasi et al., Clonal evolution 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 become FLT3-negative during treatment, but are driven to alternative pathways and the cancer progresses. Schmalbrock et al., Clonal evolution of acute myeloid leukemia with FLT3-ITD mutation under treatment with midostaurin, Blood 137(22): 3093-3104 (2021).

[0008] Kinase inhibitors that inhibit cancer driver kinases, and also downstream targets (both kinase and non-kinase targets, such as histone demethylases) and / or kinases that cooperate with the driver kinase, should be able to enhance potency and potentially reduce resistance that is generated against the kinase inhibitor. However, a common difficulty associated with such combination approaches is off-target binding, which can lead to toxicity.

[0009] Therefore, there is an unmet need for agents that can address FLT3 and other oncogenic pathways that may arise during treatment. SUMMARY OF THE INVENTION

[0010] In certain embodiments, a compound having the structure of Formula I:

[0011] [Chemical Formula] or a pharmaceutically acceptable salt thereof is provided, 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, C-N-alkyl, C-N-heteroalkyl, C-O-alkyl or C-O-heteroalkyl, R2, R3 and R4 are each independently alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl, Y is

[0012] [Chemical Formula] and wherein, n is from 1 to 5, R5 and R6 are each independently H, alkyl, or heteroalkyl, and R6 can optionally form a cyclic structure, R7 is each independently H or alkyl, Het-Ar is a heteroaromatic ring, W is NH, NMe, NEt, NCH2CH2OH, NCH2CH2OMe, O, SO or SO2, provided that Y is not NH2.

[0013] 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 has the formula IA:

[0014] [Chemistry] can have the structure of, or can be a pharmaceutically acceptable salt thereof, wherein, n2 is 1, 2 or 3, Q is, independently of each other, O, NH, substituted N, CH2, or substituted C, S, SO or SO2, each Q together forms a 6-membered, 7-membered or 8-membered ring or a bicyclic ring.

[0015] One or more of R5 and / or R6 can be Me. In certain embodiments, each R7 is, independently, H or Me. In certain embodiments, Y is,

[0016] [Chemistry] and each R6 together forms morpholine, piperidine, piperazine or pyrrolidine. In certain embodiments, Y is,

[0017] [Chemistry] and each R7 together forms a cyclic structure (e.g., here, the cyclic structure includes cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl). In certain embodiments, Y is,

[0018] [Chemistry] and each R7 together forms a cyclic structure (e.g., here, the cyclic structure includes cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl). In certain embodiments, Y is,

[0019] [Chemistry] and Het-Ar includes imidazole, oxazole, pyrazole, triazole, isoxazole, isothiazole, tetrazole, oxadiazole, thiadiazole, pyrimidine or triazine. In certain embodiments, Y is

[0020]

Chemical formula

[0021] The compound is of formula IB:

[0022]

Chemical formula

[0023] In certain embodiments, the compound is of formula II:

[0024]

Chemical formula

[0025] In certain embodiments, the compound, or a pharmaceutically acceptable salt thereof,

[0026]

Chemical formula

[0027] In certain embodiments, the compound

[0028]

Chemical formula

[0029] The compound

[0030]

Chemical formula

[0031]

Chemical formula

[0032] The compound can

[0033] [Chemical formula]

[0034] [Chemical formula]

[0035] [Chemical formula]

[0036] [Chemical formula]

[0037] [Chemical formula]

[0038] [Chemical formula]

[0039] [Chemical formula]

[0040] [Chemical formula] have the structure of.

[0041] The compound can

[0042] [Chemical formula] have the structure of, or can be its pharmaceutically acceptable salt.

[0043] The compound can

[0044]

Chem.

[0045]

Chem.

[0046]

Chem.

[0047]

Chem.

[0048]

Chem.

[0049]

Chem.

[0050]

Chem.

[0051] The compound can

[0052]

Chem.

[0053]

Chem.

[0054]

Chem.

[0055]

Chem.

[0056]

Chem.

[0057] The compound is

[0058]

Chem.

[0059]

Chem.

[0060]

Chem.

[0061] The compound is

[0062]

Chem.

[0063] The compound is

[0064]

Chem.

[0065] The compound is

[0066]

Chem.

[0067] [Chem.] can have the structure of.

[0068] PROTAC conjugates are also provided. In certain embodiments, the PROTAC conjugate has the formula X: A-L’-D (Formula X) having the chemical structure of, or a pharmaceutically acceptable salt thereof, wherein, A is any group of the compounds herein, L’ is a linker that connects A and D or is absent, D is a ubiquitin pathway protein binding site.

[0069] The use of a compound or conjugate herein, or a pharmaceutically acceptable salt of a compound or conjugate herein, in the manufacture of a drug for the treatment of a disease in a subject is also provided. In certain embodiments, the disease includes a disease in which the severity of the disease in the subject is reduced by modulation of a kinase. The disease can be, for example, cancer, diabetes, an inflammatory disease, or a neurological disease.

[0070] Pharmaceutical compositions are also provided. The pharmaceutical compositions herein can include any of a compound herein, a conjugate herein, or a pharmaceutically acceptable salt, N-oxide, hydrate, solvate, tautomer or optical isomer of a compound or conjugate; and a pharmaceutically acceptable carrier and / or diluent. The pharmaceutical composition can further include a pharmaceutically acceptable excipient.

[0071] Methods for treating a disease state (e.g., cancer) in a subject are also provided. In certain embodiments, the method comprises administering to the subject a therapeutically effective amount of a compound of the present specification, a conjugate of the present specification, or a pharmaceutically acceptable salt, N-oxide, hydrate, solvate, tautomer or optical isomer of a compound or conjugate; or a pharmaceutical composition comprising one or more of a compound of the present specification, a conjugate of the present specification, or a pharmaceutically acceptable salt, N-oxide, hydrate, solvate, tautomer or optical isomer of a compound or conjugate.

[0072] Certain embodiments of the method, compound or conjugate are

[0073]

Chemical formula

[0074] Certain embodiments of the method, compound or conjugate are

[0075]

Chemical formula

[0076] The method may further comprise administering to the subject a second therapeutic agent. The second therapeutic agent may include an effective amount of a chemotherapeutic agent, an immunotherapeutic agent or a hormonal therapeutic agent; or radiation therapy.

[0077] The cancer can be acute myeloid leukemia (AML). In certain embodiments, the cancer is 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 carcinoma, head cancer, neck cancer, thyroid cancer, kidney cancer, liver cancer, and stomach cancer.

[0078] In certain embodiments of the method, administering an effective amount to the subject inhibits CDK in the subject at an IC of about 50 nM or less (such as 50 nM or less). 50 In certain embodiments, administering an effective amount to the subject inhibits CDK in the subject at an IC of about 30 nM or less (such as 30 nM or less). 50 In certain embodiments, administering an effective amount to the subject inhibits CDK in the subject at an IC of about 30 nM or less (such as 30 nM or less).

[0079] Administering an effective amount to the subject can inhibit both FLT3 and haspin kinase with specificity in the subject, for example, where a dual inhibitor compound is administered.

[0080] The compounds and / or conjugates described herein are further provided for use in the treatment of disease states regulated by one or more kinases. For example, the disease state can be cancer, and the one or more kinases can be CDK, FLT3, and / or haspin kinase.

[0081] The disclosed embodiments, and other features, advantages, and aspects contained herein, and what they accomplish, will become apparent in light of the following detailed "Description of Embodiments for Carrying Out the Invention" of various exemplary embodiments of the present disclosure. Such "Description of Embodiments for Carrying Out the Invention" will be better understood when taken in conjunction with the accompanying drawings.

Brief Description of the Drawings

[0082]

Figure 1

Figure 2

Figure 3-1

Figure 3-2

Figure 4-1

Figure 4-2

Figure 4-3

Figure 4-4

Figure 5-1

Figure 5-2

Figure 6-1

Figure 6-2

Figure 6-3

Figure 6-4

Figure 7-1

Figure 7-2

Figure 8-1

Figure 8-2

Figure 8-3

Figure 8-4

Figure 9-1

Figure 9-2

Figure 10-1

Figure 10-2

Figure 10-3

Figure 11-1

Figure 11-2

Figure 11-3

Figure 11-4

Figure 12-1

Figure 12-2

Figure 13-1

Figure 13-2

Figure 14-1

Figure 14-2

Figure 15

Figure 16

[0083] Although the present disclosure is susceptible to various modifications and alternative forms, the exemplary embodiments thereof are shown by way of example in the drawings and are described in detail herein.

Modes for Carrying Out the Invention

[0084] Although the concepts of the present disclosure are illustrated and described in detail in the description herein, the results in the description are considered to be exemplary, not limited by the words, only exemplary embodiments are shown and described, and it is understood that all changes and modifications falling within the spirit of the present disclosure are protected.

[0085] U.S. Patent No. 11,040,973 to Sintim et al. establishes that HSD1217, 4-(3H-pyrazolo[4,3-f]quinolin-7-yl)benzamide, which contains compounds synthesized via a Doebner-Povarov type multi-component reaction, is a potent inhibitor of kinases, such as FLT3 kinase.

[0086] HSD1217 has the structure:

[0087]

Chemical formula

[0088] However, compound HSD1217 is only a moderate inhibitor of CDKs such as CDK2 and CDK12, which are important kinase targets at the forefront as potential anti-cancer targets. For example, the IC 50 for the inhibition of CDK2 by HSD1217 is 185 nM.

[0089] In certain embodiments, the present disclosure provides compounds that include either a 2-(dimethylamino)ethyl moiety or a hydroxamic acid in place of the benzamide moiety, enabling unexpected and dramatically improved kinase targeting compared to at least compound HSD1217 and other conventional CDK inhibitors.

[0090] 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, such as cancer, inflammatory diseases, and certain neurological diseases, where targeting CDKs leads to the restoration of normal cell function.

[0091] Furthermore, it has recently been reported that FMS-like tyrosine kinase-3 (FLT3) inhibitors have activity against inhibitors of other cancer-related kinases, such as dual inhibitors of FLT3 / MERTK, FLT3 / MNK, FLT3 / CDK, FLT3 / TOPK, FLT3 / PIM, or FLT3 / AURKA, where simultaneous inhibition of FLT3 and other kinase pathways may confer some advantage in overcoming 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 Chemistry 121: 105675 (2022); Goh et al., SB1317, a potent and orally 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 potently inhibits acute 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, Oncotartget 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 study in patients with relapsed / refractory acute myeloid leukemia, Blood 140(Supplement 1): 6222-6223 (2022). For example, CCT245718, a dual FLT3 / Aurora A inhibitor, can overcome FLT3-D835Y-mediated resistance. Tariq et al., CCT245718, a dual FLT3 / Aurora A inhibitor overcomes D835Y-mediated resistance to FLT3 inhibitors in acute myeloid leukemia cells, British J Cancer 125: 966-974 (2021). In another interesting report, it was shown that the frequency of resistance to the dual FLT3 / CDK4 inhibitor (AMG925) was lower than that of other FLT3 inhibitors (such as quizartinib) that did not inhibit CDK4. 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).

[0092] Despite these dual inhibitors, a dual inhibitor of FLT3 and Haspin has not been previously identified. Haspin is a novel target in cancer and is the major kinase that phosphorylates histone H3 at T3. Amoussou et al., Haspin: a promising target for the design of inhibitors as potent anticancer drugs, Drug Discovery Today 23(2): 409-415 (2018). Recent preliminary reports have suggested that reduction of Haspin via genetic means or inhibition by compounds decreased leukemia proliferation. Taking advantage of recent revelations that Haspin is overexpressed and plays an important role in proliferation in many cancers, in certain embodiments, optimized 3H-pyrazolo[4,3-f]quinoline-based compounds are provided that exhibit improved activity against both FLT3 and Haspin (i.e., 3H-pyrazolo[4,3-f]quinoline-based dual FLT3 / Haspin inhibitors).

[0093] In certain embodiments, the compound has the structure of Formula I:

[0094]

Chemical formula

[0095]

Chem.

[0096] At least one of X1 and / or X2 can 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.

[0097] At least one of R5 and / or R6 of Y can be Me. R7 of Y can each independently be H or Me. When Y is

[0098]

Chem.

[0099]

Chem.

[0100] [Chem.] In certain embodiments where it is as such, each R6 can together form piperazine. When Y is

[0101] [Chem.] In certain embodiments where it is as such, each R6 can together form pyrrolidine.

[0102] When Y is

[0103] [Chem.] In certain embodiments where it is as such, each R7 can together form a cyclic structure. The cyclic structure can include, for example, cyclopropyl. The cyclic structure can include cyclobutyl. The cyclic structure can include cyclopentyl. The cyclic structure can include cyclohexyl.

[0104] When Y is

[0105] [Chem.] In certain embodiments where it is as such, each R7 can together form a cyclic structure. The cyclic structure formed by each R7 can include, for example, cyclopropyl. The cyclic structure can include cyclobutyl. The cyclic structure can include cyclopentyl. The cyclic structure can include cyclohexyl.

[0106] In certain embodiments, Y is

[0107] [Chem.] and Het-Ar includes imidazole, oxazole, pyrazole, triazole, isoxazole, isothiazole, tetrazole, oxadiazole, thiadiazole, pyrimidine or triazine.

[0108] In certain embodiments, Y is

[0109]

Chemical formula

[0110] In certain embodiments, the compound is of formula IA:

[0111]

Chemical formula

[0112]

Chemical formula

[0113] The compound has the structure of formula IB:

[0114]

Chemical formula

[0115] In certain embodiments where the compound has the structure of Formula IB, each R7 together forms a cyclic structure. The cyclic structure can include cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In certain embodiments, at least one R8 is Me, Et or CF3.

[0116] In certain embodiments, the compound is

[0117]

Chemical formula

[0118] The compound is

[0119]

Chemical formula

[0120]

Chemical formula

[0121] In certain embodiments, the compound is

[0122]

Chemical formula

[0123]

Chemical formula

[0124]

Chemical formula

[0125]

Chemical formula

[0126]

Chem.

[0127]

Chem.

[0128]

Chem.

[0129]

Chem.

[0130] In certain embodiments, the compound

[0131]

Chem.

[0132]

Chem.

[0133]

Chem.

[0134]

Chem.

[0135]

Chem.

[0136]

Chem.

[0137] [Chemical formula] has one of the following structures or is a pharmaceutically acceptable salt.

[0138] In certain embodiments, the compound

[0139] [Chemical formula]

[0140] [Chemical formula]

[0141] [Chemical formula]

[0142] [Chemical formula]

[0143] [Chemical formula] has one of the following structures or is a pharmaceutically acceptable salt.

[0144] In certain embodiments, the compound

[0145] [Chemical formula]

[0146] [Chemical formula] has one of the following structures or is a pharmaceutically acceptable salt.

[0147] In certain embodiments, the compound is

[0148] [Chemical formula] has one of the structures of or is a pharmaceutically acceptable salt.

[0149] In certain embodiments, the compound is

[0150] [Chemical formula]

[0151] [Chemical formula]

[0152] [Chemical formula] has one of the structures of or is a pharmaceutically acceptable salt.

[0153] In certain embodiments, the compound is of formula II:

[0154] [Chemical formula] has the structure of or a salt thereof that is pharmaceutically acceptable, wherein each X1 is independently N, CH, or C-halogen, each X2 is 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, C-N-alkyl, C-N-heteroalkyl, C-O-alkyl, or C-O-heteroalkyl, Each Q is independently O, NH, substituted N, CH2, or substituted C, S, SO or SO2, and when taken together, each Q forms a 6-membered, 7-membered, 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. R9 is alkyl or heteroalkyl, provided that R9 is not H.

[0155] The compound

[0156]

Chemical formula

[0157] In certain embodiments, the compound

[0158]

Chemical formula

[0159] or is a pharmaceutically acceptable salt.

[0160]

Chemical formula

[0161]

Chemical formula

[0162] As noted above, the compound can be a CDK-specific inhibitor. In certain embodiments, the compound is a dual agent haspin and FLT3-specific inhibitor. The compound can bind, with specificity, to the CDK-receptor or to both haspin and the FLT3-receptor, thus reducing off-target interactions, for example, with growth factor receptors. "Binding with specificity", "binding with high affinity", or "specifically" or "selectively" binding, when referring to a small molecule / receptor or other binding pair, means that, under specified conditions, the specified compound, or its recognition region, binds to a particular receptor or site (e.g., the CDK inhibitory site, e.g., the ATP-binding catalytic site of the tyrosine kinase domain) and does not bind in significant amounts to other receptors present (e.g., off-target growth factor receptors). Specific binding, or binding with high affinity, can also mean, for example, binding to its target with an affinity that is often at least 20% greater, at least 25% greater, at least 50% greater, or at least 100% (2-fold) greater than the affinity of the binding compound, ligand; or the binding composition of the compound, ligand, or another receptor (e.g., growth factor receptor) that binds to the other receptor (e.g., growth factor receptor).

[0163] In a typical embodiment, a molecule that specifically binds to a target will have an affinity of at least about 10 6 liters / mol (Κ0 = 10 ~6 M), preferably at least about 50 nM, as determined, for example, by Scatchard analysis. The binding specificity of the compound to the CDK-receptor or to both haspin and the FLT3-receptor is determined using standard techniques known in the art and described herein.

[0164] In certain embodiments, N-(2-(dimethylamino)ethyl)benzamide or the hydroxamic acid derivatives herein can dramatically improve activity against kinases including CDK. Figure 1 shows the modulation of the previously known compound HSD1217 to the compounds HSH2177 and HSD1993 herein. While HSD1217 inhibits CDK2 at an IC 50 value of 185 nM, the compound HSH2177 can inhibit CDK2 at an IC 50 value of 7 nM, and the compound HSD1993 can inhibit CDK2 at an IC 50 value of 4 nM. Furthermore, the compounds HSD2177 and HSD1993 inhibit CDK12 / CyclinK at IC 50 values of 27 nM and 9 nM, respectively. Under similar experimental conditions, THZ531 and dinaciclib inhibited CDK12 at IC 50 values of 60 nM and 4 nM, respectively.

[0165] The inventors have shown previously that the "condensation" of two known kinase hinge binders, indazole and quinoline, commonly found in FDA-approved drugs such as axitinib and erlotinib, can lead to 3H-pyrazolo[4,3-f]quinoline (Figure 2A), which can bind to the hinge region of kinases. 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). Notably, the substituent at the 7-position of the 3H-pyrazolo[4,3-f]quinoline moiety can dictate which kinases are potently inhibited. For example, pyrazole substitution at the 7-position results in picomolar K dcan lead to an inhibitor that binds to a mutant FLT3 having a value, while the phenylboronic acid substituent can lead to an ultra-potent ROCK1 / 2 inhibitor. 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).

[0166] Benzothiazolyl-substituted compounds such as SHD992 can be potent CDK inhibitors, while phenolic-containing compounds such as HSD972 (see Figure 2B) are excellent haspin inhibitors (HSD972 haspin IC 50It can be (equal to 14 nM). Opoku-Temeng et al., Tetrahydro-3H-pyrazolo[4,3-a]phenanthridine-based CDK inhibitor, 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, certain other similar systems, such as the pyrazolo[3,4-h]quinoline scaffold, can 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 reticulum stress, Antioxidants 11(1): 117(2022).

[0167] Both HDS972 and HSD1169 (see Figure 2B) contain a 3H-pyrazolo[4,3-f]quinoline moiety, but they inhibit haspin or FLT3, respectively, and not both. In certain embodiments, the compounds herein include, at the 7-position, a replacement with a benzamide of the phenol moiety (see Compound HSD1217, Figure 3A), which inhibits haspin more potently than HDS972. In certain embodiments, the compounds herein are derivatives of Compound HSD1217, which has been optimized for dual FLT3 / haspin binding.

[0168] The compounds described herein may contain one or more chiral centers or, alternatively, may exist as multiple stereoisomers. In one embodiment, it is understood that the inventions described herein are not limited to any particular stereochemical requirements, and that the compounds, and compositions, methods, uses, and medicaments containing them may be optically pure or may be any of a variety of stereoisomer mixtures, including racemates, and other mixtures of enantiomers, other mixtures of diastereomers, etc. It is also understood that such mixtures of stereoisomers may contain a single stereochemical configuration at one or more chiral centers while containing a mixture of stereochemical configurations at one or more other chiral centers.

[0169] Similarly, the compounds described herein may contain geometric centers, such as cis, trans, E and Z double bonds. In another embodiment, it is understood that the inventions described herein are not limited to any particular geometric isomer requirements, and that the compounds, and compositions, and methods, uses, and medicaments containing them may be pure or may be any of a variety of geometric isomer mixtures. It is also understood that such mixtures of geometric isomers may contain a single configuration at one or more double bonds while containing a mixture of geometries at one or more other double bonds.

[0170] It is also contemplated that the compounds of the present specification may be provided in prodrug form. The term "prodrug" means an inactive derivative of a parent compound / drug that can be hydrolyzed, oxidized, or otherwise react under biological conditions (in vitro or in vivo) to provide an active compound, particularly a compound of the present specification. Prodrugs can be created to overcome one or more barriers to the effective use of the underlying active compound, such as a stability barrier and / or a possible toxicity barrier that co-exist with the active compound. Examples of prodrugs include, but are not limited to, derivatives and metabolites of compounds containing a hydrolyzable moiety such as various analogs of biolabile amides, biolabile esters, biolabile carbamates, biolabile carbonates, biolabile ureides, and biolabile phosphates. A specific prodrug of a compound having a carboxyl functional group is a lower alkyl ester of a carboxylic acid. The carboxylate ester is advantageously formed by esterifying any of the carboxylic acid moieties present on the molecule. In certain embodiments, the prodrug form of the compounds of the present specification can include a protecting group, which, in addition to protecting the compound from oxidation, etc., allows targeting of a specific site in the body of the subject (e.g., the tumor microenvironment). Prodrugs can typically 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).

[0171] PROTAC conjugate Also provided are PROTAC conjugates comprising the compounds of the present specification. The PROTAC conjugate can have the chemical structure of formula (X): A-L’-D (X) and can have In the formula, A is a compound described in this specification (for example, a kinase inhibitor described in this specification), L’ is a linker or does not exist, and D is a ubiquitin pathway protein-binding moiety.

[0172] Although protein inhibitors can block or reduce protein activity in cells, protein degradation in cells can also reduce the activity of the target protein or completely remove it. Therefore, utilizing the cellular proteolytic pathway can provide an additional means for reducing or removing protein activity. One of the major degradation pathways in cells is known as the ubiquitin-proteasome system. In this system, proteins are marked for degradation by the proteasome by ubiquitinating the proteins. Ubiquitination of proteins can be achieved by a ubiquitin pathway protein-binding moiety that binds to the protein and adds ubiquitin molecules to the protein.

[0173] To utilize this degradation pathway, PROTACs have been developed. The term "PROTAC" refers to a target protein degrading chimeric molecule that includes a ubiquitin pathway protein-binding moiety, optionally a linker, and a target moiety. A PROTAC brings together a ubiquitin pathway protein-binding moiety (such as an E3 ubiquitin ligase) with the protein that is to be targeted for degradation. To facilitate the degradation of the protein by the proteasome, a PROTAC consists of a group that binds to the ubiquitin ligase and a group that binds to the protein targeted for degradation (optionally these groups are linked by a linker). This molecular construct can bring about a ubiquitin pathway protein that binds the adjacent moieties to the protein so that it is ubiquitinated and marked for degradation.

[0174] The ubiquitin pathway protein-binding moiety is any suitable structure that recognizes and binds to a ubiquitin pathway protein. Generally, a ubiquitin pathway protein is any entity or complex that can catalyze or cause to catalyze the transfer of ubiquitin or a ubiquitin-like modified polypeptide (e.g., Nedd8, APG12, or ISG15 / UCRP) to another protein. In certain embodiments, the ubiquitin pathway protein is a ubiquitin protein ligase or E3 protein. There are at least 100 different E3 proteins encoded by the human genome. Winston et al., A Family of Mammalian F-Box Proteins, Current Biology 9(20): 1180-1182 (1999).

[0175] In certain embodiments, the ubiquitin pathway protein is a protein that is involved in the ubiquitin-like pathway that transfers a ubiquitin-like modified polypeptide (e.g., SUMO, Nedd8, APG12, or ISG15 / UCRP), or is a component of the ubiquitin-like pathway. The components of the ubiquitin-like modification pathway are typically homologs of the ubiquitin pathway. For example, the ubiquitin-like pathway for SUMO includes homologs of the ubiquitin protein activating enzyme or E1 protein, the ubiquitin protein conjugating enzyme or E2 protein, and the ubiquitin ligase or E3 protein.

[0176] In certain embodiments, the ubiquitin pathway protein-binding moiety is any suitable ligand to a ubiquitin pathway protein, such as a ubiquitin protein ligase or E3 protein or homologs thereof. In certain embodiments, the ubiquitin pathway protein-binding peptide is a domain or region of a ligand to a ubiquitin pathway protein. In certain embodiments, the ubiquitin pathway protein-binding moiety recognizes and binds to the ubiquitin pathway protein in a controlled manner.

[0177] Component A is a compound of the present specification that binds to a target protein intended to be degraded (e.g., a targeted kinase inhibitor such as CDK, FLT3, or Haspin kinase). The term "protein" includes oligopeptide sequences and polypeptide sequences of sufficient length that can bind to Component A.

[0178] The specific A group is any of the compounds described herein.

[0179] The compound and the ubiquitin pathway protein-binding moiety may be optionally linked by a linker. The linker may be any suitable linker. The linker may contain atoms selected from C, N, O, S, Si, and P; C, N, O, S, and P; or C, N, O, and S. The linker can have a backbone that ranges in length from as few as about 2 atoms in the backbone of the linker to as many as about 100 or more contiguous atoms in the backbone of the linker. The "backbone" of the linker is the shortest chain of contiguous atoms that forms a covalently linked connection between A and D. In some embodiments, the multivalent linker has a branched backbone, and each branch serves as a section of the backbone linker until it reaches a terminus.

[0180] For example, the linker can have a chain length of at least about 7 atoms. In some embodiments, the linker is at least about 10 atoms in length. In some embodiments, the linker is at least about 14 atoms in length. In some embodiments, the linker is between about 7 and about 31 atoms (e.g., about 7 - 31, 7 - about 31, or 7 - 31), between about 7 and about 24 atoms (e.g., about 7 - 24, 7 - about 24, or 7 - 24), or between about 7 and about 20 atoms (e.g., about 7 - 20, 7 - about 20, or 7 - 20). In some embodiments, the linker is between about 14 and about 31 atoms (e.g., about 14 - 31, 14 - about 31, or 14 - 31), between about 14 and about 24 atoms (e.g., about 14 - 24, 14 - about 24, or 14 - 24), or between about 14 and about 20 atoms (e.g., about 14 - 20, 14 - about 20, or 14 - 20). In some embodiments, the linker has a chain length of 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 - 15 atoms, 1 - 5 atoms, 5 - 10 atoms, 5 - 20 atoms, 10 - 40 atoms, or 25 - 100 atoms.

[0181] The linker can include at least one carbon - carbon bond and / or at least one amide bond. The linker can include one or more L - or D - configurations, natural or non - natural amino acids, PEG monomers, PEG oligomers, PEG polymers, or any combination of the foregoing. For a linker that includes one or more PEG units, all carbon and oxygen atoms of the PEG units are part of the backbone, unless otherwise specified.

[0182] In certain embodiments, the linker is a group that includes one or more covalently - linked structural units.

[0183] In certain embodiments, the linker group is an optionally substituted (poly)ethylene glycol having from 1 to about 100 ethylene glycol units, from about 1 to about 50 ethylene glycol units, from 1 to about 25 ethylene glycol units, from about 1 to 10 ethylene glycol units, from 1 to about 8 ethylene glycol units, and from 1 to 6 ethylene glycol units, from 2 to 4 ethylene glycol units, and / or is interspersed with atoms of O, N, S, P or Si, each optionally substituted, and having an optionally substituted alkyl group. In certain embodiments, the linker is substituted with each group of aryl, phenyl, benzyl, alkyl, alkylene or heterocycle. In certain embodiments, the linker is asymmetric. In certain embodiments, the linker is symmetric.

[0184] Alternatively or in addition to chain length, in some embodiments, the linker can have suitable substituents that affect hydrophobicity or hydrophilicity. Thus, for example, the linker can have hydrophobic side chain groups such as those groups of alkyl, cycloalkyl, aryl, arylalkyl, etc., each optionally substituted. If the linker contains one or more amino acids, the linker can 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.

[0185] The linker can include a spacer (e.g., conjugated with a spacer and / or including a spacer). The spacer can be any suitable spacer. The spacer of the linker can include hydrophilic, hydrophobic, amphiphilic, non-peptidic, peptidic, and / or aromatic monomers. The length of the spacer can range from 1 to 30 (e.g., 1 to 30 carbon atoms, PEG having 1 to 30 units, etc.). Examples of hydrophilic spacers include, but are not limited to, polyethylene glycol polymers and their derivatives. Examples of hydrophobic spacers include, but are not limited to, pure or mixed branched hydrocarbons, fluorocarbons, alkane, alkene, and / or alkyne polymers. Examples of amphiphilic spacers include, but are not limited to, pure or mixed phospholipids, and / or their derivatives. Examples of peptide spacers include, but are not limited to, pure or mixed single branched L- or D-configurations, essential, non-essential, natural, and non-natural amino acids and their derivatives. Examples of aromatic spacers include, but are not limited to, pure or mixed repeating quinoids.

