JP2023520330A - Small molecule inhibitor of oncogenic CHD1L with preclinical activity against colorectal cancer - Google Patents

Abstract

Treatment of CHD1L-induced cancers, including TCF transcription-induced cancers and EMT-induced cancers, using CHD1L inhibitors. A small molecule inhibitor of CHDL1 has been identified that inhibits CHD1L ATPase and inhibits CHD1L-dependent TCF transcription. CHD1L inhibitors prevent the TCF complex from binding to Wnt response elements and promoter sites. More specifically, CHD1L inhibitors induce reversal of EMT. CHD1L inhibitors are useful in the treatment of various cancers, particularly CRC and m-CRC. CHD1L-induced cancers are inter alia CRC, breast cancer, glioma, liver cancer, lung cancer or gastrointestinal (GI) cancer. Also provided are CHD1L inhibitors of Formulas I and XX as defined herein and salts thereof, as well as pharmaceutical compositions containing CHD1L inhibitors. Synergistic combinations of CHD1L inhibitors and other antineoplastic agents have also been described. [Selection figure] None

Description

Detailed description of the invention

[Cross reference to related applications]
This application claims the benefit of U.S. Provisional Patent Application Nos. 62/994,259 filed March 24, 2020 and 63/139,394 filed January 20, 2021, both of which , which are incorporated herein by reference in their entirety.

[Description of U.S. Government Grant]
This invention was made with United States Government support under Grant No. W81XWH1810142 awarded by the United States Department of Defense (DoD). The United States Government has certain rights in this invention.

[background]
Genomic integrity is maintained by conformational changes to chromatin structures that regulate the accessibility of DNA for gene expression and replication. Chromatin structure is maintained by post-translational modifications of histones and nucleosome rearrangements [Lorch et al., 2010; Kumar et al., 2016; Swygert et al., 2014]. ATP-dependent chromatin remodelers are enzymes that alter chromatin by altering histone composition and either eliminating nucleosomes or translocating them along DNA. Their activity plays a crucial role in cellular function by regulating DNA gene expression and accessibility of DNA for replication, transcription, and DNA repair [Erdel et al., 2011; Brownlee et al., 2015; ]. Dysregulation of chromatin remodeling is associated with human diseases, particularly cancer [Zhang et al., 2016; Valencia and Kadoch, 2019].

In the last decade, a chromatin remodeler called CHD1L (chromodomain helicase/ATPase DNA-binding protein 1-like), also called ALC1 (amplified in liver cancer 1), has been implicated in prominent human cancer pathologies. (Ma et al., 2008; Cheng et al., 2013]. CHD1L is involved in the upregulation of drug-resistant efflux pumps (e.g., ABCB1) [Li et al., 2019] to PARP1-mediated DNA repair [Pines et al., 2012; et al., 2017] and anti-apoptotic activity [Li et al., 2013; Hyeon et al., 2013; Su et al., 2014; Mu et al., 2015; Su et al., 2015; Li et al., 2013; He et al., 2012; , 2010].CHD1L overexpression is also associated with tumor progression and is implicated as a predictor of poor patient survival [Ji et al., 2013].The multifunctional oncogenic mechanisms of CHD1L make it an attractive therapeutic in cancer. The cancer-inducing mechanisms of CHD1L have been studied in liver cancer [Li et al., 2019], breast cancer [Wu et al., 2014], and lung cancer [Li et al., 2019], but colorectal Little is known about the pathological mechanisms associated with CHD1L in cancer (CRC).

CRC is the third most prevalent cancer diagnosed annually and the mortality rate among CRC patients is the second highest worldwide [Jemal et al., 2011]. Early detection of CRC combined with surgery and 5-fluorouracil (5FU)-based combination chemotherapy has minimally improved overall survival [Jemal et al., 2011; Fakih, 2015]. Current chemotherapy and targeted therapies are largely ineffective in metastatic CRC (mCRC), evidenced by a low 5-year overall survival rate of 11% [Jemal et al., 2011; Fakih, 2015]. There is an unmet need in the art to identify and characterize targets involved in the pathology of CRC tumor progression and metastasis.

A majority of CRC patients have mutations in the Wnt signaling pathway, resulting in abnormal T cell factor/lymphoid enhancer factor-transcription or TCF complexes [Kinzler and Voelstein, 1996; Cancer Genome Atlas, 2012]. Such mutations can lead to constitutive β-catenin migration and transactivation of TCF transcription [Clevers, 2006; Korinek et al., 1997]. The TCF complex is organized by TCF4 (also known as TCFL2), which is activated through interactions with a series of coactivators such as β-catenin, PARP1, and CREB-binding protein (CBP) [Shitashige et al., 2008]. Recently, TCF4 was shown to be a specific driver of early metastasis from both adenomas (ie, polyps) and late stage mCRC [Hyeo et al., 2013; Su et al., 2014].

TCF transcription has been reported to function as a key regulator of epithelial-mesenchymal transition (EMT) [Sanchez-Tilla et al., 2011; Zhou et al., 2016; Abraham et al., 2019]. This method increases the stemness and other malignant properties of cancer stem cells (CSCs), which can transform relatively benign epithelial tumor cells into mesenchymal cells and induce mCRC. [Chaffer et al., 2016]. Recently, alterations in several CRC driver genes were reported to be common in both primary and metastatic tumor pairs [Hu et al., 2019]. More specifically, abnormal TCF4 has been reported to be a specific driver of mCRC [Hu et al., 2019], and CRC can metastasize in early stage adenomas (i.e. polyps) (i.e. polyps [Magri and See also Bardelli, 2019], which may be triggered by TCF-induced EMT [Chaffer et al., 2016; is shown to be an inductive force.

EMT is a major inductive force in many human diseases, particularly solid tumor progression, drug and radiotherapy resistance, immune response and immunotherapy evasion, and promoting metastasis [Chaffer et al., 2016; Chaffer and Weinberg, 2011. ; Scheel and Weinberg, 2012].

Due to the importance of the Wnt signaling pathway and TCF transcription in cancer and other diseases [Clevers, 2006], small molecule drugs that inhibit the Wnt signaling pathway and TCF transcription have been tested [Lee et al., 2011; , 2012]. Well-considered therapeutic strategies include receptor targeting (eg, Frizzled); prevention of Wnt ligand secretion (eg, porcupine); inhibition of β-catenin destruction complex (eg, tankyrase); and β-catenin and coactivators. (eg, by CBP) protein-protein inhibition (PPI). To date, none of the agents targeting the Wnt/TCF pathway have been clinically approved [Lu et al., 2016], although clinical trials may be underway. In contrast, the present invention includes a novel therapeutic strategy identifying, in particular, small molecule drugs for the treatment of Wnt/TCF-induced CRC, in which CHD1L is required for TCF transcription to regulate the malignant phenotype in CRC. A new therapeutic strategy has been described that has been identified as a DNA-binding factor purported to be

For example, US Pat. No. 9,616,047 reports small molecule inhibitors of β-catenin or disruptors of the β-catenin/TCF-4 complex that are said to attenuate colon carcinogenesis. there is The inhibitors of β-catenin reported therein included esculetin and named HI-B1 to HI-B20, HI-B22 to HI-B24, HI-B26, HI-B32 and HI-B34. Compounds are mentioned and the structures of each of these are described in that patent. This patent further describes compounds which, in several general formulas therein, are identified as β-catenin inhibitors and are said to be useful in the treatment of colon carcinogenesis. This patent is incorporated herein by reference in its entirety for the structures of certain compounds therein, general formulas and variable definitions of compounds said to be useful in the present invention. The compounds identified herein are structurally different from those described in this patent.

であり、式中、変数は、Ｅｓｐａｃｅｎｅｔ英語版機械翻訳によれば以下の通り定義される：
Ｒ_１は、ｍｅｓ－トリメチルフェニル、４－メチルフェニル、４－トリフルオロメチルフェニル、ナフチル、２，３，４，５，－テトラメチルフェニル、４－メトキシフェニル、４－ｔｅｒｔ－ブチルフェニル、２，４－ジメトキシフェニル、２，５－ジメトキシフェニル又は４－フェノキシフェニルであり、
Ｒ_２は、水素、メチルアセテート、アセテート、アミノアセチル、４－ギ酸ベンジル、４－イソプロピルベンジル、４－クロロベンジル又は４－メトキシベンジルであり、
Ｒ_３は、塩素、－ＯＲａ又は－ＮＲｂＲｃであり、Ｒａは、鎖Ｃ１～３アルキル、Ｃ５～６シクロアルキル、Ｃ１～２アルコキシ、モノ－若しくはジ－Ｃ１～２アルキルアミノ、又はＣ５～６窒素含有又は酸素含有ヘテロ環式基であり、Ｒｂ及びＲｃは、それぞれＣ１～５アルキル基である。さらに詳細には、Ｒ_３は、塩素、２－ヒドロキシテトラヒドロピロリル、エタノールアミノ、２，３－ジヒドロキシ－１－メチルプロピルアミノ、２，３－ジヒドロキシプロピルアミノ、ピペラジニル、Ｎ－メチルピペラジニル、アゼピル、ピペリジニル、２－メチルプロピルアミノ、プロポキシ、メチルアミノ、エチルアミノ、シクロプロピルアミノ、１－エチルプロピルアミノ、テトラヒドロピラン－４－イルメトキシ又は２－メトキシエトキシである。参考文献は、式Ｉ－５：

の化合物も指す。 CN109761909, published May 17, 2019 (as described in its abstract in English on Espacenet) describes several N-(4-(pyrimidine-4-amino)phenyl)sulfones of certain formulas. Amide compounds or salts have been reported to inhibit Hsp90-Cdc37 (heat shock protein Hsp90 and its auxiliary chaperone Cdc37) interactive client protein expression and be useful in the treatment or prevention of various diseases mediated by Hsp90 signaling channels. are reporting that there are The formula given in this published application is

where the variables are defined as follows according to the Espacenet English machine translation:
R ₁ is mes-trimethylphenyl, 4-methylphenyl, 4-trifluoromethylphenyl, naphthyl, 2,3,4,5-tetramethylphenyl, 4-methoxyphenyl, 4-tert-butylphenyl, 2, 4-dimethoxyphenyl, 2,5-dimethoxyphenyl or 4-phenoxyphenyl,
R ₂ is hydrogen, methyl acetate, acetate, aminoacetyl, 4-benzyl formate, 4-isopropylbenzyl, 4-chlorobenzyl or 4-methoxybenzyl;
R ₃ is chlorine, —ORa or —NRbRc and Ra is chain C1-3 alkyl, C5-6 cycloalkyl, C1-2 alkoxy, mono- or di-C1-2 alkylamino, or C5-6 nitrogen is an oxygen-containing or oxygen-containing heterocyclic group, and Rb and Rc are each C1-5 alkyl groups. More particularly R ₃ is chlorine, 2-hydroxytetrahydropyrrolyl, ethanolamino, 2,3-dihydroxy-1-methylpropylamino, 2,3-dihydroxypropylamino, piperazinyl, N-methylpiperazinyl, azepyr, piperidinyl, 2-methylpropylamino, propoxy, methylamino, ethylamino, cyclopropylamino, 1-ethylpropylamino, tetrahydropyran-4-ylmethoxy or 2-methoxyethoxy. Reference is made to Formula I-5:

Also refers to compounds of

This published application is hereby incorporated by reference in its entirety for the structures of certain compounds therein, the general formulas and variable definitions of the compounds said to be useful in the present invention. Structures disclosed in this published application may be excluded from any formula in this application.

The present invention examined the clinicopathological properties of CHD1L in CRC and the results herein indicate that CHD1L is a druggable target involved in TCF transcription. A mechanism for CHD1L-mediated TCF transcription is also proposed herein. Small-molecule inhibitors of CHD1L that can prevent TCF transcription, reverse EMT, and other malignant properties in various cell models, including tumor organoids and patient-derived tumor organoids (PDTO), are disclosed herein. identified in the book. Some of the CHD1L inhibitors identified herein have drug-in vivo pharmacokinetic (PK) and pharmacodynamic (PD) profiles that are important for translational development into treatments for CRC and other cancers. It exhibits similar pharmacological properties.

[overview]
The present invention relates to treatment of CHD1L-induced cancers, more particularly TCF transcription-induced cancers, more particularly EMT-induced cancers. CHD1L is found to be an essential component of the TCF transcription complex. A small molecule inhibitor of CHDL1 has been identified that inhibits CHD1L ATPase and inhibits CHD1L-dependent TCF transcription. CHD1L inhibitors are thought to prevent the TCF complex from binding to Wnt response elements and promoter sites. More specifically, CHD1L inhibitors induce reversal of EMT. CHD1L inhibitors are useful in the treatment of various cancers, particularly CRC and m-CRC. With particular reference to CRC, CHD1L inhibitors are shown in embodiments to inhibit cancer stem cell (CSC) stemness and invasive potential. In embodiments, the CHD1L inhibitor induces cytotoxicity in CRC PDTO. In specific embodiments, the CHD1L-induced cancer is CRC, breast cancer, glioma, liver cancer, lung cancer or gastrointestinal (GI) cancer. In a specific embodiment, the TCF transcription-induced cancer is CRC, including mCRC. In a specific embodiment, the EMT-induced cancer is CRC, including mCRC.

The present invention provides a method of treating CHD1L-induced cancers, more particularly TCF transcription-induced cancers, more particularly EMT-induced cancers, particularly CRC and mCRC, including GI cancers, comprising: administering to a patient in need thereof an amount of a CHD1L inhibitor effective in inhibiting CHD1L, effective in inhibiting TCF transcriptional abnormalities, or effective in inducing EMT reversal provide a way. In embodiments, the CHD1L inhibitor is a compound of any one of Formulas I-XX or XXX-XVII. More particularly, the present invention provides methods of inhibiting TCF transcriptional abnormalities, particularly TCF transcriptional abnormalities in CRC, by administering an effective amount of a CHD1L inhibitor. More particularly, the present invention provides methods of inducing EMT reversal, particularly EMT reversal in CRC or mCRC, by administering an effective amount of a CHD1L inhibitor. The present invention provides a method of inhibiting cancer stem cell (CSC) stemness and/or invasive potential, particularly cancer stem cell (CSC) stemness and/or invasive potential in CRC, by administering an effective amount of a CHD1L inhibitor. I will provide a. The present invention treats cancerous tumors of CHD1L-induced cancers, or TCF transcription-induced cancers or EMT-induced cancers, particularly cancerous tumors in CRC, by administering an effective amount of a CHD1L inhibitor. provide a way to

In embodiments, the CHD1L inhibitor is selective for inhibition of CHD1L. In embodiments, the CHD1L inhibitor herein is not a PARP inhibitor. In embodiments, the CHD1L inhibitors herein are not inhibitors of topoisomerase. In particular, CHD1L inhibitors herein are not inhibitors of DNA topoisomerase. In particular, CHD1L inhibitors herein are not inhibitors of type IIα topoisomerase. In embodiments, the CHD1L inhibitors herein are not inhibitors of β-catenin, particularly inhibitors as described in US Pat. No. 9,616,047. In embodiments, the CHD1L inhibitors herein are not inhibitors of Hsp90-Cdc37 interactive client protein expression, particularly those described in CN109761909.

The present invention provides CHD1L inhibition, inhibition of TCF transcriptional abnormalities, to patients in need of prevention of tumor growth, invasion and/or metastasis in CHD1L-induced cancers, TCF transcription-induced cancers, or EMT-induced cancers. Tumor growth, invasion in CHD1L-induced cancers, TCF transcription-induced cancers, or EMT-induced cancers by administering an effective amount or an amount effective to reverse EMT of a CHD1L inhibitor of the present invention. and/or methods of preventing metastasis are also provided. In specific embodiments, the tumor is a solid tumor. In embodiments, the tumor is a tumor associated with GI cancer. In embodiments, the tumor is a tumor associated with CRC. In embodiments, the tumor is an mCRC associated tumor.

In a specific embodiment, the invention provides a method of treating CRC, including mCRC, comprising administering to a patient in need thereof an amount of a CHD1L inhibitor effective to inhibit CHD1L. I will provide a. In a specific embodiment, the invention provides a method of treating CRC, including mCRC, comprising administering to a patient in need thereof an amount of a CHD1L inhibitor effective to inhibit TCF transcriptional abnormalities. Provide a method that includes In a specific embodiment, the invention provides a method of treating CRC, including mCRC, comprising administering to a patient in need thereof an amount of a CHD1L inhibitor effective to induce reversal of EMT. Provide a method that includes

In a specific embodiment, the invention provides a method of inducing apoptosis in cancer cells comprising contacting the cancer cells with an effective amount of a CHD1L inhibitor. In embodiments, the CHD1L inhibitor is provided in an amount effective to inhibit TCF transcriptional abnormalities. In embodiments, the CHD1L inhibitor is provided in an amount effective to induce reversal of EMT. In embodiments, the cancer cells are CRC cancer cells. In embodiments, the cancer cells are mCRC cancer cells. In embodiments, the method is applied in vivo. In embodiments, the method is applied in vivo in a patient. In embodiments, the method is applied in vitro.

In embodiments of the methods herein comprising administering a CHD1L inhibitor, the CHD1L inhibitor is administered by any known method and dosing schedule to obtain the desired benefit. In embodiments, administration is oral administration. In embodiments, administration is by intravenous injection.

The present invention provides a method of treating drug-resistant cancer, comprising administering to a patient in need thereof an amount of a CHD1L inhibitor effective to inhibit CHD1L, inhibit TCF transcriptional aberrations, or induce reversal of EMT. Also provided are methods comprising administering in combination with known therapies that have rendered cancer resistant. In a specific embodiment, the treatments the cancer has become resistant to are conventional chemotherapy and other targeted therapies. In a specific embodiment, the invention provides a method of increasing the efficacy of a DNA-damaging drug in cancer comprising treating the cancer with a combination of a DNA-damaging drug and a CHD1L inhibitor, A method is provided wherein CHD1L is administered in an effective amount to reduce resistance to In embodiments, the DNA damaging agent is a topoisomerase inhibitor. In particular, DNA damaging agents are DNA topoisomerase inhibitors. In particular, DNA damaging agents are type IIα topoisomerase inhibitors. In particular the DNA damaging agent is etoposide or teniposide. In particular, the DNA damaging drug is SN38 or a prodrug thereof. In embodiments, the DNA damaging agent is a thymidylate synthase inhibitor. In embodiments, the thymidylate synthase inhibitor is a folic acid analogue. In embodiments, the thymidylate synthase inhibitor is a nucleotide analogue. In specific embodiments, the thymidylate synthase inhibitor is raltitrexed, pemetrexed, nolatrexed or ZD9331. In certain embodiments, the DNA damaging agent is 5-fluorouracil or capecitabine.

In embodiments, the cancer is a CDH1L-induced cancer. In embodiments, the cancer is a TCF transcription-induced cancer. In embodiments, the cancer is an EMT-induced cancer. In embodiments, the treatment is treatment of CRC. In embodiments, the treatment is treatment of mCRC. In embodiments, the DNA damaging agent and the CHD11 inhibitor are administered by any known method and on a dosing schedule suitable to achieve a combination therapy benefit. In embodiments, the CHD1L inhibitor is administered orally and the DNA damaging agent is administered by any known method and schedule of administration. In embodiments, the CHD1L inhibitor is administered prior to administration of the DNA damaging agent. In embodiments, the CHD1L inhibitor is administered prior to administration of the DNA damaging agent and optionally administered after administration of the DNA damaging agent. In embodiments, the CHD1L inhibitor is administered orally prior to the intravenous administration of the DNA damaging agent and optionally administered orally after the intravenous administration of the DNA damaging agent.

The present invention provides a method of treating a CHD1L-induced cancer, a TCF transcription-induced cancer, or an EMT-induced cancer, comprising administering to a patient in need thereof Methods are provided comprising administering an amount of a CHD1L inhibitor effective to induce reversal in combination with alternative treatment methods for cancer. In embodiments, the cancer is a GI cancer, or more particularly a CRC cancer, more particularly mCRC. In embodiments, the alternative treatment method is one of 5-fluorouracil, 5-fluorouracil in combination with folinic acid (also known as leucovorin), a topoisomerase inhibitor, or a cytotoxic or antineoplastic agent or multiple doses. In embodiments, the CHD1L inhibitor is administered in combination with 5-fluorouracil, or in combination with 5-fluorouracil and folinic acid. In embodiments, the CHD1L inhibitor is administered in combination with a topoisomerase inhibitor, particularly irinotecan (a prodrug of SN38, also known as camptothecin) or any other known prodrug of SN38. In embodiments, combination therapy using a CHD1L inhibitor and a topoisomerase inhibitor exhibits at least additive activity against cancer. In embodiments, combination therapy with a CHD1L inhibitor and a topoisomerase inhibitor exhibits synergistic activity (greater than additive activity) against cancer.

In embodiments, the CHD1L inhibitor is administered in combination with a cytotoxic or antineoplastic agent, particularly a platinum-based antineoplastic agent, more particularly cisplatin, carboplatin or oxaliplatin. In embodiments, combination therapy using a CHD1L inhibitor and a platinum-based antineoplastic agent exhibits at least additive activity against cancer. In embodiments, combination therapy with a CHD1L inhibitor and a platinum-based antineoplastic agent exhibits synergistic activity (greater than additive activity) against cancer. In embodiments, the platinum-based anti-neoplastic agent and the CHD11 inhibitor are administered by any known method and on a dosing schedule suitable to achieve a combination therapy benefit. In embodiments, the CHD1L inhibitor is administered orally and the platinum-based neoplastic agent is administered by any known dosing method and dosing schedule. In embodiments, the CHD1L inhibitor is administered prior to administration of the platinum-based neoplastic agent. In embodiments, the CHD1L inhibitor is administered prior to administration of the platinum-based antineoplastic agent and optionally administered after administration of the platinum-based antineoplastic agent. In embodiments, the CHD1L inhibitor is administered orally prior to intravenous administration of the platinum-based neoplastic agent and optionally administered orally after intravenous administration of the platinum-based neoplastic agent.

In embodiments, a CHD1L inhibitor is administered in combination with a chemotherapeutic regimen for the treatment of GI cancers, particularly CRC and mCRC. In embodiments, the CHD1L inhibitor is administered in combination with a chemotherapy regimen named FOLFOX. In embodiments, the CHD1L inhibitor is administered in combination with a chemotherapy regimen named FOLFIRI. In embodiments, the chemotherapeutic regimen and the CHD11 inhibitor are administered by any known method and on a dosing schedule suitable to achieve a combination therapy benefit. In embodiments, the CHD1L inhibitor is administered orally and the chemotherapy regimen is administered by any known method and schedule of administration. In embodiments, the CHD1L inhibitor is administered prior to administration of the chemotherapy regimen. In embodiments, the CHD1L inhibitor is administered prior to administration of the chemotherapeutic regimen and optionally administered after administration of the chemotherapeutic regimen. In embodiments, the CHD1L inhibitor is administered orally prior to the intravenous administration of the PARP inhibitor and optionally administered orally after the intravenous administration of the PARP inhibitor.

The present invention provides methods of treating cancers sensitive to poly(ADP-ribose) polymerase I (PARPI), wherein a CHD1L inhibitor is used in combination with a PARP inhibitor. In embodiments, an amount of a CHD1L inhibitor effective to inhibit CHD1L, inhibit TCF transcriptional aberrations, or induce reversal of EMT is used in combination with an amount of a PARP inhibitor effective to treat cancer. , at least improve the effectiveness of cancer treatment. In embodiments, combination therapy using a CHD1L inhibitor and a PARP inhibitor exhibits at least additive activity against cancer. In embodiments, combination therapy with a CHD1L inhibitor and a PARP inhibitor exhibits synergistic activity (greater than additive activity) against cancer. In embodiments, the cancer is a cancer that is amenable to treatment with a PARP inhibitor. In embodiments, the cancer is a cancer that is refractory to treatment with a PARP inhibitor or that has become refractory to treatment with a PARP inhibitor. In embodiments, the cancer is a cancer that is susceptible to treatment with a PARP inhibitor, or that has become resistant to treatment with a PARP inhibitor, and has a CHD1L-induced cancer, a TCF-induced cancer or an EMT-induced cancer. It is. In embodiments, the cancer is a homologous recombination deficient cancer (see, eg, Zhou et al., BioRxiv 2020). In embodiments, the cancer to be treated is a PARP inhibitor sensitive cancer, more particularly breast or ovarian cancer. In specific embodiments, the cancer is BRCA-deficient breast cancer or ovarian cancer. In embodiments, the cancer treated is GI cancer, CRC or mCRC. In embodiments, combination therapy with a CHD1L inhibitor and a PARP inhibitor reverses cancer resistance to treatment with a PARP inhibitor. In embodiments, the PARP inhibitor is olaparib, veliparib or talozoparib. In embodiments, the PARP inhibitor is rucaparib or niraparib. The present invention also provides a method of treating cancer comprising administering an amount of a PARP inhibitor effective to treat cancer in combination with administering an amount of a CHD1L inhibitor effective to inhibit CHD1L. . In embodiments, the PARP inhibitor and the CHD11 inhibitor are administered by any known method and on a dosing schedule suitable to achieve the benefits of combination therapy. In embodiments, the CHD1L inhibitor is administered orally and the PARP inhibitor is administered by intravenous injection. In embodiments, both the CHD1L inhibitor and the PARP inhibitor are administered by intravenous injection. In embodiments, the CHD1L inhibitor is administered prior to administration of the PARP inhibitor. In embodiments, the CHD1L inhibitor is administered prior to administration of the PARP inhibitor and optionally administered after administration of the PARP inhibitor. In embodiments, the CHD1L inhibitor is administered after administration of the PARP inhibitor. In embodiments, the CHD1L inhibitor is administered orally prior to the intravenous administration of the PARP inhibitor and optionally administered orally after the intravenous administration of the PARP inhibitor.

The present invention is a method of identifying CHD1L inhibitors that inhibit CHD1L-dependent TCF transcription by determining whether selected compounds inhibit CHD1L ATPase, as described in the Examples herein. A method is also provided that includes the step of determining. In a specific embodiment, inhibition of cat-CHD1L ATPase is determined. In embodiments, compounds that exhibit a % inhibition of 30% or greater are selected as inhibiting CHD1L ATPase. In embodiments, compounds that exhibit a % inhibition of 80% or greater are selected as inhibiting CHD1L ATPase. In specific embodiments, the CHD1L inhibitor exhibits an IC ₅₀ of less than 10 μM in a dose-response assay against CHD1L ATPase, particularly cat-CHD1L ATPase. In specific embodiments, the CHD1L inhibitor exhibits an IC ₅₀ of less than 5 μM in a dose-response assay against CHD1L ATPase, particularly cat-CHD1L ATPase. In a specific embodiment, the CHD1L inhibitor exhibits an IC ₅₀ of less than 3 μM in a dose-response assay against CHD1L ATPase, particularly cat-CHD1L ATPase. In specific embodiments, CHD1L inhibitors exhibit an _IC50 of less than 5 μM. In specific embodiments, CHD1L inhibitors exhibit an IC ₅₀ of less than 3 μM. In specific embodiments, CHD1L inhibitors exhibit an IC ₅₀ of less than 1 μM.

In specific embodiments, CHD1L inhibitors reduce TCF transcription in 2D cancer cell models, particularly using one or more CRC cell lines, such as those described in the Examples herein. is evaluated for inhibition of In a specific embodiment, inhibition of TCF transcription is determined using a TOPflash reporter construct, more particularly a TOPflash luciferase reporter construct described herein. In a specific embodiment, inhibition of TCF transcription by CHD1L inhibitors in the cell model is dose dependent. In a specific embodiment, inhibition of TCF transcription by CHD1L inhibitors in cell models is dose dependent in the range of 1-50 μM. In a specific embodiment, the CHD1L inhibitor exhibits % TCF transcription normalized to 75% or less cell viability at 20 μM. In a specific embodiment, the CHD1L inhibitor exhibits % TCF transcription normalized to 50% or less cell viability at 40 μM. In a specific embodiment, CHD1L inhibitors exhibit dose-dependent inhibition of TCF transcription with an _IC50 of less than 10 μM assayed using the TOPflash reporter in cancer cell lines. In a specific embodiment, CHD1L inhibitors exhibit dose-dependent inhibition of TCF transcription with an _IC50 of less than 5 μM assayed in cancer cell lines using the TOPflash reporter. In a specific embodiment, CHD1L inhibitors exhibit dose-dependent inhibition of TCF transcription with an _IC50 of less than 3 μM assayed in cancer cell lines using the TOPflash reporter. In embodiments, the cancer cell line is a CRC cancer cell, breast cancer cell, glioma cell, liver cancer cell, lung cancer cell or GI cancer cell. In embodiments, the cancer cell line is a CRC cancer cell line. In a specific embodiment, the CRC cancer cell line is SW620.