[0186] In some embodiments, the linker is formed via click chemistry / click-chemistry-derived synthetic methods. One of ordinary skill in the art will understand that the terms "click chemistry" and "click-chemistry-derived" generally refer to a class of small molecule reactions commonly used in conjugation and enable the matching of substrates for selection on specific molecules. Click chemistry describes a way to produce products following examples in nature, which also produces substances by combining small molecule units, rather than a single specific reaction. In many applications, click reactions combine biomolecules and reporter molecules. Click chemistry is not limited to biological conditions, and the concept of "click" reactions has been used in pharmacological applications and various biomimetic applications. However, they have been made useful for the detection, localization, and characterization of biomolecules, which is worthy of note.

[0187] The click reaction can occur in one pot, is typically not hindered by water, can produce minimal by-products, and is "spring-loaded" with a high thermodynamic driving force that quickly and irreversibly drives it to a high yield of a single reaction product, with high reaction specificity (in some cases, with both regioselectivity and stereoselectivity). These qualities make the click reaction suitable for the task of isolating and targeting molecules in a complex biological environment. Thus, in such an environment, the product needs to be physiologically stable and any by-products need to be non-toxic (e.g., for in vivo systems).

[0188] Salt The compounds and conjugates can be presented as pharmaceutically acceptable salts. The term "pharmaceutically acceptable salts" refers to salts whose counterions can be used in pharmaceuticals. In various embodiments, such salts include: 1) acid addition salts that can be obtained by reacting the free base of the parent compound with inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as 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, or 2) salts formed when the acidic protons present in the parent compound are replaced by metal ions, such as alkali metal ions, alkaline earth metal ions, or aluminum ions, or coordinated with organic bases such as ethanolamine, diethanolamine, triethanolamine, trimethamine, N-methylglucamine, etc., but are not limited thereto. Pharmaceutically acceptable salts are well known to those skilled in the art, and any such pharmaceutically acceptable salts are contemplated in the context of the embodiments described herein.

[0189] Pharmaceutically acceptable salts can be synthesized from parent compounds containing basic or acidic moieties by conventional chemical methods. In some examples, such salts can be prepared by reacting the free acid or free base forms of these compounds with a stoichiometric amount of an appropriate base or acid in water, or in an organic solvent, or in a mixture of the two, and generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. A listing of suitable salts can be found in Remington’s Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, the disclosure of which is hereby incorporated by reference.

[0190] In various embodiments, suitable acid addition salts are formed from acids that form non-toxic salts. Exemplary examples include acetates, aspartates, benzoates, besylates, bicarbonates / carbonates, bisulfates / sulfates, borates, camsylates, citrates, edisylates, esylates, formates, fumarates, gluceptates, glucuronates, hexafluorophosphates, hibenzates, hydrochlorides / chlorides, hydrobromides / bromides, hydroiodides / iodides, isethionates, lactates, malates, maleates, malonates, mesylates, methyl sulfates, naphthylates, 2-napsylates, nicotinates, nitrates, orotates, oxalates, palmitates, pamoates, phosphates / hydrogen phosphates / dihydrogen phosphates, saccharates, stearates, succinates, tartrates, tosylates, and trifluoroacetates.

[0191] In various embodiments, suitable base salts are formed from bases that form non-toxic salts. Exemplary examples include arginine salts, benzathine salts, calcium salts, choline salts, diethylamine salts, diolamine salts, glycine salts, lysine salts, magnesium salts, meglumine salts, olamine salts, potassium salts, sodium salts, tromethamine salts, and zinc salts. Hemisalts of acids and bases may also be formed, for example, hemisulfates and hemicalcium salts.

[0192] In each embodiment herein, it will be understood that the formulations include not only all pharmaceutically acceptable salts of the compounds and conjugates, but also any and all hydrates and / or solvates of the compound formulations or their salts. The term "solvate" means a compound or its salt that further includes a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces. Where the solvent is water, the solvate is a hydrate.

[0193] Certain functional groups, such as hydroxy, amino and other groups, are found to form complexes and / or coordination compounds with water and / or various solvents in diverse physical forms of the compound. Thus, the formulations are understood to include and represent these various hydrates and / or solvates.

[0194] Compositions, Routes of Administration and Dosages Further provided are compositions comprising the above-mentioned compound (e.g., a compound of formula I, formula IA, formula IB or formula II) or 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 component including the compound or conjugate. Compositions can be prepared from isolated compounds or conjugates, or from salts, solutions, hydrates, solvates, and other forms of the compound and / or conjugate.

[0195] The term "pharmaceutically acceptable carrier" means one or more compatible solid or liquid fillers, diluents or encapsulating substances suitable for administration to humans or other vertebrates. The term "carrier" refers to an organic or inorganic, natural or synthetic component with which the active ingredient is combined to facilitate administration. The carrier may be an excipient. The choice of carrier can depend on factors such as the particular method of administration, the effect of the carrier on solubility and stability, and the nature of the dosage form. Pharmaceutically acceptable compositions suitable for the delivery of the compounds described herein, and methods for their preparation, can be found, for example, in Remington: The Science & Practice of Pharmacy, 21st edition (Lippincott Williams & Wilkins, 2005).

[0196] The components of the composition can also be combined with the compound in a manner such that there is no interaction that would substantially impair the desired pharmaceutical efficacy.

[0197] The composition can include, for example, Cremophor, polysorbate, nanoparticles, polymers, or hydrogels. In certain embodiments, the pharmaceutical composition includes a plurality of compounds and a pharmaceutically acceptable carrier. In certain embodiments, the pharmaceutical composition further includes at least one additional pharmaceutically active agent. This at least one additional pharmaceutically active agent can be an agent useful for the treatment of cancer.

[0198] The pharmaceutical composition can be prepared by combining one or more compounds or conjugates with a pharmaceutically acceptable carrier and optionally one or more additional components (such as pharmaceutically active ingredients). The formulation can be administered in a pharmaceutically acceptable solution that contains pharmaceutically acceptable concentrations of salts, buffers, preservatives, compatible carriers, adjuvants, and optionally other therapeutic components as defined.

[0199] Examples of pharmaceutically acceptable carriers can include any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, as well as combinations thereof. The carrier can be suitable for parenteral administration. Examples of pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the immediate preparation of sterile injectable solutions or dispersions. Examples of such carriers (or excipients) include, but are not limited to, polymers such as calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polyethylene glycol. One or more other active agents can also be incorporated into the pharmaceutical composition.

[0200] The composition can be formulated as a liquid, such as a suspension or solution. Liquid formulations can include water, ethanol, polyethylene glycol, propylene glycol, methylcellulose, or a suitable oil, as well as one or more emulsifying agents and / or suspending agents. Liquid formulations can be prepared by reconstitution of a solid.

[0201] Pharmaceutical preparations (for example, for parenteral administration) include aqueous solutions of active compounds in water-soluble form. In addition, suspensions of the active compounds can be prepared as suitable oily injection suspensions. Aqueous suspensions can contain the compound, alone or in further combination with one or more other active agents, in an additive mixture with suitable excipients. Excipients include suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum and acacia gum; dispersing 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 heptadecaethyleneoxycetanol; condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols 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 contain one or more preservatives such as ascorbic acid or ethyl, n-propyl, or p-hydroxybenzoate, and one or more colorants. In certain embodiments, the aqueous suspension can further contain a suitable lipophilic solvent or vehicle, such as a fatty oil like sesame oil, or can further contain a synthetic fatty acid ester such as ethyl oleate or a triglyceride, or liposomes. Optionally, the suspension can also contain a suitable stabilizer or an agent that increases the solubility of compounds that enable the preparation of highly concentrated solutions.

[0202] Alternatively, the pharmaceutical composition can be in powder form for constitution with a suitable vehicle, for example sterile pyrogen-free water, before use. Dispersible powders and granules suitable for the preparation of aqueous suspensions by addition of water can be provided with the active ingredient in admixture with a suspending agent, dispersing agent or wetting agent, and one or more preservatives. Additional excipients, for example colorants, may also be present.

[0203] Suitable emulsifying agents include naturally occurring gums such as acacia gum or tragacanth gum; naturally occurring phospholipids such as soybean lecithin; and esters, such as sorbitan monooleate, containing partial esters derived from fatty acids and hexitol anhydrides, and condensation products of the partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. Isotonic agents, such as sugars, polyhydric alcohols such as mannitol, sorbitol, or sodium chloride, can be included in the composition. Prolonged absorption of the injectable composition can be achieved by including in the composition one or more absorption delaying agents such as monostearates and gelatin.

[0204] For use in therapy or treatment, an effective amount of the compound or composition can be administered to a subject by any method that delivers the compound as desired. Administration of the composition can be achieved 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, mucosal (e.g., topical administration to the eye), inhalation, and topical administration.

[0205] Colorants and / or flavoring agents may be included. For example, the compound can be formulated (such as by encapsulation in liposomes or microspheres) and then incorporated into an edible product such as a refrigerated beverage that further contains colorants and flavoring agents.

[0206] Exemplary formats for oral administration include, but are not limited to, tablets, capsules, elixirs, syrups, and the like.

[0207] In certain embodiments, the compound may be administered directly into the bloodstream, into muscle, or into internal organs. Suitable routes for such parenteral administration include intravenous, intraarterial, intraperitoneal, subarachnoid, epidural, intracerebroventricular, intraurethral, intrasternal, intracranial, intratumoral, intramuscular, intranasal, and subcutaneous. Suitable means for parenteral administration include needle syringes (including micro needles), needleless syringes, and infusion techniques. Where it is desirable to deliver the compound and / or composition systemically, the compound and / or composition can be formulated for parenteral administration by injection, such as by bolus injection or continuous infusion. Formulations for injection can be provided in unit dosage form, for example, in ampoules, or in multi-dose containers with added preservatives. The composition can take the form of a suspension, solution, or emulsion in an oily vehicle or an aqueous vehicle and can contain formulating agents such as suspending agents, stabilizers, and / or dispersing agents.

[0208] Parenteral formulations are typically aqueous solutions that can contain a carrier or excipient, such as salts, carbohydrates, and buffers (preferably at a pH of 3-9), but more preferably, for some applications, they are formulated as a sterile non-aqueous solution or in a dry form that will be used in conjunction with a suitable vehicle such as sterile pyrogen-free water.

[0209] Liquid formulations can be adapted for parenteral administration of the compound. For example, the preparation of parenteral formulations under aseptic conditions by lyophilization under aseptic conditions can be readily achieved 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 such as the incorporation of solubilizing agents.

[0210] Formulations for parenteral administration can be formulated for immediate and / or modified release. The compounds can be administered in a sustained release formulation, for example, in a composition containing a sustained release polymer. The compounds can be prepared with a carrier that will protect it against rapid release, for example, a controlled release formulation including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, polylactic acid and poly(lactic-co-glycolic) copolymer (PGLA) can be used. Methods for preparing such formulations are generally known to those skilled in the art.

[0211] Sterile injectable solutions can be prepared by incorporating the compound, alone or in further combination with one or more other active agents, in a suitable solvent having, as required, one or a combination of the ingredients described above in the required amounts and then filtering the solution aseptically. Typically, dispersions are prepared by incorporating the compound in a sterile vehicle containing a dispersion medium and any additional ingredients described above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze drying, which yield a powder of the active ingredient plus any additional desired ingredients from a previously aseptically filtered solution thereof, or these ingredients may be aseptically filtered together.

[0212] The pharmaceutical compositions can be formulated as 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 (for example, glycerol, propylene glycol, and liquid polyethylene glycol, etc.), and suitable mixtures thereof. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, or in the case of dispersions, by the maintenance of the required particle size by the use of surfactants.

[0213] The compound, or the pharmaceutical composition comprising the compound, may be administered continuously, where appropriate.

[0214] Methods of treatment and use A method of treating a kinase-mediated disease is provided. In certain embodiments, the kinase-mediated disease is cancer. The method can comprise administering to a subject (e.g., a subject experiencing or at risk of experiencing a kinase-mediated disease such as cancer) an effective amount of any of the above compounds, the above conjugates, pharmaceutically acceptable salts, N-oxides, hydrates, solvates, tautomers or optical isomers of the compound or conjugate, or the pharmaceutical compositions herein. In certain embodiments, the compound has the structure of formula I:

[0215]

Chemical formula

[0216]

Chemical formula

[0217] In certain embodiments, the compound is of formula IA:

[0218]

Chemical formula

[0219]

Chemical formula

[0220] In certain embodiments, the compound is of formula IB:

[0221]

Chemical formula

[0222] In certain embodiments, the compound is of formula II:

[0223]

Chemical formula

[0224] The kinase-mediated disease can 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 can be acute myeloid leukemia (AML). The cancer can be chronic myeloid leukemia. The cancer can be gastric carcinoma. The cancer can be drug-resistant cancer. The cancer can 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 carcinoma, head cancer, neck cancer, thyroid cancer, kidney cancer, liver cancer and stomach cancer.

[0225] In certain embodiments, the compound or conjugate administered to the subject comprises compound HSH2177. In certain embodiments, the compound or conjugate administered to the subject comprises compound HSH2177, and administering an effective amount to the subject results in an IC of about 50 nM or less (e.g., 50 nM or less) 50In this case, CDK in the subject is inhibited. In certain embodiments, the compound or conjugate administered to the subject comprises compound 205. In certain embodiments, the compound or conjugate administered to the subject comprises compound 205, and administering an effective amount to the subject inhibits both FLT3 and Haspin kinase with specificity in the subject.

[0226] In certain embodiments of the method, administering an effective amount to the subject is an IC of about 50 nM or less (such as 50 nM or less). 50 In this case, CDK in the subject is inhibited (such as with specificity). In certain embodiments of the method, administering the compound or pharmaceutical composition is an IC of about 30 nM or less (such as 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 less than 5 nM). 50 In this case, CDK is inhibited (such as with specificity). Administering an effective amount to the subject can inhibit both FLT3 and Haspin (such as with the specificity of a combination of two agents). In certain embodiments, administering an effective amount of compound 205 or an analog thereof is, for example, an IC of about 50 nM or less. 50 (such as an IC of 50 nM or less). 50 ) In this case, both FLT3 and Haspin kinase are inhibited in combination of two agents.

[0227] The terms "treat", "treating", "treated" or "treatment" (in terms of a disease state or disorder) are approaches for obtaining beneficial or desired results, including clinical results, preferably clinical results, and include one or more of the following, but are not limited thereto: improving the condition associated with the disease, curing the disease, reducing the severity of the disease, or otherwise suppressing the disease, slowing the progression of the disease, alleviating one or more symptoms associated with the disease, reducing the severity of the disease, suppressing the onset of the disease, suppressing the progression of the disease, improving the quality of life of a person suffering from the disease, extending the survival rate, and / or prophylactic treatment or preventive treatment.

[0228] As used herein, "effective amount" refers to any amount of a compound or conjugate in the context of use in a treatment, which, when administered as part of a desired dosage regimen (to a subject, e.g., a human), is sufficient to achieve a desired biological effect (e.g., alleviating symptoms, improving a condition, or slowing the onset of a disease state, in accordance with clinically acceptable standards for the disorder or condition to be treated or for cosmetic purposes, e.g., in a reasonable benefit / risk ratio applicable to any medical treatment). It refers to an amount of the compound or conjugate in a preparation that is sufficient to achieve the desired biological effect.

[0229] In certain embodiments, the method may further comprise administering to the subject a second therapeutic agent 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 therapeutic agent. In certain embodiments, the second therapeutic agent may comprise radiation therapy.

[0230] It is understood that in the methods described herein, the individual components or combinations of co-administered agents can be administered by any suitable means, contemporaneously, simultaneously, sequentially, separately, or in a single pharmaceutical formulation. Where the co-administered compounds or compositions are administered in separate dosage forms, the number of dosages administered per day for each compound may be the same or different. The compounds, conjugates, and / or compositions can be administered via the same or different routes of administration, both to each other and / or as the second therapeutic agent. The compounds, conjugates, or compositions can be administered according to concurrent or alternating regimens, simultaneously, at the same time or at different times, in divided or single form, during the course of the therapy.

[0231] Selected from among a variety of active compounds, in combination with the teachings provided herein by comparing factors such as efficacy, relative bioavailability, patient weight, severity of side effects, and method of administration, an effective prophylactic or therapeutic regimen can be planned that does not cause substantial unwanted toxicity but is effective for treating a particular subject. The effective amount for any particular application can vary depending on factors such as the disease or condition being treated, the particular compound being administered, the size of the subject, or the severity of the disease or condition. One of ordinary skill in the art can determine the effective amount of a particular compound and / or other therapeutic agent from an empirical perspective without the need for undue experimentation.

[0232] For example, for any compound, the effective amount can first be determined from an animal model. The effective dose can also be determined from human data for compounds tested in humans and for compounds known to exhibit similar pharmacological activity such as other related active agents. Higher doses may be required for parenteral administration. The dose applied can be adjusted based on relative bioavailability and the efficacy of the compound being administered. Adjusting the dose to achieve maximum efficacy based on the methods described above and other methods well known in the art is well within the ability of one of ordinary skill in the art.

[0233] The maximum dose, i.e., the safest dose based on some medical judgment, can be used. Multiple administrations per day can be used to achieve an appropriate systemic level of the compound. The appropriate systemic level can be determined, for example, by measuring the peak of the patient or the sustained plasma level of the drug.

[0234] "Dose" and "dosage" can be used interchangeably herein.

[0235] Generally, the oral dosage of the compound is about 0.01 milligrams / kg per day to 1,000 milligrams / kg per day in human subjects. Oral dosages in the range of 0.5 to 50 milligrams / kg in one or multiple administrations per day can produce therapeutic results. The dosage can be appropriately adjusted to achieve the desired local or systemic drug level depending on the method of administration. For example, intravenous administration can vary at dosages one to several digits lower per day. If the response in the subject is insufficient at such dosages, higher dosages (or higher effective dosages by different, more localized delivery routes) can be utilized to the extent tolerated by the patient. Multiple administrations per day are contemplated to achieve an appropriate systemic level of the compound.

[0236] For clinical use, any compound or conjugate can be administered in an amount equal to or equivalent to 0.2 to 2,000 milligrams (mg) of the compound per kilogram (1 kg) of the subject's body weight per day. The compound can be administered in an amount equal to or equivalent to 2 to 2,000 mg of the compound per 1 kg of the subject's body weight per day. The compound can be administered in an amount equal to or equivalent to 20 to 2,000 mg of the compound per 1 kg of the subject's body weight per day. The compound can be administered in an amount equal to or equivalent to 50 to 2,000 mg of the compound per 1 kg of the subject's body weight per day. The compound can be administered in an amount equal to or equivalent to 100 to 2,000 mg of the compound per 1 kg of the subject's body weight per day. The compound can be administered in an amount equal to or equivalent to 200 to 2,000 mg of the compound per 1 kg of the subject's body weight per day. Where a precursor or prodrug of the compound is to be administered, it is administered in an amount equivalent to the amount of the compound that delivers the amount of the compound described above, i.e., in an amount sufficient to deliver the amount of the compound described above.

[0237] The formulation of the compound or a pharmaceutically acceptable salt thereof can be administered to a human subject in an effective amount. A typical dosage range can be from about 0.01 micrograms / kg to about 2 mg / kg of body weight per day. The dosage of the agent to be administered will probably depend on variables such as the type and extent of the disorder, the overall health of the particular subject, the particular compound being administered, the excipients used to formulate the compound, and the route of its administration. Using conventional experimentation, the dosage and frequency of dosing for any particular compound or a pharmaceutically acceptable salt thereof can be optimized.

[0238] The compound or a pharmaceutically acceptable salt thereof may be administered at a concentration in the range from about 0.001 micrograms / kg to more than about 500 mg / kg. For example, the concentration may be 0.001 micrograms / kg, 0.01 micrograms / kg, 0.05 micrograms / kg, 0.1 micrograms / kg, 0.5 micrograms / kg, 1.0 micrograms / kg, 10.0 micrograms / kg, 50.0 micrograms / kg, 100.0 micrograms / kg, 500 micrograms / 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, 40.0 mg / kg, 45.0 mg / kg, 50.0 mg / kg, 60.0 mg / kg, 70.0 mg / kg, 80.0 mg / kg, 90.0 mg / kg, 100.0 mg / kg, 150.0 mg / kg, 200.0 mg / kg, 250.0 mg / kg, 300.0 mg / kg, 350.0 mg / kg, 400.0 mg / kg, 450.0 mg / kg to more than about 500.0 mg / kg, or any incremental value thereof. It is understood that all values between these values and all ranges between these ranges are meant to be included.

[0239] The compound or a pharmaceutically acceptable salt thereof may be administered at a dosage in the range of about 0.2 milligrams / kg / day to more than about 100 mg / kg / day. For example, the dosage may 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 10 mg / kg / day, 0.25 mg / kg / day to 7.5 mg / kg / day, 0.25 mg / kg / day to 5 mg / kg / day, 0.5 mg / kg / day to 50 mg / kg / day, 0.5 mg / kg / day to 25 mg / kg / day, 0.5 mg / kg / day to 20 mg / kg / day, 0.5 mg / kg / day to 15 mg / kg / day, 0.5 mg / kg / day to 10 mg / kg / day, 0.5 mg / kg / day to 7.5 mg / kg / day, 0.5 mg / kg / day to 5 mg / kg / day, 0.75 mg / kg / day to 50 mg / kg / day, 0.75 mg / kg / day to 25 mg / kg / day, 0.75 mg / kg / day to 20 mg / kg / day, 0.75 mg / kg / day to 15 mg / kg / day, 0.75 mg / kg / day to 10 mg / kg / day, 0.75 mg / kg / day to 7.5 mg / kg / day, 0.75 mg / kg / day to 5 mg / kg / day, 1.0 mg / kg / day to 50 mg / kg / day, 1.0 mg / kg / day to 25 mg / kg / day, 1.0 mg / kg / day to 20 mg / kg / day, 1.0 mg / kg / day to 15 mg / kg / day, 1.0 mg / kg / day to 10 mg / kg / day, 1.0 mg / kg / day to 7.5 mg / kg / day, 1.0 mg / kg / day to 5 mg / kg / day, 2 mg / kg / day to 50 mg / kg / day, 2 mg / kg / day to 25 mg / kg / day, 2 mg / kg / day to 20 mg / kg / day, 2 mg / kg / day to 15 mg / kg / day, 2 mg / kg / day to 10 mg / kg / day, 2 mg / kg / day to 7.5 mg / kg / day, or 2 mg / kg / day to 5 mg / kg / day.

[0240] The compound may be administered at a dosage in the range of about 0.25 milligrams / kg / day to about 25 mg / kg / day. For example, the dosage may 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, 4.25 mg / kg / day, 4.5 mg / kg / day, 4.75 mg / kg / day, 5 mg / kg / day, 5.5 mg / kg / day, 6.0 mg / kg / day, 6.5 mg / kg / day, 7.0 mg / kg / day, 7.5 mg / kg / day, 8.0 mg / kg / day, 8.5 mg / kg / day, 9.0 mg / kg / day, 9.5 mg / kg / day, 10 mg / kg / day, 11 mg / kg / day, 12 mg / kg / day, 13 mg / kg / day, 14 mg / kg / day, 15 mg / kg / day, 16 mg / kg / day, 17 mg / kg / day, 18 mg / kg / day, 19 mg / kg / day, 20 mg / kg / day, 21 mg / kg / day, 22 mg / kg / day, 23 mg / kg / day, 24 mg / kg / day, 25 mg / kg / day, 26 mg / kg / day, 27 mg / kg / day, 28 mg / kg / day, 29 mg / kg / day, 30 mg / kg / day, 31 mg / kg / day, 32 mg / kg / day, 33 mg / kg / day, 34 mg / kg / day, 35 mg / kg / day, 36 mg / kg / day, 37 mg / kg / day, 38 mg / kg / day, 39 mg / kg / day, 40 mg / kg / day, 41 mg / kg / day, 42 mg / kg / day, 43 mg / kg / day, 44 mg / kg / day, 45 mg / kg / day, 46 mg / kg / day, 47 mg / kg / day, 48 mg / kg / day, 49 mg / kg / day, or 50 mg / kg / day.

[0241] The compound, a pharmaceutically acceptable salt thereof, or a precursor thereof may be administered at a concentration in the range of 0.01 micromoles to 500 micromoles or more. For example, the dosage may be 0.01 micromoles, 0.02 micromoles, 0.05 micromoles, 0.1 micromoles, 0.15 micromoles, 0.2 micromoles, 0.5 micromoles, 0.7 micromoles, 1.0 micromoles, 3.0 micromoles, 5.0 micromoles, 7.0 micromoles, 10.0 micromoles, 15.0 micromoles, 20.0 micromoles, 25.0 micromoles, 30.0 micromoles, 35.0 micromoles, 40.0 micromoles, 45.0 micromoles, 50.0 micromoles, 60.0 micromoles, 70.0 micromoles, 80.0 micromoles, 90.0 micromoles, 100.0 micromoles, 150.0 micromoles, 200.0 micromoles, 250.0 micromoles, 300.0 micromoles, 350.0 micromoles, 400.0 micromoles, 450.0 micromoles to greater than about 500.0 micromoles, or any incremental value thereof. It is understood that all values between these values, and all ranges between these ranges, are meant to be included.

[0242] The compound, or a pharmaceutically acceptable salt thereof, or a precursor thereof, may be administered at a concentration in the range of 0.10 micrograms / mL to 500.0 micrograms / mL. For example, the concentration may be 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 g / mL, 250.0 micrograms / mL, 300.0 micrograms / mL, 350.0 micrograms / mL, 400.0 micrograms / mL, 450.0 micrograms / mL to about 500.0 micrograms / mL or more, or any incremental value thereof. It is understood that all values between these values and all ranges between these ranges are meant to be included.

[0243] The formulation may be administered in a pharmaceutically acceptable solution that may contain, as prescribed, a pharmaceutically acceptable concentration of a salt, buffer, preservative, compatible carrier, adjuvant, and optionally other therapeutic ingredients. For use in therapy, an effective amount of the compound may be administered to a subject by any method that delivers the compound to the desired surface. Administration of the pharmaceutical composition can be achieved 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., into a tumor or abscess), mucosal (e.g., topical administration to the eye), inhalation, and topical administration.

[0244] For intravenous and other parenteral routes of administration, the compounds can be formulated as lyophilized preparations, as lyophilized preparations of liposome-intercalated or encapsulated active compounds, as lipid complexes in aqueous suspensions, or as salt complexes. Lyophilized formulations are generally reconstituted immediately prior to administration in a suitable aqueous solution, for example, sterile water or physiological saline.

[0245] For oral administration, the compounds can be immediately formulated by combining the active compound with a pharmaceutically acceptable carrier well known in the art. Such carriers enable the compound to be formulated as 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 optionally be obtained by grinding the resulting mixture and, if desired, processing the mixture of granules after adding suitable auxiliaries to obtain the core of the tablet or dragee and as solid excipients. Suitable excipients include, specifically, sugars containing fillers such as lactose, sucrose, mannitol or sorbitol; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethyl cellulose and / or polyvinyl pyrrolidone (PVP). If desired, disintegrants can be added, for example, cross-linked PVP, agar, or alginic acid or its salts, such as sodium alginate. Optionally, oral formulations can also be formulated in physiological saline or buffer solutions, for example, ethylenediaminetetraacetic acid (EDTA) for neutralizing the internal acidic state, or administered without any carrier.

[0246] Also to be considered is the oral dosage form of the compound. The compound can be chemically modified to be effective for oral delivery of the derivative. Generally, the chemical modification to be considered is the attachment of at least one moiety to the compound itself, which moiety (a) inhibits acid hydrolysis and (b) enables uptake from the stomach or intestine into the bloodstream. Further desirable is an increase in the overall stability of the compound and an increase in the 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, N.Y., pp. 367-383 (1981); Newmark et al., J Appl Biochem 4:185-189 (1982). Other polymers that can be used are poly-1,3-dioxolane and poly-1,3,6-thioxocane. For pharmaceutical use, as indicated above, polyethylene glycol moieties are preferred.

[0247] The site of release of the compounds or pharmaceutically acceptable salts thereof described herein can be the stomach, small intestine (e.g., duodenum, jejunum or ileum), or large intestine. One of ordinary skill in the art has available formulations that will not dissolve in the stomach but will release the substance in the duodenum or somewhere in the intestine. Release can be by protection of the compound or by release of the compound or pharmaceutically acceptable salts thereof beyond the gastric environment, e.g., in the intestine, to avoid the deleterious effects of the gastric environment.

[0248] To ensure complete gastric resistance, a coating that is at least impermeable to pH 5.0 may be essential. Examples of more common inert ingredients used as enteric coatings are cellulose trimellitate acetate (CAT), hydroxypropylmethylcellulose phthalate (HPMCP), HPMCP50, HPMCP55, polyvinyl acetate phthalate (PVAP), Eudragit L30D, Aquateric, cellulose acetate phthalate (CAP), Eudragit L, Eudragit S, and shellac. These coatings can be used as a mixed film.