In a specific embodiment, CHD1L inhibitors are assessed for their ability to reverse or inhibit EMT. In a specific embodiment, CHD1L inhibitors are assessed for their ability to reverse EMT in tumor organoids. In embodiments, reversal or inhibition of EMT is assessed in vimentin-expressing tumor organoids, and a dose-dependent decrease in vimentin expression indicates reversal or inhibition of EMT. In embodiments, reversal of EMT is assessed in tumor organoids expressing E-cadherin, and a dose-dependent increase in E-cadherin expression indicates reversal or inhibition of EMT. In embodiments, reversal of EMT is assessed in tumor organoids expressing E-cadherin, vimentin, or both, wherein a dose-dependent decrease in vimentin and a dose-dependent increase in E-cadherin expression indicate reversal or inhibition of EMT. In a specific embodiment, dose-dependent reversal or inhibition of EMT is measured over compound concentrations of 0.1-100 μM. In a specific embodiment, dose-dependent reversal of EMT is measured over compound concentrations from 0.3 to 50 μM.

In a specific embodiment, CHD1L inhibitors are assessed for their ability to inhibit clonogenic colony formation, a well-established assay for measuring stemness of cancer stem cells. In embodiments, cells are pretreated with a selected concentration of a CHD1L inhibitor prior to plating. In embodiments, cells are cultured at a low density such that only CSCs form colonies over 10 days in culture. In embodiments, cells are pretreated for 12-36 hours. In embodiments, cells are pretreated for 24 hours. In embodiments, cells are pretreated with a CHD1L inhibitor at concentrations ranging from 0.5-50 μM, along with appropriate controls. In embodiments, the CHD1L inhibitor exhibits 40% or greater inhibition of clonogenic colony number at a CHD1L concentration of 40 μM compared to a no compound control. In embodiments, the CHD1L inhibitor exhibits 40% or greater inhibition of clonogenic colony number at a CHD1L concentration of 20 μM compared to a no compound control. In embodiments, the CHD1L inhibitor exhibits 40% or greater inhibition of clonogenic colony numbers at a CHD1L concentration of 2 μM compared to a no compound control. In embodiments, the inhibition of clonogenic colony formation continues over 10 days in culture.

In a specific embodiment, the CHD1L inhibitor is further assessed for loss of invasive capacity employing any known method, particularly employing the methods described in the Examples herein.

In specific embodiments, CHD1L inhibitors are further assessed for anti-tumor activity, as measured by induction of cytotoxicity in tumor organoids. In embodiments, cells are treated with a selected concentration (1-100 μM) of a CHD1L inhibitor for a selected period of time (eg, 24-96 hours, preferably 72 hours). In embodiments, the cytotoxicity is any of a variety of cytotoxic reagents known in the art, e.g., small molecules that enter damaged cells and exhibit a measurable change upon entry (e.g., CellTox). (Promega, Madison, Wis.) or IncuCyteCytotox reagent (Sartorius, France)). In embodiments, cytotoxicity is measured by measuring LDH (lactate dehydrogenase) released from dead cells.

In embodiments, CHD1L inhibitors useful in the therapeutic methods herein are inhibitors of Formulas I-XX, and XXX-XLII, or pharmaceutically acceptable salts or solvates thereof. In embodiments, the present invention provides novel compounds of any formula herein, particularly novel compounds of formulas I-XX, XXXV-XLII, or salts or solvates thereof. In embodiments, the CHD1L inhibitor is of formula I. In embodiments, the CHD1L inhibitor is of Formula XX. In embodiments, the CHD1L inhibitor is of Formulas I-IX, Xi-XiX, XX, or XXXV-XLII.

In specific embodiments, the methods of the invention employ a CHD1L inhibitor selected from one or more of Compounds 1-73, or pharmaceutically acceptable salts or solvates thereof. Two or more CHD1L inhibitors can be employed in combination in the methods herein. In specific embodiments, the CHD1L inhibitor employed in the present invention is selected from one or more of compounds 6-39 or pharmaceutically acceptable salts thereof. In specific embodiments, the CHD1L inhibitor employed in the methods of the invention is selected from one or more of compounds 40-51 or pharmaceutically acceptable salts or solvates thereof. In specific embodiments, the CHD1L inhibitor employed in the methods of the invention is selected from one or more of compounds 52-68 or pharmaceutically acceptable salts or solvates thereof. In specific embodiments, the CHD1L inhibitor employed in the methods of the invention is selected from one or more of compounds 70-73 or pharmaceutically acceptable salts or solvates thereof. In a specific embodiment, the CHD1L inhibitor employed in the methods of the invention is compound 6, 8, 52, 54, 56, 61, 62, 65 or 66 or a pharmaceutically acceptable salt or solvate thereof It is a thing. In a specific embodiment, the CHD1L inhibitor employed in the methods of the invention is compound 6, 8 or a pharmaceutically acceptable salt or solvate thereof. In a specific embodiment, the CHD1L inhibitor employed in the methods of the invention is compound 52, 54 or a pharmaceutically acceptable salt or solvate thereof. In a specific embodiment, the CHD1L inhibitor employed in the methods of the invention is compound 22, 23, 26 or 27, or a pharmaceutically acceptable salt thereof.

In specific embodiments, the methods of the invention employ CHD1L inhibitors of formula XX, including all embodiments described herein for formula XX. The present invention also provides novel compounds of Formula XX, their salts, and pharmaceutical compositions comprising such compounds and salts.

The present invention is also directed to the CHD1L inhibitors of the invention and pharmaceutically acceptable compositions comprising any such inhibitors. In embodiments, the present invention is directed to any novel compound, or a pharmaceutically acceptable salt or solvate thereof, described herein by formula. In particular, the present invention provides CHD1L inhibitors and pharmaceutically acceptable salts thereof as described in the formulas herein, with the exception that the CHD1L inhibitors are not compounds 1-8 or salts or solvates thereof. for In particular, the present invention is directed to CHD1L inhibitors described in the formulas herein and pharmaceutically acceptable salts thereof, with the exception that the CHD1L inhibitors are not compounds 1-9 or salts thereof. . In embodiments, the present invention provides compounds 9-39, 40-68, 69-73 or pharmaceutically acceptable salts or solvates thereof, or pharmaceutically acceptable compounds comprising such compounds or salts or solvates. Any acceptable composition is contemplated. In embodiments, the invention provides compounds 10-39 or 40-73 or pharmaceutically acceptable salts or solvates thereof, or pharmaceutically acceptable compositions comprising such compounds or salts or solvates. Any of In embodiments, the present invention provides any of compounds 52-73 or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutically acceptable composition comprising such a compound or salt or solvate. set to target. In embodiments, CHD1L inhibitors of the invention have a Clog P of 5 or less. In embodiments, CHD1L inhibitors of the invention have a Clog P of 3-4.

In specific embodiments, the invention is directed to the following compounds and methods herein employing these compounds for the treatment of cancer, particularly CRC and mCRC: Compounds 52-73; Compound 52 or 53; compound 54, 55 or 67; or compound 57, 58 or 59; or pharmaceutically acceptable salts or solvates thereof; compound 8, compound 52, compound 53, compound 54, compound 55, compound 56; any one of compound 57, compound 58, compound 59, compound 61, compound 62, compound 65, compound 66, or compound 67;

The present invention also provides therapeutic agents for cancer, particularly cancer, CHD1L-induced cancer, TCF-induced cancer, or EMT-induced cancer, particularly GI cancer, and more particularly therapeutic agents for CRC or mCRC. It also relates to the use of CHD1L inhibitors in manufacturing. The present invention provides CHD1L herein for use in the treatment of cancer, CHD1L-induced cancers, TCF-induced cancers, or EMT-induced cancers, particularly GI cancers, more particularly CRC or mCRC. Further relates to inhibitors.

Other embodiments and aspects of the invention will be readily apparent to one of ordinary skill in the art upon review of the drawings, detailed description, and examples herein.

Validation of CHD1L inhibitors identified from HTS. (FIG. 1A) cat-CHD1L ATPase C ₅₀ dose response with hits 1-7. Mean _IC50 values were calculated from three independent experiments and representative graphs are shown. (FIG. 1B) Inhibition of TCF transcription was measured using SW620, HCT-16, and DLD1CHD1L-OE cells with a TOPflash reporter and three doses over 24 hours. Validation of CHD1L inhibitors identified from HTS. (FIG. 1A) cat-CHD1L ATPase C ₅₀ dose response with hits 1-7. Mean _IC50 values were calculated from three independent experiments and representative graphs are shown. (FIG. 1B) Inhibition of TCF transcription was measured using SW620, HCT-16, and DLD1CHD1L-OE cells with a TOPflash reporter and three doses over 24 hours. A CHD1L inhibitor reverses the EMT and malignant phenotype in CRC. Dose response of CHD1L inhibitors modulating EMT as measured by high-content imaging of VimPro-GFP reporter downregulation (FIG. 2A) and EcadPro-RFP reporter upregulation (FIG. 2B). Mean _EC50 values ± SEM are calculated from three independent experiments. (FIG. 2C) CSC stemness measured by clonogenic colony formation after pretreatment with CHD1L inhibitor in DLD1CHD1L-OE and HCT-116 cells. (FIG. 2D) Inhibition of invasive capacity of HCT-116 cells after CHD1L inhibitor treatment. Welch t-test statistical analysis was used to determine significance. * = P < 0.05, ** = P < 0.01, *** = P < 0.001, **** = P < 0.0001. A CHD1L inhibitor reverses the EMT and malignant phenotype in CRC. Dose response of CHD1L inhibitors modulating EMT as measured by high-content imaging of VimPro-GFP reporter downregulation (FIG. 2A) and EcadPro-RFP reporter upregulation (FIG. 2B). Mean _EC50 values ± SEM are calculated from three independent experiments. (FIG. 2C) CSC stemness measured by clonogenic colony formation after pretreatment with CHD1L inhibitor in DLD1CHD1L-OE and HCT-116 cells. (FIG. 2D) Inhibition of invasive capacity of HCT-116 cells after CHD1L inhibitor treatment. Welch t-test statistical analysis was used to determine significance. * = P < 0.05, ** = P < 0.01, *** = P < 0.001, **** = P < 0.0001. A CHD1L inhibitor reverses the EMT and malignant phenotype in CRC. Dose response of CHD1L inhibitors modulating EMT as measured by high-content imaging of VimPro-GFP reporter downregulation (FIG. 2A) and EcadPro-RFP reporter upregulation (FIG. 2B). Mean _EC50 values ± SEM are calculated from three independent experiments. (FIG. 2C) CSC stemness measured by clonogenic colony formation after pretreatment with CHD1L inhibitor in DLD1CHD1L-OE and HCT-116 cells. (FIG. 2D) Inhibition of invasive capacity of HCT-116 cells after CHD1L inhibitor treatment. Welch t-test statistical analysis was used to determine significance. * = P < 0.05, ** = P < 0.01, *** = P < 0.001, **** = P < 0.0001. A CHD1L inhibitor reverses the EMT and malignant phenotype in CRC. Dose response of CHD1L inhibitors modulating EMT as measured by high-content imaging of VimPro-GFP reporter downregulation (FIG. 2A) and EcadPro-RFP reporter upregulation (FIG. 2B). Mean _EC50 values ± SEM are calculated from three independent experiments. (FIG. 2C) CSC stemness measured by clonogenic colony formation after pretreatment with CHD1L inhibitor in DLD1CHD1L-OE and HCT-116 cells. (FIG. 2D) Inhibition of invasive capacity of HCT-116 cells after CHD1L inhibitor treatment. Welch t-test statistical analysis was used to determine significance. * = P < 0.05, ** = P < 0.01, *** = P < 0.001, **** = P < 0.0001. Compound 6 induces apoptosis in CRC cell lines and PDTO. (FIG. 3A) Time course assessment of E-cadherin expression induction using the Ecad-ProRFP reporter assay and time course assessment of cytotoxicity induction using the Certox™ Green Cytotoxicity Assay (Promega, Madison, Wis.). (FIG. 3B) Annexin V-FITC staining analysis of apoptosis after 12 h treatment with SN-38 and 6. (FIG. 3C) Cytotoxicity of 6 in PDTO CRC102 using the CellTiter-Blue® Cell Viability Assay (Promega, Madison, Wis.). Mean _EC50 values ± standard deviations are calculated from 6 independent experiments. A representative graph with a representative PDTO inset is shown. Welch t-test statistical analysis is used to determine significance. * = P < 0.05, ** = P < 0.01, *** = P < 0.001, **** = P < 0.0001. Compound 6 induces apoptosis in CRC cell lines and PDTO. (FIG. 3A) Time course assessment of E-cadherin expression induction using the Ecad-ProRFP reporter assay and time course assessment of cytotoxicity induction using the Certox™ Green Cytotoxicity Assay (Promega, Madison, Wis.). (FIG. 3B) Annexin V-FITC staining analysis of apoptosis after 12 h treatment with SN-38 and 6. (FIG. 3C) Cytotoxicity of 6 in PDTO CRC102 using the CellTiter-Blue® Cell Viability Assay (Promega, Madison, Wis.). Mean _EC50 values ± standard deviations are calculated from 6 independent experiments. A representative graph with a representative PDTO inset is shown. Welch t-test statistical analysis is used to determine significance. * = P < 0.05, ** = P < 0.01, *** = P < 0.001, **** = P < 0.0001. Compound 6 induces apoptosis in CRC cell lines and PDTO. (FIG. 3A) Time course assessment of E-cadherin expression induction using the Ecad-ProRFP reporter assay and time course assessment of cytotoxicity induction using the Certox™ Green Cytotoxicity Assay (Promega, Madison, Wis.). (FIG. 3B) Annexin V-FITC staining analysis of apoptosis after 12 h treatment with SN-38 and 6. (FIG. 3C) Cytotoxicity of 6 in PDTO CRC102 using the CellTiter-Blue® Cell Viability Assay (Promega, Madison, Wis.). Mean _EC50 values ± standard deviations are calculated from 6 independent experiments. A representative graph with a representative PDTO inset is shown. Welch t-test statistical analysis is used to determine significance. * = P < 0.05, ** = P < 0.01, *** = P < 0.001, **** = P < 0.0001. Accumulation of compound 6 in SW620 xenograft tumors. Compound 6 was administered to athymic nude mice QD by intraperitoneal injection for 5 days and accumulation in SW620 xenograft tumors was measured. Proposed mechanism of action for CHD1L-mediated TCF transcription. CHD1L is activated through binding TCF complex members PARP1 and TCF4 [Abbott et al., 2020]. (1) Once activated, CHD1L targets hindered WREs that are localized on chromatin. (2) Chromatin remodeling and DNA transfer occur to expose WRE sites. (3) The TCF complex binds exposed WRE promoted by CHD1L and promotes the EMT gene and other genes associated with mCRC. CHD1L ATPase inhibitors effectively prevent step 1 and cause reversal of EMT and other malignant properties of CRC. Evaluation of compound 8. (FIG. 6A) Compound 8 exhibits potent, low μM, dose-dependent inhibition of TCF transcription based on TOPFlash reported over a 24 hour time course in 2D SW260 cell cultures. Compound 8 effectively reversed EMT over 72 h in dual reporter SW620 tumor organoids, evidenced by downregulation of vimentin (Fig. 6B) and dose-dependent upregulation of E-cadherin promoter activity (Fig. 6C). . Compound 8 significantly inhibits clonogenic colony formation over 10 days after 24 hours pretreatment (Fig. 6D) and significantly inhibits HCT116 invasion ability over 48 hours (Fig. 6E). Student's t-test indicates *=P≦0.05. Evaluation of compound 8. (FIG. 6A) Compound 8 exhibits potent, low μM, dose-dependent inhibition of TCF transcription based on TOPFlash reported over a 24 hour time course in 2D SW260 cell cultures. Compound 8 effectively reversed EMT over 72 h in dual reporter SW620 tumor organoids, evidenced by downregulation of vimentin (Fig. 6B) and dose-dependent upregulation of E-cadherin promoter activity (Fig. 6C). . Compound 8 significantly inhibits clonogenic colony formation over 10 days after 24 hours pretreatment (Fig. 6D) and significantly inhibits HCT116 invasion ability over 48 hours (Fig. 6E). Student's t-test indicates *=P≦0.05. Evaluation of compound 8. (FIG. 6A) Compound 8 exhibits potent, low μM, dose-dependent inhibition of TCF transcription based on TOPFlash reported over a 24 hour time course in 2D SW260 cell cultures. Compound 8 effectively reversed EMT over 72 h in dual reporter SW620 tumor organoids, evidenced by downregulation of vimentin (Fig. 6B) and dose-dependent upregulation of E-cadherin promoter activity (Fig. 6C). . Compound 8 significantly inhibits clonogenic colony formation over 10 days after 24 hours pretreatment (Fig. 6D) and significantly inhibits HCT116 invasion ability over 48 hours (Fig. 6E). Student's t-test indicates *=P≦0.05. Evaluation of compound 8. (FIG. 6A) Compound 8 exhibits potent, low μM, dose-dependent inhibition of TCF transcription based on TOPFlash reported over a 24 hour time course in 2D SW260 cell cultures. Compound 8 effectively reversed EMT over 72 h in dual reporter SW620 tumor organoids, evidenced by downregulation of vimentin (Fig. 6B) and dose-dependent upregulation of E-cadherin promoter activity (Fig. 6C). . Compound 8 significantly inhibits clonogenic colony formation over 10 days after 24 hours pretreatment (Fig. 6D) and significantly inhibits HCT116 invasion ability over 48 hours (Fig. 6E). Student's t-test indicates *=P≦0.05. Evaluation of compound 8. (FIG. 6A) Compound 8 exhibits potent, low μM, dose-dependent inhibition of TCF transcription based on TOPFlash reported over a 24 hour time course in 2D SW260 cell cultures. Compound 8 effectively reversed EMT over 72 h in dual reporter SW620 tumor organoids, evidenced by downregulation of vimentin (Fig. 6B) and dose-dependent upregulation of E-cadherin promoter activity (Fig. 6C). . Compound 8 significantly inhibits clonogenic colony formation over 10 days after 24 hours pretreatment (Fig. 6D) and significantly inhibits HCT116 invasion ability over 48 hours (Fig. 6E). Student's t-test indicates *=P≦0.05. Survival of colorectal cancer tumor organoids after treatment with exemplary CHDIL inhibitors. The figure shows a representative graph showing percent viability as a function of log concentration of the indicated compound. (FIG. 7A) treatment with compound 6.9; (FIG. 7B) treatment with compound 6.11; The _IC50 , sometimes the average _IC50 , is given in each figure. Viability data for some exemplary compounds are listed in Table 3. Survival of colorectal cancer tumor organoids after treatment with exemplary CHDIL inhibitors. The figure shows a representative graph showing percent viability as a function of log concentration of the indicated compound. (FIG. 7A) treatment with compound 6.9; (FIG. 7B) treatment with compound 6.11; The _IC50 , sometimes the average _IC50 , is given in each figure. Viability data for some exemplary compounds are listed in Table 3. Assessment of CHD1L-mediated DNA repair and 'on-target' effects of CHD1L inhibitor 6 alone and in combination with irinotecan (a prodrug of SN38). CHD1L is known to be essential for PARP-1-mediated DNA repair and causes resistance to DNA-damaging chemotherapy [Ahel et al., 2009; Tsuda et al., 2017]. DLD1 CRC cells with low levels of CHD1L expression (DLD1 empty vector, EV) were used compared to DLD1 cells engineered to overexpress CHD1L (DLD1 overexpressed, OE). FIG. 8A is a Western blot comparing CHD1L expression in DLD1(EV) and DLD1(OE) given control expression of α-tubulin in these cells. FIG. 8B shows a graph of γ-H2AX intensity (vs. DMSO) of compound alone, SN38 alone, and the combination of the two in DLD1 empty vector cells and DLD1 overexpressing cells. Compound 6 alone does not induce significant DNA damage and does not synergize with SN38 in DLD1 cells with low expression of CHD1L. This graph shows the 'on-target' effect of CHD1L inhibitors synergistic with SN38 in inducing DNA damage in CHD1L-overexpressing DLD1 cells. Assessment of CHD1L-mediated DNA repair and 'on-target' effects of CHD1L inhibitor 6 alone and in combination with irinotecan (a prodrug of SN38). CHD1L is known to be essential for PARP-1-mediated DNA repair and causes resistance to DNA-damaging chemotherapy [Ahel et al., 2009; Tsuda et al., 2017]. DLD1 CRC cells with low levels of CHD1L expression (DLD1 empty vector, EV) were used compared to DLD1 cells engineered to overexpress CHD1L (DLD1 overexpressed, OE). FIG. 8A is a Western blot comparing CHD1L expression in DLD1(EV) and DLD1(OE) given control expression of α-tubulin in these cells. FIG. 8B shows a graph of γ-H2AX intensity (vs. DMSO) of compound alone, SN38 alone, and the combination of the two in DLD1 empty vector cells and DLD1 overexpressing cells. Compound 6 alone does not induce significant DNA damage and does not synergize with SN38 in DLD1 cells with low expression of CHD1L. This graph shows the 'on-target' effect of CHD1L inhibitors synergizing with SN38 in inducing DNA damage in CHD1L-overexpressing DLD1 cells. Synergy studies with an exemplary CHD1L inhibitor and irinotecan (a prodrug of SN38). (FIG. 9A) Synergy study with compounds 6 and 6.3 in SW620 colorectal cancer (CRC) tumor organoids. (FIG. 9B) Synergy study with compound 6.9 in SW620 colorectal cancer (CRC) tumor organoids. (FIG. 9C) Synergy study with compound 6.11 in SW620 colorectal cancer (CRC) tumor organoids. The combination of SN38 with 6 and 6.3 is 50-fold and 150-fold more potent than SN38 alone in killing colonic SW620 tumor organoids, respectively. The combination of SN38 with 6.9 and 6.11 are both over 100 times more potent than SN38 alone. Each of compounds 6, 6.3, 6.9 and 6.11 show synergy with irinotecan (and SN38) in killing SW620 tumor organoids. Synergy studies with an exemplary CHD1L inhibitor and irinotecan (a prodrug of SN38). (FIG. 9A) Synergy study with compounds 6 and 6.3 in SW620 colorectal cancer (CRC) tumor organoids. (FIG. 9B) Synergy study with compound 6.9 in SW620 colorectal cancer (CRC) tumor organoids. (FIG. 9C) Synergy study with compound 6.11 in SW620 colorectal cancer (CRC) tumor organoids. The combination of SN38 with 6 and 6.3 is 50-fold and 150-fold more potent than SN38 alone in killing colonic SW620 tumor organoids, respectively. The combination of SN38 with 6.9 and 6.11 are both over 100 times more potent than SN38 alone. Each of compounds 6, 6.3, 6.9 and 6.11 show synergy with irinotecan (and SN38) in killing SW620 tumor organoids. Synergy studies with an exemplary CHD1L inhibitor and irinotecan (a prodrug of SN38). (FIG. 9A) Synergy study with compounds 6 and 6.3 in SW620 colorectal cancer (CRC) tumor organoids. (FIG. 9B) Synergy study with compound 6.9 in SW620 colorectal cancer (CRC) tumor organoids. (FIG. 9C) Synergy study with compound 6.11 in SW620 colorectal cancer (CRC) tumor organoids. The combination of SN38 with 6 and 6.3 is 50-fold and 150-fold more potent than SN38 alone in killing colonic SW620 tumor organoids, respectively. The combination of SN38 with 6.9 and 6.11 are both over 100 times more potent than SN38 alone. Each of compounds 6, 6.3, 6.9 and 6.11 show synergy with irinotecan (and SN38) in killing SW620 tumor organoids. In vivo synergy study of compound 6 in combination with irinotecan in mice. FIG. 10 controls tumor volume (fold) of SW620 tumor xenografts as a function of days (up to 28 days) of treatment with Compound 6 alone (2), irinotecan alone (3) or their combination (4). (1) includes graphs representing comparisons. A data table showing the data statistical significance is also provided. The combination of irinotecan and Compound 6 significantly inhibits colonic SW620 tumor xenografts to near zero tumor volume within 28 days of treatment compared to the single-agent treatment group. The in vivo synergy of the CHD1L inhibitor compound 6 and irinotecan continues after treatment. FIG. 11 contains graphs representing tumor volume (fold) of SW620 tumor xenografts as a function of days (up to 41 days) of treatment with irinotecan alone (1) or a combination of Compound 6 and irinotecan (2). A data table showing the data statistical significance is also provided. The combination of irinotecan and compound 6 significantly inhibits colonic SW620 tumors to near zero tumor volume past the last treatment (day 28) compared to irinotecan alone. Within 2 weeks of the last irinotecan-only treatment, tumor volume increased beyond that of the last treatment, signifying tumor recurrence. In contrast, the combination maintained smaller tumor volumes. Compound 6 alone and in combination with irinotecan significantly increases survival of CRC tumor-bearing mice compared to vehicle and irinotecan alone. Figure 12 shows % survival as a function of time from the last treatment on day 28 to 52 days after the last treatment with compound 6 alone (2), irinotecan alone (3) or their combination (4) compared to the control (1 ) compared to . A data table showing the data statistical significance is also provided. Survival was significantly higher with combination treatment compared to single dose compound or control. In vivo synergy study of compound 6.11 in combination with irinotecan in mice. FIG. 13 shows tumor volume (fold) of SW620 tumor xenografts as a function of days (up to 20 days) of treatment with compound 6.11 alone (2), irinotecan alone (3) or their combination (4). compared to control (1). A data table showing the data statistical significance is also provided. The combination of irinotecan and compound 6.11 significantly inhibits colonic SW620 tumor xenografts to near zero tumor volume within 20 days of treatment compared to irinotecan alone. The in vivo synergy of the CHD1L inhibitor compound 6.11 and irinotecan continues after treatment. FIG. 14 includes graphs representing tumor volume (fold) of SW620 tumor xenografts as a function of days (up to 41 days) of treatment with irinotecan alone (1) or a combination of Compound 6 and irinotecan (2). Treatment was stopped on day 33 (Tx release). The combination of irinotecan and compound 6.11 significantly inhibits colorectal SW620 tumors past the last treatment (day 33) compared to irinotecan alone. In vivo synergy between CHD1L inhibitor 6.11 and irinotecan significantly increases survival benefit. The combination of compound 6.11 and irinotecan significantly increases survival of CRC tumor-bearing mice compared to vehicle and irinotecan alone. Figure 15 shows % survival as a function of time from the last treatment on day 33 to 50 days after the last treatment with compound 6 alone (2), irinotecan alone (3) or their combination (4) compared to the control (1 ) compared to . A data table showing the data statistical significance is also provided. Survival was significantly higher with combination treatment compared to irinotecan alone or control.

[Detailed description]
The present invention relates generally to the characterization of CHD1L, a relatively new oncogene, as a tumorigenic factor associated with poor prognosis and survival in CRC. A new biological function of CHD1L as a DNA binder of the TCF transcription complex required to promote TCF-induced EMT and other malignant properties has been revealed. Abbott et al., 2020 and the supplementary information to this paper available from the journal website (mct.aacrjournals.org) describe some of the experiments and data presented herein, each incorporated herein by reference in its entirety. incorporated into.

CHD1L is amplified (Chr1q21) and overexpressed in many types of cancer [Ma et al., 2008; Cheng et al., 2013]. CHD1L overexpression has been characterized as a marker of poor prognosis and metastasis in many cancers [Ma et al., 2008; Cheng et al., 2008; Hyeon et al., 2013; Su et al., 2014]. Although the collected literature demonstrating CHD1L as an oncogene and driver of malignant cancers is compelling, previous studies and hypotheses that CHD1L is a potential oncogene as a molecular target in CRC are rigorous. Saga is tested herein. In a computational transcriptome analysis, data obtained over 15 years from a large cohort of 585 CRC patients were reported [Marisa et al., 2013]. Strikingly, CHD1L-low patients lived significantly longer than CHD1L-high patients, and CHD1L expression correlated with poor survival. Using the same cohort, Marisa et al., 2013 identified 6 different subtypes for improved clinical stratification of CRC, CHD1L is ubiquitously expressed in all 6 subtypes, We demonstrate its potential as a therapeutic target for CRC. CHD1L is also correlated with tumor-lymph node-metastasis, with increasing expression progressing from NO (no regional extension) to N3 (distant region extension). Analyzing transcriptome data from a UCCC patient cohort (n=25), CHD1L expression was found to be significantly correlated with stage IV and mCRC. Literature reports and studies herein reveal that CHD1L is an oncogene that promotes malignant CRC, and its high expression correlates with poor prognosis and survival in CRC patients.

A novel biological function of CHD1L as a DNA binder of the TCF transcription complex required to promote TCF-induced EMT and other malignant properties is revealed. Using HTS drug discovery, the first known inhibitors of CHD1L, including PDTO, that show good pharmacological efficacy in cell-based models of CRC were identified and characterized. CHD1L inhibitors effectively prevent CHD1L-mediated TCF transcription and cause reversal of EMT and other malignant properties including CSC stemness and invasive potential. Of note, CHD1L inhibitor 6 exhibits the ability to induce cell death consistent with reversal of EMT and induction of truncated E-cadherin-mediated extrinsic apoptosis through death receptors. Furthermore, compound 6 synergizes with SN38 (ie, irinotecan) and exhibits potent DNA damage induction compared to SN38 alone, consistent with inhibition of PARP1/CHD1L-mediated DNA repair. A CHD1L inhibitor with drug-like physicochemical properties and favorable in vivo PK/PD properties without acute hepatotoxicity has been identified.