[0249] The coating, or coating mixture, can also be used in tablets where protection against the stomach is not intended. This includes sugar coatings, or coatings that make the tablet easier to swallow. Capsules can consist of a hard shell (such as gelatin) for the delivery of dry therapeutic agents (e.g., powders); for liquid forms, a soft gelatin shell can be used. The shell material of cachets can be thick starch or other edible paper. For pills, lozenges, molded tablets or tablet powders, wet granulation techniques can be used.

[0250] As noted above, the compound or its pharmaceutically acceptable salts can be included in the formulation as a fine multitude of microparticles in the form of granules, or pellets with a particle size of about 1 mm. The formulation of materials for capsule administration can also be a powder, a lightly compressed plug, or even a tablet.

[0251] Oral pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, as well as soft-sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredient in a mixture with a filler such as lactose, a binder such as starch, and / or a lubricant such as talc or magnesium stearate, and optionally a stabilizer. In soft capsules, the active compound can be dissolved or suspended in a suitable liquid such as fatty oil, liquid paraffin, or liquid polyethylene glycol. In addition, a stabilizer may be added. Microspheres formulated for oral administration may also be used. Such microspheres have been well defined in the art. All formulations for oral administration should be in a dosage suitable for such administration.

[0252] For oral administration, the composition can take the form of tablets or lozenges formulated by conventional methods.

[0253] For topical administration, the compound can be formulated as a solution, gel, ointment, cream, suspension, 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, and those designed for transdermal, transmucosal, oral, or pulmonary administration.

[0254] For administration by inhalation, the compounds can conveniently be delivered in the form of an aerosol spray presentation from a pressurized pack or a nebulizer, with the use of a suitable propellant, for example dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. For example, gelatin capsules and cartridges for use in an inhaler or insufflator can be formulated to contain a powder mix of the compound with a suitable powder base such as lactose or starch.

[0255] Also contemplated is pulmonary delivery of the compound (or its salt). The compound is inhaled and delivered to the mammalian lung, crossing the alveolar 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-gamma and tumor necrosis factor alpha), and Platz et al., U.S. Patent No. 5,284,656 (granulocyte colony stimulating factor, incorporated herein by reference). Methods and compositions for pulmonary delivery of agents for systemic effect are described in U.S. Patent No. 5,451,569 to Wong et al., issued September 19, 1995 (specifically, the disclosure thereof regarding the methods and compositions is incorporated herein by reference).

[0256] What is contemplated for use are a wide range of mechanical devices designed for pulmonary delivery of therapeutic products, including but not limited to nebulizers, metered dose inhalers, and powder inhalers, all of which are familiar to those skilled in the art.

[0257] Transnasal delivery of pharmaceutical compositions is also contemplated. Transnasal delivery enables a route for the pharmaceutical composition into the bloodstream directly after administration of the therapeutic product to the nose without the need for the product to deposit in the lungs. Formulations for transnasal delivery include those having dextran or cyclodextran.

[0258] When it is desirable to deliver the compounds systemically, they can be formulated for parenteral administration by injection, for example, bolus injection or continuous infusion. Injectable formulations can be provided in unit dosage form, for example, in ampoules, or in multi-dose containers together with an added preservative. The composition can take the form of suspensions, solutions or emulsions in oily vehicles or aqueous vehicles, and can contain formulating agents such as suspending agents, stabilizers and / or dispersing agents.

[0259] Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compound in water-soluble form. In addition, suspensions of the active compound can be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. 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 enable preparation of highly concentrated solutions.

[0260] Alternatively, the active compound can be in powder form for constitution prior to use with a suitable vehicle, for example, sterile pyrogen-free water.

[0261] The compound can also be formulated in rectal or vaginal compositions, such as suppositories or retention enemas, which contain conventional suppository bases such as cocoa butter or other glycerides, for example.

[0262] In addition to the formulations described above, the compound can also be formulated as a depot preparation. Such long-acting formulations may be formulated with suitable polymeric or hydrophobic materials (such as emulsions in acceptable oils) or ion exchange resins, or as a somewhat poorly soluble derivative, for example as a somewhat poorly soluble salt.

[0263] The pharmaceutical composition may also contain suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycol.

[0264] Suitable forms of liquid or solid pharmaceutical preparations are, for example, microencapsulated, entrapped, coated on microscopic gold particles, contained in liposomes, nebulized, aerosol, pellets for implantation into the skin, or dried on sharp objects for wounding into the skin, aqueous solutions for inhalation or physiological saline solutions. The pharmaceutical composition also includes granules, powders, tablets, coated tablets, (micro)capsules, suppositories, syrups, emulsions, suspensions, creams, drops, or preparations with sustained release of the active compound, in which excipients, as well as additives and / or adjuvants, such as disintegrants, binders, coating agents, swelling agents, lubricants, flavoring agents, sweetening agents or solubilizing agents, are customarily used as described above. The pharmaceutical composition is suitable for use in various drug delivery systems. For a brief review of methods for drug delivery, see Langer R, Science 249:1527-1533 (1990).

[0265] The disintegrant may be included in the formulation of the therapeutic agent in a solid dosage form. Materials used as disintegrants include, but are not limited to, starches, such as commercially available starch-based disintegrants including Explotab. Sodium glycolate starch, Amberlite, sodium carboxymethyl cellulose, ultramylopectin, sodium alginate, gelatin, orange peel, acid carboxymethyl cellulose, natural sponges and bentonite can all be used. Another form of disintegrant is an insoluble cation exchange resin. Powdered gums may be used as disintegrants and as binders, examples of which include powdered gums such as agar, karaya or tragacanth. Alginate and its sodium salts are also useful as disintegrants.

[0266] Binders can be used to hold the compounds together to form hard tablets, and materials from natural products such as acacia, tragacanth, starch and gelatin can be mentioned. Others include methylcellulose (MC), ethylcellulose (EC) and carboxymethylcellulose (CMC). Both PVP and hydroxypropylmethylcellulose (HPMC) can be used in an alcohol solution to granulate the therapeutic agent.

[0267] Lubricants may be included in the formulation of the therapeutic agent to prevent sticking during the formulation process. Lubricants can be used as a layer between the therapeutic agent and the die wall, including, but not limited to, stearic acid including its magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils and waxes. Soluble lubricants can also be used, such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycols of various molecular weights, Carbowax 4000 and 6000.

[0268] A flow promoter may be added to the formulation to improve the flow properties of the drug and assist in rearrangement during compression. Examples of flow promoters include starch, talc, calcined silica, and aluminum silicate hydrate.

[0269] To assist in dissolving the therapeutic agent in an aqueous environment, a surfactant may be added as a wetting agent. Examples of surfactants may include anionic detergents such as sodium lauryl sulfate, sodium dioctyl sulfosuccinate, and sodium dioctyl sulfonate. Examples of cationic detergents that may be used include benzalkonium chloride and benzethonium chloride. Potential nonionic detergents that may be included in the formulation as surfactants include lauromacrogol 400, polyoxyl 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 may be present in the formulation of the compound or its derivative either alone or as a mixture in different ratios.

[0270] The compound can be administered per se (neat) or in the form of a pharmaceutically acceptable salt. When used in medicine, the salt should be pharmaceutically acceptable, although pharmaceutically unacceptable salts may sometimes be conveniently used to prepare the pharmaceutically acceptable salts thereof. Such salts include, but are not limited to, those 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. Also, such salts can be prepared as alkali metal salts or alkaline earth salts, such as the sodium, potassium, or calcium salts of carboxylic acid groups.

[0271] The volume of the compound can be diluted or increased with an inert substance. These diluents may include carbohydrates, especially mannitol, a-lactose, lactose anhydrous, cellulose, sucrose, modified dextran and starch. Certain inorganic salts can also be used as fillers including calcium phosphate, magnesium carbonate and sodium chloride. Some commercially available diluents are Fast-Flo, Emdex, STA-Rx1500, Emcompress and Avicell.

[0272] Suitable buffers include acetic acid and salts (1 - 2% w / v), citric acid and salts (1 - 3% w / v), boric acid and salts (0.5 - 2.5% w / v), and phosphoric acid and 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).

[0273] The pharmaceutical composition contains an effective amount of the compound described herein and one or more other therapeutic agents optionally contained in a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" means one or more compatible solid fillers or liquid fillers, diluents or encapsulating substances suitable for administration to humans or other vertebrates. The term "carrier" refers to natural or synthetic, organic or inorganic components with which the active ingredient is combined to facilitate its application. The components of the pharmaceutical composition can also be combined with the compound in a manner such that there is no interaction that would substantially impair the desired efficacy of the pharmaceutical.

[0274] Specifically, although this compound is exemplified, the therapeutic agent is not limited thereto and can be provided in particles. As used herein, "particles" means nanoparticles or microparticles (or in some cases larger particles) that can be incorporated into the whole or part of a compound, or other therapeutic agents described herein. The particles may contain a therapeutic agent in a core surrounded by a coating including, but not limited to, enteric coatings. The therapeutic agent can also be dispersed throughout the particle. The therapeutic agent can also be adsorbed into the particle. The particles may be of any order of release kinetics, including zero-order release, first-order release, second-order release, delayed release, sustained release, immediate release, and any combination thereof. In addition to the therapeutic agent, the particles may include any of these materials used as defined in the art of pharmacy and drug technology, including, but not limited to, erodible, non-erodible, biodegradable, or non-biodegradable substances, or combinations thereof. The particles may be microcapsules containing a compound in solution or in a semi-solid state. The particles may actually be of any shape.

[0275] Both non-biodegradable polymer substances and biodegradable polymer substances can be used in the manufacture of particles for delivering therapeutic agents. Such polymers may be natural polymers or synthetic polymers. The polymer is selected based on the period over which release is desired. Examples of bioadhesive polymers for a particular subject include the biodegradable hydrogels described in Sawhney et al., Macromolecules 26:581-587 (1993), the teachings of which are specifically incorporated herein by reference. These include polyhyaluronic acid, casein, gelatin, gluten, polyanhydrides, polyacrylic acid, alginate, chitosan, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), and poly(octadecyl acrylate).

[0276] The compound can be incorporated into a controlled release system. The term "controlled release" is intended to refer to any drug-containing formulation in which the method and profile of drug release from the formulation are controlled. This refers to both immediate release and non-immediate release formulations, and non-immediate release formulations include, but are not limited to, sustained release formulations and delayed release formulations. The term "sustained release" (also referred to as "extended release") is used in its conventional meaning to refer to a drug formulation that can provide a gradual release of a drug over a long period of time, resulting in a substantially constant blood level of the drug over a long period of time. The term "delayed release" is used in its conventional meaning to refer to a drug formulation in which there is a time delay between administration of the formulation and release of the drug from the formulation. "Delayed release" may or may not be involved in the gradual release of the drug over a long period of time, and thus may or may not be "sustained release".

[0277] The use of long-term sustained-release implants can be particularly suitable for the treatment of chronic conditions. "Long-term" release, as used herein, means that the implant is configured and arranged to deliver therapeutic levels of the active ingredient for at least 7 days and up to 30 - 60 days. Long-term sustained-release implants are well known to those skilled in the art and include some of the release systems described above.

[0278] Depending on the route of administration, a wide range of acceptable dosages are contemplated herein, including dosages that fall within the range of about 10 6 ~10 11 viral particles (VP) / kg. The dosage may be single or divided and may be administered according to a wide range of protocols, including q.d. (once daily), b.i.d. (twice daily), t.i.d (three times daily), or even once every other day, once a week, once a month, once a quarter, etc. In each of these cases, it is understood that the effective amount described herein corresponds to an instance of administration or, alternatively, is determined by the dosing protocol and corresponds to the total daily, weekly, monthly, or quarterly dosage.

[0279] In addition to the exemplary dosage and dosing protocols described herein, an effective amount of any one or mixture of the compounds described herein can be determined by the attending diagnostician or physician using known techniques and / or by observing results obtained in similar circumstances. In determining the effective amount or dosage, a number of factors are considered by the attending diagnostician or physician, including the mammalian species, including humans, its size, age and general health, the particular disease or disorder involved, the degree or severity of the disease or disorder, the response of the individual patient, the particular compound administered, the method of administration, the characteristics of the bioavailability of the preparation administered, the dosage regimen selected, the use of concomitant medications, and other relevant circumstances, but are not limited thereto.

[0280] In certain embodiments, there is provided the use of a compound, conjugate, or a pharmaceutically acceptable salt of a compound or conjugate in the manufacture of a medicament for the treatment of a disease in a subject. The compound or conjugate can be any of the compounds or conjugates herein. The disease in the subject can be a disease in which modulation of a kinase reduces the severity of such disease in the subject or otherwise treats such disease in the subject. The disease can be any disease state described herein. In certain embodiments, the disease is cancer, diabetes, an inflammatory disease, or a neurological disease.

[0281] Any of the compounds and / or conjugates herein can be for use in the treatment of a disease state modulated by one or more kinases. The disease state can be cancer, and the one or more kinases can be CKD, FLT3, and / or haspin kinase.

[0282] One of ordinary skill in the art will recognize that many modifications may be made to the specific embodiments described above. The embodiments should not be limited to the specific limitations described. Other embodiments may be possible.

[0283] Although the compounds, conjugates, and pharmaceutical compositions have been exemplified and described in detail in the foregoing description, it is to be considered that the same are illustrative and not restrictive in character, that only certain embodiments have been shown and described, and that all changes and modifications falling within the spirit of the invention are desired to be protected.

[0284] It is intended that the gist of the present compound, the present composition, and the present method be defined by the scope of the following claims. However, the present disclosure may be practiced otherwise than as specifically described and exemplified without departing from its gist or scope. Those skilled in the art will understand that various alternatives to the embodiments described herein may be utilized in practicing the claims without departing from the gist and scope defined in the following claims.

[0285] Any use of the chapter headings is intended to assist in reading this document and is not to be construed as limiting. Further, information related to the chapter headings may be within or outside of that particular chapter.

[0286] All publications, patents, patent application publications, magazine articles, textbooks, and other publications referred to in this document are indicative of the level of those skilled in the art to which this disclosure pertains. All such publications are incorporated herein by reference to the same extent as if each individual publication were specifically and individually indicated to be incorporated by reference. In the event of inconsistent usage between this document and the documents so incorporated by reference, the usage in the incorporated reference should be considered complementary to the usage in this document, and in case of irreconcilable inconsistencies, the usage in this document prevails.

[0287] A variety of techniques and mechanisms will sometimes account for the binding or linkage between two components. Words such as attached, linked, coupled, bonded, and similar terms with their inflected forms are used interchangeably unless it is specified that there are differences or unless it is otherwise clear from the context. These words and expressions do not necessarily mean a direct bond but include a bond through an intervening component. It should be noted that the bond between two components does not necessarily mean a direct, unhindered bond, because a variety of other components can exist between the two specifically noted components. As a result, a bond does not necessarily mean a direct, unhindered bond unless otherwise specified.

[0288] Certain Definitions As used herein, the following terms and phrases will have the meanings set forth below. 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.

[0289] The term "about" allows for a degree of variability within a value or range, e.g., within 10%, 5%, or 1% of a stated value or of a stated limitation of a range. In the present disclosure, the term "substantially" can allow for a degree of variability within a value or range, e.g., within 90%, 95%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more of a stated value or of a stated limitation of a range.

[0290] The terms "a", "an", or "the" are used to include one or more than one unless the context clearly dictates otherwise. The term "or" is used to refer to a non-exclusive "or" unless otherwise specified. Additionally, it is understood that phraseology or terminology used herein that is not otherwise defined is for illustrative purposes only and not for purposes of limitation.

[0291] The term "or" is used to mean a non-exclusive "or" unless otherwise specified. In addition, it is to be understood that phraseology or terminology utilized herein and not otherwise defined is for illustrative purposes only and not for purposes of limitation.

[0292] As used herein, the term "substituted" refers to a functional group in which one or more hydrogen atoms contained therein are replaced by one or more non-hydrogen atoms. The terms "functional group" or "substituent", as used herein, refer to a group that can be on or substituted on a molecule. Examples of substituents or functional groups include, but are not limited to, halogen (e.g., F, Cl, Br, and I); oxygen atoms in groups such as hydroxyl group, alkoxy group, aryloxy group, aralkyloxy group, oxo (carbonyl) group, carboxyl group including carboxylic acid, carboxylate, and carboxylate ester; sulfur atoms in groups such as thiol group, alkyl group, and arylsulfide group, sulfoxide group, sulfone group, sulfonyl group, and sulfonamide group; nitrogen atoms in groups such as amine, azide, hydroxylamine, cyano, nitro, N-oxide, hydrazide, and enamine; and other heteroatoms in various other groups.

[0293] As used herein, the term "alkyl" refers to a group having from 1 to about 20 carbon atoms (C1 - C 20 ), from 1 to 12 carbon atoms (C1 - C 12) refers to substituted or unsubstituted, linear and branched alkyl groups and cycloalkyl groups having 1 to 8 carbon atoms (C1 - C8), or in some embodiments, 1 to 6 carbon atoms (C1 - C6). Examples of linear alkyl groups include those having 1 to 8 carbon atoms, such as methyl, ethyl, n - propyl, n - butyl, n - pentyl, n - hexyl, n - heptyl, and n - octyl groups. Examples of branched alkyl groups include, but are not limited to, isopropyl, isobutyl, sec - butyl, t - butyl, neopentyl, isopentyl, and 2,2 - dimethylpropyl groups. As used herein, the term "alkyl" encompasses n - alkyl, isoalkyl, and anteisoalkyl groups, as well as other branched chain forms of alkyl. Representative substituted alkyl groups can be substituted one or more times with any of the groups listed herein, such as amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

[0294] The term "alkenyl", as used herein, refers to substituted or unsubstituted, linear and branched divalent alkenyl groups 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 some embodiments, 2 to 4 carbon atoms (C2 - C4) and at least one carbon - carbon double bond. Examples of linear alkenyl groups include those 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.

[0295] An alkynyl group is a fragment containing an open point of attachment on a carbon atom, which is considered to form when a hydrogen atom bonded to a triply bonded carbon is removed from the molecule of an alkyne. The term "hydroxyalkyl", as used herein, refers to an alkyl group as defined herein that is substituted with at least one hydroxyl (-OH) group.

[0296] The term "cycloalkyl", as used herein, refers to a substituted or unsubstituted, cyclic alkyl group, such as, but not limited to, each of the groups cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. In some embodiments, the cycloalkyl group can have 3 to about 8 to 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 can have 3 to 6 carbon atoms (C3 - C6). The cycloalkyl group further includes polycyclic cycloalkyl groups such as, but not limited to, each of the groups norbornyl, adamantyl, bornyl, camphyl, isocampphyl and carenyl, and fused rings such as, but not limited to, decalinyl.

[0297] As used herein, the term "acyl" refers to a group in which the radical contains a carbonyl moiety attached through a carbonyl carbon atom. The carbonyl carbon atom is also attached to another carbon atom, which can be part of a moiety such as an alkyl, aryl, aralkylcycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl group, whether substituted or unsubstituted. In the special case where the carbonyl carbon atom is attached to hydrogen, the group is a "formyl" group, and the acyl group as a term is defined herein. The acyl group can contain from 0 to about 12 to 40, 6 to 10, 1 to 5, or 2 to 5 additional carbon atoms attached to the carbonyl group. The acryloyl group is one example of an acyl group. The acyl group can also contain heteroatoms within the meaning of this specification. The nicotinoyl group (pyridyl-3-carbonyl) is one example of an acyl group within the meaning of this specification. Other examples include the groups of acetyl, benzoyl, phenylacetyl, pyridylacetyl, cinnamoyl, and acryloyl. When the group containing the carbon atom attached to the carbonyl carbon atom contains a halogen, the group is named a "haloacyl" group. One example is the trifluoroacetyl group.

[0298] As used herein, the term "aryl" refers to a substituted or unsubstituted, cyclic aromatic hydrocarbon that does not contain a heteroatom in the ring. Thus, aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenylene, anthracenyl, and naphthyl groups. In some embodiments, the aryl group contains from about 6 to about 14 carbon (C6 - C 14 ) or 6 to 10 carbon atoms (C6 - C 10) contains. The aryl group may be unsubstituted or substituted as defined herein. Representative substituted aryl groups can be mono-substituted or more than mono-substituted, for example, 2-, 3-, 4-, 5- or 6-substituted phenyl, or 2-8 substituted naphthyl groups, but are not limited thereto, and these can be substituted with carbon or non-carbon groups such as those listed herein.

[0299] As used herein, the terms “aralkyl” and “arylalkyl” refer to an alkyl group as defined herein in which a hydrogen bond or a carbon bond of the alkyl group is replaced by a bond to an aryl group as defined herein. Representative aralkyl groups include the benzyl group and the phenylethyl group, and fused (cycloalkylaryl) alkyl groups such as 4-ethyl-indanyl. An aralkenyl group is an alkenyl group as defined herein in which a hydrogen bond or a carbon bond of the alkyl group is replaced by a bond to an aryl group as defined herein.

[0300] As used herein, the term “heterocyclyl” refers to a substituted or unsubstituted, aromatic or non-aromatic cyclic compound containing three or more ring members, one or more of which are heteroatoms such as, but not limited to, B, N, O and S. Thus, heterocyclyl can be cycloheteroalkyl or heteroaryl, or in the case of polycyclic, can be any combination thereof. In some embodiments, the heterocyclyl group contains from 3 to about 20 ring members, while other such groups have from 3 to about 15 ring members. In some embodiments, the heterocyclyl group includes heterocyclyl groups having 3 to 8 carbon atoms (C3-C8), 3 to 6 carbon atoms (C3-C6) or 6 to 8 carbon atoms (C6-C8).

[0301] A heteroaryl ring is one embodiment of a heterocyclyl group. The term "heterocyclyl group" includes fused ring species including those containing fused aromatic and non-aromatic groups. Representative heterocyclyl groups include, but are not limited to, pyrrolidinyl, azetidinyl, piperidinyl, piperazinyl, morpholinyl, chromanyl, indolinonyl, isoindolinonyl, furanyl, pyrrolidinyl, pyridinyl, pyrazinyl, pyrimidinyl, triazinyl, thiophenyl, tetrahydrofuranyl, pyrrolyl, oxazolyl, oxadiazolyl, imidazolyl, triazolyl, tetrazolyl, benzoxazolinyl, benzthiazolinyl, and benzimidazolinyl groups.

[0302] As used herein, the term "heterocyclylalkyl" 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 heterocyclyl group as defined herein. Representative heterocyclylalkyl groups include, but are not limited to, furan-2-ylmethyl, furan-3-ylmethyl, pyridin-3-ylmethyl, tetrahydrofuran-2-ylmethyl, and indol-2-ylpropyl.

[0303] As used herein, the term "heteroarylalkyl" 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.

[0304] As used herein, the term "alkoxy" refers to an oxygen atom linked to an alkyl group, including cycloalkyl groups 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 alkoxy include, but are not limited to, isopropoxy, sec-butoxy, tert-butoxy, isopentyloxy, isohexyloxy, etc. Examples of cyclic alkoxy include, but are not limited to, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, etc. The alkoxy group can further contain a double bond or a triple bond and can also contain a heteroatom. For example, an aryloxy group is an alkoxy group within the meaning of this specification. A methoxyethoxy group is also an alkoxy group within the meaning of this specification, similar to a methylenedioxy group in the context where two adjacent atoms of the structure are substituted by them.

[0305] As used herein, the term "amine" refers to primary, secondary, and tertiary amines having, for example, the formula N(group)3, where 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 alkylamine, arylamine, alkylarylamine; R2NH (where each R is independently selected from, for example, dialkylamine, diarylamine, aralkylamine, heterocyclylamine, etc.); and R3N (where R is independently selected from, for example, trialkylamine, dialkylarylamine, alkyldiarylamine, triarylamine, etc.). The term "amine" also includes ammonium ions as used herein.

[0306] As used herein, the term "amino group" refers to substituents of the form -NH2, -NHR, -NR2, -NR3 + wherein each R is independently selected and is in its respective protonated form, except for NR3 which cannot be protonated+ Excluding. Thus, any compound substituted with an amino group can be considered an amine. The "amino group" can be a primary, secondary, tertiary, or quaternary amino group within the meaning of this specification. The "alkylamino" group includes each of the monoalkylamino, dialkylamino, and trialkylamino groups.

[0307] The terms "halo", "halogen", or "halide" group, when used herein, by themselves or as part of another substituent, unless otherwise stated, mean each atom of fluorine, chlorine, bromine, or iodine.

[0308] The term "haloalkyl" group, when used herein, includes mono-haloalkyl groups, poly-haloalkyl groups, where all halo atoms may be the same or different, and includes per-haloalkyl groups in which all hydrogen atoms are replaced by halogen atoms such as 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.

[0309] The term "optionally substituted" or "optional substituent", when used herein, means that the group under consideration is either unsubstituted or substituted with one or more of the specified substituents. When the group under consideration is substituted with more than one substituent, the substituents may be the same or different. When the terms "independently", "independently is", and "independently selected from" are used, it means that the groups under consideration may be the same or different. Certain of the terms defined herein may occur more than once in a structure, and upon such occurrence, each term will be defined independently of the others.

[0310] The terms "patient" and "subject" are used interchangeably and include human patients, laboratory animals such as rodents (e.g., mice, rats or hamsters), rabbits, monkeys, chimpanzees, companion animals such as dogs, cats or rabbits, agricultural animals such as cows, horses, pigs, sheep, goats, livestock animals raised for food production, or wild animals that have been captured, such as bears, pandas, lions, tigers, leopards, elephants, zebras, giraffes, gorillas, dolphins or whales. The patient to be treated is preferably a mammal, specifically a human.

[0311] The terms "protein", "polypeptide" and "peptide" refer to compounds containing amino acids joined via peptide bonds and are used interchangeably.

Examples

[0312] The following examples serve to illustrate the present disclosure. The examples are not intended to limit the scope of the "claims" in any way.

[0313] [Example 1] Synthesis of CDK inhibitor compounds Unless otherwise specified, all reagents and solvents were purchased from commercial sources and used as received. All reactions were carried out in screw-cap vials unless otherwise described. 1H NMR spectra and 13C NMR spectra were obtained in (CD3)2SO as solvent using a 500 MHz or 800 MHz spectrometer with Me4Si as the internal standard. Chemical shifts were reported in parts per million (δ) and calibrated using the remaining non-deuterated solvent as the internal reference.

[0314] Data for 1H NMR spectra are reported as follows: chemical shift (δ ppm) (multiplicity, coupling constant (Hz), integration). Multiplicities are reported as follows: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, or combinations thereof.

[0315] Method A, general In a 250 mL round-bottom flask equipped with a stir bar, 5-aminoindazole (10 mmol) and an appropriate aldehyde, (4-formylbenzoic acid or 4-formylbenzonitrile) (10 mmol) were dissolved in 150 mL of 200-proof ethanol. The mixture was refluxed at 120 °C for 2 hours, and then an appropriate ketone (20 mmol) and a catalytic amount of hydrochloric acid were added. This was stirred at reflux for an additional 12 hours. The reaction product was concentrated and purified in 40% ethyl acetate and 60% hexane.

[0316] Method B, general In a 50 mL round-bottom flask equipped with a stir bar, 4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzonitrile (850 mg, 2.53 mmol) was added, and NaOH (532 mg, 13 mmol) was dissolved in ethanol (4 mL) and water (8 mL). This was added to the round-bottom flask and refluxed for 12 hours. After about 1 hour, the mixture was completely dissolved. After completion of the reaction, the solvent was evaporated under reduced pressure, diluted with water, and acidified to pH 5 using HCl. This was then stirred for 30 minutes to confirm that the pH was stable. The product was then recovered via vacuum filtration.

[0317] Method C, general In a 20 mL vial equipped with a stir bar, an acid (0.5 mmol) was dissolved in N,N-dimethylformamide (DMF) (2 mL) and N,N-diisopropylethylamine (DIPEA) (12.5 equivalents). Bis(2-oxo-3-oxazolidinyl)phosphine chloride (2.7 equivalents) was added to this solution and stirred at 50 °C for 30 minutes, and then the corresponding amine (0.5 mmol) was added and stirred at 50 °C overnight. The DMF was concentrated under reduced pressure and then purified in 15% methanol and 85% ethyl acetate.

[0318] Method D, general In a 20 mL vial equipped with a stir bar, an acid (0.5 mmol), 4-dimethylaminopyridine (12 mg), N-methylmorpholine (1 mmol), and (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazol[4,5-b]pyridinium 3-oxide hexafluorophosphate, hexafluorophosphate azabenzotriazolium tetramethyluronium) were dissolved in DMF (2 mL) and stirred at room temperature. After 30 minutes, 1-methylpiperazine (2 mmol) was added and stirred overnight at room temperature. The DMF was concentrated under reduced pressure and the reaction product was purified with 25% methanol and 75% ethyl acetate.

[0319] Method E, general In a microwave vial, an acid (0.5 mmol), fluoro-N,N,N’,N’-bis(tetramethylene)formamidinium hexafluorophosphate (0.6 mmol), DIPEA (1.75 mmol) and dichloromethane (2 mL) were stirred at room temperature for 30 minutes. Then, an amine (0.38 mmol) was added and stirred overnight at 80 °C in a sand bath. The reaction mixture was concentrated and purified with 10% methanol and 90% ethyl acetate.

[0320] Method F, general In a 20 mL vial equipped with a stir bar, an acid intermediate (0.5 mmol) and hexafluorophosphate azabenzotriazolium tetramethyluronium were sealed with a septum, evacuated to air, and an N2 balloon was added. Anhydrous DMF (5 mL) and DIPEA (1.2 mL) were added to this reaction vial. This was heated at 100 °C for 10 minutes, then an amine (1 mmol) was added to the reaction and stirred overnight at 100 °C. The reaction mixture was concentrated and purified via silica gel chromatography.

[0321] Method G, general (Synthesis of compound HSH3165) 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 mixture was concentrated to obtain the pure product, compound HSH3165.

[0322] Method H, general To a solution of compound HSH3107 (0.5 mmol) in anhydrous tetrahydrofuran (THF) (720 μL) was added lithium aluminum hydride (1.5 mmol, 1 M in THF) at 0 °C. The mixture was heated to 50 °C for 2 hours. It was cooled to 0 °C, worked up with water, and then stirred at room temperature for 3 minutes.

[0323] The solid was filtered through a bed of celite, washed with THF, dried over sodium sulfate, concentrated under reduced pressure, and purified via silica gel column chromatography [hexane:ethyl acetate (30:70 - 0:100)].

[0324] Intermediate:

[0325] Synthesis of 4-(8,9,10,11-tetrahydro-3H-8,11-methanopyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid:

[0326]

Chemical formula

[0327] In a 250 mL round-bottom flask equipped with a stir 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. This was stirred at reflux for an additional 12 hours. The precipitate was washed with ethanol and taken for the next step without further purification.

[0328] 1 1H NMR (800 MHz, (CD3)2SO) δ 8.75 (s, 1H), 8.12 (dd, J = 20.4, 7.8 Hz, 2H), 7.97 (d, J = 7.9 Hz, 2H), 7.86 (q, J = 9.3 Hz, 2H), 2.95 (s, 1H), 2.18 (dd, J = 27.2, 13.6 Hz, 2H), 1.83 (d, J = 8.9 Hz, 1H), 1.67 (d, J = 9.0 Hz, 1H), 1.60 (s, 1H), 1.35 (d, J = 10.6 Hz, 1H), 1.22 (t, J = 9.9 Hz, 1H). 13 13C 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 C 22 H 18 N3O2[M+H] + Calculated value for 356.1393, found 356.1392.

[0329] 4-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzonitrile

[0330]

Chemical formula

[0331] Synthesized using Method A: off-white solid (1.5 g, 10 mmol, 46.3%). 11H NMR (500 MHz, (CD3)2SO) δ 8.55 (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 = 6.1 Hz, 2H), 2.04 - 1.97 (m, 2H), 1.79 - 1.72 (m, 2H). 13 13C 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 C 21 H 17 N4[M+H] + Calculated value for 325.1444, found 325.1448.

[0332] 4-(9-Methyl-3H-pyrazolo[4,3-f]quinolin-7-yl)benzonitrile

[0333]

Chemical Structure

[0334] Synthesized using Method A: off-white solid (0.840 g, 10 mmol, 30%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.61 - 8.53 (m, 1H), 8.44 - 8.36 (m, 2H), 8.13 (s, 1H), 7.97 - 7.88 (m, 4H), 2.93 (s, 3H). 1313C 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 C 18 H 13 N4[M + H] + Calculated value for 285.1135, measured value 285.1136.

[0335] 4-(9-Cyclopropyl-3H-pyrazolo[4,3-f]quinolin-7-yl)benzonitrile

[0336]

Chemical Structure

[0337] Synthesized using Method A: off-white solid (0.485 g, 10 mmol, 14.7%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.80 (s, 1H), 8.40 (d, J = 8.3 Hz, 2H), 8.00 - 7.86 (m, 5H), 2.62 (m, 1H), 1.39 - 1.24 (m, 2H), 1.00 (q, J = 5.3, 4.8 Hz, 2H). 13 13C 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 C 20 H 15 N4[M + H] + Calculated value for 311.1291, measured value 311.1293.

[0338] 4-(1-Methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzonitrile

[0339]

Chemical formula

[0340] Synthesized using Method A: off-white solid (402 mg, 2 mmol, 59.4%). 1 H NMR (500 MHz, (CD3)2SO) δ 7.88 (d, J = 8.1 Hz, 2H), 7.71 (d, J = 7.9 Hz, 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 C 18 H 14 N3O2[M+H] + Calculated value for 339.1604, measured value 339.1605.

[0341] 3-Fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzonitrile

[0342]

Chemical formula

[0343] Synthesized using Method A: off-white solid (1.42 g, 10 mmol, 41.5%). 11H NMR (500 MHz, (CD3)2SO) δ 8.56 (s, 1H), 7.94 (dd, J = 9.5, 1.7 Hz, 1H), 7.88 (d, J = 9.1 Hz, 1H), 7.85 - 7.78 (m, 2H), 7.68 (t, J = 7.5 Hz, 1H), 3.36 (t, J = 6.6 Hz, 2H), 2.60 (t, J = 6.2 Hz, 2H), 2.06 - 1.92 (m, 2H), 1.85 - 1.71 (m, 2H). 13 13C NMR (126 MHz, (CD3)2SO) δ 160.31 (d, 1 J = 248 Hz), 150.62, 143.80, 142.73, 138.94, 136.47, 134.29 (d, 2 J = 16 Hz), 133.27, 129.92, 129.44, 129.26, 122.57, 120.29 ( 2 J = 26 Hz), 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 C 21 H 16 N4[M + H] + calculated value 343.1353, measured value 343.1352.

[0344] 2-Fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzonitrile

[0345]

Chemical Structure

[0346] Synthesized using Method A: off-white solid (0.220 g, 5 mmol, 12.9%). 11H NMR (500 MHz, (CD3)2SO) δ 8.59 (s, 1H), 8.00 (t, J = 7.4 Hz, 1H), 7.91 - 7.77 (m, 2H), 7.73 (d, J = 10.4 Hz, 1H), 7.63 (d, J = 8.0 Hz, 1H), 3.35 (s, 2H), 2.80 (t, J = 6.2 Hz, 2H), 2.09 - 1.97 (m, 2H), 1.82 - 1.73 (m, 2H). 13 13C NMR (126 MHz, (CD3)2SO) δ 163.68 (d, 1 J = 255 Hz), 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 C 21 H 16 N4[M+H] + calculated value 343.1353, measured value 343.1352.

[0347] 3-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzonitrile

[0348]

Chemical Structure

[0349] Synthesized using Method A: off-white solid (0.665 g, 10 mmol, 20.2%). 11H NMR (500 MHz, (CD3)2SO) δ 8.54 (s, 1H), 7.98 (s, 1H), 7.91 - 7.82 (m, 3H), 7.80 (d, J = 9.1 Hz, 1H), 7.67 (t, J = 7.8 Hz, 1H), 2.74 (t, J = 6.2 Hz, 2H), 2.01 - 1.91 (m, 2H), 1.73 (m, 2H). 13 13C 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 C 21 H 17 N4[M+H] + Calculated value: 325.1448, measured value: 325.1449.

[0350] 3-Methoxy-4-(8,9,10,11-tetrahydro-3H-naphtho[1,2-e]indazol-7-yl)benzonitrile

[0351]

Chemical Structure

[0352] Synthesized using Method A: off-white solid (0.255 g, 10 mmol, 7.21%). 11H NMR (500 MHz, (CD3)2SO) δ 8.55 (s, 1H), 7.85 (d, J = 9.0 Hz, 1H), 7.80 (s, 1H), 7.59 (s, 1H), 7.51 (dd, J = 7.6, 1.4 Hz, 1H), 7.41 (d, J = 7.7 Hz, 1H), 3.78 (s, 3H), 3.34 (d, J = 16.9 Hz, 2H), 2.59 (s, 1H), 2.42 (d, J = 16.5 Hz, 1H), 1.98 (dd, J = 13.7, 6.5 Hz, 2H), 1.84 - 1.68 (m, 2H). 13 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 C 22 H 19 N4O [M+H] + Calculated value for 355.1553, measured value 355.1553.

[0353] 3-Methoxy-4-(8,9,10,11-tetrahydro-3H-naphtho[1,2-e]indazol-7-yl)benzonitrile

[0354]

Chemical Structure

[0355] Synthesized using Method A: off-white solid (0.719 g, 6.14 mmol, 33.1%). 11H NMR (500 MHz, (CD3)2SO) δ 8.51 (s, 1H), 7.84 (q, J = 8.9 Hz, 2H), 7.77 (d, J = 7.8 Hz, 1H), 7.38 (s, 1H), 7.23 (d, J = 7.9 Hz, 1H), 3.96 (s, 3H), 3.28 (t, J = 6.7 Hz, 2H), 2.74 (t, J = 6.3 Hz, 2H), 2.05 - 1.90 (m, 2H), 1.81 - 1.67 (m, 2H). 13 13C 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 C 22 H 19 N4O [M+H] + Calculated value for 355.1553, measured value 355.1552.

[0356] 4-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid

[0357]

Chemical Structure

[0358] Synthesized using Method B: off-white solid (0.812 g, 2 mmol, 93.6%). 11H NMR (500 MHz, (CD3)2SO) δ 8.75 (s, 1H), 8.26 (d, J = 9.2 Hz, 1H), 8.19 (d, J = 9.2 Hz, 1H), 8.15 - 8.11 (m, 2H), 7.85 - 7.80 (m, 2H), 3.46 (t, J = 6.5 Hz, 2H), 2.76 (t, J = 6.1 Hz, 2H), 2.03 (dd, J = 6.0, 2.9 Hz, 2H), 1.83 - 1.76 (m, 2H). 13 13C 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 C 21 H 18 N3O2[M+H] + Calculated value for C

[0359] 4-(9-Methyl-3H-pyrazolo[4,3-f]quinolin-7-yl)benzoic acid

[0360]

Chemical Structure

[0361] Synthesized using Method B: off-white solid (0.462 g, 3 mmol, 50.8%). 1 1H NMR (500 MHz, (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). 1313C 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 C 18 H 14 N3O2[M+H] + Calculated value for 304.1081, measured value 304.1081.

[0362] 4-(9-Cyclopropyl-3H-pyrazolo[4,3-f]quinolin-7-yl)benzoic acid

[0363]

Chemical formula

[0364] Synthesized using Method B: off-white solid (0.464 g, 2.2 mmol, 64.1%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.90 (s, 1H), 8.36 (d, J = 8.2 Hz, 2H), 8.21 (d, J = 9.2 Hz, 1H), 8.13 - 8.07 (m, 3H), 7.98 (s, 1H), 2.74 (d, J = 5.4 Hz, 1H), 1.39 (dd, J = 8.5, 2.4 Hz, 2H), 1.13 (dd, J = 5.5, 2.2 Hz, 2H). 13 13C NMR (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 C 20 H 16 N3O2[M+H] + Calculated value for 330.1237, measured value 330.1238.

[0365] 4-(3,8,9,10,11,12-Hexahydrocyclohepta[c]pyrazolo[4,3-f]quinolin-7-yl)benzoic acid

[0366]

Chem.

[0367] Synthesized using Method A: off-white solid (0.522 g, 4 mmol, 36.5%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.65 (s, 1H), 8.04 (d, J = 7.8 Hz, 2H), 7.84 (d, J = 9.2 Hz, 1H), 7.77 (d, J = 9.1 Hz, 1H), 7.58 (d, J = 7.8 Hz, 2H), 3.50 (d, J = 7.2 Hz, 2H), 2.93 (d, J = 8.4 Hz, 2H), 1.84 (d, J = 9.5 Hz, 4H), 1.59 (d, J = 7.8 Hz, 2H). 13 C 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 C 22 H 20 N3O2[M+H] + Calculated value for 358.1550, found 358.1550.

[0368] 4-(3,8,10,11-Tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzoic acid

[0369]

Chem.

[0370] Synthesized using Method A: off-white solid (0.220 g, 0.76 mmol, 21.2%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.68 (s, 1H), 8.13 - 7.99 (m, 4H), 7.76 (d, J = 8.4 Hz, 2H), 4.77 (s, 2H), 4.17 (t, J = 5.8 Hz, 2H), 3.49 (d, J = 5.9 Hz, 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 C 20 H 16 N3O3[M+H] + Calculated value for 346.1186, found 346.1187.

[0371] 4-(9,9-Dioxide-3,8,10,11-tetrahydropyrazolo[4,3-f]thiopyrano[3,4-c]quinolin-7-yl)benzonitrile

[0372]

Chemical Structure

[0373] Synthesized using Method A: off-white solid (1.68 g, 4.75 mmol, 89.7%). 11H NMR (800 MHz, (CD3)2SO) δ 8.74 (s, 1H), 8.12 (d, J = 8.1 Hz, 2H), 8.05 (d, J = 9.0 Hz, 1H), 7.93 (d, J = 9.0 Hz, 1H), 7.73 (d, J = 8.1 Hz, 2H), 4.52 (s, 2H), 4.04 (d, J = 6.7 Hz, 2H), 3.72 (d, J = 6.5 Hz, 2H). 13 13C 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 C 20 H 16 N3O4S [M+H] + Calculated value for C19H16N3O4S [M+H]+ 394.0856, found 394.0856.

[0374] 4-(3,8,9,10-Tetrahydrocyclopenta[c]pyrazolo[4,3-f]quinolin-7-yl)benzoic acid

[0375]

Chemical Structure

[0376] Synthesized using Method A: off-white solid (518 mg, 4 mmol, 39.3%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.48 (s, 1H), 8.12 - 8.08 (m, 2H), 8.01 - 7.93 (m, 4H), 3.43 (t, J = 7.6 Hz, 2H), 3.21 (t, J = 7.5 Hz, 2H), 2.29 - 2.18 (m, 2H). 1313C 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 C 20 H 16 N3O2[M+H] + Calculated value for 330.1237, measured value 330.1236.

[0377] 4-(8,9-Dihydro-3H-cyclobuta[c]pyrazolo[4,3-f]quinolin-7-yl)benzoic acid

[0378]

Chem.

[0379] Synthesized using Method A: off-white solid (0.030 g, 3 mmol, 3.2%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.37 (d, J = 0.9 Hz, 1H), 8.29 - 8.25 (m, 2H), 8.11 - 8.06 (m, 2H), 7.91 - 7.83 (m, 2H), 3.77 (dd, J = 5.3, 2.6 Hz, 2H), 3.70 - 3.64 (m, 2H). 13 13C 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 C 19 H 14 N3O2[M+H] + Calculated value for 316.1080, measured value 316.1080.

[0380] 4-(5-Fluoro-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid

[0381]

Chem.

[0382] Synthesized using Method A: off-white solid (0.145 g, 1.38 mmol, 29.1%). 1 H NMR (800 MHz, (CD3)2SO) δ 8.62 (s, 1H), 8.07 (d, J = 7.7 Hz, 2H), 7.71 (dd, J = 23.9, 8.8 Hz, 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 (201 MHz, (CD3)2SO) δ 167.51, 157.66 ( 1 J = 250 Hz), 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 C 21 H 17 FN3O2[M+H] + Calculated value for 362.1299, measured value 362.1300.

[0383] 3-Fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid

[0384]

Chem.

[0385] Synthesized using Method B: off-white solid (1.58 g, 4.44 mmol, 98.5%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.64 (s, 1H), 8.06 (d, J = 22.2 Hz, 3H), 7.72 (d, J = 11.2 Hz, 1H), 7.63 (d, J = 8.1 Hz, 1H), 3.31 (t, J = 6.6 Hz, 2H), 2.76 (t, J = 6.2 Hz, 2H), 2.04 - 1.91 (m, 2H), 1.76 (q, J = 6.0 Hz, 2H). 13 C NMR (126 MHz, (CD3)2SO) δ 167.55, 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 C 21 H 17 FN3O2[M+H] + Calculated value for 362.1299, measured value 362.1300.

[0386] 5-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)thiophene-2-carboxylic acid

[0387]

Chemical Structure

[0388] Synthesized using Method A: off-white solid (0.545 g, 4 mmol, 39.0%). 11H NMR (500 MHz, (CD3)2SO) δ 8.58 (s, 1H), 7.94 (dd, J = 9.1, 0.9 Hz, 1H), 7.87 (d, J = 9.2 Hz, 1H), 7.76 (d, J = 3.8 Hz, 1H), 7.62 (d, J = 4.0 Hz, 1H), 3.30 (t, J = 6.5 Hz, 2H), 3.02 (t, J = 6.2 Hz, 2H), 2.01 - 1.92 (m, 2H), 1.86 - 1.78 (m, 2H). 13 13C 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 C 19 H 16 N3O2S [M+H] + Calculated value for 350.0958, found 350.0960.

[0389] 3-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid

[0390]

Chemical Structure

[0391] Synthesized using Method B: off-white solid (0.453 g, 2 mmol, 65.9%). 11H NMR (500 MHz, (CD3)2SO) δ 8.72 (s, 1H), 8.20 (t, J = 1.8 Hz, 1H), 8.13 - 8.02 (m, 3H), 7.91 (m, 1H), 7.70 (t, J = 7.7 Hz, 1H), 3.45 (t, J = 6.5 Hz, 2H), 2.77 (t, J = 6.2 Hz, 2H), 2.09 - 1.99 (m, 2H), 1.79 (m, 2H). 13 13C 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 C 21 H 18 N3O2[M+H] + Calculated value for 344.1393, measured value 344.1394.

[0392] 3-Methoxy-4-(8,9,10,11-tetrahydro-3H-naphtho[1,2-e]indazol-7-yl)benzoic acid

[0393]

Chemical formula

[0394] Synthesized using Method B: off-white solid (0.193 g, 0.75 mmol, 68.2%). 11H NMR (500 MHz, (CD3)2SO) δ 8.79 (s, 1H), 8.23 (d, J = 9.2 Hz, 1H), 8.14 (d, J = 9.2 Hz, 1H), 7.78 - 7.72 (m, 2H), 7.61 (d, J = 7.7 Hz, 1H), 3.50 (d, J = 6.8 Hz, 2H), 2.62 (t, J = 6.3 Hz, 2H), 2.07 - 1.98 (m, 2H), 1.85 - 1.76 (m, 2H). 13 13C 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 C 22 H 21 N3O3[M+H] + Calculated value for 374.1499, found 374.1497.

[0395] 2-Methoxy-4-(8,9,10,11-tetrahydro-3H-naphtho[1,2-e]indazol-7-yl)benzoic acid

[0396]

Chemical Structure

[0397] Synthesized using Method B: off-white solid (814 mg, 2.31 mmol, 94.3%). 11H NMR (800 MHz, (CD3)2SO) δ 8.52 (d, J = 17.7 Hz, 1H), 7.86 (d, J = 9.0 Hz, 1H), 7.84 - 7.80 (m, 1H), 7.78 (d, J = 7.7 Hz, 1H), 7.39 (d, J = 1.5 Hz, 1H), 7.23 (dd, J = 7.7, 1.6 Hz, 1H), 3.96 (s, 3H), 3.23 (m, 2H), 2.72 (t, J = 6.2 Hz, 2H), 1.95 (m, 2H), 1.77 - 1.68 (m, 2H). 13 13C 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 C 22 H 21 N3O3[M+H] + Calculated value for 374.1499, measured value 374.1497.

[0398] tert-Butyl 4-(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoyl)piperazine-1-carboxylate

[0399]

Chem.

[0400] Synthesized using Method C: off-white solid (100 mg, 0.5 mmol, 39.1%). 11H NMR (800 MHz, (CD3)2SO) δ 8.59 (s, 1H), 7.93 - 7.78 (m, 2H), 7.64 (dd, J = 8.7, 2.9 Hz, 2H), 7.52 (dd, J = 8.1, 2.6 Hz, 2H), 3.70 - 3.54 (m, 2H), 3.54 - 3.38 (m, 5H), 2.80 (t, J = 6.4 Hz, 2H), 2.01 (m, 2H), 1.76 (m, 2H), 1.42 (s, 9H). 13 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 C 30 H 34 N5O3[M+H] + Calculated value for 512.2656, measured value 512.2655.

[0401] N-(2-(Dimethylamino)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH2177)

[0402]

Chemical Structure

[0403] Synthesized using Method C: off-white solid (30 mg, 0.5 mmol, 14.65%) 11H 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), 3.40 - 3.27 (m, 7H), 2.77 (t, J = 6.1 Hz, 2H), 2.39 - 2.25 (m, 7H), 2.13 (s, 3H), 2.00 (m, 4.4, 2.8 Hz, 2H), 1.74 (m, 2H), 1.68 (q, J = 7.0 Hz, 2H). 13 13C 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 C 25 H 28 N5O [M+H] + calculated value 414.2288, found 414.2285.

[0404] N-(3-(4-Methylpiperazin-1-yl)propyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH2195)

[0405]

Chemical Structure

[0406] Synthesized using Method C: off-white solid (10 mg, 0.5 mmol, 4.1%). 11H 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 C 29 H 35 N6O [M+H] + Calculated value for C25H29N6O [M+H]+ 483.2867, found 483.2868.

[0407] N-(Cyclopropylmethyl)-4-(8,9,10,11-tetrahydro-3H-naphtho[1,2-e]indazol-7-yl)benzamide (Compound HSH3001)

[0408] [Chemical formula]

[0409] Synthesized using Method C: off-white solid (20 mg, 0.5 mmol, 10.1%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.64 (t, J = 5.7 Hz, 1H), 8.57 (s, 1H), 8.04 - 7.91 (m, 2H), 7.84 (q, J = 12.4, 10.6 Hz, 2H), 7.69 - 7.57 (m, 2H), 3.18 (t, J = 6.2 Hz, 2H), 2.76 (t, J = 6.3 Hz, 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). 1313C 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 C 25 H 25 N4O [M+H] + Calculated value for 397.2023, measured value 397.2021.

[0410] N-(3-(Piperidin-1-yl)propyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3004)

[0411]

Chemical Structure

[0412] Synthesized using Method C: off-white solid (23 mg, 0.5 mmol, 9.8%). 1 1H 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 C 29 H 34 N5O [M+H] + Calculated value for 468.2758, measured value 468.2759.

[0413] N-(2-(Dimethylamino)ethyl)-4-(8,9,10,11-tetrahydro-3H-8,11-methanopyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3014)

[0414]

Chem.

[0415] Synthesized using Method F: off-white solid (10 mg, 0.5 mmol, 4.7%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.69 (m, 1H), 8.47 (d, J = 4.4 Hz, 1H), 8.28 (d, J = 8.3 Hz, 1H), 8.02 (d, J = 7.9 Hz, 2H), 7.91 (d, J = 8.0 Hz, 2H), 7.84 (m, 1H), 7.29 (m, 1H), 4.36 (s, 1H), 3.75 (d, J = 3.2 Hz, 1H), 2.87 (t, J = 6.5 Hz, 2H), 2.53 (s, 6H), 2.21 - 2.14 (m, 2H), 1.85 (d, J = 8.9 Hz, 1H), 1.71 (d, J = 8.8 Hz, 1H), 1.36 (t, J = 9.5 Hz, 1H), 1.23 (t, J = 9.4 Hz, 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 C 26 H 28 N5O [M+H] + Calculated value 426.2289, measured value 426.2287.

[0416] N-(2-Methoxyethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3016)

[0417]

Chem.

[0418] Synthesized using Method C: off-white solid (53 mg, 0.5 mmol, 26.25%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.61 (t, J = 5.1 Hz, 1H), 8.56 (s, 1H), 7.98 - 7.94 (m, 2H), 7.84 (d, J = 4.9 Hz, 2H), 7.68 - 7.61 (m, 2H), 3.51 - 3.43 (m, 4H), 3.32 (d, J = 4.0 Hz, 2H), 3.28 (s, 3H), 2.76 (t, J = 6.1 Hz, 2H), 1.99 (m, 2H), 1.73 (m, 2H). 13 C 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 C 24 H 25 N4O2[M+H] + Calculated value for 401.1972, measured value 401.1970.

[0419] 4-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-(thiazol-2-yl)benzamide (Compound HSH3017)

[0420] [Chemical]

[0421] Synthesized using Method C: off-white (16 mg, 0.5 mmol, 7.5%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.59 (s, 1H), 8.20 (d, J = 8.0 Hz, 2H), 7.92 - 7.83 (m, 2H), 7.73 (d, J = 8.0 Hz, 2H), 7.57 (d, J = 3.6 Hz, 1H), 7.29 (d, J = 3.7 Hz, 1H), 2.80 (t, J = 6.0 Hz, 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 C 24 H 20 N5OS [M+H] + Calculated value for 426.1383, found 426.1379.

[0422] N-Cyclopropyl-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3019)

[0423] [Chemical]

[0424] Synthesized using Method C: off-white solid (25.2 mg, 0.5 mmol, 13.2%). 11H NMR (500 MHz, (CD3)2SO) δ 8.56 (s, 1H), 8.52 (d, J = 4.3 Hz, 1H), 7.92 (d, J = 7.9 Hz, 2H), 7.84 (t, J = 9.7 Hz, 2H), 7.62 (d, J = 7.9 Hz, 2H), 3.34 (s, 2H), 2.88 (m, 1H), 2.74 (t, J = 6.1 Hz, 2H), 2.03 - 1.94 (m, 2H), 1.79 - 1.68 (m, 2H), 0.70 (m, 2H), 0.60 (m, 2H). 13 13C 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 C 24 H 23 N4O [M+H] + Calculated value for 383.1866, measured value 383.1865.

[0425] N-(3-Methoxypropyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3088)

[0426]

Chemical Structure

[0427] Synthesized using Method F: off-white solid (13 mg, 0.5 mmol, 6.3%). 11H NMR (500 MHz, (CD3)2SO) δ 8.55 (t, J = 9.3 Hz, 1H), 7.95 - 7.91 (m, 2H), 7.88 - 7.82 (m, 2H), 7.65 - 7.60 (m, 2H), 7.52 (t, J = 5.5 Hz, 1H), 3.24 (s, 3H), 3.21 (s, 2H), 3.15 (m, 2H), 2.76 (t, J = 6.1 Hz, 2H), 2.00 (m, 2H), 1.76 (m, 6H). 13 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 C 25 H 27 N4O2[M+H] + calculated value 415.2129, measured value 415.2129.

[0428] (4-Methylpiperazin-1-yl)(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)methanone (Compound HSH3107)

[0429]

Chemical Structure

[0430] Synthesized using Method D: off-white solid (510 mg, 4 mmol, 30%). 11H NMR (500 MHz, (CD3)2SO) δ 8.57 (s, 1H), 7.86 (d, J = 9.0 Hz, 1H), 7.82 (s, 1H), 7.61 (d, J = 7.9 Hz, 2H), 7.47 (d, J = 8.0 Hz, 2H), 3.53 (s, 4H), 3.34 (t, J = 6.8 Hz, 2H), 2.79 (t, J = 6.3 Hz, 2H), 2.39 - 2.32 (m, 4H), 2.22 (s, 3H), 2.07 - 1.98 (m, 2H), 1.77 (m, 2H). 13 13C 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 C 26 H 28 N5O [M+H] + Calculated value: 426.2289, Measured value: 426.2290.

[0431] N-(2-(Diethylamino)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3124)

[0432]

Chemical Structure

[0433] Synthesized using Method F: Off-white solid (66 mg, 0.5 mmol, 30%). 11H NMR (800 MHz, (CD3)2SO) δ 8.59 (s, 1H), 8.39 (t, J = 5.7 Hz, 1H), 7.99 - 7.91 (m, 2H), 7.87 (d, J = 9.0 Hz, 1H), 7.83 (d, J = 9.0 Hz, 1H), 7.68 - 7.63 (m, 2H), 3.40 - 3.32 (m, 4H), 2.78 (t, J = 6.2 Hz, 2H), 2.60 (t, J = 7.2 Hz, 2H), 2.53 (q, J = 7.2 Hz, 4H), 2.04 - 2.00 (m, 2H), 1.78 - 1.73 (m, 2H), 1.01 - 0.97 (m, 6H). 13 13C 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 C 27 H 32 N5O [M+H] + calculated value 442.2601, measured value 442.2600.

[0434] N-(2-(Pyridin-2-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3130)

[0435]

Chemical Structure

[0436] Synthesized using Method F: off-white solid (32 mg, 0.5 mmol, 14.3%). 11H NMR (500 MHz, (CD3)2SO) δ 8.67 (m, 1H), 8.58 (s, 1H), 8.51 - 8.43 (m, 2H), 7.88 (m, 4H), 7.64 (d, J = 8.2 Hz, 2H), 7.32 - 7.26 (m, 2H), 3.57 (q, J = 6.7, 6.7, 6.6 Hz, 2H), 2.90 (t, J = 7.1, 7.1 Hz, 2H), 2.76 (t, J = 6.1, 6.1 Hz, 2H), 2.05 - 1.96 (m, 2H), 1.73 (m, 2H). 13 13C 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 C 28 H 26 N5O [M+H] + Calculated value for 448.2132, found 448.2130.