Based on the data presented herein, a mechanism of action of EMT mediated by CHD1L, inducing TCF and involved in CRC tumor progression and metastasis is presented (FIG. 5). In this mechanism, the TCF complex specifically recruits CHD1L to dynamically regulate transgene expression. Central to this mechanism, CHD1L binds to the nucleosome hindered WRE when directed by the TCF complex through protein interactions with PARP1 and TCF4. Importantly, PARP1 is characterized as a major cellular activator of CHD1L through macrodomain binding that releases autoinhibition [Lehmann et al., 2017; Gottschalk et al., 2009]. Furthermore, PARP1 is a required component of the TCF complex that interacts with β-catenin and TCF4 [Idogawa et al., 2005]. The mechanism therefore suggests that CHD1L is recruited by the TCF complex and activated by PARP1 and TCF4. Once activated, CHD1L exposes the WRE by nucleosome translocation, facilitating binding of the TCF complex to the WRE and transcription of malignant genes that promote EMT. CHD1L inhibitors have a unique mechanism of action by inhibiting CHD1L ATPase activity that prevents exposure of WRE to the TCF complex and inhibits transcription of TCF target genes that are associated with EMT, particularly mCRC. .

Small molecule inhibitors of CHD1L described herein were identified in screens based on their inhibition of CHD1L ATPase activity. Several inhibitors identified exhibit drug-like physicochemical properties, favorable in vivo PK/PD properties, and no acute hepatotoxicity. Such inhibitors are effective as therapeutics against CRC and mCRC (metastatic CRC), among other CHD1L-induced cancers.

Active compounds of the invention can be further assessed for application to a given therapeutic use using well-known methods for assessing druggability. The term "drugability" relates to the pharmaceutical properties of administration, distribution, metabolism and excretion of a contemplated drug. Draggability is assessed in various ways in the art. For example, the “Lipinski Rule of 5” for determining drug-like properties in molecules related to in vivo absorption and permeability can be applied [Lipinski et al., 2001; Ghose et al., 1999].

The present invention provides methods of combination therapy in which administration of a CDH1L inhibitor is combined with administration of one or more anti-cancer agents that are not CDH1L inhibitors. In embodiments, the other anti-cancer agent is a topoisomerase inhibitor, a platinum-based antineoplastic agent, a PARP inhibitor, or a combination of two or more of such inhibitors and agents. In embodiments, combination therapy combines administration of a CDH1L inhibitor with a topoisomerase inhibitor. In embodiments, combination therapy combines administration of a CDH1L inhibitor with a platinum-based anti-neoplastic agent. In embodiments, combination therapy combines administration of a CDH1L inhibitor with a PARP inhibitor. In embodiments, combination therapy combines administration of a CDH1L inhibitor and a topoisomerase inhibitor with administration of a PARP inhibitor. In embodiments, combination therapy combines administration of a CDH1L inhibitor with chemotherapy to treat a particular cancer. In embodiments herein, combinations of CDH1L inhibitors and other anti-neoplastic agents exhibit synergistic activity together.

In embodiments herein, therapy employing CDH1L can be combined with radiotherapy appropriate for a given cancer.

Various PARP inhibitors are known in the art [see, e.g., Rouleau et al., 2010; Yi et al., 2019; Zhou et al., 2020; Wahlberg et al., 2012; D'Andrea, 2018]. Each of these references is incorporated herein by reference for a description of PARP inhibitors, mechanisms of action of PARP inhibitors, cancers treated using PARP inhibitors, and resistance to PARP inhibitors. incorporated into. In specific embodiments herein, PARP-resistant cancers are treated with a combination of a CDH1L inhibitor and a PARP inhibitor.

Various topoisomerase inhibitors are known in the art and have been clinically employed (eg Hevener, 2018). This reference is incorporated by reference in its entirety for descriptions of types of topoisomerase inhibitors, specific topoisomerase inhibitors, mechanisms of topoisomerase inhibition, cancers treated using topoisomerase inhibitors, and combination therapies using topoisomerase inhibitors. incorporated herein as In embodiments, topoisomerase inhibitors useful in the methods herein include camptothecin and its prodrugs, irinotecan, topotecan, berotecan, indothecan, or indimitecan. In embodiments, topoisomerase inhibitors useful in the methods herein include etoposide or teniposide. In embodiments, topoisomerase inhibitors useful in the methods herein include namitecan, silatecan, bolsaroxine, aldoxorubicin, becatecarin, or edotecarin.

A variety of platinum-based anti-neoplastic agents (also called platins) are known in the art and have been adopted clinically or in clinical trials [eg, Wheate et al., 2010]. This reference provides information on types of platinum-based anti-neoplastic agents, specific platinum-based anti-neoplastic agents, mechanisms of action of such agents, cancers treated using such agents, and platinum-based anti-neoplastic agents are incorporated herein in their entirety for their description of combination therapy. In embodiments, platinum-based antineoplastic agents useful in the methods herein include cisplatin, carbonplatin, oxaliplatin, nedaplatin, donkeyplatin, or heptaplatin. In embodiments, platinum-based antineoplastic agents include satraplatin or picoplatin. Platinum-based anti-neoplastic agents can be liposome-encapsulated (eg, Lipoplatin™) or conjugated to nanopolymers (eg, ProLindac™).

Various thymidylate synthase inhibitors are known in the art and have been clinically employed, particularly in the treatment of CRC [Papamichael, 2009; Lehman, 2002]. Thymidylate synthase inhibitors include folic acid analogs and nucleotide analogs. In specific embodiments, the thymidylate synthase inhibitor is raltitrexed, pemetrexed, nolatrexed or ZD9331. In more specific embodiments, the thymidylate synthase inhibitor is 5-fluorouracil or capecitabine.

The present invention provides CHD1L inhibitors of the formula:

の化合物又はその塩若しくは溶媒和物が挙げられる
［式中、
Ｂ環は、１、２又は３個の５員又は６員環を有するヘテロアリール環又は環系であり、これらのいずれか２個又は３個は、縮合環とすることができ、環は、炭素環式、ヘテロ環式、アリール又はヘテロアリール環であり、環の少なくとも１個は、ヘテロアリールであり、
Ｂ環において、Ｘはそれぞれ、Ｎ又はＣＨから独立的に選択され、少なくとも１つのＸはＮであり、
Ｒ_Ｐは、第一級若しくは第二級アミン基［－Ｎ（Ｒ_２）（Ｒ_３）］であり、又は－（Ｍ）_ｘ－Ｐ基であり、Ｐは、－Ｎ（Ｒ_２）（Ｒ_３）又はアリール若しくはヘテロアリール基であり、ｘは、０又は１であって、Ｍの有無を示し、Ｍは、任意選択で置換されているリンカー－（ＣＨ_２）_ｎ－又は－Ｎ（Ｒ）（ＣＨ_２）_ｎ－であり、ｎはそれぞれ、独立的に１～６の（１と６を含む）整数であり、
Ｙは、－Ｏ－、－Ｓ－、－Ｎ（Ｒ_１）－、－ＣＯＮ（Ｒ_１）－、－Ｎ（Ｒ_１）ＣＯ－、－ＳＯ_２Ｎ（Ｒ_１）－、又は－Ｎ（Ｒ_１）ＳＯ_２－からなる群から選択される２価の原子又は基であり、
Ｌ_１は、任意選択で置換されており、飽和であるか、又は二重結合を含む（ｃｉｓでも、ｔｒａｎｓでもよい）任意選択の１～４個の炭素のリンカーであり、ｘは、０又は１であって、Ｌ_１の有無を示し、
Ａ環は、１、２又は３個の環を有する炭素環式若しくはヘテロ環式環又は環系であり、これらのうちの２個又は３個は縮合されていてもよく、各環は、３～１０個の炭素原子を有し、１～６個のヘテロ原子を任意選択で有し、各環は、任意選択で飽和、不飽和又は芳香族であり、
Ｚは、Ｒ’基で置換されている少なくとも１個の窒素を含む２価の基であり、
実施形態において、Ｚは、－Ｎ（Ｒ’）－、－ＣＯＮ（Ｒ’）－、－Ｎ（Ｒ’）ＣＯ－、－ＣＳＮ（Ｒ’）－、－Ｎ（Ｒ’）ＣＳ－、－Ｎ（Ｒ’）ＣＯＮ（Ｒ’）－、－ＳＯ_２Ｎ（Ｒ’）－、－Ｎ（Ｒ’）ＳＯ_２－、－ＣＨ（ＣＦ_３）Ｎ（Ｒ’）－、－Ｎ（Ｒ’）ＣＨ（ＣＦ_３）－、－Ｎ（Ｒ’）ＣＨ_２ＣＯＮ（Ｒ’）ＣＨ_２－、－Ｎ（Ｒ’）ＣＯＣＨ_２Ｎ（Ｒ’）ＣＨ_２－、

から選択される２価の基であり、又は２価のＺ基は、少なくとも１個の窒素環員を有する５員若しくは６員のヘテロ環式環、例えば

を含み、
Ｌ_２は、任意選択で置換されており、飽和であるか、又は二重結合を含む（ｃｉｓでも、ｔｒａｎｓでもよい）任意選択の１～４個の炭素のリンカーであり、ｚは、０又は１であって、Ｌ_１の有無を示し、
Ｒは、水素、脂肪族基、カルボシクリル基、アリール基、ヘテロシクリル基及びヘテロアリール基からなる群から選択され、これらの基のそれぞれは、任意選択で置換されており、
Ｒ’はそれぞれ、水素、脂肪族基、カルボシクリル基、アリール基、ヘテロシクリル基及びヘテロアリール基からなる群から独立的に選択され、これらの基のそれぞれは、任意選択で置換されており、
Ｒ_１は、水素、脂肪族基、カルボシクリル基、アリール基、ヘテロシクリル基及びヘテロアリール基からなる群から選択され、これらの基のそれぞれは、任意選択で置換されており、
Ｒ_２及びＲ_３は、水素、脂肪族基、カルボシクリル基、アリール基、ヘテロシクリル基及びヘテロアリール基からなる群から独立的に選択され、これらの基のそれぞれは、任意選択で置換されており、或いは
Ｒ_２とＲ_３はそれらが結合しているＮと一緒になって、任意選択で置換されている５員～１０員の飽和、部分不飽和又は芳香族環のヘテロ環式環を形成し、
Ｒ_Ａ及びＲ_Ｂは、それぞれ指示されたＡ及びＢ環又は環系上の水素又は１～１０個の非水素置換基を表し、Ｒ_Ａ及びＲ_Ｂ置換基は、水素、ハロゲン、ヒドロキシル、シアノ、ニトロ、アミノ、一置換又は二置換アミノ（－ＮＲ_ＣＲ_Ｄ）、アルキル、アルケニル、シクロアルキル、シクロアルケニル、アリール、ヘテロシクリル、アルコキシ、アシル、ハロアルキル、－ＣＯＯＲ_Ｃ、－ＯＣＯＲ_Ｃ、－ＣＯＮＲ_ＣＲ_Ｄ、－ＯＣＯＮＲ_ＣＲ_Ｄ、－ＮＲ_ＣＣＯＲ_Ｄ、－ＳＲ_Ｃ、－ＳＯＲ_Ｃ、－ＳＯ_２Ｒ_Ｃ、及び－ＳＯ_２ＮＲ_ＣＲ_Ｄから独立的に選択され、アルキル、アルケニル、シクロアルキル、シクロアルケニル、アリール、ヘテロシクリル、アルコキシ、及びアシルは、任意選択で置換されており、
Ｒ_Ｃ及びＲ_Ｄはそれぞれ、水素、アルキル、アルケニル、シクロアルキル、シクロアルケニル、ヘテロシクリル、アリール、又はヘテロアリールから選択され、これらの基のそれぞれは、１個又は複数のハロゲン、アルキル、アルケニル、ハロアルキル、アルコキシ、アリール、ヘテロアリール、ヘテロシクリル、アリール置換アルキル、又はヘテロシクリル置換アルキルで任意選択で置換されており、
Ｒ_Ｈは、任意選択で置換されているアリール又はヘテロアリール基であり、
任意選択の置換は、１個又は複数のハロゲン、ニトロ、シアノ、アミノ、モノ又はジ－Ｃ１～Ｃ３アルキル置換アミノ、Ｃ１～Ｃ３アルキル、Ｃ２～Ｃ４アルケニル、Ｃ３～Ｃ６シクロアルキル、Ｃ３～Ｃ６－シクロアルケニル、Ｃ１～Ｃ３ハロアルキル、Ｃ６～Ｃ１２アリール、Ｃ５～Ｃ１２ヘテロアリール、Ｃ３～Ｃ１２ヘテロシクリル、Ｃ１～Ｃ３アルコキシ、Ｃ１～Ｃ６アシル、－ＣＯＯＲ_Ｅ、－ＯＣＯＲ_Ｅ、－ＣＯＮＲ_ＥＲ_Ｆ、－ＯＣＯＮＲ_ＥＲ_Ｄ、－ＮＲ_ＥＣＯＲ_Ｆ、－ＳＲ_Ｅ、－ＳＯＲ_Ｅ、－ＳＯ_２Ｒ_Ｅ、及び－ＳＯ_２ＮＲ_ＥＲ_Ｆでの置換を含み、アルキル、アルケニル、シクロアルキル、シクロアルケニル、アリール、ヘテロシクリル、アルコキシ、及びアシルは、任意選択で置換されており、
Ｒ_Ｅ及びＲ_Ｆはそれぞれ、水素、Ｃ１～Ｃ３アルキル、Ｃ２～Ｃ４アルケニル、Ｃ３～Ｃ６シクロアルキル、Ｃ３～Ｃ６－シクロアルケニル、Ｃ１～Ｃ３ハロアルキル、Ｃ６～Ｃ１２アリール、Ｃ５～Ｃ１２ヘテロアリール、Ｃ３～Ｃ１２ヘテロシクリル、Ｃ１～Ｃ３アルコキシ、Ｃ１～Ｃ６アシルから選択され、これらの基のそれぞれは、１個又は複数のハロゲン、ニトロ、シアノ、アミノ、モノ又はジ－Ｃ１～Ｃ３アルキル置換アミノ、Ｃ１～Ｃ３アルキル、Ｃ２～Ｃ４アルケニル、Ｃ３～Ｃ６シクロアルキル、Ｃ３～Ｃ６－シクロアルケニル、Ｃ１～Ｃ３ハロアルキル、Ｃ６～Ｃ１２アリール、Ｃ５～Ｃ１２ヘテロアリール、Ｃ３～Ｃ１２ヘテロシクリル、Ｃ１～Ｃ３アルコキシ、及びＣ１～Ｃ６アシルで任意選択で置換されている］。 Compounds useful in the methods of the present invention include formula I

or a salt or solvate thereof [wherein
Ring B is a heteroaryl ring or ring system having 1, 2 or 3 5- or 6-membered rings, any 2 or 3 of which can be fused rings, the ring a carbocyclic, heterocyclic, aryl or heteroaryl ring wherein at least one of the rings is heteroaryl;
in the B ring, each X is independently selected from N or CH, at least one X is N;
R _P is a primary or secondary amine group [—N(R ₂ )(R ₃ )] or a —(M) _x —P group, where P is —N(R ₂ )( R ₃ ) or an aryl or heteroaryl group, x is 0 or 1, indicating the presence or absence of M, and M is an optionally substituted linker —(CH ₂ ) _n — or —N( R)(CH ₂ ) _n —, where each n is independently an integer from 1 to 6 (inclusive);
Y is -O-, -S-, -N(R ₁ )-, -CON(R ₁ )-, -N(R ₁ )CO-, -SO ₂ N(R ₁ )-, or -N( R ₁ ) a divalent atom or group selected from the group consisting of SO ₂ —;
L ₁ is an optional 1-4 carbon linker (which can be cis or trans) that is optionally substituted, saturated, or contains a double bond, and x is 0 or 1, indicating the presence or absence of L ₁ ,
Ring A is a carbocyclic or heterocyclic ring or ring system having 1, 2 or 3 rings, 2 or 3 of which may be fused, each ring having 3 -10 carbon atoms, optionally having 1-6 heteroatoms, each ring optionally being saturated, unsaturated or aromatic;
Z is a divalent group containing at least one nitrogen substituted with an R'group;
In embodiments, Z is -N(R')-, -CON(R')-, -N(R')CO-, -CSN(R')-, -N(R')CS-, - N(R')CON(R')-, _-SO2N (R')-, -N(R') _SO2- , -CH( _CF3 )N(R')-, -N(R' )CH(CF ₃ )—, —N(R′)CH ₂ CON(R′)CH ₂ —, —N(R′)COCH ₂ N(R′)CH ₂ —,

or the divalent Z group is a 5- or 6-membered heterocyclic ring having at least one nitrogen ring member, such as

including
L ₂ is an optional 1-4 carbon linker (which can be cis or trans) that is optionally substituted, saturated, or contains a double bond, and z is 0 or 1, indicating the presence or absence of L ₁ ,
R is selected from the group consisting of hydrogen, aliphatic groups, carbocyclyl groups, aryl groups, heterocyclyl groups and heteroaryl groups, each of which groups are optionally substituted;
each R' is independently selected from the group consisting of hydrogen, aliphatic groups, carbocyclyl groups, aryl groups, heterocyclyl groups and heteroaryl groups, each of which groups is optionally substituted;
R ₁ is selected from the group consisting of hydrogen, aliphatic groups, carbocyclyl groups, aryl groups, heterocyclyl groups and heteroaryl groups, each of which groups are optionally substituted;
R ₂ and R ₃ are independently selected from the group consisting of hydrogen, aliphatic groups, carbocyclyl groups, aryl groups, heterocyclyl groups and heteroaryl groups, each of these groups being optionally substituted; or R ₂ and R ₃ together with the N to which they are attached form an optionally substituted 5- to 10-membered saturated, partially unsaturated or aromatic heterocyclic ring; ,
R _A and R _B represent hydrogen or from 1 to 10 non-hydrogen substituents on the indicated A and B rings or ring systems, respectively, where R _A and R _B substituents are hydrogen, halogen, hydroxyl, cyano , nitro, amino, mono- or disubstituted amino (-NR _C R _D ), alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, alkoxy, acyl, haloalkyl, -COOR _C , -OCOR _C , -CONR _C R _D , —OCONR _C R _D , —NR _C COR _D , —SR _C , —SOR _C , —SO ₂ R _C and —SO ₂ NR _C R _D independently selected from alkyl, alkenyl, cycloalkyl , cycloalkenyl, aryl, heterocyclyl, alkoxy, and acyl are optionally substituted;
R _C and R _D are each selected from hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl, each of these groups being one or more halogen, alkyl, alkenyl, haloalkyl optionally substituted with , alkoxy, aryl, heteroaryl, heterocyclyl, aryl-substituted alkyl, or heterocyclyl-substituted alkyl;
_RH is an optionally substituted aryl or heteroaryl group;
Optional substitutions are one or more of halogen, nitro, cyano, amino, mono- or di-C1-C3 alkyl substituted amino, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6- Cycloalkenyl, C1-C3 haloalkyl, C6-C12 aryl, C5-C12 heteroaryl, C3-C12 heterocyclyl, C1-C3 alkoxy, C1-C6 acyl, -COOR _E , -OCOR _E , -CONR _E R _F , -OCONR _{Alkyl , alkenyl , cycloalkyl , cycloalkenyl , aryl , _} heterocyclyl, alkoxy, and acyl are optionally substituted;
R _E and R _F are each hydrogen, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6-cycloalkenyl, C1-C3 haloalkyl, C6-C12 aryl, C5-C12 heteroaryl, C3 -C12 heterocyclyl, C1-C3 alkoxy, C1-C6 acyl, each of these groups being selected from one or more halogen, nitro, cyano, amino, mono- or di-C1-C3 alkyl-substituted amino, C1- C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6-cycloalkenyl, C1-C3 haloalkyl, C6-C12 aryl, C5-C12 heteroaryl, C3-C12 heterocyclyl, C1-C3 alkoxy, and C1- optionally substituted with C6 acyl].

In an embodiment of Formula I,
R is selected from hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl, each of which groups are optionally substituted;
each R′ is independently selected from hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl, each of which groups are optionally substituted;
R ₁ -R ₃ are independently selected from hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl, each of which groups is optionally substituted;
one or more of R ₁ -R ₃ is cycloalkyl-substituted alkyl, such as cyclopropylmethyl, cyclopentylmethyl, or cyclohexylmethyl;
R is hydrogen or C1-C3 alkyl;
each R′ is independently hydrogen or C1-C3 alkyl;
R ₁ is hydrogen or C1-C3 alkyl;
R ₂ and R ₃ are independently selected from hydrogen or C1-C3 alkyl, or R ₂ and R ₃ together with the N to which they are attached are a 5- to 7-membered saturated heterocyclic ring to form

In specific embodiments of formula I,
the A ring is optionally substituted phenyl;
the A ring is optionally substituted naphthyl;
the B ring is optionally substituted pyridyl;
ring B is optionally substituted pyrimidyl;
ring B is optionally substituted pyrazinyl;
the B ring is optionally substituted triazinyl;
ring B is optionally substituted quinazolinyl;
ring B is optionally substituted pteridinyl;
ring B is optionally substituted quinolinyl;
ring B is optionally substituted isoquinolinyl;
the B ring is optionally substituted naphthyridinyl;
ring B is optionally substituted pyridopyrimidyl;
ring B is optionally substituted pyrimidopyridyl;
ring B is optionally substituted pyranopyridyl;
ring B is optionally substituted pyranopyrimidyl;
the B ring is an optionally substituted purine;
Ring A is optionally substituted phenyl and Ring B is optionally substituted pyrimidinyl, or Ring A is optionally substituted phenyl and Ring B is an optionally substituted pteridinyl.

Preferred A and B ring substitutions include one or more of C1-C3 alkyl, C3-C7 cycloalkyl, C4-C10 cycloalkyl substituted alkyl, C2-C4 alkenyl, C1-C3 alkoxy, C1-C3 acyl, halogen, It includes hydroxyl, C1-C3 haloalkyl, mono- or di-substituted phenyl or mono- or di-substituted benzyl. More specific A and B ring substitutions include methyl, ethyl, isopropyl, cyclopropyl, cyclopropylmethyl, methoxy, ethoxy, phenyl, benzyl, halophenyl, halobenzyl, Cl, Br, F, CF ₃ —, HO—, CF ₃ O—, CH ₃ CO— and CHCO are included.

In a more specific embodiment, ring B has the structure shown in Scheme 4, formula RBI, X ¹ and X ² are selected from CH and N, and at least one of X ¹ and X ² is N, X ³ -X ⁶ are selected from CH, CH ₂ , O, S, N and NH, and the B ring is saturated, unsaturated or aromatic depending on the selection of X ¹ -X ⁶ R _B represents an optional substitution as defined for Formula I. In embodiments, R _B represents hydrogen and the B ring is unsubstituted. In embodiments, R 1 _B represents one or more halogen, C1-C3 alkyl, C1-C3 acyl, C1-C3 alkoxy. In embodiments, _RB represents one or more of F, Cl or Br, methyl, ethyl, acetyl or methoxy, or combinations thereof. In embodiments of Formula I, the B ring is selected from any of RB2-RB5 shown in Scheme 4.

In an embodiment of Formula I,
x is 1, L ₁ is -(CH ₂ ) _n -, n is 1 or 2,
x is 0, L ₁ is absent,
y is 1, L ₂ is —(CH ₂ ) _n —, n is 1 or 2,
y is 1, L ₂ is -CH=CH-,
y is 1, L ₂ is trans-CH=CH-,
both x and y are 0, and
x is 1, y is 0, L ₁ is -(CH ₂ ) _n -, n is 1 or 2,
y is 1, x is 0, L ₂ is -(CH ₂ ) _n -, n is 1 or 2, or both x and y are 1, L ₂ and L ₁ are -(CH ₂ ) _n -, where n is 1 or 2.

In an embodiment of Formula I,
Y is -O-, -S-, -N(R ₁ )-, -CON(R ₁ )-, or -N(R ₁ )CO-;
Y is -N(R ₁ )-, -CON(R ₁ )-, or -N(R ₁ )CO-;
R ₁ is hydrogen, C1-C3 alkyl or C1-C3 haloalkyl, especially C1-C3 fluoroalkyl,
R ₁ is hydrogen, a methyl group or CF ₃ —;
R ₁ is hydrogen,
Y is -N(R ₁ )-, -CON(R ₁ )-, or -N(R ₁ )CO-, R ₁ is hydrogen, methyl or CF ₃ -;
Y is -NH-, -CONH-, or -NHCO-,
Y is -N(R ₁ )-, R ₁ is hydrogen, C1-C3 alkyl or C1-C3 haloalkyl, especially C1-C3 fluoroalkyl, or Y is -N(R ₁ )-, R ₁ is hydrogen, methyl or CF ₃ —.

In an embodiment of Formula I, x and y are both 0 and Y is -N(R ₁ )-. In embodiments of Formula I, both x and y are 0 and Y is -NH-.

であり、
Ｚは、

であり、
Ｚは、

であり、
Ｒ’は、水素、Ｃ１～Ｃ６アルキル又はＣ１～Ｃ３ハロアルキル、特にＣ１～Ｃ３フルオロアルキルであり、
Ｒ’は、水素又はＣ１～Ｃ３アルキルであり、
Ｒ’は、水素、メチル又はＣＦ_３－であり、
Ｒ’は、水素又はメチルであり、
Ｒ’は水素であり、
Ｚは、－Ｎ（Ｒ’）－、－ＣＯＮ（Ｒ’）－、又は－Ｎ（Ｒ’）ＣＯ－であり、Ｒ’は、水素又はメチルであり、
Ｚは、－ＣＯＮ（Ｒ’）－又は－Ｎ（Ｒ’）ＣＯ－であり、Ｒ’は、水素又はメチルであり、
Ｚは－Ｎ（Ｒ’）ＣＯＮ（Ｒ’）－であり、Ｒ’は両方とも水素であり、
Ｚは、

であり、Ｒ’は水素であり、或いは
Ｚは、

であり、Ｒ’は水素である。 In an embodiment of Formula I,
Z is -N(R')-, -CON(R')-, or -N(R')CO-;
Z is —CH(CF ₃ )N(R′)—;
Z is -SO ₂ N(R')-,
Z is -N(R')CON(R')-,
Z is -N(R')CH ₂ CON(R')CH ₂ -;
Z is

and
Z is

and
Z is

and
R' is hydrogen, C1-C6 alkyl or C1-C3 haloalkyl, especially C1-C3 fluoroalkyl,
R' is hydrogen or C1-C3 alkyl,
R′ is hydrogen, methyl or CF ₃ —;
R' is hydrogen or methyl,
R' is hydrogen;
Z is -N(R')-, -CON(R')-, or -N(R')CO-, R' is hydrogen or methyl;
Z is -CON(R')- or -N(R')CO-, R' is hydrogen or methyl,
Z is -N(R')CON(R')- and both R' are hydrogen;
Z is

and R' is hydrogen, or Z is

and R' is hydrogen.

In an embodiment of Formula I,
x is 0;
x is 1, L ₂ is -(CH ₂ ) _n -, n is 1 to 3,
y is 0, x is 1, L ₂ is -(CH ₂ ) _n -, n is 1 to 3,
x is 0, Z is -N(R')-, -CON(R')-, or -N(R')CO-;
x is 0, Z is -N(R')-, -CON(R')-, or -N(R')CO-, R' is hydrogen or methyl,
x is 0, Z is -CON(R')-,
x is 0, Z is -CON(R')-, R' is hydrogen or methyl,
x is 0, Z is -CON(R')-, R' is hydrogen,
x is 1, L ₂ is —(CH ₂ ) _n —, n is 1 to 3, Z is —N(R ₁ )—, —CON(R′)—, or —N (R′) CO—,
x is 1, L ₂ is -(CH ₂ ) _n -, n is 1 to 3, Z is -CON(R')-,
x is 1, L ₂ is -CH ₂ -, Z is -CON(R')-,
x is 1, L ₂ is -CH ₂ -CH ₂ -, Z is -CON(R')-,
x is 0 or 1, L ₂ if present is -CH ₂ - or -CH ₂ -CH ₂ -, Z is -CON(R')-,
x is 0 or 1, L ₂ if present is —CH ₂ — or —CH ₂ —CH ₂ —, Z is —CON(R′)— and R′ is hydrogen ,
y is 0, x is 0 or 1, L ₂ when present is -CH ₂ - or -CH ₂ -CH ₂ -, Z is -CON(R')-, R' is hydrogen;
y is 0, Y is -N(R ₁ )-, x is 0 or 1, if L ₂ is present it is -CH ₂ - or -CH ₂ -CH ₂ -; Z is -CON(R')-, or y is 0, Y is -N(R ₁ )-, R ₁ is hydrogen, x is 0 or 1, L ₂ is When present it is -CH ₂ - or -CH ₂ -CH ₂ -, Z is -CON(R') and R' is hydrogen.