[0437] N-(3-Morpholinopropyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3146)

[0438]

Chemical Structure

[0439] Synthesized using Method F: off-white solid (92.9 mg, 0.5 mmol, 39.5%). 11H NMR (500 MHz, (CD3)2SO) δ 8.57 (t, J = 5.4 Hz, 2H), 7.96 - 7.90 (m, 2H), 7.85 (q, J = 9.0 Hz, 2H), 7.67 - 7.61 (m, 2H), 3.56 (t, J = 4.6 Hz, 5H), 3.30 (d, J = 6.5 Hz, 3H), 2.77 (t, J = 6.1 Hz, 2H), 2.34 (d, J = 6.7 Hz, 6H), 2.00 (m, 2H), 1.72 (m, 4H). 13 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 C 28 H 32 N5O2[M+H] + Calculated value for 470.2550, measured value 470.2548.

[0440] N-(2-(Piperidin-1-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3117)

[0441]

Chemical Structure

[0442] Synthesized using Method F: off-white solid (10 mg, 0.5 mmol, 4.3%). 11H NMR (800 MHz, DMSO-d6) δ 8.59 (s, 1H), 8.40 (t, J = 5.7 Hz, 1H), 7.95 (d, J = 8.1 Hz, 2H), 7.87 (d, J = 9.0 Hz, 1H), 7.82 (d, J = 9.2 Hz, 1H), 7.65 (d, J = 8.1 Hz, 2H), 3.42 (q, J = 6.4 Hz, 2H), 3.33 (t, J = 6.8 Hz, 2H), 2.78 (t, J = 6.3 Hz, 2H), 2.47 (t, J = 7.1 Hz, 2H), 2.44 - 2.34 (m, 4H), 2.01 (p, J = 6.4 Hz, 2H), 1.75 (p, J = 6.1 Hz, 2H), 1.50 (p, J = 5.6 Hz, 4H), 1.38 (q, J = 6.2 Hz, 2H). 13 13C 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 C 28 H 32 N5O [M+H] + Calculated value: 454.2601, Measured value: 454.2600.

[0443] N-(2-(Dimethylamino)ethyl)-4-(3,8,9,10-tetrahydrocyclopenta[c]pyrazolo[4,3-f]quinolin-7-yl)benzamide (Compound HSH3159)

[0444]

Chemical Structure

[0445] Synthesized using method F: off-white solid (10.8 mg, 0.25 mmol, 10.8%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.95 (t, J = 5.6 Hz, 1H), 8.51 (s, 1H), 8.07 (d, J = 8.1 Hz, 2H), 7.99 (d, J = 8.1 Hz, 2H), 7.90 (q, J = 9.2 Hz, 2H), 3.67 (q, J = 5.8 Hz, 2H), 3.50 (t, J = 7.6 Hz, 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 C 24 H 26 N5O[M+H] + Calculated value for 400.2132, found 400.2131.

[0446] N-(Cyanomethyl)-4-(3,8,9,10-tetrahydrocyclopenta[c]pyrazolo[4,3-f]quinolin-7-yl)benzamide (Compound HSH3160)

[0447]

Chemical Structure

[0448] Synthesized using method F: off-white solid (12.5 mg, 0.25 mmol, 13.6%). 11H NMR (500 MHz, (CD3)2SO) δ 9.30 (t, J = 5.5 Hz, 1H), 8.47 (s, 1H), 8.01 (d, J = 1.2 Hz, 4H), 7.93 (d, J = 10.0 Hz, 1H), 7.87 (d, J = 9.0 Hz, 1H), 4.35 (d, J = 5.3 Hz, 2H), 3.49 (t, J = 7.6 Hz, 2H), 3.29 (t, J = 7.5 Hz, 2H), 2.27 (m, 2H). 13 13C 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 C 22 H 18 N5O [M+H] + Calculated value: 368.1506, Measured value: 368.1504.

[0449] tert-Butyl (2-(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide)ethyl)carbamate (Compound HSH3150)

[0450]

Chemical Structure

[0451] Synthesized using Method F: off-white solid (170 mg, 0.5 mmol, 70%). 11H NMR (500 MHz, (CD3)2SO) δ 8.56 (s, 1H), 8.41 (s, 1H), 7.95 (d, J = 8.0 Hz, 2H), 7.84 (d, J = 10.1 Hz, 2H), 7.63 (d, J = 7.9 Hz, 2H), 6.78 (s, 1H), 3.34 (dt, J = 13.1, 6.4 Hz, 4H), 3.15 (q, J = 5.7 Hz, 2H), 2.76 (q, J = 10.7, 8.4 Hz, 2H), 2.00 (h, J = 9.1, 7.3 Hz, 2H), 1.75 (h, J = 9.6, 7.7 Hz, 2H), 1.38 (s, 9H). 13 13C NMR (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 C 28 H 32 N5O3[M+H] + Calculated value for 486.2499, found 486.2494.

[0452] N-(2-Aminoethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3165)

[0453]

Chemical Structure

[0454] Synthesized using Method G: off-white solid (85 mg, 0.245 mmol, 89%). 11H 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 C 23 H 24 N5O [M+H] + Calculated value for 386.1975, measured value 386.1973.

[0455] N-(2-(Methylsulfonyl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3175)

[0456]

Chemical Structure

[0457] Synthesized using Method E: off-white solid (6.7 mg, 0.5 mmol, 3%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.81 (t, J = 5.6 Hz, 1H), 8.57 (s, 1H), 7.99 - 7.88 (m, 2H), 7.85 (q, J = 9.2 Hz, 2H), 7.70 - 7.60 (m, 2H), 3.76 - 3.63 (m, 2H), 3.41 (t, J = 6.9 Hz, 2H), 3.05 (s, 3H), 2.76 (t, J = 6.1 Hz, 2H), 2.00 (m, 2H), 1.74 (m, 2H). 1313C 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 C 24 H 25 N4O3S [M+H] + Calculated value for 449.1642, measured value 449.1642.

[0458] N-(2-Sulfamoylethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3184)

[0459]

Chemical formula

[0460] Synthesized using Method F: off-white solid (10 mg, 0.5 mmol, 2.23%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.58 (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 = 6.5 Hz, 2H), 3.29 (dd, J = 8.1, 6.2 Hz, 2H), 2.77 (t, J = 6.2 Hz, 2H), 2.07 - 1.98 (m, 2H), 1.76 (m, 2H). 1313C 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 C 23 H 24 N5O3S [M+H] + Calculated value for 450.1594, found 450.1591.

[0461] (4-Isopropylpiperazin-1-yl)(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)methanone (Compound HSH3182)

[0462]

Chem.

[0463] Synthesized using Method F: off-white solid (56 mg, 0.5 mmol, 24.6%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.56 (s, 1H), 7.91 - 7.77 (m, 2H), 7.68 - 7.57 (m, 2H), 7.47 (d, J = 8.0 Hz, 2H), 3.51 (s, 4H), 2.78 (t, J = 6.1 Hz, 2H), 2.67 (m, 1H), 2.46 (d, J = 5.3 Hz, 3H), 2.01 (m, 2H), 1.81 - 1.71 (m, 2H), 0.97 (d, J = 6.5 Hz, 6H). 1313C 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 C 28 H 32 N5O [M+H] + Calculated value for 454.2601, measured value 454.2598.

[0464] N-(Prop-2-yn-1-yl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3199)

[0465]

Chemical formula

[0466] Synthesized using Method F: off-white solid (14 mg, 0.2 mmol, 18.4%).( 1 1H 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 C 24 H 21 N4O [M+H] + Calculated value for 381.1710, measured value 381.1709.

[0467] N-(1-((Dimethylamino)methyl)cyclopropyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3192)

[0468]

Chem.

[0469] Synthesized using Method F: off-white solid (18.2 mg, 0.5 mmol, 8.3%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.59 (d, J = 9.7 Hz, 2H), 7.98 - 7.90 (m, 2H), 7.84 (d, J = 11.5 Hz, 2H), 7.65 - 7.55 (m, 2H), 2.75 (t, J = 6.1 Hz, 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 = 4.8 Hz, 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 C 27 H 30 N5O [M+H] + Calculated value for 440.2444, found 440.2443.

[0470] N-(2-(Dimethylamino)ethyl)-4-(3,8,9,10,11,12-hexahydrocyclohepta[c]pyrazolo[4,3-f]quinolin-7-yl)benzamide (Compound HSH3194)

[0471]

Chem.

[0472] Synthesized using Method F: off-white solid (13.5 mg, 0.5 mmol, 6.3%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.62 (s, 1H), 8.32 (q, J = 5.7, 4.9 Hz, 1H), 7.96 - 7.90 (m, 2H), 7.84 (dd, J = 9.1, 0.9 Hz, 1H), 7.78 (d, J = 9.1 Hz, 1H), 7.60 - 7.51 (m, 2H), 3.54 (t, J = 4.9 Hz, 2H), 3.41 (td, J = 6.7, 5.4 Hz, 2H), 3.00 - 2.96 (m, 2H), 2.22 (s, 6H), 1.93 - 1.82 (m, 4H), 1.63 (m, 2H). 13 C NMR (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 C 26 H 30 N5O [M+H] + Calculated value for 428.2444, found 428.2442.

[0473] N-(2-(Dimethylamino)ethyl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide (Compound HSH3195)

[0474]

Chem.

[0475] Synthesized using Method F: off-white solid (57 mg, 0.5 mmol, 27.4%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.59 (s, 1H), 8.34 (t, J = 5.6 Hz, 1H), 7.98 - 7.83 (m, 4H), 7.67 - 7.64 (m, 2H), 4.78 (d, J = 1.5 Hz, 2H), 4.16 (t, J = 5.9 Hz, 2H), 3.43 - 3.37 (m, 4H), 2.46 (d, J = 6.8 Hz, 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 C 24 H 26 N5O2[M+H] + Calculated value for 416.2081, found 416.2080.

[0476] 4-(8,9-Dihydro-3H-cyclobuta[c]pyrazolo[4,3-f]quinolin-7-yl)-N-(2-(dimethylamino)ethyl)benzamide (Compound HSH3197)

[0477]

Chem.

[0478] Synthesized using method F: off-white solid (9 mg, 0.095 mmol, 24.5%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.39 - 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, 5.5 Hz, 2H), 2.52 (t, J = 6.8 Hz, 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 C 23 H 24 N5O [M+H] + Calculated value for 386.1975, found value 386.1974.

[0479] N-(2-(Pyrrolidin-1-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3180)

[0480]

Chemical Structure

[0481] Synthesized using method F: off-white solid (9 mg, 0.5 mmol, 4.1%). 11H NMR (500 MHz, (CD3)2SO) δ 8.56 (s, 1H), 8.34 (t, J = 5.7 Hz, 1H), 7.98 - 7.93 (m, 2H), 7.87 - 7.78 (m, 2H), 7.66 - 7.61 (m, 2H), 3.43 (d, J = 5.8 Hz, 2H), 3.32 (t, J = 6.6 Hz, 2H), 2.76 (t, J = 6.1 Hz, 2H), 2.62 (t, J = 6.9 Hz, 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 13C NMR (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 C 27 H 30 N5O [M+H] + calculated value 440.2444, found value 440.2443.

[0482] (4-(3,8,9,10,11,12-Hexahydrocyclohepta[c]pyrazolo[4,3-f]quinolin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3181)

[0483]

Chemical Structure

[0484] Synthesized using Method F: off-white solid (74 mg, 0.4 mmol, 42.1%). 11H NMR (800 MHz, (CD3)2SO) δ 8.63 (s, 1H), 7.85 (d, J = 9.0 Hz, 1H), 7.79 (s, 1H), 7.54 (d, J = 8.0 Hz, 2H), 7.48 (d, J = 8.1 Hz, 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 13C 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 C 27 H 30 N5O [M+H] + Calculated value for 440.2445, found 440.2444.

[0485] (4-(5-Fluoro-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3183)

[0486]

Chemical Structure

[0487] Synthesized using Method F: off-white solid (37 mg, 0.38 mmol, 22%). 11H NMR (800 MHz, (CD3)2SO) δ 8.55 (s, 1H), 7.71 (d, J = 10.0 Hz, 1H), 7.64 (d, J = 7.6 Hz, 2H), 7.50 (d, J = 7.0 Hz, 2H), 3.65 (s, 2H), 3.30 (s, 2H), 2.79 (d, J = 6.7 Hz, 2H), 2.34 (d, J = 68.6 Hz, 4H), 2.20 (d, J = 6.3 Hz, 3H), 1.99 (q, J = 6.6, 6.1 Hz, 2H), 1.74 (q, J = 6.2 Hz, 2H). 13 13C NMR (201 MHz, (CD3)2SO) δ 169.18, 157.78 ( 1 J = 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 C 26 H 27 FN5O [M+H] + calculated value 444.2194, found 444.2196.

[0488] Piperidin-1-yl(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)methanone (Compound HSH3185)

[0489]

Chemical Structure

[0490] Synthesized using Method F: off-white solid (14 mg, 0.5 mmol, 6.8%). 11H NMR (800 MHz, (CD3)2SO) δ 8.57 (s, 1H), 7.91 - 7.83 (m, 2H), 7.63 (d, J = 8.1 Hz, 2H), 7.47 (d, J = 8.0 Hz, 2H), 3.61 (s, 2H), 3.37 (t, J = 6.7 Hz, 4H), 2.81 (t, J = 6.2 Hz, 2H), 2.02 (td, J = 6.1, 3.3 Hz, 2H), 1.81 - 1.72 (m, 2H), 1.67 - 1.62 (m, 2H), 1.55 (m, 4H). 13 13C 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 C 26 H 27 N4O [M+H] + calculated value 411.2179, found 411.2178.

[0491] N-Cyclohexyl-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3186)

[0492]

Chem.

[0493] Synthesized using Method F: off-white solid (13 mg, 0.5 mmol, 6.5%). 11H NMR (800 MHz, (CD3)2SO) δ 8.59 (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, 2.4 Hz, 2H), 3.82 (tt, J = 7.5, 4.0 Hz, 1H), 3.27 (d, J = 5.7 Hz, 2H), 2.85 - 2.73 (m, 2H), 2.03 (q, J = 6.2 Hz, 2H), 1.87 (d, J = 11.4 Hz, 2H), 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 13C 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 C 27 H 29 N4O [M+H] + calculated value 425.2336, measured value 425.2336.

[0494] 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)

[0495]

Chem.

[0496] Synthesized using Method F: off-white solid (13.8 mg, 0.5 mmol, 5.7%). 11H NMR (500 MHz, (CD3)2SO) δ 8.56 (s, 1H), 7.86 (d, J = 9.1 Hz, 1H), 7.81 (d, J = 9.8 Hz, 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, 7.4 Hz, 4H), 2.79 (t, J = 6.1 Hz, 2H), 2.05 - 1.97 (m, 5H), 1.79 - 1.73 (m, 2H). 13 13C 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 C 27 H 28 N5O2[M+H] + Calculated value for 454.2237, measured value 454.2241.

[0497] (4-Methylpiperazin-1-yl)(5-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)thiophen-2-yl)methanone (Compound HSH3200)

[0498]

Chemical Structure

[0499] Synthesized using Method F: off-white solid (17 mg, 0.5 mmol, 7.9%). 11H NMR (800 MHz, (CD3)2SO) δ 8.56 (s, 1H), 7.88 (d, J = 9.0 Hz, 1H), 7.83 - 7.76 (m, 1H), 7.57 (d, J = 3.9 Hz, 1H), 7.40 (d, J = 3.8 Hz, 1H), 3.68 (t, J = 5.2 Hz, 4H), 3.34 (s, 2H), 3.11 (t, J = 6.2 Hz, 2H), 2.39 (t, J = 5.2 Hz, 4H), 2.23 (s, 3H), 2.05 - 2.00 (m, 2H), 1.89 - 1.85 (m, 2H). 13 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 C 24 H 26 N5OS [M+H] + Calculated value for 432.1852, measured value 432.1853.

[0500] (4-Methylpiperazin-1-yl)(3-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)methanone (Compound HSH3201)

[0501]

Chemical Structure

[0502] Synthesized using Method F: off-white solid (20 mg, 0.5 mmol, 9.4%). 11H NMR (800 MHz, (CD3)2SO) δ 8.57 (s, 1H), 7.91 - 7.80 (m, 2H), 7.65 (d, J = 7.8 Hz, 1H), 7.55 (d, J = 7.2 Hz, 2H), 7.44 (d, J = 7.8 Hz, 1H), 3.78 - 3.55 (m, 4H), 2.79 (t, J = 6.3 Hz, 2H), 2.43 - 2.30 (m, 5H), 2.19 (s, 3H), 2.04 - 1.96 (m, 2H), 1.81 - 1.71 (m, 2H). 13 13C 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 C 26 H 28 N5O [M+H] + Calculated value for 426.2289, measured value 426.2290.

[0503] N-(Cyanomethyl)-4-(9-methyl-3H-pyrazolo[4,3-f]quinolin-7-yl)benzamide (Compound HSH3202)

[0504]

Chemical Structure

[0505] Synthesized using Method F: off-white solid (23 mg, 0.5 mmol, 13.5%). 11H NMR (800 MHz, (CD3)2SO) δ 9.26 (m, 1H), 8.60 (s, 1H), 8.41 (d, J = 8.7 Hz, 2H), 8.19 (d, J = 9.0 Hz, 1H), 8.05 (d, J = 8.7 Hz, 2H), 8.01 - 7.91 (m, 2H), 4.36 (q, J = 5.6 Hz, 2H), 2.97 (d, J = 8.4 Hz, 3H). 13 13C NMR (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 C 20 H 16 N5O [M+H] + Calculated value for 342.1349, measured value 342.1353.

[0506] N-(Cyanomethyl)-3-fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3203)

[0507]

Chemical formula

[0508] Synthesized using Method F: off-white solid (70 mg, 0.5 mmol, 35%). 11H NMR (500 MHz, (CD3)2SO) δ 8.59 (s, 1H), 7.88 (d, J = 9.1 Hz, 1H), 7.81 (d, J = 9.1 Hz, 1H), 7.51 (t, J = 7.6 Hz, 1H), 7.35 (m, 2H), 3.35 (t, J = 6.5 Hz, 2H), 2.64 (t, J = 6.3 Hz, 2H), 2.52 (s, 2H), 2.04 - 1.96 (m, 2H), 1.79 (dd, J = 7.5, 4.2 Hz, 2H). 13 13C NMR (126 MHz, (CD3)2SO) δ 169.03, 158.32 ( 1 J = 245.7), 151.69, 143.97, 142.42, 140.00, 139.02 ( 3 J = 7.56 Hz), 134.97, 131.90, 130.08, 129.76 ( 2 J = 16.4 Hz), 129.44, 123.46, 122.44, 116.21, 115.64, 114.58 ( 2 J = 23.9 Hz), 34.73, 29.47, 27.38, 22.59, 22.19.

[0509] (2-Fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3204)

[0510]

Chem.

[0511] Synthesized using Method F: Yellow solid (18 mg, 0.25 mmol, 8%). 11H NMR (500 MHz, (CD3)2SO) δ 8.57 (s, 1H), 7.86 (d, J = 9.0 Hz, 1H), 7.81 (s, 1H), 7.47 (t, J = 5.1 Hz, 3H), 3.68 (s, 2H), 2.81 (t, J = 6.4 Hz, 2H), 2.42 - 2.27 (m, 5H), 2.22 (d, J = 7.7 Hz, 4H), 2.01 (m, 2H), 1.82 - 1.71 (m, 2H). 13 13C NMR (126 MHz, (CD3)2SO) δ 164.36, 161.15, 158.69 (d, 1 J = 247 Hz), 154.77, 144.56 (d, 3 J = 7.56 Hz), 143.85, 142.72, 142.45, 129.56, 129.22, 128.81, 126.08, 123.87 (d, 2 J = 17.6 Hz), 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 C 26 H 27 FN5O [M+H] + Calculated value for 444.2194, found 444.2194.

[0512] (3-Fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3205)

[0513]

Chemical Structure

[0514] Synthesized using Method F: off-white solid (56 mg, 0.5 mmol, 25.3%). 11H NMR (500 MHz, (CD3)2SO) δ 8.58 (s, 1H), 7.88 (d, J = 9.1 Hz, 1H), 7.82 (s, 1H), 7.52 (t, J = 7.6 Hz, 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 = 6.2 Hz, 2H), 2.46 (s, 2H), 2.28 (s, 3H), 2.00 (p, J = 6.5 Hz, 2H), 1.80 (tq, J = 9.2, 6.2, 4.4 Hz, 2H). 13 13C NMR (126 MHz, (CD3)2SO) δ 167.89, 160.36 (d, 1 J = 245.7 Hz), 158.41, 151.64, 143.87, 142.44, 138.89, 138.35 (d, 3 J = 7.56 Hz), 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 C 26 H 27 FN5O [M+H] + Calculated value for 444.2194, measured value 444.2193.

[0515] (2-Methoxy-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3206)

[0516]

Chemical Structure

[0517] Synthesized using method F: off-white solid (44 mg, 0.5 mmol, 19.31%). 1 H NMR (800 MHz, (CD3)2SO) δ 8.61 - 8.53 (m, 1H), 7.87 (dd, J = 9.0, 4.2 Hz, 1H), 7.84 (s, 1H), 7.26 (dd, J = 7.6, 4.2 Hz, 1H), 7.22 (d, J = 4.1 Hz, 1H), 7.15 (dd, J = 7.9, 3.8 Hz, 1H), 3.84 (d, J = 4.4 Hz, 3H), 3.72 - 3.64 (m, 1H), 3.61 (d, J = 16.7 Hz, 1H), 3.33 - 3.28 (m, 2H), 3.25 - 3.17 (m, 2H), 2.80 (q, J = 6.1, 5.6 Hz, 2H), 2.34 (m, 3H), 2.26 - 2.22 (m, 1H), 2.21 (d, J = 4.3 Hz, 3H), 2.01 (m, 2H), 1.76 (m, J = 6.1 Hz, 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 C 27 H 30 N5O2[M+H] + Calculated value 456.2394, measured value 456.2394.

[0518] N-(1-(Dimethylamino)propan-2-yl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 3207)

[0519] [Chemical]

[0520] Synthesized using Method F: off-white solid (31 mg, 0.5 mmol, 14.5%). 1 H NMR (800 MHz, (CD3)2SO) δ 8.57 (d, J = 8.7 Hz, 1H), 8.08 (d, J = 8.3 Hz, 1H), 7.94 (d, J = 7.9 Hz, 2H), 7.88 - 7.80 (m, 2H), 7.62 (d, J = 8.0 Hz, 2H), 4.22 - 4.15 (m, 1H), 2.76 (t, J = 6.2 Hz, 2H), 2.51 - 2.47 (m, 2H), 2.47 (dd, J = 12.2, 7.4 Hz, 1H), 2.28 (dd, J = 12.2, 6.7 Hz, 1H), 2.20 (d, J = 5.4 Hz, 6H), 2.03 - 1.99 (m, 2H), 1.79 - 1.73 (m, 2H), 1.18 (d, J = 6.7 Hz, 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 C 26 H 30 N5O [M+H] + Calculated value for 428.2444, measured value 428.2445.

[0521] (4-(2-Hydroxyethyl)piperazin-1-yl)(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)methanone (Compound HSH3208)

[0522] [Chemistry]

[0523] Synthesized using method F: off-white solid (27 mg, 0.75 mmol, 7.9%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.56 (s, 1H), 7.87 - 7.75 (m, 2H), 7.61 (d, J = 8.1 Hz, 2H), 7.47 (d, J = 8.1 Hz, 2H), 4.28 (t, J = 5.3 Hz, 1H), 3.63 - 3.42 (m, 6H), 3.34 (dq, J = 10.4, 5.0 Hz, 2H), 3.20 (d, J = 14.4 Hz, 2H), 2.79 (t, J = 6.2 Hz, 2H), 2.48 - 2.42 (m, 4H), 2.06 - 1.95 (m, 2H), 1.80 - 1.71 (m, 2H). 13 C NMR (126 MHz, (CD3)2SO) δ 169.30, 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.

[0524] (4-Methylpiperazin-1-yl)(4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)phenyl)methanone (Compound HSH3209)

[0525] [Chemistry]

[0526] Synthesized using method F: off-white solid (5 mg, 0.5 mmol, 4.7%). 11H NMR (800 MHz, (CD3)2SO) δ 8.60 (s, 1H), 7.92 (d, J = 9.0 Hz, 1H), 7.86 (d, J = 9.3 Hz, 1H), 7.69 - 7.62 (m, 2H), 7.53 - 7.48 (m, 2H), 4.81 (s, 2H), 4.16 (t, J = 5.9 Hz, 2H), 3.65 (s, 2H), 3.40 (t, J = 6.1 Hz, 4H), 2.47 - 2.28 (m, 4H), 2.24 (s, 3H). 13 13C 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 C 25 H 26 N5O2[M+H] + Calculated value for 428.2081, measured value 428.2080.

[0527] N-(1-Methylpiperidin-4-yl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3210)

[0528]

Chemical formula

[0529] Synthesized using Method F: off-white solid (83 mg, 0.75 mmol, 25%). 11H NMR (500 MHz, (CD3)2SO) δ 8.56 (s, 1H), 8.20 (d, J = 7.6 Hz, 1H), 7.95 (d, J = 8.1 Hz, 2H), 7.88 - 7.75 (m, 2H), 7.61 (d, J = 8.1 Hz, 2H), 3.87 - 3.70 (m, 1H), 3.32 (t, J = 6.6 Hz, 2H), 2.85 - 2.69 (m, 4H), 2.19 (s, 3H), 2.08 - 1.95 (m, 4H), 1.84 - 1.59 (m, 6H).

[0530] (4-(9,9-Dioxide-3,8,10,11-tetrahydropyrazolo[4,3-f]thiopyrano[3,4-c]quinolin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3211)

[0531]

Chem.

[0532] Synthesized using Method F: off-white solid (53 mg, 0.75 mmol, 14.9%). 1 1H NMR (800 MHz, (CD3)2SO) δ 8.70 (s, 1H), 8.02 (d, J = 9.0 Hz, 1H), 7.91 - 7.84 (m, 1H), 7.68 (d, J = 8.0 Hz, 2H), 7.63 (d, J = 8.1 Hz, 2H), 4.58 (s, 2H), 4.02 (d, J = 6.8 Hz, 2H), 3.72 (t, J = 6.7 Hz, 2H), 3.38 (d, J = 35.2 Hz, 8H), 2.78 (s, 3H). 1313C 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 C 25 H 26 N5O3S [M+H] + Calculated value for 476.1751, found 476.1750.

[0533] N-(2-(Dimethylamino)ethyl)-4-(9,9-dioxide-3,8,10,11-tetrahydropyrazolo[4,3-f]thiopyrano[3,4-c]quinolin-7-yl)benzamide (Compound HSH3212)

[0534]

Chemical Structure

[0535] Synthesized using Method F: off-white solid (68 mg, 0.75 mmol, 19.6%). 1 1H NMR (800 MHz, (CD3)2SO) δ 8.71 (s, 1H), 8.50 (t, J = 4.8 Hz, 1H), 8.01 (dt, J = 9.3, 4.2 Hz, 3H), 7.92 - 7.84 (m, 1H), 7.71 - 7.61 (m, 2H), 4.51 (s, 2H), 4.01 (d, J = 9.4 Hz, 2H), 3.71 (d, J = 9.0 Hz, 2H), 3.42 (p, J = 6.9 Hz, 2H), 2.48 - 2.40 (m, 2H), 2.21 (d, J = 3.9 Hz, 6H). 1313C 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 C 24 H 26 N5O3S [M+H] + Calculated value for 464.1750, found 464.1751.

[0536] 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)

[0537] [Chemical formula]

[0538] Synthesized using Method F: off-white solid (99 mg, 0.75 mmol, 29.9%). 1 1H NMR (500 MHz,, (CD3)2SO) δ 8.57 (s, 1H), 8.38 (t, J = 5.7 Hz, 1H), 8.03 - 7.93 (m, 2H), 7.90 (d, J = 9.2 Hz, 1H), 7.84 (d, J = 9.1 Hz, 1H), 7.71 - 7.61 (m, 2H), 4.78 (s, 2H), 4.14 (t, J = 5.9 Hz, 2H), 3.44 (t, J = 6.3 Hz, 2H), 3.39 (q, J = 6.9, 6.0 Hz, 2H), 2.63 (t, J = 6.9 Hz, 2H), 2.58 - 2.48 (m, 4H), 1.68 (h, J = 3.2 Hz, 4H). 1313C NMR (126 MHz, in (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 C 26 H 28 N5O2[M+H] + Calculated value for 442.2237, measured value 442.2238.

[0539] Piperazin-1-yl(4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)methanone (Compound HSH3215)

[0540]

Chem.

[0541] Synthesized using Method G: off-white solid (37 mg, 0.26 mmol, 34.6%). 1 1H NMR (800 MHz, in (CD3)2SO) δ 8.59 (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 C 25 H 26 N5O [M+H] + Calculated value for 412.2132, measured value 412.2133.

[0542] N-(2-(Dimethylamino)ethyl)-5-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)thiophene-2-carboxamide (Compound HSH3216)

[0543]

Chem.

[0544] Synthesized using Method F: off-white solid (35 mg, 0.5 mmol, 16.7%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.54 (s, 1H), 8.28 (t, J = 5.8 Hz, 1H), 7.85 (d, J = 9.1 Hz, 1H), 7.77 (d, J = 9.1 Hz, 1H), 7.72 (d, J = 4.1 Hz, 1H), 7.57 (d, J = 4.1 Hz, 1H), 3.36 (q, J = 6.8 Hz, 2H), 3.30 (s, 2H), 3.08 (t, J = 6.1 Hz, 2H), 2.43 (t, J = 6.8 Hz, 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 C 23 H 26 N5OS [M+H] + Calculated value 420.1852, found value 420.1850.