In an embodiment of Formula I,
R 1 _P contains at least one nitrogen, or when R 1 _P is -(M)xP and x=0, P is -N(R ₂ )(R ₃ ); or P is a heterocyclic or heteroaryl group having at least one ring N, or R _P is —(M)xP, x=1 and M=—(CH ₂ ) _n , then P is —N(R ₂ )(R ₃ ), or P is a heterocyclic or heteroaryl group having at least one ring N;

In embodiments of Formula I, R _P is
—N(R ₂ )(R ₃ ),
-(M)-N(R ₂ )(R ₃ ) [M is an optionally substituted linker -(CH ₂ ) _n - or -N(R)(CH ₂ ) _n - and n is each independently an integer from 1 to 6 (inclusive) and R is hydrogen or an optionally substituted alkyl group having 1 to 3 carbon atoms],
-(M)-N(R ₂ )(R ₃ ) [M is an optionally substituted linker -(CH ₂ ) _n -, and each n is independently from 1 to 6 (1 and 6) and R is hydrogen or an optionally substituted alkyl group having 1 to 3 carbon atoms];
-(M)-N(R ₂ )(R ₃ ) [M is an optionally substituted linker -(CH ₂ ) _n -, and each n is independently from 1 to 6 (1 and 6) and R is hydrogen or an optionally substituted alkyl group having from 1 to 3 carbon atoms, the optional substitution being one or more halogen or one or substitution with multiple C1-C3 alkyl groups],
-(M)-N(R ₂ )(R ₃ ), [M is optionally substituted -N(R)(CH ₂ ) _n -, and each n is independently 1-6 is an integer (including 1 and 6) of and R is hydrogen or an optionally substituted alkyl group having 1 to 3 carbon atoms],
-(M)-N(R ₂ )(R ₃ ) [M is optionally substituted -N(R)(CH ₂ ) _n - and each n is independently from 1 to 6 is an integer (including 1 and 6) and R is H or an optionally substituted alkyl group having 1 to 3 carbon atoms, the optional substitution being one or more halogen or substitution with one or more C1-C3 alkyl groups],
-(M)-N(R ₂ )(R ₃ ), where M is an optionally substituted linker -(CH ₂ ) ⁿ -, n is 1, 2 or 3];
-(M)-N(R ₂ )(R ₃ ) [M is an optionally substituted linker -N(R)(CH ₂ ) _n - and n is 1, 2 or 3 ],
-(M)-N(R ₂ )(R ₃ ) [M is -(CH ₂ ) _n - and n is 1, 2 or 3],
-(M)-N(R ₂ )(R ₃ ) [M is -N(R)(CH ₂ ) _n - and n is 1, 2 or 3],
—(M) _x —P group [P is an aryl or heteroaryl group, x is 0 or 1 and indicates the presence or absence of M, M is an optionally substituted linker-(CH ₂ ) _n - or -N(R)(CH ₂ ) _n -, where each n is independently an integer from 1 to 6 (inclusive) and R is H or from 1 to 3 is an optionally substituted alkyl group having carbon atoms of
-(M)-P group [P is an aryl or heteroaryl group and M is an optionally substituted linker -(CH ₂ ) _n - or -N(R)(CH ₂ ) _n - and each n is independently an integer from 1 to 6 (inclusive) and R is H or an optionally substituted alkyl group having 1 to 3 carbon atoms] ,
-(M)-P group [P is an aryl or heteroaryl group, M is an optionally substituted linker -N(R)(CH ₂ ) _n -, and each n is independently is an integer from 1 to 6 (inclusive) and R is H or an optionally substituted alkyl group having 1 to 3 carbon atoms],
-(M)-P group [P is an aryl or heteroaryl group, M is an optionally substituted linker -(CH ₂ ) _n -, n is each independently 1-3 is an integer (including 1 and 3) of and R is H or an optionally substituted alkyl group having 1 to 3 carbon atoms],
and
P is optionally substituted phenyl or naphthyl,
P is optionally substituted phenyl or naphthyl, the optional substitution being with one or more halogen, C1-C3 alkyl, C1-C3 alkoxy or C1-C3 haloalkyl;
P is an optionally substituted heteroaryl group having a 5- or 6-membered ring or two fused 5- or 6-membered rings;
P is an optionally substituted heteroaryl group having a 5- or 6-membered ring or two fused 5- or 6-membered rings and having 1-3 nitrogen ring members;
R ₂ in R _P is hydrogen (ie —N(R ₂ )(R ₃ ) is a primary amine group);
both R ₂ and R ₃ in R _P are groups other than hydrogen (i.e., —N(R ₂ )(R ₃ ) is a secondary amine group);
R ₂ is hydrogen, R ₃ is an optionally substituted 3-8 membered cycloalkyl group,
R ₂ is hydrogen, R ₃ is a C1-C3 alkyl group substituted with a 3-8 membered cycloalkyl group,
R ₂ is hydrogen, R ₃ is an optionally substituted aryl group having 6 to 12 carbon atoms,
R ₂ is hydrogen, R ₃ is an optionally substituted heteroaryl group having 6 to 12 carbon atoms and 1 to 3 heteroatoms (N, O, or S);
R ₂ is hydrogen, R ₃ is an optionally substituted heteroaryl group having 6-12 carbon atoms and 1-3 ring nitrogens;
R ₂ and R ₃ together with the N to which they are attached form an optionally substituted 5- to 10-membered saturated, partially unsaturated or aromatic heterocyclic ring;
R _P is —(CH ₂ ) _n —N(R ₂ )(R ₃ ), n is 1 or 2, R ₂ and R ₃ together with the N to which they are attached, forming an optionally substituted 5- to 10-membered saturated, partially unsaturated or aromatic heterocyclic ring;
R _P is —N(R)(CH ₂ ) _n —N(R ₂ )(R ₃ ), n is 1 or 2, R is hydrogen or methyl, R ₂ and R ₃ together with the N to which they are attached form an optionally substituted 5- to 10-membered saturated, partially unsaturated or aromatic heterocyclic ring;
R _P is -MN(R ₂ )(R ₃ ), and R ₂ and R ₃ together with the N to which they are attached are optionally substituted 5- to 10-membered forming a saturated, partially unsaturated or aromatic heterocyclic ring,
R ₂ and R ₃ together with the N to which they are attached form an optionally substituted 5- to 10-membered heterocyclic ring containing no double bonds;
R _P is —N(R ₂ )(R ₃ ) and R ₂ and R ₃ together with the N to which they are attached are optionally substituted 5 not containing a double bond forming a 1- to 10-membered heterocyclic ring;
R ₂ and R ₃ together with the N to which they are attached form an optionally substituted 5- to 10-membered heterocyclic ring containing 1, 2 or 3 double bonds death,
R _P is —N(R ₂ )(R ₃ ) and R ₂ and R ₃ together with the N to which they are attached optionally contain 1, 2 or 3 double bonds forming a substituted 5- to 10-membered heterocyclic ring;
R ₂ and R ₃ together with the N to which they are attached form an optionally substituted 5- to 10-membered heteroaryl ring, or R _P is —N(R ₂ ) ( R ₃ ) and R ₂ and R ₃ together with the N to which they are attached form an optionally substituted 5- to 10-membered heteroaryl ring.

In specific embodiments of Formula I, R _P or -N(R ₂ )(R ₃ ) is
any one of R _N 1 to R _N 31 of Scheme 2;
R _N 1;
R _N 3;
R _N 2 or R _N 4;
R _N 5 or R _N 6;
R _N 7 or R _N 8;
R _N 9;
R _N 10;
R _N 11;
R _N 12;
R _N 13;
R _N 14;
R _N 15;
R _N 16;
R _N 17 or R _N 18;
R _N 19 or R _N 20;
R _N 21;
R _N 22;
R _N 23 or R _N 24
R _N 25;
R _N 26 to R _N 29;
R _N 30;
R _N 31;
R _N 1, R _N 2, R _N 3, R _N 4, R _N 11, R _N 13, or R _N 14; or unsubstituted R _N 1 to R _N 31.

である。 In embodiments of Formula I, R _H is
optionally substituted phenyl;
unsubstituted phenyl;
optionally substituted naphthyl;
unsubstituted naphthyl;
optionally substituted naphth-2-yl;
optionally substituted naphth-1-yl;
naphth-2-yl;
naphth-1-yl;
optionally substituted thiophenyl;
halogen-substituted thiophenyl;
brominated thiophenyl optionally substituted thiophen-2-yl;
halogen-substituted thiophen-2-yl;
brominated thiophen-2-yl 4-halothiophen-2-yl;
4-bromothiophen-2-yl;
optionally substituted frills;
optionally substituted fur-2-yl optionally substituted indolyl;
unsubstituted indolyl;
indol-3-yl;
indol-2-yl;
indol-1-yl;
optionally substituted pyridinopyrrolyl;
optionally substituted pyridinopyrrol-2-yl;
optionally substituted benzimidazolyl;

is.

In specific embodiments, the optional substitution of R _H is one or more of halogen, C1-C3 alkyl, C1-C3 alkoxyl, C1-C3 haloalkyl, C1-C3 fluoroalkyl, C4-C7 cycloalkylalkyl , OH, amino, C1-C6 acyl, -COOR _E , -OCORE , -CONR _E R _F , -OCONR _E R _D , -NR _E COR _F , -SR _E , _{-SOR E} , -SO ₂ R _E , and —SO ₂ NR _E R _F , where R _E and R _e are as defined above, in particular hydrogen, C1-C3 alkyl, phenyl or benzyl. More particularly, the optional substitution of R _H is one or more halogen (especially Br or Cl), C1-C3 alkyl, C1-C3 alkoxyl, C1-C3 fluoroalkyl (especially CF ₃ —) is a replacement with .

を有する
［式中、
Ｘ_１１は、ＣＨ、ＣＲ_Ｔ又はＮであり、Ｒ_Ｔは、上記の任意選択のＲ_Ｈ環置換であり、Ｒ及びＲ’は、独立的に水素、Ｃ１～Ｃ６アルキル基、Ｃ４～Ｃ１０シクロアルキルアルキル基、アリール基、ヘテロシクリル基、又はヘテロアリール基であり、これらの基のそれぞれは、任意選択で置換されている］。具体的な実施形態において、Ｒ_Ｔは、水素又はハロゲン、ＯＨ、Ｃ１～Ｃ３アルキル、Ｃ１～Ｃ３アルコキシ、若しくはＣ３～Ｃ６シクロアルキルで置換されているＣ１～Ｃ３アルキルのうちの１つ若しくは複数での置換であり、Ｒ’は、水素、Ｃ１～Ｃ３アルキル、Ｃ１～Ｃ３アルコキシ、又はＣ３～Ｃ６シクロアルキルで置換されているＣ１～Ｃ３アルキルであり、Ｒは、水素、Ｃ１～Ｃ３アルキル、Ｃ１～Ｃ３アルコキシ、又はＣ３～Ｃ６シクロアルキルで置換されているＣ１～Ｃ３アルキルである。 In embodiments, R _H is of the formula

having [wherein
X ₁₁ is CH, CR _T or N, R _T is an optional R _H ring substitution as described above, R and R′ are independently hydrogen, C1-C6 alkyl groups, C4-C10 cyclo an alkylalkyl group, an aryl group, a heterocyclyl group, or a heteroaryl group, each of which is optionally substituted]. In specific embodiments, R _T is one or more of hydrogen or C1-C3 alkyl substituted with halogen, OH, C1-C3 alkyl, C1-C3 alkoxy, or C3-C6 cycloalkyl. and R′ is hydrogen, C1-C3 alkyl, C1-C3 alkoxy, or C1-C3 alkyl substituted with C3-C6 cycloalkyl, and R is hydrogen, C1-C3 alkyl, C1 ~C3 alkoxy, or C1-C3 alkyl substituted with C3-C6 cycloalkyl.

を有する
［式中、
Ｘ_１１は、ＣＨ、ＣＲ_Ｔ又はＮであり、Ｘ_１０は、ＣＨ、ＣＲ_Ｔ又はＮであり、Ｒ_Ｔは、上記の任意選択のＲ_Ｈ環置換であり、Ｒ及びＲ’は、独立的に水素、Ｃ１～Ｃ６アルキル基、Ｃ４～Ｃ１０シクロアルキルアルキル基、アリール基、ヘテロシクリル基、又はヘテロアリール基であり、これらの基のそれぞれは、任意選択で置換されている］。具体的な実施形態において、Ｒ_Ｔは、水素又はハロゲン、ＯＨ、Ｃ１～Ｃ３アルキル、Ｃ１～Ｃ３アルコキシ、若しくはＣ３～Ｃ６シクロアルキルで置換されているＣ１～Ｃ３アルキルのうちの１つ若しくは複数での置換であり、Ｒ’は、水素、Ｃ１～Ｃ３アルキル、Ｃ１～Ｃ３アルコキシ、又はＣ３～Ｃ６シクロアルキルで置換されているＣ１～Ｃ３アルキルであり、Ｒは、水素、Ｃ１～Ｃ３アルキル、Ｃ１～Ｃ３アルコキシ、又はＣ３～Ｃ６シクロアルキルで置換されているＣ１～Ｃ３アルキルである。 In embodiments, R _H is of the formula

having [wherein
X ₁₁ is CH, _CRT or N, X ₁₀ is CH, _CRT or N, R _T is an optional R _H ring substitution as described above, R and R' are independently is hydrogen, a C1-C6 alkyl group, a C4-C10 cycloalkylalkyl group, an aryl group, a heterocyclyl group, or a heteroaryl group, each of which is optionally substituted]. In specific embodiments, R _T is one or more of hydrogen or C1-C3 alkyl substituted with halogen, OH, C1-C3 alkyl, C1-C3 alkoxy, or C3-C6 cycloalkyl. and R′ is hydrogen, C1-C3 alkyl, C1-C3 alkoxy, or C1-C3 alkyl substituted with C3-C6 cycloalkyl, and R is hydrogen, C1-C3 alkyl, C1 ~C3 alkoxy, or C1-C3 alkyl substituted with C3-C6 cycloalkyl.

In embodiments, R _H is selected from the formula in Scheme 3:
or R12-12, R12-13, R2-145, R12-15, R12-16, R12-17, R12-18, R12-19, R12-20, R12-21, R12-21 or R12-22 [p is 0] or R12-70 or R12-71 [p is 0].

の５員ヘテロ環式基から選択される
［式中、
Ｔ、Ｕ、Ｖ、及びＷは、Ｏ、Ｓ、Ｃ（Ｒ’’）（Ｒ’’）、Ｃ（Ｒ’’）－／、Ｃ（Ｒ’’）、Ｃ－／、Ｎ（Ｒ’’）、又はＮ－／から選択され、
基は、Ｔ、Ｕ、Ｖ、及びＷの選択に依存して１個又は２個の二重結合を含み、
Ｒ_Ｈ基は、式Ｉの化合物における－（Ｌ_２）ｙ－Ｚ－部分にＣ－／、Ｃ（Ｒ’’）－／、又はＮ－／を通して結合しており、
Ｒ’’は、Ｎ又はＣ上における任意選択の置換を示す］。 In embodiments, R _H has the general formula:

is selected from 5-membered heterocyclic groups of
T, U, V, and W are O, S, C(R'')(R''), C(R'')-/, C(R''), C-/, N(R''), or N-/,
the group contains one or two double bonds depending on the choice of T, U, V, and W;
the R _H group is attached to the —(L ₂ )yZ— moiety in the compound of Formula I through C—/, C(R″)—/, or N—/;
R'' indicates optional substitution on N or C].

の５員ヘテロ環式基から選択される
［式中、
Ｔは、Ｃ（Ｒ’’）、Ｃ－／、若しくはＮであり、又は
Ｕは、Ｏ、Ｓ、Ｃ（Ｒ’’）（Ｒ’’）、Ｃ（Ｒ’’）－／、Ｎ（Ｒ’’）、若しくはＮ－／であり、
Ｖは、ＣＲ’’、Ｃ－／、若しくはＮであり、
Ｗは、ＣＲ’’、Ｃ－／、Ｎであり、Ｒ_Ｈ基は、式Ｉの化合物における－（Ｌ_２）ｙ－Ｚ－部分にＣ－／、Ｃ（Ｒ’’）－／、又はＮ－／を通して結合しており、
Ｒ_Ｈ基は、式Ｉの化合物における－（Ｌ_２）ｙ－Ｚ－部分にＣ－／、Ｃ（Ｒ’’）－／、又はＮ－／を通して結合しており、
Ｒ’’は、Ｎ又はＣ上における任意選択の置換を示す］。記号「－／」は、ヘテロ環式基を本明細書の化合物において結合させている１価の結合を示す。例えば、Ｃ－／は、ヘテロ環式基を本明細書の化合物に結合させている、環炭素からの１価の結合を示す。 More specifically, R _H has the formula:

is selected from 5-membered heterocyclic groups of
T is C(R''), C-/, or N, or U is O, S, C(R'')(R''), C(R'')-/, N( R''), or N-/,
V is CR'', C-/, or N;
W is CR'', C-/, N, and the R _H group is C-/, C(R'')- _/ , or is connected through N-/,
the R _H group is attached to the —(L ₂ )yZ— moiety in the compound of Formula I through C—/, C(R″)—/, or N—/;
R'' indicates optional substitution on N or C]. The symbol "-/" indicates a monovalent bond joining heterocyclic groups in the compounds herein. For example, C-/ indicates a monovalent bond from the ring carbon that attaches the heterocyclic group to the compounds herein.

の縮合ヘテロ環式基である
［式中、
Ｕ、Ｖ及びＷは、Ｏ、Ｓ、Ｎ、Ｃ（Ｒ’’）（Ｒ’’）、Ｃ（Ｒ’’）－／、Ｃ（Ｒ’’）、Ｃ－／、Ｎ（Ｒ’’）、又はＮ－／から選択され、
Ｔ’、Ｕ’、Ｖ’及びＷは、Ｃ（Ｒ’’）、Ｃ－／、Ｎ（Ｒ’’）、又はＮ－／から選択され、
Ｒ_Ｈ基は、式Ｉの化合物における－（Ｌ_２）ｙ－Ｚ－部分に指示された環におけるＣ－／、Ｃ（Ｒ’’）－／、又はＮ－／を通して結合しており、
基は、Ｕ、Ｖ、及びＷの選択に依存して結合を含み、
Ｒ’’は、Ｎ又はＣ上における任意選択の置換を示す］。 In embodiments, R _H is of the formula:

is a fused heterocyclic group of [wherein
U, V and W are O, S, N, C(R'')(R''), C(R'')-/, C(R''), C-/, N(R'' ), or N-/,
T', U', V' and W are selected from C(R''), C-/, N(R''), or N-/;
the R _H group is attached to the -(L ₂ )yZ- moiety in the compounds of Formula I through C-/, C(R'')-/, or N-/ in the indicated ring;
The group contains a bond depending on the choice of U, V, and W;
R'' indicates optional substitution on N or C].

の縮合ヘテロ環式基である
［式中、
Ｕ及びＶは、Ｎ、Ｃ（Ｒ’’）、又はＣ－／から選択され、
Ｗは、Ｏ、Ｓ、Ｃ（Ｒ’’）（Ｒ’’）、Ｃ（Ｒ’’）－／、Ｎ（Ｒ’’）、又はＮ－／から選択され、
Ｔ’、Ｕ’、Ｖ’及びＷ’は、Ｃ（Ｒ’’）、Ｃ－／、Ｎ（Ｒ’’）、又はＮ－／から選択され、
Ｒ_Ｈ基は、式Ｉの化合物における－（Ｌ_２）ｙ－Ｚ－部分に指示された環におけるＣ－／、Ｃ（Ｒ’’）－／、又はＮ－／を通して結合しており、
Ｒ’’は、Ｎ又はＣ上における任意選択の置換を示す］。 More specifically, R _H has the formula:

is a fused heterocyclic group of [wherein
U and V are selected from N, C(R''), or C-/;
W is selected from O, S, C(R'')(R''), C(R'')-/, N(R''), or N-/;
T', U', V' and W' are selected from C(R''), C-/, N(R''), or N-/;
the R _H group is attached to the -(L ₂ )yZ- moiety in the compounds of Formula I through C-/, C(R'')-/, or N-/ in the indicated ring;
R'' indicates optional substitution on N or C].

R'' is each hydrogen, halogen, nitro, cyano, amino, mono- or di-C1-C3 alkyl-substituted amino, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6-cycloalkenyl, C1-C3 haloalkyl, C6-C12 aryl, C5-C12 heteroaryl, C3-C12 heterocyclyl, C1-C3 alkoxy, C1-C6 acyl, -COOR _E , -OCOR _E , -CONR _E R _F , -OCONR _E R _D , —NR _E COR _F , —SR _E , _—SORE , —SO ₂ R _E , and —SO ₂ NR _E R _F and are independently selected from alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, alkoxy and acyl are optionally substituted,
R _E and R _F are each hydrogen, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6-cycloalkenyl, C1-C3 haloalkyl, C6-C12 aryl, C5-C12 heteroaryl, C3 -C12 heterocyclyl, C1-C3 alkoxy, C1-C6 acyl, each of these groups being selected from one or more halogen, nitro, cyano, amino, mono- or di-C1-C3 alkyl-substituted amino, C1- C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6-cycloalkenyl, C1-C3 haloalkyl, C6-C12 aryl, C5-C12 heteroaryl, C3-C12 heterocyclyl, C1-C3 alkoxy, and C1- Optionally substituted with C6 acyl.

のいずれか１つから選択される：
［式中、
Ｒ_Ｔは、上記の任意選択のＲ_Ｈ環置換であり、Ｒ及びＲ’は、独立的に水素、Ｃ１～Ｃ６アルキル基、Ｃ４～Ｃ１０シクロアルキルアルキル基、アリール基、ヘテロシクリル基、又はヘテロアリール基であり、これらの基のそれぞれは、任意選択で置換されている］。具体的な実施形態において、Ｒ_Ｔは、水素又はハロゲン、ＯＨ、Ｃ１～Ｃ３アルキル、Ｃ１～Ｃ３アルコキシ、若しくはＣ３～Ｃ６シクロアルキルで置換されているＣ１～Ｃ３アルキルの１つ若しくは複数での置換であり、Ｒ’は、水素、Ｃ１～Ｃ３アルキル、Ｃ１～Ｃ３アルコキシ、又はＣ３～Ｃ６シクロアルキルで置換されているＣ１～Ｃ３アルキルであり、Ｒは、水素、Ｃ１～Ｃ３アルキル、Ｃ１～Ｃ３アルコキシ、又はＣ３～Ｃ６シクロアルキルで置換されているＣ１～Ｃ３アルキルである。具体的な実施形態において、Ｒ及びＲ’は、独立的に水素、Ｃ１～Ｃ３アルキル又はＣ４－Ｃ７シクロアルキルアルキルである。具体的な実施形態において、Ｒ_Ｔは、水素又は１つのハロゲン、特にＢｒでの置換を表す。 In a more specific embodiment, _RH is

is selected from one of:
[In the formula,
R _T is an optional R _H ring substitution as described above and R and R′ are independently hydrogen, C1-C6 alkyl, C4-C10 cycloalkylalkyl, aryl, heterocyclyl, or heteroaryl groups, each of which is optionally substituted]. In specific embodiments, R _T is hydrogen or substituted with one or more of C1-C3 alkyl substituted with halogen, OH, C1-C3 alkyl, C1-C3 alkoxy, or C3-C6 cycloalkyl and R′ is hydrogen, C1-C3 alkyl, C1-C3 alkoxy, or C1-C3 alkyl substituted with C3-C6 cycloalkyl, and R is hydrogen, C1-C3 alkyl, C1-C3 Alkoxy, or C1-C3 alkyl substituted with C3-C6 cycloalkyl. In specific embodiments, R and R' are independently hydrogen, C1-C3 alkyl or C4-C7 cycloalkylalkyl. In a specific embodiment, R _T represents hydrogen or substitution with one halogen, especially Br.

In embodiments, R _H is 1-3 nitrogens in the ring, 1 or 2 oxygens in the ring, sulfur or both, or 1 or 2 nitrogens and 1 oxygen in the ring or An optionally substituted 6-membered heterocyclic or heteroaryl group containing sulfur, wherein the optional substitution is defined as in Formula I. Heterocyclic groups can be unsaturated, partially unsaturated or heteroaryl groups.

In embodiments, R _H is 1-5 nitrogens in a fused ring, 1-3 oxygen, sulfur or both in a fused ring or 1-4 nitrogens and 1 or 2 in a fused ring optionally substituted fused heterocyclic or heteroaryl group having two fused 6-membered rings with oxygen, sulfur or both, wherein optional substitution is defined as in Formula I be. In more specific embodiments, fused rings have 1, 2, 3, or 4 nitrogens in the fused ring. In more specific embodiments, fused rings have 1 or 2 oxygens or sulfur in the fused ring. In more specific embodiments, fused rings have 1 or 2 nitrogens and one oxygen or sulfur in the fused ring. A fused heterocyclic group can be an unsaturated, partially unsaturated or heteroaryl group.

In specific embodiments, R _H groups are selected from phenyl, oxazinyl, pyridinyl, pyrimidinyl, shinli, pyranyl, thiazinyl, 4H-pyranyl, naphthyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl, pteridinyl, purinyl and chromanyl; _{The H} group is attached to the -(L ₂ )yZ- moiety in compounds of Formula I at any available ring position. In specific embodiments, the R _H group is attached to the -(L ₂ )yZ- moiety in compounds of formula I at a carbon in the ring.

In specific embodiments of Formula I, -Z-(L ₂ )y-R _H is a group other than -NH-SO ₂ -R _W , and R _W is mes-trimethylphenyl, 4-methylphenyl , 4-trifluoromethylphenyl, naphthyl, 2,3,4,5,-tetramethylphenyl, 4-methoxyphenyl, 4-tert-butylphenyl, 2,4-dimethoxyphenyl, 2,5-dimethoxyphenyl or 4 - is phenoxyphenyl. In specific embodiments of Formula I, -Z-(L ₂ )y- is a moiety other than -NR _X -SO ₂ -, and R _X is H, hydrogen, methyl acetate, acetate, aminoacetyl, 4-benzyl formate, 4-isopropylbenzyl, 4-chlorobenzyl or 4-methoxybenzyl. In embodiments of Formula I, -Z- is other than -NR _X -SO ₂ - and R _X is H, hydrogen, methyl acetate, acetate, aminoacetyl, 4-benzyl formate, 4-isopropylbenzyl, 4 - chlorobenzyl or 4-methoxybenzyl.

In embodiments of Formula I, R _H is other than a phenyl group or an optionally substituted phenyl group. In an embodiment of Formula I, _RH is a heterocyclic group substituted with a single halogen, especially Br.

In embodiments of Formula I, R _P or —N(R ₂ )(R ₃ ) are optionally substituted amine groups R _N 1 through R _N 31 depicted in Scheme 2. Exemplary optional substitutions of groups are illustrated in Scheme 2. The depicted R substituents may be placed at any available ring position. In part of Scheme 2, preferred alkyl is C1-C3 alkyl, acyl includes formyl, preferred acyl is C1-C6 acyl, more preferably acetyl, and hydroxyalkyl is C1-C6. C6 hydroxyalkyl, preferably -CH ₂ -CH ₂ -OH, preferred alkyl for amine group is C1-C3 alkyl, preferred alkyl for -SO ₂ alkyl is C1-C3 alkyl, Methyl is more preferred.

In specific embodiments of Formula I, —N(R ₂ )(R ₃ ) is R _N 1. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 3. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 2 or R _N 4. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 5 or R _N 6. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 7 or R _N 8. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 9. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 10. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 11. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 12. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 13. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 14. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 15. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 16. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 17 or R _N 18. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 19 or R _N 20. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 21. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 22. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 23 or R _N 24. In specific embodiments, —N(R ₂ )(R ₃ ) is R _N 25. In embodiments, —N(R ₂ )(R ₃ ) is R _N 1, R _N 2, R _N 3, R _N 4, R _N 11, R _N 13, or R _N 14. In embodiments, —N(R ₂ )(R ₃ ) are R _N 26-R _N 29. In embodiments, -N(R ₂ )(R ₃ ) is R _N 30. In embodiments, -N(R ₂ )(R ₃ ) is R _N 31.

In embodiments of Formula I, R _{1 H} are moieties R12-1 through R12-69 depicted in Scheme 3. In embodiments, R 1 _H is from R12-35 to R12-42. In embodiments, R 1 _H is any of R12-43 to R12-69. In embodiments, R 1 _H is any of R12-43 to R12-45. In embodiments, R 1 _H is any of R12-46 to R12-48. In embodiments, R 1 _H is any of R12-49 through R12-51. In embodiments, R 1 _H is any of R12-52 to R12-54. In embodiments, R 1 _H is any of R12-55 to R12-58. In embodiments, R 1 _H is any of R12-59 to R12-62. In embodiments, R 1 _H is any of R12-63 to R12-66. In embodiments, R 1 _H is any of R12-67 to R12-69. In part of Scheme 3, preferred alkyl groups are C1-C6 alkyl groups, or more preferred C1-C3 alkyl groups, preferred halogens are F, Cl and Br, and acyl includes formyl, which is preferred. Acyl is -CO-C1-C6 alkyl, more preferably acetyl, and phenyl optionally substituted with one or more halogen, alkyl or acyl.

の化合物又はそれらの塩若しくは溶媒和物が挙げられる
［式中、変数は、式Ｉに定義される通りであり、点線は、単結合又は二重結合を表す］。実施形態において、ｘは１であり、ｙは１である。実施形態において、Ｘは両方とも窒素である。実施形態において、Ｒ_Ｐは－Ｎ（Ｒ_２）（Ｒ_３）である。実施形態において、Ｌ_１及びＬ_２は－（ＣＨ_２）ｎ－であり、ｎは、独立的には１、２又は３である。実施形態において、Ｒ_Ｈは、ヘテロ環式又はヘテロアリール基である。実施形態において、Ｙは、－Ｎ（Ｒ_１）－、－ＣＯＮ（Ｒ_１）－、又は－Ｎ（Ｒ_１）ＣＯ－である。実施形態において、Ｚは、－ＣＯＮ（Ｒ’）－又は－Ｎ（Ｒ’）ＣＯ－である。実施形態において、Ｒ’は、水素、Ｃ１～Ｃ３アルキル又はＣ１～Ｃ３ハロアルキルである。実施形態において、Ｒ’は、水素、メチル又はトリフルオロメチルである。実施形態において、Ｒ_Ａは、水素、ハロゲン、Ｃ１～Ｃ３アルキル、Ｃ１～Ｃ３アルコキシル、Ｃ１～Ｃ３アシル、又はＣ１～Ｃ３ハロアルキルである。実施形態において、Ｒ’は、水素、メチル、メトキシ又はトリフルオロメチルである。実施形態において、Ｒ_４及びＲ_５は、水素、ハロゲン、Ｃ１～Ｃ３アルキル、Ｃ１～Ｃ３アルコキシル、又はＣ１～Ｃ３ハロアルキルから選択される。実施形態において、Ｒ_４とＲ_５は一緒になって、飽和、部分不飽和又はヘテロ芳香族の、５員又は６員の炭素環式又はヘテロ環式環を形成する。実施形態において、Ｒ_Ｈは、ＲＨ１～ＲＨ１２のいずれか１つである。 In specific embodiments, compounds useful in the methods herein include Formula II:

or salts or solvates thereof [wherein the variables are as defined in Formula I and the dotted line represents a single or double bond]. In embodiments, x is 1 and y is 1. In embodiments, both X are nitrogen. In embodiments, R _P is -N(R ₂ )(R ₃ ). In embodiments, L ₁ and L ₂ are -(CH ₂ )n-, where n is independently 1, 2 or 3. In embodiments, R _H is a heterocyclic or heteroaryl group. In embodiments, Y is -N(R ₁ )-, -CON(R ₁ )-, or -N(R ₁ )CO-. In embodiments, Z is -CON(R')- or -N(R')CO-. In embodiments, R' is hydrogen, C1-C3 alkyl or C1-C3 haloalkyl. In embodiments, R' is hydrogen, methyl or trifluoromethyl. In embodiments, R _A is hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, C1-C3 acyl, or C1-C3 haloalkyl. In embodiments, R' is hydrogen, methyl, methoxy or trifluoromethyl. In embodiments, R ₄ and R ₅ are selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, or C1-C3 haloalkyl. In embodiments, R ₄ and R ₅ together form a saturated, partially unsaturated or heteroaromatic 5- or 6-membered carbocyclic or heterocyclic ring. In embodiments, _RH is any one of RH1-RH12.

の化合物又はそれらの塩若しくは溶媒和物が挙げられる
［式中、変数は、式Ｉに定義される通りであり、点線は、単結合又は二重結合を表す］。 In specific embodiments, compounds useful in the methods herein include Formula III:

or salts or solvates thereof [wherein the variables are as defined in Formula I and the dotted line represents a single or double bond].

In embodiments, y is one. In embodiments, y is zero. In embodiments, both X are nitrogen. In embodiments, R _P is -N(R ₂ )(R ₃ ). In embodiments, L ₂ is -(CH ₂ ) _n -, where n is 1, 2 or 3. In embodiments, R _H is a heterocyclyl or heteroaryl group. In embodiments, Y is -N(R ₁ )-, -CON(R ₁ )-, or -N(R ₁ )CO-. In embodiments, Z is -CON(R')- or -N(R')CO-. In embodiments, R' is hydrogen, C1-C3 alkyl or C1-C3 haloalkyl. In embodiments, R' is hydrogen, methyl or trifluoromethyl. In embodiments, R _A is hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, C1-C3 acyl, or C1-C3 haloalkyl. In embodiments, R' is hydrogen, methyl, methoxy or trifluoromethyl. In embodiments, R ₄ and R ₅ are selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, or C1-C3 haloalkyl. In embodiments, R ₄ and R ₅ together form a saturated, partially unsaturated or heteroaromatic 5- or 6-membered carbocyclic or heterocyclic ring. In embodiments, _RH is any one of RH1-RH12.

の化合物又はそれらの塩若しくは溶媒和物が挙げられる
［式中、変数は、式Ｉに定義される通りであり、点線は、単結合又は二重結合を表す］。 In specific embodiments, compounds useful in the methods herein include Formula IV:

or salts or solvates thereof [wherein the variables are as defined in Formula I and the dotted line represents a single or double bond].

In embodiments, y is one. In embodiments, y is zero. In embodiments, both X are nitrogen. In embodiments, R _P is -N(R ₂ )(R ₃ ). In embodiments, L ₂ is -(CH ₂ ) _n -, where n is 1, 2 or 3. In embodiments, R _H is a heterocyclyl or heteroaryl group. In embodiments, R ₁ is hydrogen. In embodiments, R ₁ is hydrogen, methyl or trifluoromethyl. In embodiments, Z is -CON(R')- or -N(R')CO-. In embodiments, R' is hydrogen, C1-C3 alkyl or C1-C3 haloalkyl. In embodiments, R' is hydrogen, methyl or trifluoromethyl. In embodiments, R _A is hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, C1-C3 acyl, or C1-C3 haloalkyl. In embodiments, R' is hydrogen, methyl, methoxy or trifluoromethyl. In embodiments, R ₄ and R ₅ are selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, or C1-C3 haloalkyl. In embodiments, R ₄ and R ₅ together form a saturated, partially unsaturated or heteroaromatic 5- or 6-membered carbocyclic or heterocyclic ring. In embodiments, _RH is any one of RH1-RH12.

の化合物又はそれらの塩若しくは溶媒和物が挙げられる
［式中、変数は、式Ｉに定義される通りであり、点線は、単結合又は二重結合を表す］。 In specific embodiments, compounds useful in the methods herein include Formula V:

or salts or solvates thereof [wherein the variables are as defined in Formula I and the dotted line represents a single or double bond].

In embodiments, y is one. In embodiments, y is zero. In embodiments, both X are nitrogen. In embodiments, R _P is -N(R ₂ )(R ₃ ). In embodiments, L ₂ is -(CH ₂ ) _n -, where n is 1, 2 or 3. In embodiments, R _H is a heterocyclyl or heteroaryl group. In embodiments, R ₁ is hydrogen. In embodiments, R ₁ is hydrogen, methyl or trifluoromethyl. In embodiments, Rs is hydrogen, C1-C3 alkyl, optionally substituted C1-C3 alkyl, or aryl. In embodiments, R _A is hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, C1-C3 acyl, or C1-C3 haloalkyl. In embodiments, R' is hydrogen, methyl, methoxy or trifluoromethyl. In embodiments, R ₄ and R ₅ are selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, or C1-C3 haloalkyl. In embodiments, R ₄ and R ₅ together form a saturated, partially unsaturated or heteroaromatic 5- or 6-membered carbocyclic or heterocyclic ring. In embodiments, _RH is any one of RH1-RH12.

の化合物又はそれらの塩若しくは溶媒和物が挙げられる
［式中、変数は、式Ｉに定義される通りであり、点線は、単結合又は二重結合を表す］。 In specific embodiments, compounds useful in the methods herein include Formula VI:

or salts or solvates thereof [wherein the variables are as defined in Formula I and the dotted line represents a single or double bond].

In embodiments, y is one. In embodiments, y is zero. In embodiments, both X are nitrogen. In embodiments, x is 1 and -N-(CH ₂ )n- and n is 1, 2 or 3. In embodiments, y is zero. In embodiments, L ₂ is -(CH ₂ ) _n -, where n is 1, 2 or 3. In embodiments, R _H is a heterocyclyl or heteroaryl group. In embodiments, R ₁ is hydrogen. In embodiments, R ₁ is hydrogen, methyl or trifluoromethyl. In embodiments, Rs is hydrogen, C1-C3 alkyl, optionally substituted C1-C3 alkyl, or aryl. In embodiments, R _A is hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, C1-C3 acyl, or C1-C3 haloalkyl. In embodiments, R' is hydrogen, methyl, methoxy or trifluoromethyl. In embodiments, R ₄ and R ₅ are selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, or C1-C3 haloalkyl. In embodiments, R ₄ and R ₅ together form a saturated, partially unsaturated or heteroaromatic 5- or 6-membered carbocyclic or heterocyclic ring. In embodiments, _RH is any one of RH1-RH12.

の化合物又はそれらの塩若しくは溶媒和物が挙げられる
［式中、変数は、式Ｉに定義される通りであり、点線は、単結合又は二重結合を表す］。 In specific embodiments, compounds useful in the methods herein include Formula VII:

or salts or solvates thereof [wherein the variables are as defined in Formula I and the dotted line represents a single or double bond].

In embodiments, y is one. In embodiments, y is zero. In embodiments, both X are nitrogen. In embodiments, x is 1 and -N-(CH ₂ )n- and n is 1, 2 or 3. In embodiments, y is zero. In embodiments, L ₂ is -(CH ₂ ) _n -, where n is 1, 2 or 3. In embodiments, R _H is a heterocyclyl or heteroaryl group. In embodiments, R ₁ is hydrogen. In embodiments, R ₁ is hydrogen, methyl or trifluoromethyl. In embodiments, Rs is hydrogen, C1-C3 alkyl, optionally substituted C1-C3 alkyl, or aryl. In embodiments, R _A is hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, C1-C3 acyl, or C1-C3 haloalkyl. In embodiments, R' is hydrogen, methyl, methoxy or trifluoromethyl. In embodiments, R ₄ and R ₅ are selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, or C1-C3 haloalkyl. In embodiments, R ₄ and R ₅ together form a saturated, partially unsaturated or heteroaromatic 5- or 6-membered carbocyclic or heterocyclic ring. In embodiments, _RH is any one of RH1-RH12.

の化合物又はそれらの塩若しくは溶媒和物が挙げられる
［式中、変数は、式Ｉに定義される通りであり、点線は、単結合又は二重結合を表す］。Ｒ_６～Ｒ_９は、水素及び式Ｉにおいて定義されるＲ_Ａ基から独立的に選択される。Ｒ_Ｍは、縮合環上の任意選択の置換を表し、Ｒ_Ｍは、式ＩにおけるＲ_Ａの値をとる。 In specific embodiments, compounds useful in the methods herein include Formula VIII:

or salts or solvates thereof [wherein the variables are as defined in Formula I and the dotted line represents a single or double bond]. R ₆ -R ₉ are independently selected from hydrogen and R _A groups as defined in Formula I; _RM represents optional substitutions on the fused ring and _RM takes the value of _RA in Formula I.

In embodiments, y is one. In embodiments, y is zero. In embodiments, both X are nitrogen. In embodiments, x is 1 and -N-(CH ₂ )n- and n is 1, 2 or 3. In embodiments, x is zero. In embodiments, L ₂ is -(CH ₂ ) _n -, where n is 1, 2 or 3. In embodiments, R ₁ is hydrogen. In embodiments, R ₁ is hydrogen, methyl or trifluoromethyl. In embodiments, R ₇ -R ₉ are independently selected from hydrogen, C1-C3 alkyl, optionally substituted C1-C3 alkyl, or aryl. In embodiments, R ₇ -R ₉ are independently selected from hydrogen, halogen C1-C3 alkyl, C1-C3 alkoxyl, C1-C3 acyl, or C1-C3 haloalkyl. In embodiments, R ₇ -R ₉ are all hydrogen. In embodiments, R ₄ and R ₅ are selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, or C1-C3 haloalkyl. In embodiments, R ₄ and R ₅ together form a saturated, partially unsaturated or heteroaromatic 5- or 6-membered carbocyclic or heterocyclic ring. In embodiments, R 1 _M is one or more of hydrogen, halogen, C1-C3 alkyl groups, C4-C7 cycloalkylalkyl groups, or C1-C3 haloalkyl groups. In embodiments, _RM is one or more of hydrogen, halogen, especially Br, methyl or trifluoromethyl. In embodiments, _RM is hydrogen.

の化合物又はそれらの塩若しくは溶媒和物が挙げられる
［式中、変数は、式Ｉに定義される通りであり、点線は、単結合又は二重結合を表す］。Ｒ_６～Ｒ_９は、水素及び式Ｉにおいて定義されるＲ_Ａ基から独立的に選択され、Ｒ_Ｍは、式Ｉにおいて定義される任意選択の置換を表す。 In specific embodiments, compounds useful in the methods herein include Formula IX:

or salts or solvates thereof [wherein the variables are as defined in Formula I and the dotted line represents a single or double bond]. R ₆ -R ₉ are independently selected from hydrogen and R _A groups as defined in Formula I, wherein R _M represents optional substitution as defined in Formula I;

In embodiments, y is one. In embodiments, y is zero. In embodiments, both X are nitrogen. In embodiments, x is 1, M is -N-(CH ₂ )n-, and n is 1, 2 or 3. In embodiments, x is 1, M is -(CH ₂ ) _n -, and n is 1, 2 or 3. In embodiments, x is 0. In embodiments, L ₂ is -(CH ₂ ) _n -, where n is 1, 2 or 3. In embodiments, R ₁ is hydrogen. In embodiments, R ₁ is hydrogen, methyl or trifluoromethyl. In embodiments, R ₇ -R ₉ are independently selected from hydrogen, C1-C3 alkyl, optionally substituted C1-C3 alkyl, or aryl. In embodiments, R ₇ -R ₉ are independently selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, C1-C3 acyl, or C1-C3 haloalkyl. In embodiments, R ₇ -R ₉ are all hydrogen. In embodiments, R ₄ and R ₅ are selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, or C1-C3 haloalkyl. In embodiments, R ₄ and R ₅ together form a saturated, partially unsaturated or heteroaromatic 5- or 6-membered carbocyclic or heterocyclic ring. In embodiments, R 1 _M is hydrogen, halogen, C1-C3 alkyl group or C1-C3 haloalkyl group. In embodiments, _RM is hydrogen, halogen, especially Br, methyl or trifluoromethyl.

の化合物又はその塩若しくは溶媒和物を提供する
［式中、
Ｘはそれぞれ、Ｎ又はＣＨから独立的に選択され、少なくとも１つのＸはＮであり、
Ａ環は、３～１０個の炭素原子を有し、１～６個のヘテロ原子を任意選択で有し、任意選択で飽和、不飽和又は芳香族である、炭素環式又はヘテロ環式環であり、
Ｌ_１は、任意選択で置換されている、任意選択の１～３個の炭素リンカーであり、ｘは、０又は１であって、Ｌ_１の有無を示し、
Ｒ_１は、水素、アルキル基、アルケニル基、シクロアルキル基、シクロアルケニル基、ヘテロシクリル基、又はアリール基からなる群から選択され、これらの基のそれぞれは、任意選択で置換されており、
Ｒ_２及びＲ_３は、水素、アルキル、アルケニル、シクロアルキル、シクロアルケニル、アリール又はヘテロシクリルから独立的に選択され、これらの基のそれぞれは、任意選択で置換されており、或いは
Ｒ_２とＲ_３は一緒になって、任意選択で置換されている５員～８員の飽和、部分不飽和又は芳香族環のヘテロ環式環を形成し、
Ｒ_４及びＲ_５は、水素、ハロゲン、アルキル、アルケニル、シクロアルキル、シクロアルケニル、アリール又はヘテロシクリルから独立的に選択され、これらの基のそれぞれは、任意選択で置換されており、或いは
Ｒ_４とＲ_５は一緒になって、１個若しくは２個の二重結合を任意選択で含み、又は芳香族であり、１～３個のヘテロ原子を任意選択で含む、任意選択で置換されている５員又は６員環を形成し、
点線は、Ｒ_４及びＲ_５の選択に依存して単結合又は二重結合であり、
Ｒ_Ａは、水素又は指示された環上の１～１０個の置換基を表し、Ｒ_Ａ置換基は、水素、ハロゲン、ヒドロキシル、シアノ、ニトロ、アミノ、一置換若しくは二置換アミノ、アルキル、アルケニル、シクロアルキル、シクロアルケニル、アリール、ヘテロシクリル、－ＯＲ_１５、－ＣＯＲ_１５、－ＣＯＯＲ_１５、－ＯＣＯＲ_１５、－ＣＯ－ＮＲ_１６Ｒ_１７、－ＯＣＯＮＲ_１６Ｒ_１７、－ＮＲ_１６－ＣＯ－Ｒ_１５、－ＳＲ_１５、－ＳＯＲ_１５、－ＳＯ_２Ｒ_１５、－ＳＯ_２－ＮＲ_１６Ｒ_１７、Ｒ１０、－ＮＨ－ＣＯ－（Ｌ_２）_ｙ－Ｒ_１２、又は－ＮＨ－ＣＯ－（Ｌ_２）_ｙ－Ｒ_１２から独立的に選択され、Ｌ_２は、任意選択の１～６個の炭素原子のリンカー基であり、そのリンカーは任意選択で置換されており、リンカーの１個又は２個の炭素は、Ｏ又はＳで任意選択で置き換えられ、ｙは、０又は１であって、Ｌ_２の有無を示し、
Ｒ_１０は、アルキル、アルケニル、シクロアルキル、シクロアルケニル、ヘテロシクリル、又はアリールから選択され、これらの基のそれぞれは、１個又は複数のハロゲン、アルキル、アルケニル、ハロアルキル、アルコキシ、アリール、ヘテロアリール、ヘテロシクリル、アリール置換アルキル、又はヘテロシクリル置換アルキルで任意選択で置換されており、
Ｒ_１２は、シクロアルキル、シクロアルケニル、ヘテロシクリル、ヘテロアリール、若しくはアリールから選択され、これらの基のそれぞれは、任意選択で置換されており、又はＲ_１２は、シクロアルキル、シクロアルケニル、ヘテロシクリル、ヘテロアリール、又はアリールで置換されているＣ１～Ｃ３アルキルであり、これらのそれぞれは、任意選択で置換されており、任意選択の置換は、１個又は複数のハロゲン、アルキル、アルケニル、ハロアルキル、アルコキシ、アリール、ヘテロアリール、又はヘテロシクリルであり、
Ｒ_１５はそれぞれ、水素、アルキル、アルケニル、シクロアルキル、シクロアルケニル、アリール又はヘテロシクリル、アリールアルキル（アリールで置換されているアルキル基）及びヘテロシクリルアルキル、シクロアルキルアルキル、シクロアルケニルアルキルから独立的に選択され、これらの基のそれぞれは、任意選択で置換されており、
Ｒ_１６及びＲ_１７はそれぞれ、水素、アルキル、アルケニル、シクロアルキル、シクロアルケニル、アリール又はヘテロシクリル、アリールアルキル（アリールで置換されているアルキル基）及びヘテロシクリルアルキル、シクロアルキルアルキル、シクロアルケニルアルキルから独立的に選択され、これらの基のそれぞれは、任意選択で置換されており、
任意選択の置換は、１個又は複数のハロゲン、ニトロ、シアノ、アミノ、モノ又はジ－Ｃ１～Ｃ３アルキル置換アミノ、Ｃ１～Ｃ３アルキル、Ｃ２～Ｃ４アルケニル、Ｃ３～Ｃ６シクロアルキル、Ｃ３～Ｃ６－シクロアルケニル、Ｃ６～Ｃ１２アリール、及びＣ６～Ｃ１２ヘテロシクリルでの置換を含む］。 In another embodiment, the present invention provides a compound of Formula XI:

or a salt or solvate thereof [wherein
each X is independently selected from N or CH, at least one X is N;
Ring A is a carbocyclic or heterocyclic ring having 3 to 10 carbon atoms, optionally having 1 to 6 heteroatoms, optionally saturated, unsaturated or aromatic and
L ₁ is an optionally substituted 1-3 carbon linker, x is 0 or 1 to indicate the presence or absence of L ₁ ,
R ₁ is selected from the group consisting of hydrogen, alkyl groups, alkenyl groups, cycloalkyl groups, cycloalkenyl groups, heterocyclyl groups, or aryl groups, each of which groups is optionally substituted;
R ₂ and R ₃ are independently selected from hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl or heterocyclyl, each of these groups optionally substituted or R ₂ and R ₃ together form an optionally substituted 5- to 8-membered saturated, partially unsaturated or aromatic heterocyclic ring;
R ₄ and R ₅ are independently selected from hydrogen, halogen, alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl or heterocyclyl, each of which groups are optionally substituted _or R ₅ taken together optionally contains 1 or 2 double bonds, or is aromatic and optionally contains 1-3 heteroatoms, optionally substituted 5 forming a membered or six-membered ring,
the dotted line is a single or double bond depending on the choice of _R4 and _R5 ;
R _A represents hydrogen or 1 to 10 substituents on the indicated ring, where R _A substituents are hydrogen, halogen, hydroxyl, cyano, nitro, amino, mono- or disubstituted amino, alkyl, alkenyl , cycloalkyl, cycloalkenyl, aryl, heterocyclyl, -OR ₁₅ , -COR ₁₅ , -COOR ₁₅ , -OCOR ₁₅ , -CO-NR ₁₆ R ₁₇ , -OCONR ₁₆ R ₁₇ , -NR ₁₆ -CO-R ₁₅ , -SR ₁₅ , -SOR ₁₅ , -SO ₂ R ₁₅ , -SO ₂ -NR ₁₆ R ₁₇ , R10, -NH-CO-(L ₂ ) _y -R ₁₂ , or -NH-CO-(L ₂ ) _y independently selected from —R ₁₂ , L ₂ is an optional linker group of 1 to 6 carbon atoms, the linker optionally substituted, and 1 or 2 carbons of the linker is optionally replaced with O or S, y is 0 or 1, indicating the presence or absence of _L2 ,
R ₁₀ is selected from alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocyclyl, or aryl, each of these groups being one or more of halogen, alkyl, alkenyl, haloalkyl, alkoxy, aryl, heteroaryl, heterocyclyl optionally substituted with , aryl-substituted alkyl, or heterocyclyl-substituted alkyl;
R ₁₂ is selected from cycloalkyl, cycloalkenyl, heterocyclyl, heteroaryl, or aryl, each of which groups are optionally substituted, or R ₁₂ is cycloalkyl, cycloalkenyl, heterocyclyl, hetero aryl, or C1-C3 alkyl substituted with aryl, each of which is optionally substituted, where optional substitution is one or more of halogen, alkyl, alkenyl, haloalkyl, alkoxy, aryl, heteroaryl, or heterocyclyl,
each R ₁₅ is independently selected from hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl or heterocyclyl, arylalkyl (an alkyl group substituted with aryl) and heterocyclylalkyl, cycloalkylalkyl, cycloalkenylalkyl; , each of these groups is optionally substituted,
R ₁₆ and R ₁₇ are each independently hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl or heterocyclyl, arylalkyl (an alkyl group substituted with aryl) and heterocyclylalkyl, cycloalkylalkyl, cycloalkenylalkyl each of these groups is optionally substituted,
Optional substitutions are one or more of halogen, nitro, cyano, amino, mono- or di-C1-C3 alkyl substituted amino, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6- including substitution with cycloalkenyl, C6-C12 aryl, and C6-C12 heterocyclyl].

又はその塩若しくは溶媒和物を有する
［式中、変数は、式ＸＩに対して定義される通りである］。 In embodiments, the compound has Formula XII:

or a salt or solvate thereof, wherein the variables are as defined for Formula XI.

又はその塩若しくは溶媒和物を有する
［式中、変数は、式ＸＩにおいて定義される通りであり、
Ｙはそれぞれ、Ｎ又はＣＨから独立的に選択され、
Ｒ_Ｂは、水素又は指示された環上の１～１０個の置換基を表し、Ｒ_Ａ置換基は、水素、ハロゲン、ヒドロキシル、シアノ、ニトロ、アミノ、一置換若しくは二置換アミノ、アルキル、アルケニル、シクロアルキル、シクロアルケニル、アリール、ヘテロシクリル、－ＯＲ_１５、－ＣＯＲ_１５、－ＣＯＯＲ_１５、－ＯＣＯＲ_１５、－ＣＯ－ＮＲ_１６Ｒ_１７、－ＯＣＯＮＲ_１６Ｒ_１７、－ＮＲ_１６－ＣＯ－Ｒ_１５、－ＳＲ_１５、－ＳＯＲ_１５、－ＳＯ_２Ｒ_１５、－ＳＯ_２－ＮＲ_１６Ｒ_１７、又は－（Ｌ_２）_ｙ－Ｒ_１０から独立的に選択され、Ｌ_２は、任意選択の１～６個の炭素原子のリンカー基であり、そのリンカーは任意選択で置換されており、ｙは、０又は１であって、Ｌ_２の有無を示す］。 In embodiments, the compound is of Formula XIII:

or a salt or solvate thereof, wherein the variables are as defined in Formula XI;
each Y is independently selected from N or CH;
R _B represents hydrogen or 1 to 10 substituents on the indicated ring, where R _A substituents are hydrogen, halogen, hydroxyl, cyano, nitro, amino, mono- or disubstituted amino, alkyl, alkenyl , cycloalkyl, cycloalkenyl, aryl, heterocyclyl, -OR ₁₅ , -COR ₁₅ , -COOR ₁₅ , -OCOR ₁₅ , -CO-NR ₁₆ R ₁₇ , -OCONR ₁₆ R ₁₇ , -NR ₁₆ -CO-R ₁₅ , independently selected from -SR ₁₅ , -SOR ₁₅ , -SO ₂ R ₁₅ , -SO ₂ -NR ₁₆ R ₁₇ , or -(L ₂ ) _y -R ₁₀ where L ₂ is optionally 1-6 a linker group of 1 carbon atom, wherein the linker is optionally substituted, and y is 0 or 1 to indicate the presence or absence of _L2 ].

又はその塩若しくは溶媒和物を有する
［式中、変数は、式ＸＩ、ＸＩＩ又はＸＩＩＩにおいて定義される通りである］。 In embodiments, the compound has Formula XIV or XV:

or a salt or solvate thereof, wherein the variables are as defined in Formula XI, XII or XIII.

又はその塩若しくは溶媒和物を有する
［式中、変数は、式ＸＩ又はＸＶにおいて定義される通りであり、
Ｒ_１１及びＲ_１２は、水素、ハロゲン、アルキル基、アルケニル基、シクロアルキル基、シクロアルケニル基、又はヘテロシクリル基から独立的に選択され、これらの基のそれぞれは、任意選択で置換されている］。 In embodiments, the compound is of formula XVI or XVII:

or a salt or solvate thereof, wherein the variables are as defined in Formula XI or XV;
R ₁₁ and R ₁₂ are independently selected from hydrogen, halogen, alkyl groups, alkenyl groups, cycloalkyl groups, cycloalkenyl groups, or heterocyclyl groups, each of which is optionally substituted] .