[0545] N-(2-(Dimethylamino)ethyl)-3-fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3217)

[0546]

Chem.

[0547] Synthesized using Method F: off-white solid (53 mg, 0.75 mmol, 16.4%). 1 H NMR (800 MHz, (CD3)2SO) δ 8.61 (s, 1H), 8.53 (t, J = 5.6 Hz, 1H), 7.89 (d, J = 9.0 Hz, 1H), 7.83 (dd, J = 7.8, 1.8 Hz, 2H), 7.79 (m, 1H), 7.57 (t, J = 7.6 Hz, 1H), 3.44 (q, J = 6.7 Hz, 3H), 3.36 (t, J = 6.8 Hz, 2H), 2.63 (t, J = 6.4 Hz, 2H), 2.54 (t, J = 6.8 Hz, 2H), 2.28 (s, 6H), 2.02 (q, J = 6.2, 5.0 Hz, 2H), 1.84 - 1.77 (m, 2H). 13 C NMR (201 MHz, (CD3)2SO) δ 165.08, 160.01 (d, 1 J= 245 Hz), 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 C 25 H 27 FN5O [M+H] + calculated value 432.2194, found value 432.2190.

[0548] 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)

[0549]

Chem.

[0550] Synthesized using Method F: off-white solid (146 mg, 0.75 mmol, 42.5%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.59 (s, 1H), 8.46 (t, J = 5.7 Hz, 1H), 7.87 (d, J = 9.1 Hz, 1H), 7.81 (m, 2H), 7.77 (dd, J = 10.7, 1.8 Hz, 1H), 7.54 (t, J = 7.6 Hz, 1H), 3.46 - 3.38 (m, 2H), 3.34 (t, J = 6.5 Hz, 2H), 2.62 (dd, J = 8.3, 5.6 Hz, 4H), 2.51 (m, 4H), 2.05 - 1.94 (m, 2H), 1.84 - 1.73 (m, 2H), 1.68 (h, J = 3.2 Hz, 4H). 13 C NMR (126 MHz, (CD3)2SO) δ 165.12, 160.48 (d, 1 J= 245 Hz), 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 C 27 H 29 FN5O [M+H]+ Calculated value: 458.2350, measured value: 458.2350.

[0551] (2-Methoxy-4-(8,9,10,11-tetrahydro-3H-naphtho[1,2-e]indazol-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3222)

[0552] [Chemical formula]

[0553] Synthesized using Method F: off-white solid (77 mg, 0.75 mmol, 22.6%). 1 H NMR (800 MHz, (CD3)2SO) δ 8.61 - 8.53 (m, 1H), 7.87 (dd, J = 9.0, 4.2 Hz, 1H), 7.84 (s, 1H), 7.26 (dd, J = 7.6, 4.2 Hz, 1H), 7.22 (d, J = 4.1 Hz, 1H), 7.15 (dd, J = 7.9, 3.8 Hz, 1H), 3.84 (d, J = 4.4 Hz, 3H), 3.72 - 3.64 (m, 1H), 3.61 (d, J = 16.7 Hz, 1H), 3.33 - 3.28 (m, 2H), 3.25 - 3.17 (m, 2H), 2.80 (q, J = 6.1, 5.6 Hz, 2H), 2.34 (m, 3H), 2.26 - 2.22 (m, 1H), 2.21 (d, J = 4.3 Hz, 3H), 2.01 (m, 2H), 1.76 (m, J = 6.1 Hz, 2H). 1313C 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 C 27 H 30 N5O2[M+H] + Calculated value for 456.2394, found 456.2394.

[0554] (4-(9-Cyclopropyl-3H-benzo[e]indazol-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone (Compound HSH3223)

[0555] [Chemical formula]

[0556] Synthesized using Method F: off-white solid (69 mg, 0.75 mmol, 22.4%). 1 1H NMR (800 MHz, (CD3)2SO) δ 8.84 (s, 1H), 8.32 (d, J = 8.1 Hz, 2H), 7.98 - 7.91 (m, 3H), 7.52 (d, J = 8.2 Hz, 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 = 5.3 Hz, 2H). 1313C 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 C 25 H 26 N5O [M+H] + Calculated value for 412.2132, measured value 412.2132.

[0557] 7-(4-((4-Methylpiperazin-1-yl)methyl)phenyl)-8,9,10,11-tetrahydro-3H-naphtho[1,2-e]indazole (Compound HSH3225)

[0558]

Chem.

[0559] Synthesized using Method H: off-white solid (30 mg, 0.5 mmol, 14.6%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.56 (s, 1H), 7.86 - 7.76 (m, 2H), 7.48 (d, J = 8.0 Hz, 2H), 7.37 (d, J = 8.0 Hz, 2H), 3.52 (s, 2H), 3.33 (t, J = 6.6 Hz, 2H), 2.78 (t, J = 6.2 Hz, 2H), 2.42 (t, J = 4.9 Hz, 4H), 2.38 - 2.27 (m, 4H), 2.16 (s, 3H), 2.01 (m, 2H), 1.79 - 1.70 (m, 2H). 1313C 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 C 26 H 30 N5[M+H] + Calculated value for 412.2495, measured value 412.2496

[0560] (4-(Methylsulfonyl)piperazin-1-yl)(4-(3,8,10,11-tetrahydroisochromeno[5,6-e]indazol-7-yl)phenyl)methanone (Compound HSH3126)

[0561] [Chemical formula]

[0562] Synthesized using Method F: off-white solid (127 mg, 1 mmol, 25.9%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.52 (s, 1H), 7.88 (t, J = 10.6 Hz, 2H), 7.65 (d, J = 8.1 Hz, 2H), 7.54 (d, J = 8.1 Hz, 2H), 4.78 (s, 2H), 4.13 (t, J = 5.9 Hz, 2H), 3.65 (m, 4H), 3.36 (t, J = 5.9 Hz, 2H), 3.25 - 3.17 (m, 4H), 2.91 (s, 3H). 1313C NMR (126 MHz, (CD3)2SO) δ 169.50, 152.93, 144.29, 141.17, 139.43, 138.80, 135.91, 135.80, 129.50, 129.38, 127.51, 127.39, 121.39, 116.31, 115.07, 66.87, 64.42, 45.85, 35.09, 28.69.

[0563] N-(2-(Dimethylamino)ethyl)-4-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSH3127)

[0564]

Chem.

[0565] Synthesized using Method F: off-white solid (38.4 mg, 0.5 mmol, 17.9%). 1 1H NMR (800 MHz, (CD3)2SO) δ 8.28 (t, J = 5.7 Hz, 1H), 7.97 - 7.94 (m, 2H), 7.74 (d, J = 8.9 Hz, 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 = 6.9 Hz, 2H), 2.22 (s, 6H), 1.80 (h, J = 4.7, 3.3 Hz, 4H). 13 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 C 26 H30 N5O [M+H] + Calculated value 428.2444, measured value 428.2445.

[0566] 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)

[0567]

Chemical Structure

[0568] Synthesized using Method F: off-white solid (40.7 mg, 0.3 mmol, 29.92%). 1 H NMR (800 MHz, (CD3)2SO) δ 8.33 (t, J = 5.6 Hz, 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 = 6.0 Hz, 2H), 2.64 (t, J = 7.0 Hz, 2H), 2.57 - 2.51 (m, J = 2.8, 2.0 Hz, 4H), 1.84 - 1.77 (m, 4H), 1.70 (h, J = 3.1 Hz, 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 C 28 H 32 N5O [M+H] + Calculated value 454.2601, measured value 454.2603.

[0569] (4-(9-Methyl-3H-pyrazolo[4,3-f]quinolin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone

[0570]

Chem.

[0571] Synthesized using Method D: off-white solid (53 mg, 0.5 mmol, 27.5%) 1 H NMR (800 MHz, (CD3)2SO) δ 8.60 (d, J = 6.8 Hz, 1H), 8.34 (m, 2H), 8.16 (d, J = 6.9 Hz, 1H), 7.96 (dt, J = 31.9, 8.0 Hz, 2H), 7.54 (d, J = 7.8 Hz, 2H), 3.56 - 3.41 (m, 3H), 2.97 (d, J = 6.8 Hz, 4H), 2.35 (s, 4H), 2.22 (d, J = 6.9 Hz, 3H), 1.67 (d, J = 6.2 Hz, 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 C 23 H 25 N5O [M+H] + Calculated value for 386.1975, measured value 386.1974.

[0572] (4-Methylpiperazin-1-yl)(4-(3,8,9,10-tetrahydrocyclopenta[c]pyrazolo[4,3-f]quinolin-7-yl)phenyl)methanone

[0573]

Chem.

[0574] Synthesized using Method D: off-white solid (71 mg, 0.5 mmol, 34.5%) 1 H NMR (800 MHz, (CD3)2SO) δ 8.48 (s, 1H), 7.97 (d, J = 7.8 Hz, 2H), 7.93 (s, 1H), 7.88 (d, J = 8.9 Hz, 1H), 7.53 (d, J = 7.7 Hz, 2H), 3.52 (s, 2H), 3.31 (d, J = 7.4 Hz, 2H), 2.33 - 2.26 (m, 3H), 2.23 (s, 4H), 1.66 (s, 2H). 13 C 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 C 25 H 27 N5O [M+H] + Calculated value for 412.2132, found 412.2133.

[0575] (4-(1-Methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)phenyl)(4-methylpiperazin-1-yl)methanone

[0576]

Chemical Structure

[0577] Synthesized using Method D: off-white solid (43 mg, 0.5 mmol, 19.5%). 11H NMR (500 MHz, (CD3)2SO) δ 7.75 - 7.64 (m, 2H), 7.58 (d, J = 8.1 Hz, 2H), 7.46 (d, J = 8.1 Hz, 2H), 3.38 (dt, J = 15.1, 5.2 Hz, 3H), 2.84 (s, 3H), 2.77 (t, J = 6.3 Hz, 2H), 2.41 - 2.31 (m, 5H), 2.21 (s, 5H), 1.82 - 1.73 (m, 5H). 13 13C 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 C 27 H 30 N5O [M+H] + Calculated value 440.2444, found value 440.2445.

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

[0579]

Chemical Structure

[0580] Synthesized using Method D: off-white solid (111 mg, 0.5 mmol, 52.6%) 11H NMR (800 MHz, (CD3)2SO) δ 8.99 (t, J = 7.1 Hz, 1H), 8.05 - 8.01 (m, 2H), 7.89 (d, J = 8.9 Hz, 1H), 7.83 (d, J = 6.0 Hz, 2H), 7.65 (d, J = 7.9 Hz, 2H), 7.05 (s, 1H), 4.47 (d, J = 5.5 Hz, 2H), 2.78 (t, J = 6.3 Hz, 2H), 2.02 (p, J = 6.3 Hz, 2H), 1.76 (p, J = 5.8 Hz, 2H). 13 13C 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 C 25 H 23 N6O [M+H] + Calculated value 423.1927, measured value 423.1927.

[0581] N-(2-(1H-Imidazol-1-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0582]

Chem.

[0583] Synthesized using Method D: off-white solid (84 mg, 0.5 mmol, 38.5%) 11H NMR (800 MHz, (CD3)2SO) δ 8.74 (t, J = 5.5 Hz, 1H), 8.57 (s, 1H), 7.93 (d, J = 7.8 Hz, 2H), 7.89 (d, J = 8.9 Hz, 1H), 7.86 - 7.80 (m, 1H), 7.65 (d, J = 7.8 Hz, 2H), 7.63 (s, 1H), 7.18 (s, 1H), 6.89 (s, 1H), 4.22 (t, J = 6.2 Hz, 2H), 3.63 (q, J = 5.9 Hz, 2H), 2.78 (t, J = 6.2 Hz, 2H), 2.06 - 1.99 (m, 2H), 1.80 - 1.73 (m, 2H). 13 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 C 26 H 25 N6O [M+H] + calculated value 437.2084, measured value 437.2081.

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

[0585]

Chemical Structure

[0586] Synthesized using Method D: off-white solid (65 mg, 0.5 mmol, 30.8%). 11H NMR (800 MHz, (CD3)2SO) δ 9.17 (t, J = 5.9 Hz, 1H), 8.57 (s, 1H), 8.07 (d, J = 7.8 Hz, 2H), 7.89 (d, J = 9.0 Hz, 1H), 7.82 (d, J = 9.3 Hz, 1H), 7.65 (d, J = 7.8 Hz, 2H), 6.93 (s, 2H), 4.56 (d, J = 5.6 Hz, 2H), 2.77 (t, J = 6.3 Hz, 2H), 2.00 (q, J = 6.8, 6.3 Hz, 2H), 1.75 (p, J = 5.6 Hz, 2H). 13 13C 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 C 25 H 23 N6O [M+H] + Calculated value for 423.1927, measured value 423.1926.

[0587] N-((1-Methyl-1H-imidazol-2-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0588]

Chemical Structure

[0589] Synthesized using Method D: off-white solid (68 mg, 0.5 mmol, 31.2%). 11H NMR (500 MHz, (CD3)2SO) δ 8.56 (s, 1H), 8.37 (d, J = 4.3 Hz, 3H), 8.16 (d, J = 8.3 Hz, 3H), 7.04 (d, J = 13.1 Hz, 1H), 6.78 (d, J = 18.9 Hz, 1H), 4.56 (d, J = 5.3 Hz, 2H), 3.68 (s, 5H), 2.77 (t, J = 6.2 Hz, 3H), 1.88 (dp, J = 127.7, 5.7 Hz, 6H), 1.23 (dd, J = 14.7, 7.0 Hz, 2H). 13 13C 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 C 26 H 26 N6O [M+H] + Calculated value 437.2084, measured value 437.2081.

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

[0591]

Chemical Structure

[0592] Synthesized using Method D: off-white solid (79 mg, 0.5 mmol, 46.0%). 11H NMR (500 MHz, (CD3)2SO) δ 8.58 (s, 1H), 8.43 - 8.06 (m, 2H), 8.01 (d, J = 7.8 Hz, 2H), 7.84 (q, J = 9.1 Hz, 2H), 7.65 (d, J = 7.8 Hz, 2H), 4.61 (d, J = 5.6 Hz, 2H), 3.34 (d, J = 13.1 Hz, 2H), 2.78 (t, J = 6.1 Hz, 2H), 1.89 (dp, J = 125.7, 6.1 Hz, 4H). 13 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 C 24 H 22 N7O [M+H] + Calculated value 424.1880, measured value 424.1877.

[0593] N-((1H-Imidazol-2-yl)methyl)-3-fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0594]

Chem.

[0595] Synthesized using Method D: off-white solid (109 mg, 1.0 mmol, 49.5%). 11H NMR (500 MHz, (CD3)2SO) δ 8.59 (s, 1H), 7.93 - 7.78 (m, 4H), 7.57 (t, J = 7.6 Hz, 1H), 7.16 (s, 2H), 4.65 (d, J = 5.5 Hz, 2H), 3.35 (t, J = 6.5 Hz, 2H), 2.62 (t, J = 6.2 Hz, 2H), 2.04 - 1.74 (m, 4H). 13 13C NMR (126 MHz, (CD3)2SO) δ 165.6, 159.5 (d, 1 J = 252 Hz), 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 C 25 H 22 FN6O [M+H] + calculated value 441.1833, measured value 441.1830.

[0596] N-((1H-Imidazol-2-yl)methyl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide

[0597]

Chemical Structure

[0598] Synthesized using Method D: off-white solid (117 mg, 0.5 mmol, 55.2%). 11H NMR (500 MHz, (CD3)2SO) δ 8.59 (s, 1H), 8.04 (d, J = 7.9 Hz, 2H), 7.94 - 7.83 (m, 2H), 7.67 (d, J = 7.9 Hz, 2H), 6.95 (s, 2H), 4.79 (s, 2H), 4.55 (d, J = 5.6 Hz, 2H), 4.16 (t, J = 5.8 Hz, 2H), 3.41 (d, J = 11.8 Hz, 2H). 13 13C 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 C 24 H 21 N6O2[M+H] + Calculated value 425.1720, found value 425.1716.

[0599] N-((1H-Imidazol-2-yl)methyl)-4-(9,9-dioxide-3,8,10,11-tetrahydropyrazolo[4,3-f]thiopyrano[3,4-c]quinolin-7-yl)benzamide

[0600]

Chemical Structure

[0601] Synthesized using Method D: off-white solid (98 mg, 0.5 mmol, 41.5%). 11H NMR (500 MHz, (CD3)2SO) δ 8.69 (s, 1H), 8.05 (dd, J = 47.7, 8.4 Hz, 3H), 7.88 (d, J = 9.1 Hz, 1H), 7.68 (d, J = 7.8 Hz, 2H), 7.19 (s, 2H), 4.67 (d, J = 5.4 Hz, 2H), 4.47 (s, 2H), 4.02 (t, J = 6.7 Hz, 2H), 3.69 (t, J = 6.8 Hz, 2H). 13 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 C 24 H 21 N6O3S [M+H] + Calculated value 473.1390, measured value 473.1386.

[0602] N-((1H-Tetrazol-5-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0603]

Chemical Structure

[0604] Synthesis of the final compound via the "click" reaction: To a solution of alkyne (1 equiv) in DMF (2 mL) and water (0.5 mL), TMS-azide (2 equiv) was added. A catalytic amount of copper(II) sulfate 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, washed with water, and dried over sodium sulfate. It was then purified via column chromatography with 9% methanol, 1% ammonium hydroxide in water, and 90% ethyl acetate.

[0605] Off-white solid. (36 mg, 1.0 mmol, 34.4%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.79 (d, J = 3.9 Hz, 1H), 8.26 - 8.08 (m, 4H), 7.83 (dd, J = 8.1, 3.8 Hz, 2H), 4.59 (dd, J = 236.2, 5.5 Hz, 2H), 3.50 (t, J = 6.2 Hz, 2H), 2.78 (t, J = 6.2 Hz, 2H), 2.11 - 2.00 (m, 2H), 1.82 (ddt, J = 11.9, 9.4, 4.1 Hz, 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 C 23 H 21 N8O [M+H] + Calculated value for 425.1833, found 425.1831.

[0606] N-((4-Methyl-1H-imidazol-2-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0607]

Chemical Structure

[0608] Method D: off-white solid. (79 mg, 0.5 mmol, 36.2%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.58 (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 = 5.6 Hz, 2H), 2.78 (t, J = 6.2 Hz, 2H), 2.12 (d, J = 1.1 Hz, 4H), 2.05 (d, J = 16.2 Hz, 1H), 2.01 (td, J = 9.0, 7.4, 4.7 Hz, 2H), 1.77 (tq, J = 8.5, 6.0, 4.3 Hz, 2H), 1.24 (d, J = 6.8 Hz, 1H). 13 C NMR (126 MHz, (CD3)2SO) δ 166.7, 156.2, 150.0, 144.6, 144.3, 143.9, 142.5, 133.9, 131.2, 129.6, 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.

[0609] (S)-N-((tetrahydrofuran-2-yl)methyl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide

[0610] [Chemical Structure]

[0611] Synthesized using Method D: off-white solid (98 mg, 0.5 mmol, 45.7%). 11H NMR (500 MHz, (CD3)2SO) δ 8.62 (s, 1H), 8.55 (t, J = 5.8 Hz, 1H), 8.00 - 7.86 (m, 4H), 7.69 - 7.65 (m, 2H), 4.79 (d, J = 1.8 Hz, 2H), 4.16 (t, J = 5.9 Hz, 2H), 3.79 - 3.69 (m, 2H), 3.63 (td, J = 8.0, 6.7 Hz, 2H), 3.51 (dd, J = 8.5, 5.4 Hz, 2H), 3.43 (d, J = 11.9 Hz, 3H), 2.57 - 2.49 (m, 1H), 1.97 (dtd, J = 12.2, 8.0, 5.6 Hz, 1H), 1.64 (ddt, J = 12.5, 7.7, 6.1 Hz, 1H). 13 13C NMR (126 MHz, (CD3)2SO) δ 166.6, 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.

[0612] (R)-N-((Tetrahydrofuran-2-yl)methyl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide

[0613]

Chem.

[0614] Synthesized using Method D: off-white solid (121 mg, 0.5 mmol, 56.5%). 11H NMR (500 MHz, (CD3)2SO) δ 8.60 (s, 1H), 8.05 (d, J = 7.9 Hz, 2H), 7.97 - 7.90 (m, 2H), 7.86 (d, J = 9.2 Hz, 1H), 7.69 (dd, J = 17.1, 7.9 Hz, 2H), 4.79 (s, 2H), 4.15 (t, J = 5.9 Hz, 2H), 3.41 (d, J = 6.0 Hz, 3H). 13 13C NMR (126 MHz, (CD3)2SO) δ 167.5, 152.8, 144.5, 144.0, 139.6, 131.2, 129.7, 129.5, 129.4, 129.2, 127.6, 127.5, 121.6, 116.1, 71.1, 67.3, 66.8, 64.4, 42.6, 30.11, 28.7.

[0615] N-(Tetrahydro-2H-pyran-4-yl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide

[0616]

Chem.

[0617] Synthesized using Method D: off-white solid (96 mg, 0.5 mmol, 44.8%) 11H NMR (500 MHz, (CD3)2SO) δ 8.27 (d, J = 7.6 Hz, 1H), 7.94 (dd, J = 31.0, 8.4 Hz, 4H), 7.65 (d, J = 8.2 Hz, 2H), 4.78 (s, 2H), 4.16 (t, J = 5.9 Hz, 2H), 4.04 (tdd, J = 13.4, 9.3, 5.1 Hz, 1H), 3.97 - 3.83 (m, 2H), 3.41 (td, J = 11.5, 2.3 Hz, 4H), 1.81 (ddd, J = 12.4, 4.7, 2.1 Hz, 2H), 1.63 (qd, J = 11.8, 4.4 Hz, 2H). 13 13C NMR (126 MHz, (CD3)2SO) δ 165.9, 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.

[0618] N-((3-Methyloxetan-3-yl)methyl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide

[0619]

Chem.

[0620] Synthesized using Method D: off-white solid (87 mg, 0.5 mmol, 40.6%) 11H NMR (500 MHz, (CD3)2SO) δ 8.58 (t, J = 6.1 Hz, 1H), 8.06 - 7.79 (m, 2H), 7.77 - 7.58 (m, 1H), 4.80 (s, 1H), 4.52 (d, J = 5.7 Hz, 1H), 4.28 - 4.08 (m, 2H), 3.47 (dd, J = 50.8, 5.9 Hz, 2H), 3.18 (s, 3H), 1.29 (s, 1H). 13 13C NMR (126 MHz, (CD3)2SO) δ 167.3, 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.

[0621] N-(2-Hydroxy-2-methylpropyl)-4-(3,8,10,11-tetrahydropyrano[3,4-c]pyrazolo[4,3-f]quinolin-7-yl)benzamide

[0622]

Chem.

[0623] Synthesized using Method D: off-white solid (72 mg, 0.5 mmol, 34.6%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.19 (t, J = 6.1 Hz, 1H), 8.06 - 7.78 (m, 4H), 7.74 - 7.56 (m, 2H), 4.80 (s, 2H), 4.16 (t, J = 5.9 Hz, 2H), 3.36 (dd, J = 43.4, 6.0 Hz, 4H), 1.15 (d, J = 1.8 Hz, 6H). 1313C NMR (126 MHz, (CD3)2SO) δ 168.3, 154.0, 144.4, 141.6, 141.0, 139.7, 134.9, 131.7, 128.7, 127.5, 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.

[0624] General scheme for the synthesis of hydroxamic acid analogs:

[0625]

Chemical formula

[0626] General procedure for amide coupling: In a 20 mL capped vial, the acid intermediate (0.5 mmol), O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (0.7 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (1.3 equiv) were dissolved in anhydrous DMF (3 mL). Subsequently, DIPEA (5 equiv) was added. The reaction mixture was stirred at 80 °C overnight. After completion, the reaction mixture was concentrated and purified via silica gel column chromatography using ethyl acetate and hexane (30:70 to 70:30).

[0627] General procedure for the deprotection of tetrahydropyranyl ether: In a 5 mL vial, the tetrahydropyranyl protected intermediate (0.25 mmol) was dissolved in dichloromethane (1.5 mL). Subsequently, trifluoroacetic acid or hydrochloric acid (2 equiv) was added. The reaction mixture was refluxed in a capped vial for 2 h to obtain the desired deprotected product.

[0628] N-Hydroxy-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSD1993):

[0629] [Chemical formula]

[0630] According to Scheme 2, solution A was prepared by suspending the substrate ester (0.25 mmol) in methanol (2 mL), and the separated solution B was prepared by dissolving crushed potassium hydroxide (KOH) (10 equivalents) and hydroxylamine hydrochloride (5 equivalents) in methanol (4 mL), and stirred at ambient temperature for 30 minutes. The filtrate of solution B was added to solution A and stirring was continued overnight. After completion, the solvent was removed under reduced pressure, and the obtained solid was dissolved in water. The pH was adjusted to 7 with an aqueous solution of HCl (1N), the obtained precipitate was filtered, dehydrated under high vacuum, and purified by washing with ethanol and recrystallization to obtain the desired product as an off-white solid (85 mg, 24%).

[0631] 1 H NMR (500 MHz, DMSO-d6) δ 11.31 (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 = 8.3 Hz, 2H), 3.34 - 3.27 (m, 2H), 2.76 (t, J = 6.1 Hz, 2H), 2.02 - 1.94 (m, 2H), 1.78 - 1.67 (m, 2H); 13 C NMR (126 MHz, DMSO-d6) δ 164.5, 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.

[0632] 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)

[0633]

Chem.

[0634] Off-white solid (41%); 1 H NMR (500 MHz, DMSO-d6) δ 11.72 (s, 1H), 7.87 (d, J = 8.0 Hz, 2H), 7.77 - 7.67 (m, 2H), 7.62 (d, J = 7.9 Hz, 2H), 5.03 (d, J = 3.0 Hz, 1H), 4.07 (t, J = 9.7 Hz, 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 (126 MHz, DMSO-d6) δ 164.5, 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.

[0635] N-Hydroxy-4-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSD19931)

[0636]

Chem.

[0637] Off-white solid (58%); 11H NMR (500 MHz, DMSO-d6) δ 11.40 (s, 1H), 8.00 (d, J = 9.1 Hz, 1H), 7.94 (d, J = 7.9 Hz, 2H), 7.83 (d, J = 9.1 Hz, 1H), 7.73 (d, J = 7.9 Hz, 2H), 3.55 (t, J = 5.5 Hz, 2H), 2.91 (s, 3H), 2.76 (t, J = 5.8 Hz, 2H), 2.48 (t, J = 2.0 Hz, 2H), 1.89 - 1.73 (m, 4H).

[0638] 4-(9,9-Dioxide-3,8,10,11-tetrahydropyrazolo[4,3-f]thiopyrano[3,4-c]quinolin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide

[0639]

Chem.

[0640] Off-white solid (36%); 1 1H NMR (500 MHz, DMSO-d6) δ 11.77 (s, 1H), 8.68 (s, 1H), 8.00 (dd, J = 9.1, 0.9 Hz, 1H), 7.96 - 7.89 (m, 3H), 7.70 - 7.62 (m, 2H), 5.08 - 4.98 (m, 1H), 4.54 - 4.44 (m, 2H), 4.12 - 3.96 (m, 3H), 3.69 (t, J = 6.6 Hz, 2H), 3.59 - 3.51 (m, 1H), 1.80 - 1.67 (m, 3H), 1.63 - 1.48 (m, 3H); 1313C NMR (126 MHz, DMSO) δ 164.3, 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.

[0641] 4-(9,9-Dioxide-3,8,10,11-tetrahydropyrazolo[4,3-f]thiopyrano[3,4-c]quinolin-7-yl)-N-hydroxybenzamide (Compound 19932)

[0642]

Chem.

[0643] Off-white solid (61%); 1 1H NMR (500 MHz, DMSO-d6) δ 11.34 (s, 1H), 9.12 (s, 1H), 8.70 (d, J = 12.2 Hz, 1H), 8.00 (d, J = 9.0 Hz, 1H), 7.91 (d, J = 8.0 Hz, 2H), 7.64 (d, J = 8.0 Hz, 2H), 4.50 (s, 2H), 4.01 (s, 2H), 3.69 (t, J = 6.7 Hz, 2H).

[0644] 2-Fluoro-N-((tetrahydro-2H-pyran-2-yl)oxy)-5-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0645]

Chem.

[0646] Off-white solid (40%); 11H NMR (500 MHz, DMSO-d6) δ 11.49 (s, 1H), 8.59 (s, 1H), 7.92 - 7.81 (m, 1H), 7.78 - 7.72 (m, 1H), 7.69 (dd, J = 6.7, 2.3 Hz, 1H), 7.40 (t, J = 9.9, 8.5 Hz, 1H), 5.03 (s, 1H), 4.06 - 3.90 (m, 1H), 3.51 (d, J = 11.3 Hz, 1H), 3.37 - 3.32 (m, 2H), 2.78 (t, J = 6.1 Hz, 2H), 2.01 (t, J = 6.0 Hz, 2H), 1.84 - 1.65 (m, 5H), 1.53 (s, 3H).