又はその塩（若しくは溶媒和物）を有する
［式中、
Ｒ_１は、水素、アルキル基、アルケニル基、シクロアルキル基、シクロアルケニル基、ヘテロシクリル基、又はアリール基からなる群から選択され、これらの基のそれぞれは、任意選択で置換されており（置換を定義する必要がある）、
Ｒ_２とＲ_３は一緒になって、１個若しくは２個の二重結合を含むことができるか、又は芳香族とすることができる、任意選択で置換されている５員又は６員のヘテロ環式環を形成し、
Ｒ_４及びＲ_５は、水素、ハロゲン、アルキル基、アルケニル基、シクロアルキル基、シクロアルケニル基、又はヘテロシクリル基から独立的に選択され、これらの基のそれぞれは、任意選択で置換されており、或いは
Ｒ_４とＲ_５は一緒になって、１個若しくは２個の二重結合を含むことができるか、又は芳香族とすることができる、任意選択で置換されている５員又は６員ヘテロ環式環を形成し、
点線は、Ｒ_４及びＲ_５の選択に依存して単結合又は二重結合であり、
Ｒ_６～Ｒ_９は、水素、ハロゲン、アルキル基、アルケニル基、シクロアルキル基、シクロアルケニル基、又はヘテロシクリル基から独立的に選択され、これらの基のそれぞれは、任意選択で置換されており、
Ｌは、任意選択の１～６個の原子リンカー基であり、ｘは、１又は０であって、Ｌ基の有無を示し、
Ｒ_１０は、アルキル基、アルケニル基、シクロアルキル基、シクロアルケニル基、ヘテロシクリル基、又はアリール基から選択され、これらの基のそれぞれは、任意選択で置換されている］。 In embodiments, the compound is of Formula XVIII:

or having a salt (or solvate) thereof [wherein
R ₁ is selected from the group consisting of hydrogen, alkyl groups, alkenyl groups, cycloalkyl groups, cycloalkenyl groups, heterocyclyl groups, or aryl groups, each of which groups are optionally substituted (without substitution must be defined),
R ₂ and R ₃ taken together are an optionally substituted 5- or 6-membered hetero hetero which can contain 1 or 2 double bonds or can be aromatic forming a cyclic ring,
_R4 and _R5 are independently selected from hydrogen, halogen, alkyl groups, alkenyl groups, cycloalkyl groups, cycloalkenyl groups, or heterocyclyl groups, each of which groups are optionally substituted; or R ₄ and R ₅ together are an optionally substituted 5- or 6-membered hetero, which can contain 1 or 2 double bonds or can be aromatic; forming a cyclic ring,
the dotted line is a single or double bond depending on the choice of _R4 and _R5 ;
R ₆ -R ₉ are independently selected from hydrogen, halogen, alkyl groups, alkenyl groups, cycloalkyl groups, cycloalkenyl groups, or heterocyclyl groups, each of which groups are optionally substituted;
L is an optional 1-6 atom linker group, x is 1 or 0, indicating the presence or absence of the L group;
R ₁₀ is selected from an alkyl group, an alkenyl group, a cycloalkyl group, a cycloalkenyl group, a heterocyclyl group, or an aryl group, each of which is optionally substituted].

For example, L is a 2-6 atom linker group (eg, -CH ₂ -O-, -CH ₂ -CH ₂ -O-, -O-CH ₂ -, -O-CH ₂ -CH ₂ -, -CO-NH-, -NH-CO-, -CH ₂ -CO-NH-, -CH ₂ -CH ₂ -CO-NH-).

又はその塩（若しくは溶媒和物）を有する
［式中、
Ｒ_１～Ｒ_９は以上に定義した通りであり、点線は、Ｒ_４及びＲ_５の選択に依存している単結合又は二重結合を表し、
ｙは、０又は１～３に及ぶ（１と３を含む）整数であり、
Ｒ_１０は、アルキル基、アルケニル基、シクロアルキル基、シクロアルケニル基、ヘテロシクリル基、又はアリール基から選択され、これらの基のそれぞれは、任意選択で置換されている］。 In embodiments, the compound is of Formula XIX:

or having a salt (or solvate) thereof [wherein
R ₁ -R ₉ are as defined above, the dashed lines represent single or double bonds depending on the choice of R ₄ and R ₅ ;
y is 0 or an integer ranging from 1 to 3 (inclusive);
R ₁₀ is selected from an alkyl group, an alkenyl group, a cycloalkyl group, a cycloalkenyl group, a heterocyclyl group, or an aryl group, each of which is optionally substituted].

の化合物及びその塩又は溶媒和物である
［式中、Ｒ_１～Ｒ_９は、水素又は任意選択の置換基を表し、Ｒ_１０は、効力に関連していると考えられる部分であり、Ｒ_Ｎは、溶解度などの物理化学的特性に関連していると考えられる部分である］。実施形態において、Ｒ_５は、代謝安定性に関連していると考えられる、水素以外の置換基である。具体的な実施形態において、Ｒ_５は、ハロゲン、特にＦ又はＣｌ、Ｃ１～Ｃ３アルキル基、特にメチル基である。実施形態において、Ｒ_４は、水素以外の置換基であり、特にＣ１～Ｃ３アルキル基であり、さらに詳細にはメチル基である。具体的な実施形態において、Ｒ_５はＦであり、Ｒ_４はメチルである。実施形態において、Ｒ_６～Ｒ_９は、水素、Ｃ１～Ｃ３－アルキル、ハロゲン、ヒドロキシル、Ｃ１～Ｃ３アルコキシ、ホルミル、又はＣ１～Ｃ_３アシルから選択される。実施形態において、Ｒ_６～Ｒ_９のうちの１つ又は２つは、水素以外の部分である。実施形態において、Ｒ_６～Ｒ_９のうちの１つは、ハロゲン、特にフッ素である。具体的な実施形態において、Ｒ_６～Ｒ_９のすべては、水素である。実施形態において、Ｒ_Ｎは、アミノ部分－Ｎ（Ｒ_２）（Ｒ_３）である。具体的な実施形態において、Ｒ_Ｎは、任意選択で置換されているヘテロ環式基であり、別のヘテロ原子（Ｎ、Ｓ又はＯ）を任意選択で含む５員～７員環を有する。実施形態において、Ｒ_Ｎは、任意選択で置換されているピロリジン－１－イル、ピペリジン－１－イル、アゼパン－１－イル、ピペラジン－１－イル、又はモルホリノである。実施形態において、Ｒ_Ｎは、Ｃ１～Ｃ３アルキル、ホルミル、Ｃ１～Ｃ３アシル（特に、アセチル）、ヒドロキシル、ハロゲン（特に、Ｆ又はＣｌ）、ヒドロキシＣ１～Ｃ３アルキル（特に、－ＣＨ_２－ＣＨ_２－ＯＨ）から選択される１つの置換基で置換されている。実施形態において、Ｒ_Ｎは、非置換のピロリジン－１－イル、ピペリジン－１－イル、アゼパン－１－イル、ピペラジン－１－イル、又はモルホリノである。 In embodiments, the CDH1L inhibitor has Formula XX:

and salts or solvates thereof, wherein R ₁ -R ₉ represent hydrogen or optional substituents, R ₁₀ is a moiety believed to be associated with efficacy, and R _N is the moiety thought to be related to physicochemical properties such as solubility]. In embodiments, R ₅ is a substituent other than hydrogen that is believed to be relevant for metabolic stability. In specific embodiments, R ₅ is halogen, especially F or Cl, a C1-C3 alkyl group, especially a methyl group. In embodiments, R ₄ is a substituent other than hydrogen, particularly a C1-C3 alkyl group, more particularly a methyl group. In a specific embodiment, _R5 is F and _R4 is methyl. In embodiments, R ₆ -R ₉ are selected from hydrogen, C1-C3-alkyl, halogen, hydroxyl, C1-C3 alkoxy, formyl, or C1- _C3 acyl. In embodiments, one or two of R ₆ -R ₉ are moieties other than hydrogen. In embodiments, one of R ₆ -R ₉ is halogen, especially fluorine. In specific embodiments, all of R ₆ -R ₉ are hydrogen. In embodiments, R _N is an amino moiety -N(R ₂ )(R ₃ ). In specific embodiments, R _N is an optionally substituted heterocyclic group having a 5- to 7-membered ring optionally containing another heteroatom (N, S or O). In embodiments, R _N is optionally substituted pyrrolidin-1-yl, piperidin-1-yl, azepan-1-yl, piperazin-1-yl, or morpholino. In embodiments, R _N is C1-C3 alkyl, formyl, C1-C3 acyl (especially acetyl), hydroxyl, halogen (especially F or Cl), hydroxy C1-C3 alkyl (especially -CH ₂ -CH ₂ —OH). In embodiments, R _N is unsubstituted pyrrolidin-1-yl, piperidin-1-yl, azepan-1-yl, piperazin-1-yl, or morpholino.

In embodiments, R ₁₀ is -NRy-CO-(L ₂ )y-R ₁₂ or -CO-NRy--(L ₂ )y-R ₁₂ , y is 0 or 1, and any Indicates the presence or absence of _L2 , an optional 1-6 carbon atom linker group, wherein the linker is optionally substituted, wherein 1 or 2 carbons of the linker are O, NH, NRy or S and Ry is hydrogen or alkyl of 1-3 carbons and R ₁₂ is an aryl group, cycloalkyl group, heterocyclic group, or heteroaryl group, each of which is , optionally replaced. In embodiments, y is one. L ₂ is -(CH ₂ )p- and p is 0-3. In embodiments, R ₁₂ is thiophen-2-yl, thiophen-3-yl, furan-2-yl, furan-3-yl, pyrrol-2-yl, pyrrol-3-yl, oxazol-4-yl, oxazol-5-yl, oxazol-2-yl, indol-2-yl, indol-3-yl, benzofuran-2-yl, benzofuran-3-yl, benzo[b]thiophen-2-yl, benzo[b] thiophen-3-yl, isobenzofuran-1-yl, isoindol-1-yl, or benzo[c]thiophen-1-yl. In embodiments, R ₁ is hydrogen or methyl. In embodiments, R ₁₂ is thiophen-2-yl, furan-2-yl, pyrrol-2-yl, oxazol-4-yl, indol-2-yl, benzofuran-2-yl, or benzo[b]thiophene -2-yl. In embodiments, R ₁₂ is thiophen-2-yl or indol-2-yl. In embodiments, R ₁ is hydrogen or methyl.

In a more general embodiment of Formula XX,
R ₁ is selected from the group consisting of hydrogen, alkyl groups, alkenyl groups, cycloalkyl groups, cycloalkenyl groups, heterocyclyl groups, or aryl groups, each of which groups is optionally substituted;
R _N is —NR ₂ R 3 and R ₂ and _{R 3 are} independently selected from hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl or heterocyclyl, each of these groups optionally substituted or R ₂ and R ₃ taken together form a heterocyclic ring of an optionally substituted 5- to 8-membered saturated, partially unsaturated or aromatic ring;
R ₄ -R ₉ are hydrogen, halogen, hydroxyl, cyano, nitro, amino, mono- or dialkyl-substituted amino, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted cycloalkyl, optionally substituted cycloalkenyl, optionally substituted aryl, optionally substituted heterocyclyl, -OR ₁₅ , -COR ₁₅ , -COOR ₁₅ , -OCOR ₁₅ , -CO- independently selected from NR ₁₅ R ₁₆ , —OCONR ₁₅ R ₁₆ , —NR ₁₅ —CO—R ₁₆ , —SR ₁₅ , —SOR ₁₅ , —SO ₂ R ₁₅ and —SO ₂ —NR ₁₅ R ₁₆ ;
R ₁₀ is -NRy-CO-(L ₂ )y-R ₁₂ , -CO-NRy-(L ₂ )y-R ₁₂ and L ₂ is an optional 1-6 carbon atom linker wherein the linker is optionally substituted, wherein 1 or 2 carbons of the linker are optionally replaced with O or S, y is 0 or 1, the presence or absence of _L2 indicates
R ₁₂ is selected from cycloalkyl, cycloalkenyl, heterocyclyl, heteroaryl, or aryl, each of which groups are optionally substituted, or R ₁₂ is cycloalkyl, cycloalkenyl, heterocyclyl, hetero aryl, or C1-C3 alkyl substituted with aryl, each of which is optionally substituted, where optional substitution is one or more of halogen, alkyl, alkenyl, haloalkyl, alkoxy, aryl, heteroaryl, or heterocyclyl,
R ₁₅ and R ₁₆ are each independently selected from hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl or heterocyclyl, arylalkyl and heterocyclylalkyl, cycloalkylalkyl, cycloalkenylalkyl, each of these groups being , optionally replaced by
Optional substitutions are one or more of halogen, nitro, cyano, amino, mono- or di-C1-C3 alkyl substituted amino, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6- cycloalkenyl, C6-C12 aryl, C6-C12 heterocyclyl, -OR ₁₇ , -COR ₁₇ , -COOR ₁₇ , -OCOR ₁₇ , -CO-NR ₁₇ R ₁₈ , -OCONR ₁₇ R ₁₈ , -NR ₁₇ -CO-R ₁₈ , —SR ₁₇ , —SOR ₁₇ , —SO ₂ R ₁₇ , and —SO ₂ —NR ₁₇ R ₁₈ , wherein R ₁₇ and R ₁₈ are independently hydrogen or C ₁ -C ₆ alkyl. be.

In embodiments of Formula XX, R _N is an optionally substituted cyclic amine group selected from any of R _N 1-R _N 31 (Scheme 2). Exemplary optional substitutions of groups are illustrated in Scheme 2. The depicted R substituents may be placed at any available ring position. In part of Scheme 2, preferred alkyl is C1-C3 alkyl, acyl includes formyl, preferred acyl is C1-C6 acyl, more preferably acetyl, and hydroxyalkyl is C1-C6. C6 hydroxyalkyl, preferably -CH ₂ -CH ₂ -OH, preferred alkyl for amine group is C1-C3 alkyl, preferred alkyl for -SO ₂ alkyl is C1-C3 alkyl, Methyl is more preferred.

In specific embodiments of formula XX, R _N is R _N 1. In specific embodiments, R _N is R _N3 . In specific embodiments, _RN is _RN2 or _RN4 . In specific embodiments, _RN is _RN5 or _RN6 . In specific embodiments, _RN is _RN7 or _RN8 . In specific embodiments, _RN is _RN9 . In specific embodiments, R _N is R _N 10. In specific embodiments, R _N is R _N 11. In specific embodiments, R _N is R _N 12. In specific embodiments, R _N is R _N 13. In specific embodiments, R _N is R _N 14. In specific embodiments, R _N is R _N 15. In specific embodiments, R _N is R _N 16. In specific embodiments, R _N is R _N 17 or R _N 18. In specific embodiments, R _N is R _N 19 or R _N 20. In specific embodiments, R _N is R _N 21. In specific embodiments, R _N is R _N22 . In specific embodiments, _RN is _RN23 or _RN24 . In specific embodiments, R _N is R _N25 . In embodiments, R _N is R _N 1, R _N 2, R _N 3, R _N 4, R _N 11, R _N 13, or R _N 14. In embodiments, R _N is R _N 26-R _N 29. In embodiments, _RN is _RN30 . In embodiments, _RN is _RN31 .

In an embodiment of Formula XX, R ₁₂ is optionally substituted thienyl, thienylmethyl, furyl, furylmethyl, indolyl or methylindolyl. In embodiments, R ₁₂ is moieties R12-1 through R12-69 depicted in Scheme 3. In part of Scheme 3, preferred alkyl groups are C1-C6 alkyl groups, or more preferred C1-C3 alkyl groups, preferred halogens are F, Cl and Br, and acyl includes formyl, which is preferred. Acyl is -CO-C1-C6 alkyl, more preferably acetyl, and phenyl optionally substituted with one or more halogen, alkyl or acyl. In embodiments, R ₁₂ is a methyl, ethyl group or propyl substituted with moieties R12-1 through R12-22 depicted in Scheme 3. In embodiments, R ₁₂ is R12-1. In embodiments, R ₁₂ is R12-2. In embodiments, R ₁₂ is R12-3. In embodiments, R ₁₂ is R12-4. In embodiments, R ₁₂ is R12-5. In embodiments, R ₁₂ is R12-6. In embodiments, R ₁₂ is R12-7. In embodiments, R ₁₂ is R12-8. In embodiments, R ₁₂ is R12-9. In embodiments, R ₁₂ is R12-10. In embodiments, R ₁₂ is R12-11. In embodiments, R ₁₂ is R12-12. In embodiments, R ₁₂ is R12-13. In embodiments, R ₁₂ is R12-14. In embodiments, R ₁₂ is R12-15. In embodiments, R ₁₂ is R12-16. In embodiments, R ₁₂ is R12-17. In embodiments, R ₁₂ is R12-18. In embodiments, R ₁₂ is R12-19. In embodiments, R ₁₂ is R12-20. In embodiments, R ₁₂ is R12-21. In embodiments, R ₁₂ is R12-22. In embodiments, R ₁₂ is from R12-23 to R12-26. In embodiments, R ₁₂ is from R12-27 to R12-30. In embodiments, R ₁₂ is from R12-31 to R12-34. In embodiments, R ₁₂ is from R12-35 to R12-42. In embodiments, R12 is any of R12-43 to R12-69. In embodiments, R ₁₂ is a methyl, ethyl or propyl group substituted with moieties R12-43 through R12-69 depicted in Scheme 3. In embodiments, R12 is any of R12-43 to R12-45. In embodiments, R ₁₂ is a methyl, ethyl or propyl group substituted with moieties R12-43 to R12-45 depicted in Scheme 3. In embodiments, R12 is any of R12-46 to R12-48. In embodiments, R ₁₂ is a methyl, ethyl or propyl group substituted with moieties R12-46 to R12-48 depicted in Scheme 3. In embodiments, R12 is any of R12-49 to R12-51. In embodiments, R ₁₂ is a methyl, ethyl or propyl group substituted with moieties R12-49 through R12-51 depicted in Scheme 3. In embodiments, R12 is any of R12-52 to R12-54. In embodiments, R ₁₂ is a methyl, ethyl or propyl group substituted with moieties R12-52 to R12-54 depicted in Scheme 3. In embodiments, R12 is any of R12-55 to R12-58. In embodiments, R ₁₂ is a methyl, ethyl or propyl group substituted with moieties R12-55 to R12-58 depicted in Scheme 3. In embodiments, R12 is any of R12-59 to R12-62. In embodiments, R ₁₂ is a methyl, ethyl or propyl group substituted with moieties R12-59 through R12-62 depicted in Scheme 3. In embodiments, R12 is any of R12-63 to R12-66. In embodiments, R ₁₂ is a methyl, ethyl or propyl group substituted with moieties R12-63 to R12-66 depicted in Scheme 3. In embodiments, R12 is any of R12-67 to R12-69. In embodiments, R ₁₂ is a methyl, ethyl or propyl group substituted with moieties R12-67 to R12-69 depicted in Scheme 3. In embodiments, R12 is the moiety R12-70 or R12-71 depicted in Scheme 3.

In embodiments of Formula XX herein, R _N is an optionally substituted cyclic _amine group selected from any of R _N 1-R _N 25 (Scheme 2); is thienyl, thienylmethyl, furyl, furylmethyl, indolyl or methylindolyl. In an embodiment of Formula XX, R _N is R _N 1, R _N 2, R _N 3, R _N 4, R _N 11, R _N 13, R _N 14, or R _N 25, and R ₁₂ is thienyl. , thienylmethyl, furyl, furylmethyl, indolyl or methylindolyl. In embodiments, R ₁₀ is -NHCOR ₁₂ . In embodiments, R ₁₀ is -CONHR ₁₂ . In embodiments of Formula XX herein, R ₁₀ is —CO—NH—R ₁₂ , R _N is any one of R _N 1 through R _N 25, and R ₁₂ is R12-1 to R12-22. In embodiments of Formula XX herein, R ₁₀ is —CO—NH—R ₁₂ , R _N is any one of R _N 1 through R _N 25, and R ₁₂ is R12-1 any one of ~R12-69.

又はその塩（若しくは溶媒和物）を有する
［式中、
Ｒ_１は、水素、アルキル基、アルケニル基、シクロアルキル基、シクロアルケニル基、ヘテロシクリル基、又はアリール基からなる群から選択され、これらの基のそれぞれは、任意選択で置換されており（置換を定義する必要がある）、
Ｒ_２とＲ_３は一緒になって、１個若しくは２個の二重結合を含むことができるか、又は芳香族とすることができる、任意選択で置換されている５員又は６員ヘテロ環式環を形成し、
Ｒ_６～Ｒ_９は、水素、ハロゲン、アルキル基、アルケニル基、シクロアルキル基、シクロアルケニル基、又はヘテロシクリル基から独立的に選択され、これらの基のそれぞれは、任意選択で置換されており、
Ｙは、０又は１～３に及ぶ（１と３を含む）整数であり、
Ｒ_１０は、アルキル基、アルケニル基、シクロアルキル基、シクロアルケニル基、ヘテロシクリル基、又はアリール基から選択され、これらの基のそれぞれは、任意選択で置換されており、
Ｒ_１１及びＲ_１２は、水素、ハロゲン、アルキル基、アルケニル基、シクロアルキル基、シクロアルケニル基、又はヘテロシクリル基から独立的に選択され、これらの基のそれぞれは、任意選択で置換されている］。実施形態において、Ｒ_１０は、ＲＨ１～ＲＨ１２のいずれか１つである。 In embodiments, the compound has the formula XXX:

or having a salt (or solvate) thereof [wherein
R ₁ is selected from the group consisting of hydrogen, alkyl groups, alkenyl groups, cycloalkyl groups, cycloalkenyl groups, heterocyclyl groups, or aryl groups, each of which groups are optionally substituted (without substitution must be defined),
R ₂ and R ₃ together are an optionally substituted 5- or 6-membered heterocyclic ring which can contain 1 or 2 double bonds or can be aromatic forming a formula ring,
R ₆ -R ₉ are independently selected from hydrogen, halogen, alkyl groups, alkenyl groups, cycloalkyl groups, cycloalkenyl groups, or heterocyclyl groups, each of which groups are optionally substituted;
Y is 0 or an integer ranging from 1 to 3 (inclusive);
R ₁₀ is selected from an alkyl group, an alkenyl group, a cycloalkyl group, a cycloalkenyl group, a heterocyclyl group, or an aryl group, each of which is optionally substituted;
R ₁₁ and R ₁₂ are independently selected from hydrogen, halogen, alkyl groups, alkenyl groups, cycloalkyl groups, cycloalkenyl groups, or heterocyclyl groups, each of which is optionally substituted] . In embodiments, R ₁₀ is any one of RH1-RH12.

の化合物又はそれらの塩若しくは溶媒和物が挙げられる
［式中、変数は、式Ｉに定義される通りであり、Ｒ_６～Ｒ_９は、水素及び式Ｉにおいて定義されるＲ_Ａ基から独立的に選択され、Ｒ_Ｍは、縮合環上の任意選択の置換を表し、Ｒ_Ｍは、式ＩにおけるＲＡの値を取り、Ｗ_１は、Ｎ又はＣＨである］。 In specific embodiments, compounds useful in the methods herein include Formula XXXI:

or salts or solvates thereof, wherein the variables are as defined in Formula I and R ₆ -R ₉ are independent of hydrogen and the R _A groups defined in Formula I optionally selected, _RM represents optional substitution on the fused ring, _RM takes the value of RA in Formula I, and W ₁ is N or CH].

In embodiments, y is one. In embodiments, y is zero. In embodiments, both X are nitrogen. In embodiments, x is 1, M is -N-(CH ₂ )n-, and n is 1, 2 or 3. In embodiments, x is 1, M is -(CH ₂ ) _n -, and n is 1, 2 or 3. In embodiments, x is zero. In embodiments, L ₂ is -(CH ₂ ) _n -, where n is 1, 2 or 3. In embodiments, R ₁ is hydrogen. In embodiments, R ₁ is hydrogen, methyl or trifluoromethyl. In embodiments, R ₇ -R ₉ are independently selected from hydrogen, C1-C3 alkyl, optionally substituted C1-C3 alkyl, or aryl. In embodiments, R ₇ -R ₉ are independently selected from hydrogen, halogen C1-C3 alkyl, C1-C3 alkoxyl, C1-C3 acyl, or C1-C3 haloalkyl. In embodiments, R ₇ -R ₉ are all hydrogen. In embodiments, R ₄ and R ₅ are selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, or C1-C3 haloalkyl. In embodiments, R ₄ and R ₅ together form a saturated, partially unsaturated or heteroaromatic 5- or 6-membered carbocyclic or heterocyclic ring. In embodiments, _RM is one or more of hydrogen, halogen, C1-C3 alkyl groups or C1-C3 haloalkyl groups. In embodiments, _RM is one or more of hydrogen, halogen, especially Br, methyl or trifluoromethyl. In embodiments, _RM is hydrogen.

の化合物又はそれらの塩若しくは溶媒和物が挙げられる
［式中、変数は、式Ｉに定義される通りであり、Ｒ_Ｂは、式Ｉにおいて定義される任意選択の置換を表し、Ｒ_６～Ｒ_９は水素であるか、又は式ＩによるＲ_Ａの値を取る］。 In embodiments, compounds useful in the methods of the present invention include Formula XXXII:

or salts or solvates thereof, wherein the variables are as defined in Formula I, R _B represents an optional substitution as defined in Formula I, R ₆ to R ₉ is hydrogen or takes on the value of _RA according to Formula I].

In embodiments, y is one. In embodiments, y is zero. In embodiments, both X are nitrogen. In embodiments, x is 1, M is -N-(CH ₂ )n-, and n is 1, 2 or 3. In embodiments, x is 1, M is -(CH ₂ ) _n -, and n is 1, 2 or 3. In embodiments, x is 0. In embodiments, L ₂ is -(CH ₂ ) _n -, where n is 1, 2 or 3. In embodiments, R ₁ is hydrogen. In embodiments, R ₁ is hydrogen, methyl or trifluoromethyl. In embodiments, R ₆ -R ₉ are independently selected from hydrogen, C1-C3 alkyl, optionally substituted C1-C3 alkyl, or aryl. In embodiments, R ₆ -R ₉ are independently selected from hydrogen, halogen C1-C3 alkyl, C1-C3 alkoxyl, C1-C3 acyl, or C1-C3 haloalkyl. In embodiments, R ₇ -R ₉ are all hydrogen. In embodiments, R ₄ and R ₅ are selected from hydrogen, halogen, C1-C3 alkyl, C1-C3 alkoxyl, or C1-C3 haloalkyl. In embodiments, R 1 _B is one or more of hydrogen, halogen, C1-C3 alkyl groups or C1-C3 haloalkyl groups. In embodiments, R _B is one or more of hydrogen, halogen, especially Br, methyl or trifluoromethyl. In embodiments, R _B is hydrogen. In embodiments, R _H is a heterocyclyl or heteroaryl group. In embodiments, _RH is optionally substituted naphthyl, thiophene, indolyl, or pyridinopyrrolyl.

［式中、変数は、上記の式Ｉ～ＸＩＸに定義される通りであり、Ｘ_５は、Ｆ、Ｃｌ及びＢｒを含めてハロゲンであり、具体的な実施形態においてＢｒである］。式ＸＸＸＶ～ＸＬＩＩの具体的な実施形態において、ｙは０である。式ＸＸＸＶ～ＸＬＩＩの具体的な実施形態において、ｙは１であり、Ｌ_２は－（ＣＨ_２）ｎ－であり、ｎは、１、２又は３である。式ＸＸＸＶ～ＸＬＩＩの具体的な実施形態において、Ａ環はフェニル環であり、Ｒ_Ａは水素である。具体的な実施形態において、Ｒ_Ｐは、Ｒ_Ｎ－１～Ｒ_Ｎ－３１のいずれか１つから選択される基である。具体的な実施形態において、式ＸＬＩＩのＢ環は、スキーム４に示す式ＲＢＩのＢ環である。より具体的な実施形態において、式ＸＬＩＩのＢ環は、スキーム４のＲＢ２～ＲＢ５のＢ環である。 Compounds of Formulas XXXV-XLII are useful in the methods herein:

[wherein the variables are as defined in Formulas I-XIX above and X ₅ is halogen, including F, Cl and Br, and in specific embodiments is Br]. In specific embodiments of formulas XXXV-XLII, y is 0. In specific embodiments of formulas XXXV-XLII, y is 1, L ₂ is -(CH ₂ )n-, and n is 1, 2 or 3. In specific embodiments of formulas XXXV-XLII, the A ring is a phenyl ring and R _A is hydrogen. In specific embodiments, R _P is a group selected from any one of R _N -1 through R _N -31. In specific embodiments, the B ring of formula XLII is the B ring of formula RBI shown in Scheme 4. In a more specific embodiment, the B ring of formula XLII is the B ring of RB2-RB5 of Scheme 4.

The present invention provides respective salts, particularly pharmaceutically acceptable salts, of compounds of any of Formulas I-IX, XI-XIX, XXX-XXXII, XXXV-XLII and Formula XX below. The present invention provides respective solvates and salts, particularly pharmaceutically acceptable solvates and salts, of compounds of any of Formulas I-XIX, XXX-XXXII, XXXV, XXXV-XLII and Formula XX below do. Preferred solvates are hydrates. The present invention provides pharmaceutical compositions comprising any compound of any one of the formulas herein.

Aliphatic compounds are organic compounds containing carbon and hydrogen joined in straight, branched, or non-aromatic rings, which may contain single, double, or triple bonds. Aliphatic compounds are distinguished from aromatic compounds. As used herein, the term aliphatic group refers to a monovalent group containing carbon and hydrogen that is not aromatic. Aliphatic groups include alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, and cycloalkynyl, as well as aliphatic groups substituted with other aliphatic groups, such as alkenyl groups substituted with alkyl groups, cycloalkyl and alkyl groups substituted with groups.

The terms alkyl or alkyl group refer to monovalent radicals of straight or branched saturated hydrocarbons. Alkyl groups include straight chain and branched alkyl groups. Unless otherwise indicated, alkyl groups have from 1 to 8 carbon atoms (C1-C8 alkyl groups), preferably from 1 to 6 carbon atoms (C1-C6 alkyl groups). and more preferred are those containing 1 to 3 carbon atoms (C1-C3 alkyl groups). Alkyl groups are optionally substituted with one or more non-hydrogen substituents described herein. Exemplary alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, s-butyl, t-butyl, n-pentyl, various branched pentyls, n-hexyl, various branched Branched hexyls are included, all of which are optionally substituted, the substitution being defined elsewhere herein. Substituted alkyl groups include fully halogenated or semi-halogenated alkyl groups, such as alkyl groups in which one or more hydrogen atoms are replaced with one or more fluorine, chlorine, bromine and/or iodine atoms. be done. Substituted alkyl groups include fully fluorinated or semi-fluorinated alkyls.

A cycloalkyl group is an alkyl group having at least one 3 or more membered carbocyclic ring. Cycloalkyl groups include those having 3- to 12-membered carbon rings. Cycloalkyl groups include those having 3 to 20 carbon atoms and those having 3 to 12 carbon atoms. More specifically, cycloalkyl groups can have at least one 3- to 10-membered carbocyclic ring. A cycloalkyl group can have a single carbocyclic ring having from 3 to 10 carbons in the ring. Cycloalkyl groups are optionally substituted. Cycloalkyl groups can be bicyclic having 6 to 12 carbons. Exemplary cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl and cyclodecyl groups, among others. Bicyclic alkyl groups include fused bicyclic groups and bridged bicyclic groups. Exemplary bicycloalkyl groups include bicyclo[2.2.2]octyl, bicyclo[4.4.0]decyl (decalinyl), and bicyclo[2.2.2]heptyl (norbornyl), among others.

A cycloalkylalkyl group is an alkyl group as described herein substituted with a cycloalkyl group as described herein. More particularly, alkyl groups are methyl or ethyl groups and cycloalkyl groups are cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl groups. Cycloalkyl groups are optionally substituted. In specific embodiments, optional substitutions include one or more halogens, alkyl groups having 1-3 carbon atoms, alkoxy groups having 1-3 carbon atoms, hydroxyl and nitro groups. substitution.

The term alkylene refers to a straight or branched saturated hydrocarbon divalent group. Alkylene groups can have from 1 to 12 carbon atoms unless otherwise specified. Alkylene groups include those having 2 to 12, 2 to 8, 2 to 6, or 2 to 4 carbon atoms. As used herein, linker groups (L1) include alkylene groups, particularly unsubstituted linear alkylene groups —(CH ₂ )—, where n is 1-12, n is 1-10, n is 1-9, n is 1-8, n is 1-7, n is 1-6, n is 1-5, n is 1-4 or n is 1-3, n is 2-10, n is 2-9, n is 2-8, n is 2-7, n is 2-6, n is 2-5, or n is 2-4.

Alkoxy groups are widely discussed alkyl groups linked to oxygen (Ralkyl-O-). Alkoxy groups are monovalent.

Alkenylene groups are divalent radicals of straight or branched alkylene groups having one or more carbon-carbon double bonds. In specific embodiments, the same carbon atom is not part of two double bonds. In alkenylene groups, one or more CH ₂ —CH ₂ moieties of an alkylene group are replaced with carbon-carbon double bonds. In specific embodiments, the alkenylene group contains 2-12 carbon atoms, or, more preferably, 3-12 carbon atoms. In specific embodiments, the alkenylene group contains 1 or 2 double bonds. In specific embodiments, the alkenylene group contains 1 or 2 trans-double bonds. In specific embodiments, the alkenylene group contains one or two cis-double bonds. Exemplary alkenylene groups include:
-(CH ₂ )n-CH=CH-(CH ₂ )n- [where n is 1 to 4, more preferably 2], and -(CH ₂ )n-CH=CH-CH =CH-(CH ₂ )n- [where n is 1 to 4, more preferably 1 or 2]
are mentioned.

Alkoxyalkyl groups are alkyl groups in which one or more of the non-adjacent internal --CH ₂ -- groups have been replaced with --O--; such groups are sometimes referred to as ether groups. Alkoxyalkyl groups are monovalent. These groups can be straight-chained or branched, but straight-chained groups are preferred. Alkoxyalkyl groups include those having 2 to 12 carbon atoms and 1, 2, 3 or 4 oxygen atoms. More specifically, alkoxyalkyl groups include those having 3 or 4 carbons and one oxygen, or those having 4, 5 or 6 carbons and 2 oxygens. Each oxygen in the group is attached to a carbon in the group. A group is attached to a molecule through a bond to a carbon in the group.

An alkoxyalkylene group is a divalent alkoxyalkyl group. This group can be described as an alkylene group in which one or more of the internal --CH ₂ -- groups have been replaced by oxygen. These groups can be straight-chained or branched, but straight-chained groups are preferred. Alkoxyalkylene groups include those having from 2 to 12 carbon atoms and 1, 2, 3 or 4 oxygen atoms. More specifically, alkoxyalkylene groups include those having 3 or 4 carbons and one oxygen, or those having 4, 5 or 6 carbons and 2 oxygens. Each oxygen in the group is attached to a carbon in the group. A group is attached to a molecule through a bond to a carbon in the group. As used herein, linker groups (L1) include alkoxyalkylene groups, particularly unsubstituted linear alkoxyalkylene groups. Specific alkoxyalkylene groups include -CH ₂ -O-CH ₂ -, -CH _2- CH 2 -O-CH ₂ -CH ₂ -, -CH ₂ -CH ₂ -CH _{2 -O} -CH ₂ , among others. —CH ₂ —CH ₂ —, —CH ₂ —CH ₂ —O—CH ₂ —, —CH ₂ —O—CH ₂ —CH ₂ —, —CH ₂ —CH ₂ —O—CH ₂ —CH ₂ —O -CH ₂ -, -CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -, -CH ₂ -CH ₂ -CH ₂ -O-CH ₂ -CH ₂ -CH ₂ - O—CH ₂ — and —CH ₂ —CH ₂ —CH ₂ —O—CH ₂ —CH ₂ —CH ₂ —O—CH ₂ —CH ₂ —.

The term acyl group refers to a -CO-R group, where R is hydrogen, an alkyl or an aryl group, as described herein.

Aryl groups include monovalent groups having one or more 5- or 6-membered aromatic rings. Aryl groups can contain 1, 2 or 3 six-membered aromatic rings. Aryl groups can contain two or more fused aromatic rings. The aryl group can contain 2 or 3 fused aromatic rings. Aryl groups are optionally substituted with one or more non-hydrogen substituents. Substituted aryl groups include among others those substituted with optionally substituted alkyl or alkenyl groups. Particular aryl groups include phenyl, biphenyl, and naphthyl groups, all of which are optionally substituted as described herein. Substituted aryl groups include fully halogenated or semihalogenated aryl groups, such as aryl groups in which one or more hydrogen atoms are replaced with one or more fluorine, chlorine, bromine and/or iodine atoms. mentioned. Substituted aryl groups include fully fluorinated or semi-fluorinated aryl groups, such as aryl groups in which one or more hydrogen atoms have been replaced with one or more fluorine atoms.

Alkyl groups include arylalkyl groups in which the alkyl group is substituted with an aryl group. Arylalkyl groups include, among others, benzyl and phenethyl groups. Arylalkyl groups are optionally substituted as described herein. Substituted arylalkyl groups include those in which the aryl group is substituted with 1 to 5 non-hydrogen substituents, particularly those substituted with 1, 2 or 3 non-hydrogen substituents. Useful substituents include methyl, methoxy, hydroxy, halogen, and nitro, among others. A particularly useful substituent is one or more halogens. Specific substituents include F, Cl, and nitro.

Heterocyclic groups are monovalent groups having one or more saturated or unsaturated carbocyclic rings and containing from 1 to 3 heteroatoms (eg, N, O or S) per ring. These groups optionally contain 1, 2 or 3 double bonds. To satisfy valence requirements, ring atoms may be bonded to one or more hydrogens or substituted as described herein. One or more carbons in the heterocyclic ring may be a -CO- group. Heterocyclic groups include those having 3 to 12 carbon atoms and 1 to 6 heteroatoms, wherein 1 or 2 carbon atoms are replaced with -CO- groups. Heterocyclic groups include those having from 3 to 12 or from 3 to 10 ring atoms, up to 3 of which ring atoms can be non-carbon heteroatoms. A heterocyclic group can contain one or more rings, each of which is saturated or unsaturated. Heterocyclic groups include bicyclic and tricyclic groups. Preferred heterocyclic groups have 5- or 6-membered rings. Heterocyclic groups are optionally substituted as described herein. Specifically, heterocyclic groups may be optionally substituted with one or more alkyl groups. Heterocyclic groups include those having 5- and 6-membered rings containing 1 or 2 nitrogens and 1 or 2 double bonds. Heterocyclic groups include those having 5- and 6-membered rings with oxygen or sulfur and 1 or 2 double bonds. Heterocyclic groups include those having 5- or 6-membered rings and two different heteroatoms, eg, N and O, O and S, or N and S. Exemplary heterocyclic groups include pyrrolidinyl, piperidyl, piperazinyl, pyrrolyl, pyrrolinyl, furyl, thienyl, morpholinyl, oxazolyl, oxazolinyl, oxazolidinyl, indolyl, triazolyl, and triazinyl groups, among others.

A heterocyclylalkyl group is an alkyl group that is substituted with one or more heterocyclyl groups, the alkyl group optionally having additional substituents, and the heterocyclyl group being optionally substituted. A particular group is a heterocyclyl-substituted methyl or ethyl group.

Heteroaryl groups are monovalent groups having one or more aromatic rings, at least one ring of which contains a heteroatom (non-carbon ring atom). Heteroaryl groups include those having 1 or 2 heteroaromatic rings having 1, 2 or 3 heteroatoms, and optionally one 6-membered aromatic ring. A heteroaryl group can contain 5 to 20, 5 to 12, or 5 to 10 ring atoms. Heteroaryl groups include those having one aromatic ring containing heteroatoms and one aromatic ring containing carbon ring atoms. Heteroaryl groups include those having one or more 5- or 6-membered heteroaromatic rings and one or more 6-membered carboaromatic rings. Heteroaromatic rings can contain one or more N, O, or S atoms in the ring. Heteroaromatic rings include those with 1, 2 or 3 N, those with 1 or 2 O, and those with 1 or 2 S, or 1 or 2 or 3 Any combination of N, O or S may be mentioned. Exemplary heteroaryl groups include furyl, pyridinyl, pyrazinyl, pyrimidinyl, quinolinyl, purinyl, and indolyl groups. In specific embodiments, the heteroaryl group is an indolyl group, more particularly an indol-3-yl group.

Heteroatoms include O, N, S, P or B. More particularly heteroatoms are N, O or S. In specific embodiments, one or more heteroatoms are substituted for carbons in aromatic or carbocyclic rings. Any heteroatom in such aromatic or carbocyclic rings may be attached to H or a substituent such as an alkyl group or other substituent to satisfy valences.

A heteroarylalkyl group is an alkyl group that is substituted with one or more heteroaryl groups, the alkyl group optionally having additional substituents, and the aryl group optionally substituted. . Particular alkyl groups are methyl and ethyl groups.

The term amino group refers to the species -N(H) ₂ . The term alkylamino refers to the species —NHR″, where R″ is an alkyl group, especially an alkyl group having 1 to 3 carbon atoms. The term dialkylamino refers to the species —N(R″) ² where each R″ is independently an alkyl group, particularly an alkyl group having 1 to 3 carbon atoms.

The groups herein are optionally substituted. Most commonly, any alkyl, cycloalkyl, aryl, heteroaryl and heterocyclic group may contain one or more halogens, hydroxyl groups, nitro groups, cyano groups, isocyano groups, oxo groups, thioxo groups, azido groups , cyanate group, isocyanate group, acyl group, haloalkyl group, alkyl group, alkenyl group or alkynyl group (especially those having 1 to 4 carbons), (halogen-substituted or alkyl-substituted including), alkoxy, alkylthio, or mercapto (HS-). In specific embodiments, optional substitution is with 1-12 non-hydrogen substituents. In specific embodiments, optional substitution is with 1-6 non-hydrogen substituents. In specific embodiments, optional substitution is with 1-3 non-hydrogen substituents. In specific embodiments, optional substituents contain 6 or fewer carbon atoms. In specific embodiments, optional substitution is with one or more halogens, hydroxy groups, cyano groups, oxo groups, thioxo groups, C1-C6 unsubstituted alkyl groups, or unsubstituted aryl groups. The terms oxo and thioxo refer to replacement of a carbon atom with =O or =S to form a -CO- (carbonyl) or -CS- (thiocarbonyl) group, respectively.

Particular substituted alkyl groups include haloalkyl groups, particularly trihalomethyl groups, particularly trifluoromethyl groups. Illustrative substituted aryl groups include monohalo, dihalo, trihalo, tetrahalo and pentahalo substituted phenyl groups; monohalo, dihalo, trihalo, tetrahalo, pentahalo, hexahalo and heptahlo substituted naphthalene groups; 3- or 4-halo substituted phenyl groups; - or 4-alkyl substituted phenyl groups, 3- or 4-alkoxy substituted phenyl groups, 3- or 4-RCO substituted phenyl groups, 5- or 6-halo substituted naphthalene groups. More particularly, substituted aryl groups include acetylphenyl groups, especially 4-acetylphenyl groups; fluorophenyl groups, especially 3-fluorophenyl and 4-fluorophenyl groups; chlorophenyl groups, especially 3-chlorophenyl and 4-chlorophenyl groups. a methylphenyl group, especially a 4-methylphenyl group, and a methoxyphenyl group, especially a 4-methoxyphenyl group.

The term aromatic as applied to a cyclic group includes a ring structure containing double bonds conjugated around the entire ring structure possibly through one or more heteroatoms such as oxygen, sulfur or nitrogen atoms. point to Aryl and heteroaryl groups are examples of aromatic groups. A conjugated system of an aromatic group contains a characteristic number of electrons, eg, 6 or 10 electrons, occupying the electron orbitals that make up the conjugated system, which are typically unhybridized p-orbitals.

The term carbocyclic refers to a monovalent radical having a carbocyclic ring or ring system containing from 3 to 12 carbon atoms and which can be monocyclic, bicyclic or tricyclic. The ring does not contain any heteroatoms. Rings can be unsaturated, partially unsaturated, or saturated.

The compounds and substituents of the formulas herein are optionally substituted. A substituent refers to a single atom (e.g., a halogen atom) or group of two or more atoms covalently bonded to each other, typically in place of a hydrogen atom, on the atom(s) in a molecule. Bond to satisfy the valence requirements of the atom(s) of the molecule. Examples of substituents include alkyl groups, hydroxyl groups, alkoxy groups, acyloxy groups, mercapto groups, and aryl groups, among others. Substituents may themselves be substituted.

Substituted or substitution includes hydrogen atoms of a molecule or chemical group or moiety, such as halogen, alkyl, alkoxy, alkylthio, trifluoromethyl, acyloxy, hydroxy, mercapto, carboxy, aryloxy, aryl, arylalkyl, heteroaryl, one or more additional substituents including but not limited to amino, alkylamino, dialkylamino, morpholino, piperidino, pyrrolidin-1-yl, piperazin-1-yl, nitro, sulfato, or other R groups refers to replacement by

Carbocyclic or heterocyclic rings are optionally substituted as generally described for other groups such as alkyl and aryl groups herein. Where substitutions are present, they are typically on ring C, ring N or both. In addition, carbocyclic and heterocyclic rings can optionally include -CO-, -CO-O-, -CS- or -CS-O- moieties in the ring.

With respect to any chemical group substituted herein, i.e., containing one or more non-hydrogen substituents, such groups may be sterically impractical and/or synthetically impractical. It is understood that it does not include any substitutions or substitution patterns. Additionally, the compounds of this invention include all stereochemical isomers resulting from substitution of these compounds.

Protected derivatives of the disclosed compounds are also contemplated. A variety of suitable protecting groups for use with the disclosed compounds are disclosed in Greene and Wuts, Protective Groups in Organic Synthesis; 3rd Edition; John Wiley & Sons, New York, 1999. Generally, protecting groups are removed under conditions which will not affect the remainder of the molecule. These methods are well known in the art and include acid hydrolysis, hydrogenolysis, and the like. One preferred method is cleavage of the phosphonate ester using Lewis acidic conditions, eg TMS-Br mediated ester cleavage to remove the ester to give the free phosphonate. Another preferred method is removal of the protecting group, such as removal of the benzyl group by hydrogenolysis over palladium on carbon in a suitable solvent system such as alcohol, acetic acid, etc. or mixtures thereof. t-Butoxy-based groups, including t-butoxycarbonyl protecting groups, are removed utilizing inorganic or organic acids such as HCl or trifluoroacetic acid in a suitable solvent system such as water, dioxane and/or methylene chloride. be able to. Another exemplary protecting group suitable for protecting amino and hydroxy functional amino is trityl. Other conventional protecting groups are known and suitable protecting groups are selected by those skilled in the art with reference to Greene and Wuts, Protective Groups in Organic Synthesis; 3rd ed.; John Wiley & Sons, New York, 1999. be able to. When the amine is deprotected, the resulting salt can be easily neutralized to yield the free amine. Similarly, when an acid moiety such as a phosphonic acid moiety appears, the compound may be isolated as an acid compound or salt thereof. Protected derivatives of the compounds herein may be employed, for example, in the synthesis of compounds structurally related herein.

The present invention provides methods to promote tumor cell heterogeneity, MDR, and metastasis, novel therapeutic strategies that target TCF-induced EMT. Our structure-based drug design has generated novel potent CHD1L inhibitors that in embodiments target TCF-induced EMT. Reversal of EMT by CHD1L inhibitors can be an effective treatment when used in combination with cytotoxic chemotherapy and targeted anti-tumor agents as well as radiotherapy. These EMT-targeted agents can also sensitize both primary tumors and metastatic foci to clinically relevant therapies and potentially inhibit tumor cell metastasis.

Accordingly, one aspect of the present invention are CHD1L inhibitors that can be used to treat or prevent metastasis of a wide variety of advanced solid tumors and hematological cancers. Pharmaceutically acceptable salts, prodrugs, stereoisomers and metabolites of all CHD1L inhibitor compounds of the invention are also contemplated.

The present invention expressly includes pharmaceutically usable solvates of the compounds of the formulas herein. Specifically useful solvates are hydrates. The compounds of formula I or their salts can be solvated (eg hydrated). Solvation may occur during the manufacturing process or may occur (eg, as a result of the hygroscopic nature of the initially anhydrous compounds of the formulas herein (hydration)).

The compounds of the invention may take the form of prodrugs. Prodrugs of the compounds of this invention are useful in the methods of this invention. Any compound that is transformed in vivo to yield the biologically, pharmaceutically or therapeutically active form of the compound of the invention is a prodrug. Various examples and forms of prodrugs are well known in the art. A prodrug is an active or inactive compound that is chemically modified into an active compound through physiological effects such as hydrolysis, metabolism, etc. in vivo after administration of the prodrug to a subject. The term prodrug as used throughout this document means pharmacologically acceptable derivatives such as esters, amides and phosphates, the resulting in vivo biotransformation products of the derivatives being described herein. An active agent as defined in a compound. The prodrug preferably has excellent aqueous solubility, increased bioavailability, and is readily metabolized in vivo to the active TOP2A inhibitor. Prodrugs of the compounds described herein can be prepared by modifying functional groups present in the compound such that the modification is cleaved to the parent compound by routine manipulation or in vivo. can be done. The suitability and techniques involved in making and using prodrugs are well known to those skilled in the art. Examples of prodrugs are described, inter alia, in Design of Prodrugs, H.J. Bundgaard (Elsevier, 1985), Methods in Enzymology, Vol. 42, pp. 309-396; Widder et al. (Academic Press, 1985); Bundgaard, Chapter 5, "Design and Application of Prodrugs", H.E. Bundgaard, 113-191, 1991); Bundgaard, Advanced Drug Delivery Reviews, vol. 8, pp. 1-38 (1992); Bundgaard et al., Journal of Pharmaceutical Sciences, 77:285 (1988); and Nogrady (1985), Medicinal Chemistry A Biochemical Approach, Oxford University Press, N. ew York, pp. 388-392).

Administration of a compound or composition and administration thereof should be understood to mean providing the compound or salt thereof, a prodrug of the compound, or a pharmaceutical composition comprising the compound. The compound or composition can be administered to the patient (eg, intravenously) by another person, or can be self-administered by the subject (eg, tablets or capsules). The term "patient" refers to mammals (eg, humans and domestic animals such as dogs, cats, pigs, horses, sheep, and cows). Administration of the CHD1L inhibitors herein in combination with other agents such as alternative anticancer, antineoplastic or cancer cytotoxic agents is contemplated. For such combination administration, the two or more active ingredients are administered simultaneously so that they are found to be effective and do not interfere with the administration of any known alternative therapy to be combined with the CHD1L inhibitor. or at times separated by minutes, hours or days. Administration in combination further includes administration in the same manner and/or at a site of the patient's body or at a different site than in a different manner so as not to interfere with the administration of known alternative therapies that are also to be combined with the CHD1L inhibitor. .

A pharmaceutical composition herein comprises an effective amount of the named active ingredient to achieve the desired biological activity for a given form of administration to a given patient, optionally contains a carrier acceptable to A pharmaceutical composition comprises an amount (e.g., unit dose) of one or more of the disclosed compounds, including one or more pharmaceutically acceptable carriers, diluents, and/or adjuvants. It can be included together with non-toxic additives and can optionally include other bioactive ingredients. Such pharmaceutical compositions are prepared according to standard pharmaceutical formulation techniques, eg, Remington's Pharmaceutical Sciences, Mack Publishing Co.; Co., Easton, Pa.; (19th edition).

Pharmaceutically acceptable carriers are those carriers that are compatible with the other ingredients in the formulation and are biologically acceptable. A carrier can be solid or liquid. It is presently contemplated that the preferred carrier is a liquid carrier. A carrier includes one or more substances, which may also act as solubilizers, suspending agents, fillers, glidants, compression aids, binders, tablet disintegrating agents, or an encapsulating material. be able to. Liquid carriers can be used in preparing solutions, suspensions, emulsions, syrups and elixirs. The active ingredient can be dissolved in a pharmaceutically acceptable liquid carrier such as water (of suitable purity, e.g. pyrogen-free, sterile, etc.), an organic solvent, a mixture of both, or a pharmaceutically acceptable oil or fat. or can be suspended. Liquid carriers include, for example, solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, coloring agents, viscosity modifiers, stabilizers, or osmotic agents. of suitable excipients. Compositions for oral administration may be in liquid or solid form.

Suitable examples of liquid carriers for oral and parenteral administration include water of suitable purity, aqueous solutions (especially carboxymethylcellulose sodium solutions containing additives such as cellulose derivatives), (monohydric alcohols and polyhydric alcohols , including, for example, glycols), and their derivatives, and oils. For parenteral administration the carrier can also be an oily ester such as ethyl oleate and isopropyl myristate. Sterile liquid carriers are used in sterile liquid form compositions for parenteral administration. The liquid carrier for pressurized compositions can be halogenated hydrocarbon or other pharmaceutically acceptable propellant. Liquid pharmaceutical compositions which are sterile solutions or suspensions can be administered by, for example, intramuscular, intraperitoneal or subcutaneous injection. Sterile solutions can also be administered by intravenous injection. Compositions for oral administration may be in liquid or solid form. The carrier can also take the form of creams and ointments, pastes, and gels. Creams and ointments can be oil-in-water or water-in-oil viscous liquid or semisolid emulsions.

A "therapeutically effective amount" of a disclosed compound is a sufficient dose of the compound to achieve the desired therapeutic effect, such as an anti-tumor or anti-metastatic effect. In some examples, the therapeutically effective amount is tissue at sites of action similar to those shown to modulate TCF transcription and/or epithelial-mesenchymal transition (EMT) in tissue culture, in vitro, or in vivo. Sufficient amount to achieve medium consistency. For example, a subject may receive a dose of about 0.1 μg/kg body weight to about 1000 mg/kg body weight/day, such as a dose of about 1 μg/kg body weight to about 1000 μg/kg body weight/day, such as about 5 μg/kg body weight/day. A therapeutically effective amount of the compound such that it receives a dose of from daily to about 500 μg/kg body weight/day.

The term modulate refers to the ability of the disclosed compounds to alter the amount, extent, or rate of amelioration of a biological function, disease progression, or condition. For example, modulating can refer to the ability of a compound to induce increased or decreased angiogenesis, inhibit TCF transcription and/or EMT, or inhibit tumor metastasis or tumorigenesis.

Treatment refers to therapeutic intervention that ameliorates the signs or symptoms of a disease or condition after it begins to develop. As used herein, the term ameliorating in relation to a disease or condition refers to any observable beneficial effect of treatment. A beneficial effect may be, for example, delaying the onset of clinical symptoms of a disease in a susceptible subject, reducing the severity of some or all clinical symptoms of a disease, slowing disease progression, improving the general health or well-being of a subject. or by other parameters known in the art specific to a particular disease. The phrase treating a disease includes inhibiting the full development of a disease or condition, for example in a subject at risk for the disease or suffering from a disease such as cancer or a disease associated with an impaired immune system. . Preventing a disease or condition shows no signs of the disease, or shows only early signs of the disease, for the purpose of reducing the risk of developing the pathology or condition or reducing the severity of the pathology or condition. Refers to prophylactic administration of a composition to a subject.

All references throughout this application, including patent documents, including issued or granted patents or equivalents; patent application publications; Incorporated herein by reference in its entirety. All patents and publications mentioned in the specification are indicative of the levels of skill of those skilled in the art to which the invention pertains. The documents cited herein are hereby incorporated by reference in their entireties to indicate prior art, their filing dates, as the case may be, and this information is incorporated herein to It is intended that specific embodiments existing in the prior art can be excluded (eg, negated) accordingly. For example, when a compound is claimed, the compound known in the prior art, including some of the compounds disclosed in the references disclosed herein (particularly the referenced patent documents), is the subject of the claim. It should be understood that it is not intended to be included in

Abbott et al., 2020 and Supplementary Information therein, respectively, are incorporated herein by reference in their entirety for descriptions of biological and chemical methods useful in making and assessing the activity and properties of CHD1L inhibitors herein. incorporated into the specification.

When a group of substituents is disclosed herein, all individual members and subgroups of the group, including any isomers and enantiomers of the group members, and the substituents are used to form It is understood that classes of compounds capable of are disclosed separately. It should be understood that when a compound is claimed, it is not intended to include compounds known in the art, including compounds disclosed in the references disclosed herein. be. When a Markush group or other grouping is used herein, all individual members of the group and all possible combinations and subcombinations of the group are intended to be individually included in the invention.

When a compound is described herein such that a specific isomer, enantiomer or diastereomer of the compound is not specified, for example in a formula or chemical name, the description refers to the compound being described. It is intended to include each isomer and enantiomer (eg, cis/trans isomers, R/S enantiomers) individually or in any combination. Moreover, all isotopic variants of the compounds disclosed herein, unless otherwise specified, are intended to be included in the present invention. For example, it is understood that any one or more hydrogens in a molecule disclosed can be replaced with deuterium or tritium. Isotopic variants of molecules are generally useful as standards in molecular assays and chemical and biological studies involving molecules or their uses. Isotopic variants, including those with radioactive isotopes, may also be useful in diagnostic assays and therapeutics. Methods for making such isotopic variants are known in the art.

The molecules disclosed herein contain one or more ionizable groups [groups that can be deprotonated (eg, —COOH) or groups that can be protonated (eg, amines) or groups that can be grouped (eg, amines)]. All possible ionic forms of such molecules and their salts are intended to be individually included in the invention herein. With respect to salts of the compounds herein, one skilled in the art can select from a wide variety of available counterions an appropriate counterion for preparing the salts of the invention for a given application. For specific applications, selection of a given anion or cation for salt preparation can result in an increase or decrease in the solubility of that salt.

CHD1L inhibitors of the invention are commercially available, or can be prepared by the methods disclosed herein, or can be made by known methods, without undue experimentation. It can be prepared by routine adaptation of such methods using starting materials and reagents available. It is recognized that depending on the compound being synthesized, it may be necessary to protect potentially reactive groups in the starting materials from undesirable bonding. Useful protecting groups for various reactive groups are known in the art, for example as described in Wutts and Greene, 2007.

The compounds herein may take the form of salts, eg ammonium salts, with anionic or quaternary ammonium salts being selected. A salt can be formed by addition of an acid to the free base as is known in the art. Salts can be prepared from inorganic acids such as hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, or acetic, propionic, glycolic, pyruvic, oxalic, maleic, malonic, succinic, fumaric, tartaric, It can be formed using organic acids such as citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, N-acetylcysteine and the like.