[0647] 2-Fluoro-N-hydroxy-5-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSD19933)

[0648]

Chemical Structure

[0649] Off-white solid (52%); 1 1H NMR (500 MHz, DMSO-d6) δ 11.12 (s, 1H), 8.73 (s, 1H), 8.06 (d, J = 9.2 Hz, 1H), 7.92 (d, J = 9.1 Hz, 1H), 7.85 - 7.78 (m, 2H), 7.49 (t, J = 9.4 Hz, 1H), 3.42 (t, J = 6.4 Hz, 2H), 2.78 (t, J = 6.1 Hz, 2H), 2.12 - 1.99 (m, 2H), 1.87 - 1.73 (m, 2H).

[0650] 4-(9-Methyl-3H-pyrazolo[4,3-f]quinolin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide

[0651] [Chemistry]

[0652] Off-white solid (39%); 1 H NMR (500 MHz, DMSO-d6) δ 11.74 (s, 1H), 8.58 (s, 1H), 8.37 (d, J = 8.1 Hz, 2H), 8.21 (s, 1H), 8.00 - 7.88 (m, 4H), 5.11 - 4.92 (m, 1H), 4.09 (d, J = 11.5 Hz, 1H), 3.54 (dd, J = 9.7, 5.4 Hz, 1H), 2.97 (s, 3H), 1.73 (s, 3H), 1.60 - 1.49 (m, 3H); 13 C NMR (126 MHz, DMSO) δ 164.4, 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.

[0653] N-Hydroxy-4-(9-methyl-3H-pyrazolo[4,3-f]quinolin-7-yl)benzamide (Compound 19934)

[0654] [Chemistry]

[0655] Off-white solid (60%); 1 H NMR (500 MHz, DMSO-d6) δ 11.36 (s, 1H), 8.64 (s, 1H), 8.32 (d, J = 8.4 Hz, 2H), 8.23 (s, 1H), 8.04 - 7.96 (m, 2H), 7.94 (d, J = 8.4 Hz, 2H), 2.98 (s, 3H).

[0656] 4-(9-Cyclobutyl-3H-pyrazolo[4,3-f]quinolin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide (Compound HSD19935P)

[0657]

Chem.

[0658] Off-white solid (53%); 1 H NMR (500 MHz, methanol-d4) δ 8.57 (s, 1H), 8.25 (d, J = 8.1 Hz, 2H), 8.07 (s, 1H), 8.02 (d, J = 8.5 Hz, 1H), 7.96 (d, J = 8.1 Hz, 2H), 7.89 (d, J = 9.2 Hz, 1H), 5.11 (d, J = 2.9 Hz, 1H), 4.43 (s, 1H), 4.17 (t, J = 10.7 Hz, 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).

[0659] 4-(9-Cyclobutyl-3H-pyrazolo[4,3-f]quinolin-7-yl)-N-hydroxybenzamide (Compound HSD19935)

[0660]

Chem.

[0661] Off-white solid (55%); 11H NMR (500 MHz, DMSO-d6) δ 11.35 (s, 1H), 8.65 (s, 1H), 8.39 (d, J = 8.4 Hz, 2H), 8.19 (s, 1H), 7.97 - 7.90 (m, 4H), 4.44 (p, J = 8.1 Hz, 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).

[0662] N-Hydroxy-4-(1-methyl-9-morpholino-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSD19936)

[0663]

Chem.

[0664] Yellow solid (68%); 1 1H NMR (500 MHz, DMSO-d6) δ 11.44 (s, 1H), 11.04 (s, 1H), 7.95 (d, J = 8.1 Hz, 3H), 7.84 (d, J = 9.0 Hz, 1H), 7.73 (d, J = 7.9 Hz, 2H), 4.01 - 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).

[0665] 4-(5-Fluoro-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-hydroxybenzamide (Compound HSD19937)

[0666]

Chem.

[0667] Off-white solid (59%); 1 H NMR (500 MHz, DMSO-d6) δ 11.33 (s, 1H), 8.61 (s, 1H), 7.87 (d, J = 8.3 Hz, 2H), 7.71 (d, J = 10.0 Hz, 1H), 7.67 - 7.60 (m, 2H), 3.30 (t, J = 6.3 Hz, 2H), 2.78 (t, J = 6.1 Hz, 2H), 2.03 - 1.94 (m, 2H), 1.82 - 1.68 (m, 2H).

[0668] N-Methoxy-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSD199310)

[0669]

Chemical Structure

[0670] Off-white solid (65%); 1 H NMR (500 MHz, DMSO-d6) δ 11.83 (s, 1H), 8.57 (s, 1H), 7.97 - 7.78 (m, 4H), 7.66 (d, J = 8.2 Hz, 2H), 3.74 (s, 3H), 3.36 - 3.32 (m, 2H), 2.76 (t, J = 6.1 Hz, 2H), 2.08 - 1.90 (m, 2H), 1.82 - 1.47 (m, 2H); 13 C NMR (126 MHz, 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.

[0671] 5-(1-Methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)picolylamide

[0672]

Chem.

[0673] Off-white solid (57%); 1 H NMR (500 MHz, DMSO-d6) δ 11.90 (s, 1H), 8.79 (d, J = 2.2 Hz, 1H), 8.19 (dd, J = 8.0, 2.1 Hz, 1H), 8.09 (d, J = 8.0 Hz, 1H), 7.76 (s, 1H), 5.11 (d, J = 2.7 Hz, 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 (126 MHz, DMSO) δ 161.5, 152.8, 149.1, 149.0, 144.1, 143.1, 142.8, 141.3, 139.4, 138.5, 129.9, 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.

[0674] N-Hydroxy-5-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)picolylamide (Compound HSD199312)

[0675]

Chem.

[0676] Off-white solid (70%); 11H NMR (500 MHz, DMSO-d6) δ 11.58 (s, 1H), 8.84 (s, 1H), 8.26 (dd, J = 8.1, 2.2 Hz, 1H), 8.13 (d, J = 8.0 Hz, 1H), 7.94 (d, J = 9.2 Hz, 1H), 7.80 (d, J = 9.1 Hz, 1H), 3.64 - 3.41 (m, 2H), 2.90 (s, 3H), 2.84 - 2.74 (m, 2H), 1.99 - 1.72 (m, 4H).

[0677] 6-(1-Methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)nicotinamide

[0678]

Chem.

[0679] Off-white solid (50%); 1 1H NMR (500 MHz, DMSO-d6) δ 11.91 (s, 1H), 9.07 - 8.84 (m, 1H), 8.27 (dd, J = 8.2, 2.3 Hz, 1H), 7.91 (d, J = 8.2 Hz, 1H), 7.80 - 7.55 (m, 1H), 5.05 (s, 1H), 4.14 - 3.98 (m, 1H), 3.56 (dd, J = 10.2, 5.2 Hz, 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).

[0680] N-Hydroxy-6-(1-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)nicotinamide (HSD199313)

[0681] [Chemistry]

[0682] Off-white solid (61%); 1 H NMR (500 MHz, DMSO-d6) δ 11.52 (s, 1H), 9.03 (d, J = 2.2 Hz, 1H), 8.31 (dd, J = 8.1, 2.3 Hz, 1H), 7.96 (d, J = 8.1 Hz, 1H), 7.89 (d, J = 9.1 Hz, 1H), 7.79 (d, J = 9.1 Hz, 1H), 3.66 - 3.24 (m, 2H), 2.99 - 2.94 (m, 2H), 2.89 (s, 3H), 1.97 - 1.48 (m, 4H).

[0683] N-Hydroxy-4-(3,8,10,11-tetrahydropyrazolo[4,3-f]thiopyrano[3,4-c]quinolin-7-yl)benzamide (Compound HSD199315)

[0684] [Chemistry]

[0685] Off-white solid (56%); 1 H NMR (500 MHz, DMSO-d6) δ 11.38 (s, 1H), 8.73 (s, 1H), 8.03 (d, J = 9.1 Hz, 1H), 7.96 - 7.91 (m, 2H), 7.89 (d, J = 9.1 Hz, 1H), 7.76 - 7.70 (m, 2H), 3.88 (s, 2H), 3.68 - 3.62 (m, 1H), 3.17 (t, J = 6.1 Hz, 2H).

[0686] 4-(5-Methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide

[0687]

Chem.

[0688] Off-white solid (38%); 1 H NMR (500 MHz, DMSO-d6) δ 8.52 (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 = 6.1 Hz, 2H), 2.69 (s, 3H), 2.08 - 1.96 (m, 2H), 1.79 - 1.60 (m, 5H), 1.60 - 1.49 (m, 3H).

[0689] N-Hydroxy-4-(5-methyl-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSD199316)

[0690]

Chem.

[0691] Off-white solid (62%); 1 H NMR (500 MHz, DMSO-d6) δ 11.30 (s, 1H), 8.53 (s, 1H), 7.90 - 7.83 (m, 2H), 7.75 (s, 1H), 7.73 - 7.66 (m, 2H), 3.32 (t, J = 6.6 Hz, 2H), 2.81 (t, J = 6.1 Hz, 2H), 2.08 - 1.96 (m, 3H), 1.81 - 1.68 (m, 3H).

[0692] 4-(5-Methoxy-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-((tetrahydro-2H-pyran-2-yl)oxy)benzamide

[0693]

Chem.

[0694] Off-white solid (51%); 1 1H NMR (500 MHz, DMSO-d6) δ 11.73 (s, 1H), 8.46 (d, J = 7.2 Hz, 1H), 7.88 (d, J = 8.3 Hz, 2H), 7.65 - 7.59 (m, 2H), 7.17 (s, 1H), 5.04 (d, J = 3.0 Hz, 1H), 4.12 - 4.04 (m, 1H), 3.96 (s, 3H), 3.55 (dd, J = 9.4, 5.9 Hz, 1H), 3.38 - 3.29 (m, 2H), 2.74 (t, J = 6.0 Hz, 2H), 2.03 - 1.94 (m, 2H), 1.77 - 1.71 (m, 5H), 1.60 - 1.52 (m, 3H).

[0695] N-Hydroxy-4-(5-methoxy-8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound HSD199317)

[0696]

Chem.

[0697] Off-white solid (70%); 11H NMR (500 MHz, DMSO-d6) δ 11.35 (s, 1H), 8.52 (s, 1H), 8.00 - 7.80 (m, 2H), 7.73 - 7.56 (m, 2H), 7.29 (s, 1H), 3.99 (d, J = 1.4 Hz, 3H), 3.32 (t, J = 6.4 Hz, 2H), 2.72 (t, J = 6.1 Hz, 2H), 2.19 - 1.94 (m, 2H), 1.75 (q, J = 5.5 Hz, 2H).

[0698] [Example 2] Evaluation of CDK Inhibitor Compounds in Cancer Cells Molm-14 (human acute myeloid leukemia (AML) cell line), Molm-14 D835Y (human AML cell line with a mutation at D835Y), Molm-14 F691L (human AML cell line with a mutation at F691L, a common cause of acquired resistance to FLT3 inhibitors), SNU-16 (human gastric cancer cell line), and KCL22-IR (chronic myeloid leukemia cell line) were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin, along with L-glutamine and sodium bicarbonate. Cells were seeded at 2.1E4 cells per mL in 190 μL per well in a 96-well plate. The plate was incubated in a 5% CO2 incubator for 24 hours. The cells were then treated with the compounds of the present specification at the desired concentrations or in a dimethyl sulfoxide (DMSO) control and incubated in a 5% CO2 incubator for 72 hours.

[0699] Thereafter, the cells were treated with the CellTiter-Blue Viability Assay, incubated in a 5% CO2 incubator for 3 hours, and then the absorbance was measured using a Cytation 5 plate reader at 570 nm. The data were analyzed using GraphPad prism. Figures 3-10C show the results.

[0700] [Example 3] Analysis of the Structure Docked to the Haspin of Compound HSD1217 To optimize compounds for dual FLT3 and haspin kinase inhibition, the docking scores for some of the dual FLT3 / haspin inhibitor compounds were evaluated. The determination of the three-dimensional structure of the protein-receptor complex is an important part of structure-based drug design.

[0701] 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 protein active site. The 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 by Schrodinger. Water was removed prior to docking into haspin. The docking trial validation for haspin kinase was based off the previously generated docking pose of HSD972. Visualization was performed through PyMol and Maestro.

[0702] Analysis of the structure of compound HSD1217 docked to haspin (Figure 11A) suggested that the benzamide - NH2 moiety had a hydrogen - bonding interaction with the key residue D687 and was positioned towards the region that harbored the H651 and D649 residues bound to the ATP phosphate moiety inside. Therefore, adding a polar group such as imidazole to the benzamide with the possibility of donating a hydrogen bond to the benzamide could reach either the H651 residue or the D649 residue while maintaining the hydrogen bond with the D687 residue and bringing about further interactions to increase the ligand affinity for the protein. Since H651, D649, or D687 is important for ATP binding to haspin, it was speculated that inhibitors that associate these residues have a higher chance of being resistant to kinase mutations that could inhibit the binding of the inhibitor. Eswaran et al., Structure and functional characterization of the atypical human kinase haspin, Proceedings National Academy Science USA 106(48): 20198 - 20203 (2009).

[0703] In - silico analysis of compounds 4, 5, and 6 (analogues of compound HSD1217), each containing an imidazole moiety, suggested that all three compounds could form key interactions with at least one of these ATP - binding residues (Figures 11B - 11D). For the docking analysis of 3H - pyrazolo[4,3 - f]quinoline - based compounds to FLT3, see Dayal et al. (2021) above.

[0704] [Example 4] Kinase assay and structure - activity relationship study Using the preliminary compounds identified in Example 3, a structure-activity relationship (SAR) study was conducted to explore how modulation affects hSpin and / or FLT3 inhibition, and to identify new imidazolyl-containing pyrazolo-based compounds (described herein) that are dual FLT3 / hSpin inhibitors inhibiting both kinases at ultra-low (sub-nanomolar) concentrations.

[0705] Multicomponent reactions (MCRs) have often been utilized to rapidly synthesize a wide range of highly functionalized analogs in a single pot for constructing libraries for biological screening. Herein, the compounds were synthesized using a Doebner-like MCR followed by amidation thereafter (Scheme 3).

[0706]

Chemical formula

[0707] For the initial pilot library, amines retaining imidazole or their biological equivalents were in focus, because such heterocyclic amines (which are found in other drugs such as nilotinib) were thought to potentially facilitate ligand association with protein residues (see above). During the Doebner-like MCR, the amine and aldehyde react to form a Schiff base, which then reacts with the added ketone in the presence of an acid catalyst (HCl).

[0708] Together with the MCR product A in hand (see Scheme 3), the final amide products (5-18) were formed via amidation using a commercially available amine, HATU (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, hexafluorophosphate azabenzotriazole tetramethyluronium) and Hunig's base (DIPEA).

[0709] Using the established library of amidokinase inhibitors, kinase inhibition and growth inhibition of AML cell lines (Molm-14 with FLT3-ITD inside) were analyzed (Table 1). More specifically, haspin and FLT3 inhibition were evaluated in-house via the ADP Glo Kinase Assay (Promega, Madison, WI). All compounds were screened against 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 triple means.

[0710] Briefly, the ADP-Glo Kinase Assay Kit was used to determine the inhibition of haspin. Kinase Buffer (SignalChem Cat#K01-09; 25 mM MOPS, pH 7.2, 12.5 mM β-glycerol-phosphate, 25 mM MgCl2, 2 mM EDTA) was supplemented with dithiothreitol (DTT) (0.2 mM) and bovine serum albumin (BSA) (40 μg / mL) immediately before use. A 5 μL reaction volume containing 9 ng / μL kinase, 10 μM ATP, 50 nM inhibitor, and an appropriate peptide substrate (20 μM) was incubated for 3 hours and then quenched with an equal volume (5 μL) of GLO reagent. After 40 minutes, 10 μL of DET reagent was added and incubated for 1 hour, and then visualized on a Cytation (trademark) 5 Cell Imaging Multi-Mode Reader (BioTek).

[0711]

Table 1-1

[0712]

Table 1-2

[0713]

Table 1-3

[0714] The SAR study supported that the imidazole ring is important for the activity against both Haspin and FLT3 and that even minor modifications dramatically affect kinase inhibition. Compounds 4, 5, and 6 all contain an imidazole moiety, although differently substituted, and yet their inhibition profiles against the kinases tested are different.

[0715] Compound 6, the 2-substituted imidazole, inhibited Haspin and FLT3 (77% and 76% respectively), which was better than that of compound 5, the 1-substituted imidazole (33% and 31% respectively), and compound 4, the 4-substituted imidazole (54% and 49% respectively).

[0716] Since compound 6 showed the best inhibition of the kinases tested, the substitution pattern of compound 6 was explored to potentially improve the potency. Substituting the -NH on compound 6 with -NMe as in compound 7 decreased Haspin and FLT3 inhibition, as well as AML growth inhibition. For example, on the one hand, at 50 nM, compound 6 inhibited Haspin by 77%, while the -NMe analogue (compound 7) inhibited the same enzyme at 26%. For FLT3, the potency dropped from 76% inhibition by compound 6 at 50 nM to only 14% inhibition by compound 7. Replacing the imidazole unit in compound 6 with other 5-membered bioisosteres (triazole (compound 8), tetrazole (compound 10), or thiazole (compound 13)) gave compounds that could inhibit the growth of both kinases and AML cells, but generally less than compound 6. Extending the carbon chain length between the amide and imidazole by one unit to give compound 15 did not significantly affect the inhibition of any of the kinases.

[0717] The effect of fluorine substituents on lead compounds has always been explored because fluorine is known to be useful for modulating the properties of drugs (both target binding and general drug properties such as metabolic stability). Therefore, the benzamide moiety of compound 6 was fluorinated to obtain compounds 9 and 11, but FLT3 and Haspin kinase inhibition was not significantly affected (Table 1 comparing compounds 9 and 11 with compound 6).

[0718] Some imidazole-containing drugs, such as nilotinib, have an imidazole unit methylated at the 4-position. Therefore, the degree of methylation of the imidazole unit in compound 6, which is thought to affect kinase inhibition, was also investigated. Methylation of the imidazole unit significantly improved the potency of the action against the growth of Haspin, FLT3, and Molm-14 cells. Compound 14 (also called compound HSK205) inhibited Haspin by 97% and FLT3 by 94% only at 50 nM, supporting that compound HSK205 is a super-potent inhibitor of both FLT3 and Haspin (see below). In fact, at a concentration of only 50 nM, compound HSK205 almost completely inhibited the growth of Molm-14 (FLT3-ITD) cells (96%).

[0719] To verify the laboratory results, compound HSK205 was sent to contract research organizations Eurofins and Reaction Biology to obtain binding data for Haspin (Eurofins) and enzyme inhibition data for FLT3 (Reaction Biology).

[0720] Compound HSK205 bound to Haspin with a K d = 0.55 nM (K d < 0.17 nM of the control Haspin inhibitor CHR-6494) (Figure 12A).

[0721] In terms of FLT3 inhibition, at 10 μM ATP, compound HSK205 had an IC 50has <0.5 nM, while under similar conditions, gilteritinib has IC 50 = 0.7 nM and IC 50 <0.5 nM (Figures 12B - 12C).

[0722] These SAR studies verified previous reports that substitution at the C7 position can be used to target other kinases, that the 3H - pyrazolo[4,3 - f]quinoline moiety can target FLT3, and that the moiety is highly tunable. For example, compound HSD1169, which is also a 3H - pyrazolo[4,3 - f]quinoline - containing compound and retains the fluorinated pyrazole, is a potent inhibitor of FLT3 while being a weak inhibitor of Haspin, as the change of the C7 moiety to benzamide leads to a combination FLT3 / Haspin inhibitor.

[0723] [Example 5] Target binding and in vitro biological activity Next, immunoblotting was performed to evaluate whether compound HSK205 actually inhibits Haspin in AML cells and to confirm the inhibition of histone H3 (Thr3) phosphorylation by Haspin (Figure 13A). Briefly, Molm - 14 cells were treated with either 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 the cells were harvested 24 hours after treatment. The samples were then analyzed for phosphorylation of histone H3 (Thr3).

[0724] More specifically, cells were cultured to confluence in RPMI medium supplemented with 10% fetal bovine serum and 1× penicillin-streptomycin (Gibco™). Cells were seeded at 5,000 cells / well in a 96-well plate. After 24 hours of incubation at 37 °C in a 5% CO2 incubator, the compound diluted in the same medium was added to the indicated final concentration and incubated at 37 °C for 72 hours in a 5% CO2 incubator. Cell viability was determined using the CellTiter-Blue® Cell Viability Assay (Promega, Madison, WI), and absorbance readings were taken at 570 nm on a Cytation™ 5 Cell Imaging Multi-Mode Reader (BioTek) 3 hours after addition. Appropriate vehicle, and a cell-free control were included. GI 50 Readings were confirmed by two scientists on separate days.

[0725] Acute myeloid leukemia cells (Molm-14) were incubated with varying concentrations of the compound as specified. Cells were collected and lysed, and protein concentration was quantified against a BSA standard. After protein denaturation, the proteins were separated via SDS-PAGE electrophoresis and transferred to an NC membrane. After blocking, peroxidase activity was detected using SuperSignal West Pico reagent (Thermo Scientific) with a CCD camera LAS-4000 (Fujifilm) using an overnight incubation with specific primary antibodies and a 2-hour room temperature incubation with a peroxidase-conjugated secondary antibody. Bands were normalized using the housekeeping gene Actin. Reported values represent the mean of triplicates, and error bars represent SD. All specific antibodies were purchased from Cell Signaling.

[0726] The results showed that compound HSK205 was effective in reducing the phosphorylation of histone H3 (Thr3), with a more pronounced effect at 20 nM compared to 5 nM and 100 nM compared to 20 nM (Figure 13B). However, gilteritinib did not exhibit haspin inhibition in Molm-13 cells.

[0727] Furthermore, the FLT3 target association in Molm-14 cells was evaluated to confirm the FLT3 inhibition seen 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 led to a reduction in phosphorylated FLT3 (Figures 14A and 14B).

[0728] Many FLT3 inhibitors have been tested in the clinic, and several such as gilteritinib, midostaurin, and quizartinib have been approved, but on-target FLT mutations that reduce AML sensitivity to these drugs have been observed. See Scholl et al. (2020) above. Therefore, compound HSK205 was evaluated against Molm-14 cell lines harboring the resistance mutations FLT3-ITD-D835Y and F691L, which are commonly found in the clinic. (Figure 15). Notably, compound HSK205 had low nanomolar GI 50 values against Molm-14-ITD-F691L (2.2 nM) and Molm14-ITD-D835Y (4.3 nM).

[0729] To advance the compound for further development, a DNA intercalation assay was performed to remove compounds with high affinity for DNA. Although compound HSK205 contains polycyclic rings that can enhance DNA intercalation and unwanted toxicity, compound HSK205 did not intercalate into DNA (Figure 16). Under similar experimental conditions, mitoxantrone, a known DNA intercalator, associated with DNA (Figure 16).

[0730] [Example 6] Chemistry and Synthesis of Compound HSK205 and Its Analogs For Examples 3 - 5, unless otherwise specified, all reagents and solvents were purchased from commercial sources and used as received. 1 H and 13 1H and 13C NMR spectra were obtained in DMSO-d6 as the solvent using a 500 MHz spectrometer or an 800 MHz spectrometer with Me4Si as the internal standard. Chemical shifts were reported in parts per million (δ), and were calibrated using the remaining non-deuterated solvent as the internal reference.

[0731] 1 Data for 1H NMR spectra were reported as follows: chemical shift (δ ppm) (multiplicity, coupling constant (Hz), integration). Multiplicity was reported as: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, or combinations thereof. High-resolution mass spectra (HRMS) were obtained using electrospray ionization (ESI) technology and a TOF mass analyzer. New compounds were characterized by 1H NMR, 13C NMR, and HRMS data, and the purity of the final compounds was reported using HPLC. 1 1H NMR 13 13C NMR, and HRMS data, and the purity of the final compounds was reported using HPLC.

[0732] 4-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid (Compound 1)

[0733] In a round-bottom flask equipped with a stir bar, the appropriate indazole (1 equiv) and the appropriate aldehyde (1 equiv) were dissolved in ethanol. The mixture was refluxed at 120 °C for 2 h, then the appropriate ketone (2 equiv) and a catalytic amount of hydrochloric acid were added. This was stirred at reflux for an additional 12 h. The reaction solvent was evaporated using a rotary evaporator and purified via column chromatography with 10% methanol in ethyl acetate to afford Compound 1 as an off-white solid (2.38 g, 69.4%).

[0734] 11H NMR (500 MHz, (CD3)2SO) δ 8.75 (s, 1H), 8.26 (d, J = 9.2 Hz, 1H), 8.19 (d, J = 9.2 Hz, 1H), 8.15 - 8.11 (m, 2H), 7.85 - 7.80 (m, 2H), 3.46 (t, J = 6.5 Hz, 2H), 2.76 (t, J = 6.1 Hz, 2H), 2.03 (dd, J = 6.0, 2.9 Hz, 2H), 1.83 - 1.76 (m, 2H). 13 13C 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 C 21 H 17 N3O2[M + H] + Calculated value for 344.1393, measured value 344.1392.

[0735] 3-Fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid (Compound 2)

[0736] Following the same synthetic procedure as for Compound 1, Compound 2 was obtained as an off-white solid (636 mg, 58.7%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.64 (s, 1H), 8.06 (d, J = 22.2 Hz, 3H), 7.72 (d, J = 11.2 Hz, 1H), 7.63 (d, J = 8.1 Hz, 1H), 3.31 (t, J = 6.6 Hz, 2H), 2.76 (t, J = 6.2 Hz, 2H), 2.04 - 1.91 (m, 2H), 1.76 (q, J = 6.0 Hz, 2H). 1313C NMR (126 MHz, (CD3)2SO) δ 167.55, 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 C 21 H 16 FN3O2 [M+H] + Calculated value for 362.1299, found 362.1300.

[0737] 2-Fluoro-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzoic acid (Compound 3)

[0738] Following the same synthetic procedure as for Compound 1, Compound 3 was obtained as an off-white solid (556 mg, 51.3%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.76 (d, J = 8.6 Hz, 1H), 8.16 (d, J = 14.3 Hz, 2H), 8.04 (t, J = 7.7 Hz, 1H), 7.67 (d, J = 11.1 Hz, 1H), 7.60 (d, J = 8.1 Hz, 1H), 3.47 (t, J = 6.6 Hz, 2H), 2.80 (t, J = 6.2 Hz, 2H), 2.05 (p, J = 6.0 Hz, 2H), 1.81 (p, J = 5.8 Hz, 2H). 13 13C NMR (126 MHz, (CD3)2SO) δ 165.04, 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 ( 2J = 20 Hz), 115.32, 109.16, 30.48, 28.04, 21.99, 21.86. HRMS (ESI) m / z C 21 H 16 FN3O2[M+H] + Calculated value 362.1299, measured value 362.1301.

[0739] N-((1H-Imidazol-4-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 4)

[0740] In a 20 mL screw-cap vial equipped with a magnetic stir bar, the acid intermediate (1.3 equiv) and HATU (1.5 equiv) were sealed off by a septum, air was removed via a vacuum pump, and a nitrogen balloon was added to the reaction vessel, filling the vessel with an inert gas. To this reaction vial, anhydrous DMF (2 mL) and DIPEA (8 equiv) were added. This was heated at 60 °C for 30 minutes, and then the amine (1 equiv) was added. The reaction mixture was stirred at 60 °C overnight.

[0741] The reaction solvent was evaporated using a rotary evaporator and purified via column chromatography using 1% ammonium hydroxide, 9% methanol, and 90% ethyl acetate to obtain Compound 4 as an off-white solid (111 mg, 52.6%).

[0742] 11H NMR (500 MHz, (CD3)2SO) δ 8.81 (t, J = 5.6 Hz, 1H), 8.57 (s, 1H), 7.99 (d, J = 8.2 Hz, 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 = 5.6 Hz, 2H), 3.32 (t, J = 6.5 Hz, 2H), 2.76 (t, J = 6.1 Hz, 2H), 2.03 - 1.95 (m, 2H), 1.74 (dp, J = 9.1, 3.2, 2.6 Hz, 2H). 13 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 C 25 H 22 N6O [M+H] + calculated value 423.1928, measured value 423.1911.

[0743] N-(2-(1H-Imidazol-1-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 5)

[0744] Following the same synthetic procedure as for Compound 4, Compound 5 was obtained as an off-white solid (84 mg, 38.5%). 11H NMR (800 MHz, (CD3)2SO) δ 8.74 (t, J = 5.5 Hz, 1H), 8.57 (s, 1H), 7.93 (d, J = 7.8 Hz, 2H), 7.89 (d, J = 8.9 Hz, 1H), 7.86 - 7.80 (m, 1H), 7.65 (d, J = 7.8 Hz, 2H), 7.63 (s, 1H), 7.18 (s, 1H), 6.89 (s, 1H), 4.22 (t, J = 6.2 Hz, 2H), 3.63 (q, J = 5.9 Hz, 2H), 2.78 (t, J = 6.2 Hz, 2H), 2.06 - 1.99 (m, 2H), 1.80 - 1.73 (m, 2H). 13 13C 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 C 26 H 24 N6O [M+H] + Calculated value for 437.2084, found 437.2081.

[0745] N-((1H-Imidazol-2-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 6)

[0746] Following the same synthetic procedure as for Compound 4, Compound 6 was obtained as an off-white solid (65 mg, 30.8%). 11H NMR (800 MHz, (CD3)2SO) δ 9.17 (t, J = 5.9 Hz, 1H), 8.57 (s, 1H), 8.07 (d, J = 7.8 Hz, 2H), 7.89 (d, J = 9.0 Hz, 1H), 7.82 (d, J = 9.3 Hz, 1H), 7.65 (d, J = 7.8 Hz, 2H), 6.93 (s, 2H), 4.56 (d, J = 5.6 Hz, 2H), 2.77 (t, J = 6.3 Hz, 2H), 2.00 (q, J = 6.8, 6.3 Hz, 2H), 1.75 (p, J = 5.6 Hz, 2H). 13 13C NMR (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 C 25 H 22 N6O [M+H] + Calculated value 423.1927, measured value 423.1926.

[0747] 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)

[0748] Following the same synthetic procedure as for Compound 4, Compound 7 was obtained as a pale yellow solid (68 mg, 31.2%). 11H NMR (500 MHz, (CD3)2SO) δ 8.56 (s, 1H), 8.37 (d, J = 4.3 Hz, 3H), 8.16 (d, J = 8.3 Hz, 3H), 7.04 (d, J = 13.1 Hz, 1H), 6.78 (d, J = 18.9 Hz, 1H), 4.56 (d, J = 5.3 Hz, 2H), 3.68 (s, 3H), 3.58 (s, 2H) 2.77 (t, J = 6.2 Hz, 3H), 1.88 (dp, J = 127.7, 5.7 Hz, 6H), 1.23 (dd, J = 14.7, 7.0 Hz, 2H). 13 13C 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 C 26 H 25 N6O [M+H] + Calculated value for 437.2084, found 437.2081.

[0749] 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)

[0750] Following the same synthetic procedure as for Compound 4, Compound 8 was obtained as an off-white solid (79 mg, 37.4%). 11H NMR (500 MHz, (CD3)2SO) δ 8.58 (s, 1H), 8.43 - 8.06 (m, 2H), 8.01 (d, J = 7.8 Hz, 2H), 7.84 (q, J = 9.1 Hz, 2H), 7.65 (d, J = 7.8 Hz, 2H), 4.61 (d, J = 5.6 Hz, 2H), 3.34 (d, J = 13.1 Hz, 2H), 2.78 (t, J = 6.1 Hz, 2H), 1.89 (dp, J = 125.7, 6.1 Hz, 4H). 13 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 C 24 H 21 N7O [M+H] + calculated value 424.1880, measured value 424.1877.

[0751] 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)

[0752] Following the same synthetic procedure as for Compound 4, Compound 9 was obtained as an off-white solid (109 mg, 49.5%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.59 (s, 1H), 7.93 - 7.78 (m, 4H), 7.57 (t, J = 7.6 Hz, 1H), 7.16 (s, 2H), 4.65 (d, J = 5.5 Hz, 2H), 3.35 (t, J = 6.5 Hz, 2H), 2.62 (t, J = 6.2 Hz, 2H), 2.04 - 1.74 (m, 4H). 1313C NMR (126 MHz, (CD3)2SO) δ 165.63, 161.85, 159.52 ( 1 J = 246.4 Hz), 151.59, 151.13, 145.34, 144.18, 143.97, 142.50, 136.45, 131.98, 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 C 25 H 21 FN6O [M+H] + Calculated value 441.1833, found 441.1830.

[0753] N-((1H-Tetrazol-5-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 10)

[0754] To a solution of alkyne (1 equiv) in DMF (2 mL) and water (0.5 mL) was added TMS-azide (2 equiv). Catalytic amounts of copper(II) sulfate pentahydrate and sodium ascorbate were added. The reaction mixture was stirred at 100 °C overnight. The reaction solvent was evaporated 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 with 90% ethyl acetate and 10% hexane to afford an off-white solid (36 mg, 34%).

[0755] 11H NMR (500 MHz, (CD3)2SO) δ 9.43 (dt, J = 33.1, 5.5 Hz, 1H), 8.79 (d, J = 3.9 Hz, 1H), 8.33 - 8.08 (m, 4H), 7.83 (dd, J = 8.1, 3.9 Hz, 2H), 4.82 (d, J = 5.6 Hz, 2H), 3.50 (t, J = 6.2 Hz, 2H), 2.78 (t, J = 6.2 Hz, 2H), 2.15 - 1.98 (m, 2H), 1.82 (ddt, J = 11.9, 9.4, 4.1 Hz, 2H). 13 13C 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 C 23 H 20 N8O [M+H] + calculated value 425.1833, found 425.1831.

[0756] 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)

[0757] Following the same synthetic procedure as for Compound 4, Compound 11 was obtained as an off-white solid (87 mg, 39.5%). 11H NMR (500 MHz, (CD3)2SO) δ 8.82 (q, J = 4.9 Hz, 1H), 8.55 (s, 1H), 7.83 (dq, J = 22.1, 9.0, 7.4 Hz, 3H), 7.50 (t, J = 10.4 Hz, 2H), 6.97 (s, 2H), 4.55 (d, J = 5.6 Hz, 2H), 3.25 (t, J = 6.7 Hz, 2H), 2.76 (t, J = 6.1 Hz, 2H), 1.96 (p, J = 6.5, 6.1 Hz, 2H), 1.72 (p, J = 5.7 Hz, 2H). 13 13C NMR (126 MHz, (CD3)2SO) δ 164.00, 161.89, 160.52 ( 1 J = 249.4 Hz), 154.65, 149.62, 145.80, 145.73, 145.06, 143.72, 142.71, 138.50, 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 C 25 H 21 FN6O [M+H] + Calculated value for 441.1873, found 441.1837.

[0758] N-((1H-Pyrrol-3-yl)methyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 12)

[0759] Following the same synthetic procedure as for Compound 4, Compound 12 was obtained as an off-white solid (77 mg, 36.5%). 11H NMR (500 MHz, (CD3)2SO) δ 8.47 (s, 1H), 8.01 (d, J = 8.0 Hz, 2H), 7.79 (q, J = 9.1 Hz, 2H), 7.60 (d, J = 7.9 Hz, 2H), 6.74 (t, J = 2.1 Hz, 1H), 6.67 (q, J = 2.4 Hz, 1H), 6.08 (q, J = 2.3 Hz, 1H), 4.41 (d, J = 5.6 Hz, 2H), 3.53 (p, J = 6.6 Hz, 2H), 3.17 (t, J = 6.4 Hz, 2H), 1.89 (q, J = 7.6, 6.3 Hz, 2H), 1.63 (p, J = 6.1, 5.7 Hz, 2H). 13 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 C 26 H 23 N5O [M+H] + Calculated value 422.1981, measured value 422.1971.

[0760] 4-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-(thiazol-2-ylmethyl)benzamide (Compound 13)

[0761] Following the same synthetic procedure as for Compound 4, Compound 13 was obtained as an off-white solid (65 mg, 29.8%). 11H NMR (500 MHz, (CD3)2SO) δ 9.40 (t, J = 6.0 Hz, 1H), 8.57 (s, 1H), 8.07 - 7.98 (m, 2H), 7.90 - 7.78 (m, 2H), 7.73 (d, J = 3.3 Hz, 1H), 7.72 - 7.65 (m, 2H), 7.61 (d, J = 3.3 Hz, 1H), 4.80 (d, J = 5.9 Hz, 2H), 3.33 (s, 2H), 2.78 (t, J = 6.1 Hz, 2H), 2.05 - 1.96 (m, 2H), 1.84 - 1.71 (m, 2H). 13 13C NMR (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 C 25 H 21 N5OS [M+H] + calculated value 440.1545, found 440.1547.

[0762] 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, otherwise known as Compound HSK205)

[0763]

Chemical Structure

[0764] Following the same synthetic procedure as for Compound 4, Compound 14 was obtained as an off-white solid (89 mg, 40.7%). 11H NMR (500 MHz, (CD3)2SO) δ 9.01 (t, J = 5.7 Hz, 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 = 5.7 Hz, 2H), 2.77 (t, J = 6.1 Hz, 2H), 2.09 (d, J = 1.0 Hz, 3H), 2.07 (s, 1H), 2.01 (dd, J = 7.8, 3.9 Hz, 2H), 1.97 (s, 1H), 1.75 (dd, J = 7.6, 4.2 Hz, 2H). 13 13C 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 C 26 H 24 N6O [M+H] + Calculated value for 437.2079, found 437.2083.

[0765] N-(2-(1H-Imidazol-2-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide (Compound 15)

[0766] Following the same synthetic procedure as for Compound 4, Compound 15 was obtained as an off-white solid (73 mg, 33.4%). 11H NMR (500 MHz, (CD3)2SO) δ 8.63 (t, J = 5.6 Hz, 1H), 8.57 (s, 1H), 7.93 - 7.89 (m, 2H), 7.87 - 7.77 (m, 2H), 7.64 (d, J = 8.3 Hz, 2H), 7.16 (d, J = 21.5 Hz, 1H), 6.90 (d, J = 10.4 Hz, 1H), 4.21 (t, J = 6.1 Hz, 2H), 3.63 (q, J = 5.9 Hz, 2H), 3.42 (q, J = 6.0 Hz, 2H), 2.76 (t, J = 6.1 Hz, 2H), 1.99 (ddd, J = 15.1, 7.5, 4.5 Hz, 2H), 1.73 (ddd, J = 9.0, 7.2, 4.2 Hz, 2H). 13 13C NMR (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 C 26 H 24 N6O [M+H] + calculated value 437.2084, measured value 437.2070.

[0767] 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

[0768]

Chemical formula

[0769] Following the same synthetic procedure as for compound HSK205, the compound was obtained as a pale yellow solid (119 mg, 52.9%).1 1H NMR (500 MHz, (CD3)2SO) δ 8.01 (d, J = 7.9 Hz, 2H), 7.79 - 7.65 (m, 2H), 7.61 (d, J = 8.0 Hz, 2H), 6.65 (s, 1H), 4.48 (d, J = 4.9 Hz, 2H), 3.40 (s, 2H), 2.85 (d, J = 5.8 Hz, 3H), 2.75 (s, 2H), 2.10 (s, 3H), 1.77 (s, 4H). 13 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 C 27 H 26 N6O [M+H] + Calculated value for 451.2241, found 451.2230.

[0770] 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

[0771]

Chemical formula

[0772] Following the same synthetic procedure as for compound HSK205, the compound was obtained as an off-white solid (98 mg, 39.3%). 11H NMR (500 MHz, (CD3)2SO) δ 8.43 (d, J = 7.0 Hz, 2H), 7.95 - 7.87 (m, 2H), 7.81 (dd, J = 8.9, 2.5 Hz, 1H), 5.31 (dq, J = 7.0, 4.8 Hz, 1H), 4.09 - 4.00 (m, 2H), 3.90 (d, J = 4.9 Hz, 2H), 3.48 (dd, J = 17.5, 7.1 Hz, 1H), 3.38 (ddd, J = 15.6, 9.1, 6.4 Hz, 1H), 3.30 - 3.18 (m, 2H), 2.28 (s, 3H), 2.05 - 1.97 (m, 3H), 1.86 - 1.75 (m, 3H). 13 13C NMR (201 MHz, (CD3)2SO) δ 165.89, 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.

[0773] 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

[0774]

Chem.

[0775] Following the same synthetic procedure as for compound HSK205, the compound was obtained as a yellow solid (87 mg, 37.1%). 11H NMR (500 MHz, (CD3)2SO) δ 8.74 (s, 1H), 7.98 (d, J = 9.2 Hz, 2H), 7.92 - 7.79 (m, 3H), 7.12 - 7.07 (m, 1H), 4.49 (s, 2H), 4.01 (ttd, J = 7.9, 6.0, 4.2 Hz, 1H), 3.90 (d, J = 6.1 Hz, 2H), 3.39 - 3.25 (m, 2H), 3.18 (ddd, J = 15.6, 8.8, 6.6 Hz, 2H), 1.95 - 1.79 (m, 3H). 13 13C NMR (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 C 26 H 23 FN6O2[M+H] + Calculated value for 471.1967, found 471.1963.

[0776] 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

[0777]

Chemical Structure

[0778] Following the same synthetic procedure as for compound HSK205, the compound was obtained as an off-white solid (69 mg, 30.4%). 11H NMR (500 MHz, (CD3)2SO) δ 8.72 (s, 1H), 7.94 (d, J = 8.9 Hz, 1H), 7.90 - 7.81 (m, 2H), 7.09 - 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 13C NMR (201 MHz, (CD3)2SO) δ 166.57, 166.51, 161.57, 159.58, 153.97, 153.91, 150.38, 144.34, 142.00, 141.52, 135.30, 135.27, 135.21, 132.37, 132.31, 131.28, 129.46, 129.35, 126.23, 125.52, 125.49, 124.22, 124.19, 120.45, 116.80, 115.62, 114.27, 114.09, 113.63, 36.61, 28.38, 27.08, 22.70, 21.62, 14.20. HRMS (ESI) m / z C 26 H 23 FN6O [M+H] + calculated value 455.1927, measured value 455.1936.

[0779] 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

[0780]

Chemical Structure

[0781] Following the same synthetic procedure as for compound HSK205, the compound was obtained as a pale yellow solid (70 mg, 30.6%). 11H NMR (500 MHz, (CD3)2SO) δ 8.68 (s, 1H), 7.97 (d, J = 9.2 Hz, 2H), 7.83 - 7.74 (m, 4H), 6.45 (dt, J = 8.4, 0.9 Hz, 1H), 4.40 (s, 2H), 4.21 (s, 2H), 3.60 (t, J = 0.9 Hz, 2H), 3.31 - 3.21 (m, 4H), 2.04 - 1.91 (m, 2H), 1.86 (s, 3H). 13 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 C 27 H 26 N6O [M+H]+ + Calculated value for 451.2253, found 451.2251.

[0782] 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

[0783]

Chemical Structure

[0784] Following the same synthetic procedure as for compound HSK205, the compound was obtained as a white solid (109 mg, 46.7%). 11H NMR (500 MHz, (CD3)2SO) δ 8.64 (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 = 5.6 Hz, 2H), 4.07 (s, 3H), 2.79 (t, J = 6.2 Hz, 2H), 2.11 (d, J = 1.0 Hz, 3H), 2.07 (s, 1H), 2.02 (dd, J = 7.6, 3.9 Hz, 2H), 1.94 (s, 1H), 1.71 (dd, J = 7.2, 4.0 Hz, 2H).

[0785] N-(1-(4-Methyl-1H-imidazol-2-yl)ethyl)-4-(8,9,10,11-tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)benzamide

[0786]

Chemical Structure

[0787] Following the same synthetic procedure as for compound HSK205, the compound was obtained as a pale yellow solid (101 mg, 44.8%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.75 (s, 1H), 7.98 - 7.86 (m, 4H), 7.81 (d, J = 9.1 Hz, 2H), 6.82 (dd, J = 6.1, 0.9 Hz, 1H), 5.04 (dq, J = 7.0, 4.8 Hz, 1H), 3.31 - 3.21 (m, 4H), 1.93 - 1.84 (m, 4H), 1.82 (d, J = 4.8 Hz, 3H), 1.81 - 1.74 (m, 3H). 1313C NMR (201 MHz, (CD3)2SO) δ 165.83, 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.

[0788] 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

[0789]

Chem.

[0790] Following the same synthetic procedure as for compound HSK205, the compound was obtained as a bright orange solid (23 mg, 9.9%). 1 1H NMR (500 MHz, (CD3)2SO) δ 7.96 - 7.87 (m, 4H), 7.80 (d, J = 9.3 Hz, 2H), 6.85 (dd, J = 6.1, 0.6 Hz, 1H), 5.01 (dq, J = 6.6, 4.2 Hz, 1H), 3.34 (d, J = 13.5 Hz, 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). 1313C NMR (201 MHz, (CD3)2SO) δ 165.79, 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.

[0791] 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

[0792]

Chem.

[0793] Following the same synthetic procedure as for compound HSK205, the compound was obtained as an off-white solid (84 mg, 36.2%). 1 1H NMR (500 MHz, (CD3)2SO) δ 8.61 (s, 1H), 7.99 - 7.92 (m, 2H), 7.92 - 7.85 (m, 2H), 7.81 (d, J = 8.8 Hz, 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). 1313C NMR (201 MHz, (CD3)2SO) δ 166.23, 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.

[0794] 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

[0795]

Chemical Structure

[0796] Following the same synthetic procedure as for compound HSK205, the compound was obtained as a white solid (109 mg, 45.6%). 1 1H NMR (500 MHz, (CD3)2SO) δ 7.98 - 7.88 (m,2H), 7.91 - 7.85 (m, 2H), 7.46 - 7.41 (m, 2H), 3.35 (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). 13 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 C29 H 30 N6O [M+H] + Calculated value: 479.2411, measured value: 479.2422.

[0797] 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

[0798]

Chemical formula

[0799] Following the same synthetic procedure as for compound HSK205, the compound was obtained as a yellow solid (43 mg, 18.5%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.07 - 7.96 (m, 4H), 7.80 (d, J = 9.2 Hz, 2H), 6.23 (d, J = 4.9 Hz, 1H), 4.48 (s, 2H), 3.39 (d, J = 13.1 Hz, 2H), 3.29 - 3.20 (m, 4H), 2.70 (q, J = 7.1 Hz, 2H), 1.93 - 1.82 (m, 2H), 1.75 (s, 3H), 1.28 (t, J = 6.6 Hz, 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 C 28 H 28 N6O [M+H] + Calculated value: 465.2347, measured value: 465.2340.

[0800] 4-(8,9,10,11-Tetrahydro-3H-pyrazolo[4,3-a]phenanthridin-7-yl)-N-((4-(trifluoromethyl)-1H-imidazol-2-yl)methyl)benzamide

[0801]

Chemical formula

[0802] Following the same synthetic procedure as for compound HSK205, the compound was obtained as a yellow-orange solid (217 mg, 88.4%). 1 H NMR (500 MHz, (CD3)2SO) δ 8.66 (s, 1H), 7.99 - 7.87 (m, 4H), 7.81 (d, J = 9.1 Hz, 2H), 6.72 (dq, J = 4.1, 2.3 Hz, 1H), 4.57 (s, 2H), 3.32 - 3.21 (m, 4H), 1.92 - 1.83 (m, 4H). 13 C NMR (201 MHz, (CD3)2SO) δ 165.89, 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, 128.52, 126.95, 126.48, 126.02, 124.37, 122.19, 120.02, 119.66, 117.85, 116.66, 116.35, 116.32, 116.29, 116.26, 113.46, 36.87, 27.86, 27.39, 22.62, 21.75. HRMS (ESI) m / z C 26 H 21 F3N6O [M+H] + Calculated value 491.1781, measured value 491.1785.

[0803] 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

[0804]

Chem.

[0805] Following the same synthetic procedure as for compound HSK205, the compound was obtained as an off-white solid (197 mg, 78.1%). 1 H NMR (500 MHz, (CD3)2SO) δ 7.97 - 7.87 (m, 4H), 7.80 (d, J = 8.8 Hz, 2H), 6.68 (dq, J = 4.0, 1.8 Hz, 1H), 4.49 (s, 2H), 3.18 (d, J = 10.9 Hz, 2H), 3.27 - 3.20 (m, 4H), 1.85 (s, 3H), 1.83 - 1.76 (m, 2H). 13 C NMR (201 MHz, (CD3)2SO) δ 165.91, 153.98, 150.16, 150.13, 150.09, 150.03, 144.21, 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 C 27 H 23 F3N6O [M+H] + Calculated value for 505.1925, found 505.1918.

[0806] 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

[0807]

Chem.

[0808] 1 H NMR (500 MHz, (CD3)2SO) δ 9.01 (t, J = 5.7 Hz, 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 = 5.7 Hz, 2H), 2.76 (t, J = 6.3 Hz, 2H), 2.61 (s, 3H), 2.09 (d, J = 1.0 Hz, 3H), 2.05 (s, 1H), 2.01 (dd, J = 7.8, 3.9 Hz, 2H), 1.94 (s, 1H), 1.77 (dd, J = 7.5, 4.1 Hz, 2H).

[0809] N-((4-methyl-1H-imidazol-2-yl)methyl)-4-(3H-pyrazolo[4,3-f]quinolin-7-yl)benzamide

[0810]

Chem.

[0811] 11H NMR (500 MHz, (CD3)2SO) δ 8.99 (t, J = 5.5 Hz, 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 = 5.6 Hz, 2H), 2.08 (d, J = 1.1 Hz, 3H).

[0812] 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

[0813]

Chemical Structure

[0814] 1 1H NMR (500 MHz, (CD3)2SO) δ 9.00 (t, J = 5.6 Hz, 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 = 5.6 Hz, 2H), 3.89 (d, J = 8.1 Hz, 2H), 3.65 (d, J = 8.2 Hz, 2H), 2.77 (t, J = 6.1 Hz, 2H), 2.61 (s, 3H), 2.09 (d, J = 1.1 Hz, 3H), 2.04 (s, 1H), 2.01 (dd, J = 7.7, 3.6 Hz, 2H), 1.91 (s, 1H), 1.72 (dd, J = 7.7, 4.2 Hz, 2H).

[0815] 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

[0816]

Chem.

[0817] 1 H NMR (500 MHz, (CD3)2SO) δ 8.59 (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 = 5.5 Hz, 2H), 3.71 (dt, J = 10.8, 2.8 Hz, 2H), 3.62 (dt, J = 10.8, 6.5 Hz, 2H), 2.77 (t, J = 6.1 Hz, 2H), 2.61 (s, 3H), 2.09 (d, J = 1.1 Hz, 3H), 2.04 (s, 1H), 2.01 (dd, J = 7.7, 3.6 Hz, 2H), 1.97-1.88 (m, 5H), 1.72 (dd, J = 7.7, 4.2 Hz, 2H).

Claims

1. Formula I: 【Chemical 1】 A compound having the structure of, Or a pharmaceutically acceptable salt thereof, wherein, X １ Are each independently N, CH, or C-halogen, X ２ Are each independently N, CH, C-halogen, C-CH ｎ１ X ｍ (Wherein n1 + m = 3 and X is halogen), C-OCF ３ C-OCHF ２ C-OMe, C-Me, C-Et, C-tBu, C-iPr, C-N-alkyl, C-N-heteroalkyl, C-O-alkyl or C-O-heteroalkyl, R ２ R ３ And R ４ Are each independently alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl, Y is, 【Chemical 2】 And in the formula, n is 1 to 5, R ５ And R ６ Are each independently H, alkyl, or heteroalkyl, R ６ Can optionally form a cyclic structure, R ７ Are each independently H or alkyl, Het-Ar is a heteroaromatic ring, W is NH, NMe, NEt, NCH ２ CH ２ OH, NCH ２ CH ２ OMe, O, SO or SO ２ And Y is not NH ２ A compound of.

2. X １and / or X ２ at least one of which is CF, CHF ２ CF ３ or CHF ３ The compound according to claim 1, which is **Claim 3** Formula IA: **Chemical Formula 3** The compound according to claim 1 having the structure of or a pharmaceutically acceptable salt thereof, wherein n ２ is 1, 2 or 3, Q is each independently O, NH, substituted N, CH ２ , or substituted C, S, SO or SO ２ is each Q together forms a 6-membered, 7-membered or 8-membered ring or bicyclic ring. A compound **Claim 4** R ５ and / or R ６ One or more of which is Me, the compound according to claim 1. **Claim 5** R ７ is each independently H or Me, the compound according to claim 1. **Claim 6** Y is **Chemical Formula 4** and each R ６ together forms morpholine, piperidine, piperazine or pyrrolidine, the compound according to claim 1. **Claim 7** Y is **Chemical Formula 5** and each R ７ together forms a cyclic structure, the compound according to claim 1. **Claim 8** The compound according to claim 7, wherein the cyclic structure comprises cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

9. Y is 【Chemical Formula 6】 and each R ７ together form a cyclic structure, the compound according to claim 1.

10. The cyclic structure contains cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, the compound according to claim 9.

11. Y is 【Chemical Formula 7】 and Het-Ar contains imidazole, oxazole, pyrazole, triazole, isoxazole, isothiazole, tetrazole, oxadiazole, thiadiazole, pyrimidine or triazine, the compound according to claim 1.

12. Y is 【Chemical Formula 8】 and R ５ is Me, the compound according to claim 1.

13. Formula IB: 【Chemical Formula 9】 having the structure of, the compound according to claim 3, or a pharmaceutically acceptable salt thereof, wherein o is 0 to 2, each R ８ is independently H, alkyl or heteroalkyl, compound.

14. Each R ７ together form a cyclic structure, the compound according to claim 13.

15. The cyclic structure contains cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, the compound according to claim 14.

16. At least one R ８is Me, Et or CF ３ The compound according to any one of claims 13 to 15.

17. Formula II: 【Chemical Formula 10】 A compound having the structure of, or a pharmaceutically acceptable salt thereof, wherein, X １ is each independently N, CH, or C-halogen, X ２ is each independently N, CH, C-halogen, C-CH ｎ１ X ｍ (wherein n1 + m = 3 and X is halogen), C-OCF ３ C-OCHF ２ C-OMe, C-Me, C-Et, C-tBu, C-iPr, C-N-alkyl, C-N-heteroalkyl, C-O-alkyl or C-O-heteroalkyl, Q is each independently O, NH, substituted N, CH ２ , or substituted C, S, SO or SO ２ is, and when each Q is taken together, forms a 6-membered, 7-membered or 8-membered ring or a bicyclic ring, n ３ is 1, 2 or 3, R ４ is alkyl, cycloalkyl, heteroalkyl or heterocycloalkyl, W is O, NH, or NMe, R ９ is alkyl or heteroalkyl, provided that R ９ is not H, a compound.

18. 【Chemical Formula 11】 The compound according to any one of claims 1 to 17 having the structure of, or a pharmaceutically acceptable salt thereof.

19. 【Chemical Formula 12】 The compound according to claim 1 or 3 having the structure, or a pharmaceutically acceptable salt thereof.

20. 【Chemical Formula 13-1】 【Chemical Formula 13-2】 The compound according to claim 2 or 3 having the structure.

21. 【Chemical Formula 14-1】 【Chemical Formula 14-2】 【Chemical Formula 14-3】 【Chemical Formula 14-4】 【Chemical Formula 14-5】 【Chemical Formula 14-6】 【Chemical Formula 14-7】 【Chemical Formula 14-8】 The compound according to claim 1 having the structure.

22. 【Chemical Formula 15】 The compound according to claim 1 having the structure, or a pharmaceutically acceptable salt thereof.

23. 【Chemical Formula 16-1】 【Chemical Formula 16-2】 【Chemical Formula 16-3】 【Chemical Formula 16-4】 【Chemical Formula 16-5】 【Chemical Formula 16-6】 【Chemical Formula 16-7】 The compound according to claim 1 having the structure.

24. 【Chemical Formula 17-1】 【Chemical Formula 17-2】 【Chemical Formula 17-3】 【Chemical Formula 17-4】 【Chemical 17-5】 The compound according to claim 1, having the structure of .

25. 【Chemical 18-1】 【Chemical 18-2】 【Chemical 18-3】 The compound according to claim 1, having the structure of .

26. 【Chemical 19】 The compound according to claim 1 or 3, having the structure of .

27. 【Chemical 20】 The compound according to claim 1 or 3, having the structure of .

28. 【Chemical 21-1】 【Chemical 21-2】 The compound according to claim 1 or 3, having the structure of .

29. Formula X: A-L'-D (Formula X) A PROTAC conjugate having the chemical structure of , or a pharmaceutically acceptable salt thereof, wherein A is a group of the compound according to any one of claims 1 to 28, L' is a linker that binds A and D or does not exist, D is a ubiquitin pathway protein binding site, PROTAC conjugate.

30. Use in the manufacture of a drug for the treatment of a disease in a subject of the compound according to any one of claims 1 to 28, the conjugate according to claim 29, or a pharmaceutically acceptable salt of said compound or conjugate, wherein the severity of the disease in the subject is reduced by modulation of a kinase.

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

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

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

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

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

36. The method according to claim 34, wherein said compound or conjugate comprises the compound according to claim 19.

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

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

39. The method according to any one of claims 34 to 37, wherein the cancer is 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, stomach cancer, head cancer, neck cancer, thyroid cancer, kidney cancer, liver cancer and stomach cancer.

40. Administering an effective amount to the subject inhibits CDK in the subject at an IC of about 50 nM or less (e.g., 50 nM or less). ５０ The method according to claim 35.

41. The IC ５０ is about 30 nM or less (e.g., 30 nM or less). The method according to claim 40.

42. Administering an effective amount to the subject inhibits both FLT3 and haspin kinase with specificity in the subject. The method according to claim 36.

43. The compound according to any one of claims 1 to 28 or the conjugate according to claim 29 for use in the treatment of a disease state regulated by one or more kinases.

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

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