In specific embodiments, compounds of the invention may contain one or more negatively charged groups (free acids), which may be in the form of salts. Exemplary salts of free acids formed with inorganic bases include alkali metal salts (eg, Li ⁺ , Na ⁺ , K ⁺ ), alkaline earth metal salts (eg, Ca ²⁺ , Mg ²⁺ ), Toxic heavy metal salts and ammonium salts (NH ₄ ⁺ ) and substituted ammonium salts (N(R') ₄ ⁺ [wherein R' is hydrogen, alkyl, or substituted alkyl, i.e., methyl, ethyl, or hydroxyethyl; Examples include trimethylammonium salts, triethylammonium salts, and triethanolammonium salts), salts in the form of cations such as lysine arginine, N-ethylpiperidine, and piperidine, but are not limited thereto. The compounds of the invention may also exist in zwitterionic form. The compounds herein may take the form of pharmaceutically acceptable salts, which retain the biological utilities and properties of the free base or free acid, and which are biologically or otherwise undesirable. No salt refers.

The scope of the invention as described and claimed includes the racemates of the compounds as well as their individual enantiomers and non-racemic mixtures. The compounds of the invention may contain one or more asymmetric carbon atoms and therefore the compounds can exist in different stereoisomeric forms. The compounds can be, for example, racemates or optically active forms. Optically active forms can be obtained by resolution of racemates or by asymmetric synthesis. In preferred embodiments of the invention, the enantiomers of the invention exhibit + (positive) specific rotation. Preferably, the (+) enantiomer is substantially free of the corresponding (-) enantiomer. An enantiomer that is substantially free of the corresponding enantiomer therefore refers to a compound that is isolated or separated by a separation technique or prepared free of the corresponding enantiomer. "Substantially free" means that the compound is made up of a significantly greater proportion of one enantiomer. In preferred embodiments, the compound is made up of at least about 90% by weight of the preferred enantiomer. In another embodiment of the invention, the compound is made up of at least about 99% by weight of the preferred enantiomer. The preferred enantiomer can be isolated from the racemic mixture by any method known to those skilled in the art, including high performance liquid chromatography (HPLC) and chiral salt formation and crystallization, or described herein. [see, eg, Jacques et al., 1981; Wilen et al., 1977; Eliel, 1962; Wilen, 1972].

The compounds of the invention and their salts can exist in their tautomeric forms, in which hydrogen atoms are rearranged to other parts of the molecule such that chemical bonds between the atoms of the molecule are rearranged. It should be understood that all tautomeric forms that may exist are included within the scope of the present invention.

Any formulation, compound or combination of components described or exemplified herein can be used to practice the invention, unless otherwise stated. Specific names of compounds are intended to be exemplary, as it is known that one skilled in the art can name the same compounds differently. When a compound is described herein such that no specific isomer or enantiomer of the compound is specified, for example in a formula or chemical name, the description refers to each isomer of the compound described. and enantiomers individually or in any combination.

Those skilled in the art will appreciate that methods, alternative therapies, starting materials, and synthetic methods other than those specifically exemplified can be employed in the practice of this invention without resort to undue experimentation. All art-known functional equivalents of any such methods, device elements, starting materials, and synthetic methods are intended to be included in this invention. Whenever a range, e.g., a temperature range, a time range, or a composition range is described herein, all intermediate and subranges and all individual values subsumed within a given range are hereby included. shall be included in the invention.

The singular terms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. Similarly, the word "or" shall include "and" unless the context clearly dictates otherwise. The term "comprises" means "includes." Also, "comprising A or B" means including A or B, or A and B, unless the context clearly indicates otherwise. It is further to be understood that all molecular weight or molecular mass values given for compounds are approximate and are given for illustrative purposes. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. In addition, the materials, methods, and examples are illustrative only and not limiting.

As used herein, "comprising" is synonymous with "including," "containing," or "characterized by" and is inclusive or open-ended; Do not exclude additional undescribed elements or method steps. As used herein, "consisting of" excludes any element, step, or material not specified in the claim component. As used herein, “consisting essentially of” does not exclude materials or steps that do not materially affect the basic novel character of the claim. Any reference herein to the term "comprising", particularly in describing a component of a composition or in describing an element of a device, refers to a composition consisting essentially of, or consisting of, the recited component or element. and methods. The invention illustratively described herein may suitably be practiced in the absence of any element(s), limitation(s) not specifically disclosed herein.

Without wishing to be bound by any particular theory, there may be some discussion herein of underlying beliefs or understandings regarding the present invention. Regardless of the ultimate correctness of any mechanistic explanation or hypothesis, it is recognized that embodiments of the invention can still be implemented and useful.

The terms and expressions that have been employed herein are used as terms of description and not as terms of limitation, as such terms and expressions are used to set forth and describe the It is recognized that various modifications are possible within the scope of the invention as claimed without any intention to exclude any equivalents of the features or portions thereof. Thus, while the present invention has been specifically disclosed in terms of preferred embodiments and optional features, modifications and variations of the concepts disclosed herein may be made by those skilled in the art and such modifications and variations may include: , are considered to be within the scope of the present invention.

[Example]
Example 1 Clinicopathologic Characterization of CHD1L in CRC Patients CHD1L expression correlates with poor prognosis in several cancers, but only limited information is known about the pathology of CHD1L in CRC. is. This example describes pathogenic characterization and pathological mechanisms for CHD1L in CRC patients. Clinicopathological characteristics of 585 CRC patients were analyzed on the basis of CHD1L expression by the Cartes d'Identite des Tumeurs (CIT) program (GEO: GSE39582) [Marisa et al., 2013]. These properties are summarized in Abbott et al., 2020, Supplementary Information.

Additional data for this example can be found in Abbott et al., 2020 and its Supplementary Information. Follow-up information is available for all patients in the 15-year CIT cohort. Across patient cohorts, high CHD1L expression is associated with lower OS (P=0.0167) and median survival (MS) of 8.8 years in high CHD1L patients. Median survival was not reached in the low CHD1L cohort, as 72% (115/159) patients were censored and 26% (42/159) patients died. Patient data were evaluated using the TNM staging system. Because stage I and IV patients have a high chance of survival or death, respectively, survival of stage II and III CRC patients was assessed. In stage II and III CRC, high CHD1L expression was associated with lower OS (P=0.0191) and 11-year MS, again not reaching median survival in the low CHD1L cohort. Survival was analyzed based on CHD1L expression for each stage of CRC. Stage II patients showed a significant difference in survival (P=00319) at 11 years MS, and no significant difference was observed for stage I, III or IV patients. Analysis of CHD1L expression showed significant differences in expression by cancer stage. Stage I and II colorectal cancer patients versus stage III and IV patients were evaluated and showed a significant increase in CHD1L expression in stages III and IV versus the early cohort (P=0.0051). Analysis of CHD1L expression on the basis of lymph node metastasis suggests that CHD1L is overexpressed in patients with increased regional lymph node metastasis (N1 P=0.0128 compared to N0 , N2 P=0.05). For the N3 cohort, trends in CHD1L expression were the same, but due to the limited number of patient samples available, significance was not determined. No significant differences in CHD1L expression based on tumor size, metastasis or site were found.

Evaluation of CHD1L in CRC Molecular Subtypes CHD1L expression and six molecular subtypes of CRC: C1 (immune system down, n=116), C2 (mismatch repair deficient, n=104), C3 (KRAS mutant, n=75). ), C4 (CSC, n=59), C5 (activated WNT pathway, n=152), and C6 (chromosomal instability normal, n=60) [Marisa et al., 2013]. There is a significant difference in CHD1L expression among the 6 molecular subtypes (P<0.001). CHD1L expression is high in C5, C4 and C3 and low in C2 and C6. The C2 subtype is associated with reduced WNT signaling pathways and is defective in mismatch repair. The C4 and C6 subtypes are associated with poor recurrence-free survival compared to other subtypes. The C4 subtype is associated with increased CSC stemness and the C5 subtype is associated with activated WNT signaling and deregulated EMT pathways. Reduced CHD1L expression in the C2 (mismatch repair deficient) subtype is consistent with its known function in DNA damage response [Ahel et al., 2009]. Moreover, CHD1L expression was lower in patients deficient in mismatch repair than in non-deficient patients (P<0.001). CHD1L expression was also higher in patients with KRAS mutations (P=0.049). Expression of CHD1L in the C3, C4, and C5 molecular subtypes prompted further investigation of the function of CHD1L expression in the EMT, CSC stemness, and WNT/TCF pathways.

CHD1L expression correlates with Wnt/TCF-associated genes Using a smaller cohort of CRC patients (n=26) from the UCCC GI Tumor Tissue Bank, a similar trend was observed, and similar to the larger CIT cohort, CHD1L Expression was significantly correlated with late and metastatic CRC compared to early and primary CRC (Abbott et al., 2020, Supplementary Information). Expression is quantified as FPKM (number of fragments/kilobase of exon/million fragments mapped). Metastatic tumor samples had significantly more CHD1L than primary tumors. Moreover, CHD1L levels were higher in stage IV compared to the stage II/III patient cohort. When CHD1L expression was analyzed with genes involved in the KEGG WNT pathway using Spearman's correlation, significant positive correlations were found in 65 out of 125 genes. Among these are type IIα topoisomerase (TOP2A) (r = 0.65, P = 0.004 [Zhou et al., 2016; Abraham et al., 2019] and TCF4 (r = 0.61, P = 0.0012) ( There were well-established genes involved in TCF-mediated transcription, such as Abbott et al., Supplementary Fig. 2. Genes had a correlation value of P<0.05. Log ₂ normalization and quantification by million fragments mapped).

A significant positive correlation was observed between the known CSC markers CD44 (r=0.43, P=0.038), LGR5 (r=0.55, P=0.0075) and CHD1L. A significant correlation was observed for TOP2A (r=404 0.1275, P=0.0020) and TCF4 (r=0.1050, P=0.011) when comparing the CIT cohort with the UCCC cohort. Consistent with this result, TOP2A has been shown to be a required component of the TCF complex and promote EMT in CRC [Zhou et al., 2016; Abraham et al., 2019]. Therefore, CHD1L appears to be involved in TCF transcription and EMT in CRC patients.

Example 2: CHD1L Mediates TCF Transcription in CRC Based on the correlation between CHD1L and TCF complex members, CHD1L may have a mechanistic role in TCF transcription. To assess this role, we utilized the SW620 cell line with high endogenous CHD1L expression and the DLD1 cell line with low endogenous CHD1L expression. Additional data for this example can be found in Abbott et al., 2020 and its Supplementary Information. Small hairpin RNA (shRNA) was used to knock down CHD1L in SW620 cells (SW620 ^CHD1L-KD ). CHD1L was overexpressed in DLD1 cells (DLD1 ^CHD1L-OE ). Using a TOPflash luciferase reporter [Morin et al., 1997; Zhou et al., 2016] transfected into SW620 ^CHD1L-KD or DLD1 ^CHD1L-OE , overexpression of CHD1L resulted in a significant increase in TCF transcription (P<0.0001 ) (Abbott et al., 2020). Conversely, SW620 ^CHD1L-KD cells showed a significant decrease in TCF transcription (P=0.0006). These results indicate that CHD1L is a potential factor directly involved in TCF transcription. Morin et al., 1997 and Zhou et al., 2016 are each incorporated herein by reference in their entirety for their description of TOPflash reporters and assays employing them.

CHD1L directly interacts with the TCF transcription complex Activation of TCF transcription is a dynamic process involving shedding of corepressor proteins, binding of coactivator proteins, and remodeling of chromatin spatial arrangement [Lorch et al. 2010, Shitashige et al., 2008]. Co-immunoprecipitation (Co-IP) studies with TCF4 were performed, revealing that CHD1L directly binds to the TCF complex [Abbott et al., 2020].

CHD1L has been well characterized as a binding partner with PARP1 in the DNA damage response [Pines, 2012; Ahel et al., 2009]. PARP1 is also a component of the TCF complex that binds to TCF4 and β-catenin [Idogawa et al., 2005]. The results herein demonstrate that CHD1L binds to the TCF complex, presumably through interactions between TCF4 and PARP1.

To further characterize CHD1L as a component of the TCF complex, chromatin immunoprecipitation (ChIP) of CHD1L onto the TCF complex WNT response element (WRE) was performed in SW620 cells [Abbott et al., 2020]. CHD1L was enriched in c-Myc, vimentin, slug, LEF1, and N-cadherin WREs, further supporting that CHD1L functions directly with the TCF complex. Taken together, the data implicate CHD1L as a critical component of TCF transcription.

CHD1L-mediated TCF transcription promotes EMT and CSC stemness in CRC Previously, TCF transcription was characterized as a key regulator of EMT in CRC [Zhou et al., 2016]. Moreover, CHD1L localizes to the WRE of the EMT effector gene [Abbott et al., 2020]. Therefore, biomarker expression in SW620 ^CHD1L-KD and DLD1 ^CHD1L-OE cells was measured to determine whether knockdown or overexpression of CHD1L modulates EMT. Knockdown of CHD1L induces reversal of EMT, decreasing vimentin and Slug while increasing E-cadherin expression [Abbott et al., 2020]. Conversely, EMT is induced in DLD1 ^CHD1L-OE cells, evidenced by decreased E-cadherin and increased vimentin and Slug expression [Abbott et al., 2020]. These results indicate that CHD1L is an EMT effector gene involved in promoting the mesenchymal phenotype in CRC. A hallmark of EMT is increased CSC stemness.

A clonogenic colony formation assay [Abbott et al., 2020] was performed to characterize the effect of CHD1L expression on stemness [Franken et al., 2006]. CSC stemness, as measured by colony formation, was increased in DLD1 ^CHD1L-OE cells (P=0.0001) and decreased in SW620 ^CHD1L-KD cells (P=0.002).

Example 3 Identification of Small Molecule Inhibitors of CHD1L As established in Examples 1 and 2 herein, CHD1L is a driver of TCF-mediated EMT. Based on this, assays to identify small molecule inhibitors of CHD1L are described herein. The drug discovery goal was to target CHD1L DNA translocation or interaction with DNA, which is dependent on CHD1L's catalytic domain ATPase activity [Ryan and Owen-Hughes, 2011; Flaus et al., 2011].

CHD1L belongs to the SNF2 (sucrose non-fermentative 2) ATPase superfamily of chromatin remodelers [Abbott et al., 2020 and its supplementary information], which contains a bileaflet ATPase domain. CHD1L also has a unique macrodomain compared to other chromatin remodelers that promotes an autoinhibitory state through interactions between the macrodomain and the ATPase domain. [Lehmann et al., 2017; Gottschalk et al., 2009]. However, the macrodomain binds to PARP1, the major activator of CHD1L, and relieves autoinhibition [Lehmann et al., 2017; Gottschalk et al., 2009].

The method of Lehmann et al., 2017 was used to purify full-length CHD1L (fl-CHD1L) and the catalytic ATPase domain (cat-CHD1L). [Abbott et al., 2020 and supplementary information]. The protein construct was used for recombinant expression and purification of CHD1L and in vitro HTS was performed as exemplified in Abbott et al., 2020. SDS page gel showed purified cat-CHD1L (68 kD) and fl-CHD1L (101 kD). Enzyme kinetics of cat-CHD11 versus fl-CHD1L were compared. cat-CHD1L provides a robust ATPase assay compared to fl-CHD1L, consistent with reports from Lehman et al., 2017. Therefore, in order to identify direct inhibitors of CHD1L ATPase, we examined the context of TCF transcription, including phosphate-binding proteins that fluoresce upon binding cat-CHD1L, c-Myc DNA, ATP, and inorganic phosphate (Pi). An exemplary high-throughput screening (HTS) assay is described therein.

This assay was validated and a pilot screen was preliminarily formed for clinically important kinase inhibitors [Abbott et al., 2020 and supplementary information]. No hits were found in a pilot screen, demonstrating that CHD1L is not a useful target for kinase inhibitors. Once validated, primary HTS was preformed using 20,000 compounds from the Life Chemicals Diversity Set screened at 20 μM in 1% DMSO with 10 mM EDTA as a positive control [ Abbott et al., 2020 and supplementary information]. Screening provided robust statistics, with mean Z' factor values of 0.57±0.06 across 64 plates. The average compound activity was 92.3%±17.8. As a result, the hit limit was set to 3 standard deviations from the mean at 39% ATPase activity. This strict hit limit identified 64 hits, of which 53 hits were confirmed for recombinant CHD1L ATPase activity.

Example 4: Exemplary Inhibitors A subset of the 7 confirmed hits (compounds 1-7, see Scheme 1) representing various pharmacophores with greater than 50% inhibition against cat-CHD1L ATPase was purchased. bottom. Compounds 1-7 were subjected to dose-response studies against cat-CHD1L ATPase, validating these hits as potent CHD1L inhibitors with activity between 900 nM and 5 μM (FIG. 1A). Structures of additional exemplary compounds 8-73 are set forth in Scheme 1, where SEM represents the protecting group trimethylsilylethoxymethyl. Note that in some cases in Scheme 1, additional compound numbers that may be employed in tables and figures herein or in Abbott et al., 2020 and its Supplementary Information are listed in brackets.

Compounds 1-7 were tested for their ability to inhibit TCF transcription in HCT116, SW620, and DLD1 ^CHD1L-OE cells using the TOPflash reporter system (FIG. 1B). Compounds 1-3 were not shown to have significant activity in cells. Compound 4 was shown to have moderate activity in cells with no dose-dependent inhibition of TCF activity. However, compounds 5-7 show excellent dose-dependent activity against TCF transcription in all three CRC cell lines. Of note, in DLD1 ^CHD1L-OE cells, doses as low as 2 μM of 5-7 showed reduced inhibition of TCF transcription. This is evidence of intracellular CHD1L target binding.

CHD1L inhibitors reverse EMT and malignant traits in CRC After validating hits 5-7 against CHD1L-mediated TCF transcription, these compounds were evaluated for their ability to reverse EMT and other malignant traits in CRC. E-cadherin and vimentin are putative biomarkers for epithelial and mesenchymal phenotypes, respectively [McDonald et al., 2015]. E-cadherin loss and vimentin increase are also clinical biomarkers of poor prognosis [Yun et al., 2014; Richardson et al., 2012; Dhanasekaran et al., 2001; Kashiwagi et al., 2010; Toiyama et al., 2013]. In view of the above, lentiviral promoter-inducible reporters for E-cadherin (pCDH1-EcadPro-RFP) and vimentin (pCDH1-VimPro-GFP) were developed to accurately report E-cadherin and vimentin protein expression, respectively. [Zhou et al., 2016; Abraham et al., 2019]. SW620 cells transduced with EcadPro-RFP or VimPro-GFP were cultured as tumor organoids for 72 hours and reached 600 μm in diameter. Tumor organoids were treated with compounds 5-7 for an additional 72 hours to determine the 50% effective concentration (EC ₅₀ ) that modulates promoter activity. A high-content analysis algorithm based on 3D confocal images 507 was used to quantify changes in promoter expression (FIGS. 2A-2B) [Zhou et al., 2016; Abraham et al., 2019].

Compounds 5-7 potentiated vimentin promoter activity with EC ₅₀ values of 15.6±1.7 μM (5), 4.7±1.5 μM (6), and 12.8±1.3 μM (7). adjusted downward to Conversely, E-cadherin promoter activity was upregulated with EC ₅₀ values of 11.9±0.3 μM (5), 11.4±0.3 μM (6), and 28±0.003 μM (7). . Representative images showing reversal of EMT by Compound 6 in SW620 tumor organoids as measured by EMT reporter assay are shown in Abbott et al., 2020. These results indicate that a small molecule inhibitor of CHD1L reverses TCF-induced EMT in CRC.

To confirm that CHD1L inhibitors reverse EMT, protein expression of two additional putative biomarkers of EMT, slug (mesenchyme) and zona obturata-1 (ZO-1, epithelium), was assessed. Changes in Slug and ZO-1 are considered major criteria for EMT [Zeisberg and Neilson, 2009]. SW620 tumor organoids treated with CHD1L inhibitors downregulate Slug and upregulate ZO-1, further showing reversal of EMT. Western blot analysis demonstrating protein expression changes for additional EMT biomarkers Slug and ZO1 is presented in Abbott et al., 2020.

EMT is characterized by increased CSC stemness and cellular infiltration. Therefore, the ability of compounds 5-7 to inhibit migration and invasion in HCT-116 and DLD1 ^CHD1L-OE cells was tested. All three compounds showed significant inhibition of CSC stemness (Fig. 2C). However, compounds 5 and 6 show more potent dose-dependent inhibition. Note that DLD1 ^CHD1L-OE cells form twice as many colonies as HCT-116 cells and have moderate CHD1L expression. This finding is consistent with the carcinogenic and tumorigenic properties of CHD1L. HCT-116 cells with uniform scratches embedded in 50% Matrigel® matrix (Corning Life Sciences, Corning, NY) were then used to irrigate the cells at the indicated concentrations. of CHD1L inhibitor and invasion was monitored for 72 hours. Compounds 5-7 showed dose-dependent inhibition of invasion (Fig. 2D), with compound 6 showing the most potent activity.

Example 5: CHD1L Inhibitory Efficacy of DNA-damaging Drugs CHD1L is known to function in PARP1-mediated DNA damage response repair and is a mechanism of increased drug resistance to DNA-damaging chemotherapy [Li et al. 2019; Ahel et al., 2009; Gottschalk et al., 2009]. For example, drug resistance to cisplatin in lung cancer was observed in cells overexpressing CHD1L. Cisplatin efficacy was restored after CHD1L knockdown. [Li Y. et al., 2019]. Furthermore, knockdown of CHD1L alone does not increase DNA damage. [Ahel D et al., 2009]. To determine whether CHD1L inhibitors can increase the efficacy of DNA damaging drugs against low CHD1L expressing DLD1 cells transduced with empty vector (DLD1CHD1L-EV) and overexpressing DLD1CHD1L-OE in CRC cells Therefore, compound 6 was evaluated as a single agent alone and in combination with SN-38 (the active pharmacophore of the prodrug irinotecan), oxaliplatin, and etoposide. To assess DNA damage, immunofluorescent phosphorylation of H2AX (γ-H2AX), a biomarker of DNA-damaging chemotherapy [Ahel D et al., 2009], is shown in Abbott et al., 2020 and its Supplementary Information. measured as Compound 6 alone did not show significant DNA damage when cells were treated at 10 μM and γ-H2AX activity was measured, consistent with previously reported CHD1L knockdown studies [Ahel D et al., 2009]. However, combination treatment with compound 6 in DLD1CHD1L-OE cells synergized with etoposide (10 μM) and SN-38 (1 μM) and significantly increased DNA damage compared to etoposide and SN-38 alone. . Only the combination of etoposide and Compound 6 showed significant synergy in DLD1CHD1L-EV cells. No synergy was observed with oxaliplatin under the experimental conditions used. Nevertheless, SN-38 (ie, irinotecan) combination therapy, called FOLFIRI, is the standard of care in the treatment of CRC. Thus, the enhancement of DNA damage produced by combining compound 6 with SN-38 supports the hypothesis that CHD1L inhibitors can increase the efficacy of DNA-damaging chemotherapy, the standard treatment for CRC.

Example 6: CHD1L inhibitors reverse EMT prior to induction of cell death CHD1L was reported to confer anti-apoptotic activity by inhibiting the activation of caspase-dependent apoptosis [Li et al., 2013; Sun et al., 2016]. Moreover, reversal or inhibition of EMT is known to restore the apoptotic activity of cancer cells [Lu et al., 2014]. To determine whether CHD1L inhibitors reverse EMT prior to induction of cell death, we monitored E-cadherin expression by EcadPro-RFP reporter activity and measured cytotoxicity using the Certox™ Green assay. was measured. Cells were treated with CHD1L inhibitor for 72 hours and imaged every 2 hours. Compound 6 showed a significant increase in E-cadherin expression prior to induction of cytotoxicity against DMSO (Fig. 3A).

To determine whether CHD1L inhibitors can induce apoptosis in CRC, Western blots from SW620 tumor organoids were performed and E-cadherin was found to be cleaved after treatment with 5 and 6. [Abbott et al., 2020]. E-cadherin cleavage is a marker of apoptosis [Steinhusen et al., 2001].

The more potent CHD1L inhibitor 6 showed an increase in cleaved PARP1, cleaved caspase 8, and cleaved caspase 3 relative to the DMSO control [Abbott et al., 2020]. These results indicate that compound 6 induces extrinsic apoptosis consistent with E-cadherin-mediated apoptosis through death receptors [Lu et al., 2014].

To further characterize the apoptotic activity of CHD1L inhibitors, Annexin-V staining over 12 hours in SW620 cells was examined. Compound 6 induced significant apoptosis compared to DMSO alone and had activity similar to the positive control SN-38, the active metabolite of irinotecan (Fig. 3B).

CHD1L inhibitors are effective on patient-derived tumor organoids (PDTO). The use of PDTO in preclinical drug development has been established as a predictive in vitro cellular model for clinical efficacy. [Drost J and Clevers H, 2018]. After establishing the ability of compound 6 to reverse EMT and induce apoptosis using a cell line-based model, the efficacy of compound 6 was obtained from the University of Colorado Cancer Center (UCCC) gastrointestinal (GI) tissue bank. was evaluated in PDTO generated from the patient sample CRC102 (Fig. 3C). Consistent with results in CRC cell lines, compound 6 exhibited potent cytotoxicity in PDTO with an _EC50 of 11.6±2 μM.

Example 7 In Vitro and In Vivo PK, PD, and Hepatotoxicity of Exemplary Inhibitors Compound 6 To assess in silico the drug-like potency and properties of Compound 6, in vitro and in vivo PK studies were conducted to CLogP, aqueous solubility, stability in microsomes, and PK in CD-1 mice were assessed.

Table 1 provides a summary of the in vivo and in vitro pharmacokinetic parameters of Compound 6. Consensus LogP (CLogP) values were obtained using the SwissADME web tool [Daina et al., 2017]. Compound 6 was administered by intraperitoneal injection to athymic nude mice QD for 5 days to measure accumulation in SW620 xenograft tumors (Fig. 4) and assess histopathology of hepatotoxicity. Representative H&E-stained photomicrograph sections (5x magnification) of livers in vehicle- and compound 6-treated animals are shown in Abbott et al., 2020. The images show normal hepatocyte cord and lobular architecture, with no hepatocyte degeneration, necrosis, hyperplasia, or parenchymal inflammation. Compound 6 is relatively stable to hepatic metabolic enzymes, has an excellent balance of lipophilicity (CLogP=3.2) and aqueous solubility, and excellent PK pharmacokinetics when administered to CD-1 mice. Compound 6 reached high drug plasma concentrations C _Max (approximately 30,000 ng/mL) and AUC (approximately 80,000 ng/mL/h), with a half-life (T _1/2λ ) is relatively long at 3 hours.

In early studies, compound 6 had a half-life in liver microsomes of less than 20 minutes. In subsequent similar in vitro liver microsomal half-life experiments performed with different liver microsomal preparations (data not shown), compound 6 exhibited a longer half-life of 67 min, and compound 6.3 ( ) exhibited an improved in vitro half-life of 98 minutes, and compound 6.11 (over 6) exhibited an even improved in vitro half-life of 130 minutes. Early half-life studies with Compound 6 were performed in different liver microsomal preparations and were not comparable to later in vitro microsomal half-life experiments. The results of a second series of in vitro and in vivo half-life measurements are set forth in Table 2 and include data for some additional compounds indicated.

A second acute in vivo experiment was performed using the maximum tolerated dose of 6 (50 mg/kg) by intraperitoneal administration once daily for 5 days to athymic nude mice. The goals of this experiment were to (1) determine whether compound 6 causes acute liver toxicity, (2) determine whether it accumulates in VimPro-GFP SW620 xenograft tumors, and (3) detect PD effects. was to decide. Compound 6 accumulates in SW620 tumors at a concentration of 10,533±5,579 ng/mL (n=4). As expected, when comparing the ratio of compound 6 accumulation in tissue/plasma, 2.7-fold more accumulation was found in liver compared to tumor (Figure 4). However, there was no apparent hepatotoxicity resulting from Compound 6 at dose and schedule (Table 3). Overall, there were no significant histological differences between the livers of vehicle-treated or Compound 6-treated mice. The primary histological changes observed were minimal fibrosis and inflammation of the liver capsule in both vehicle- and Compound 6-treated animals. This suggests a very low degree of subclinical peritonitis, consistent with accompanying intraperitoneal drug administration.

According to the accumulation of compound 6 in tumors, the PD effect on tumor tissue was measured by Western blot analysis, showing significant downregulation of the mesenchymal markers vimentin, vimentin reporter (VimPro-GFP), and Slug [Abbott et al. , 2020]. Although not statistically significant, upregulation of the epithelial marker ZO-1 and induction of cleaved caspase 3 (a putative biomarker of apoptosis) were also observed. Taken together, these findings of PD effects by compound 6 demonstrate reversal of EMT and apoptosis in vivo, consistent with compound 6's in vitro cell-based anti-tumor activity. Compound 6 exhibits good PK drug-like properties and the ability to alter EMT and induce apoptosis in vivo without observed hepatotoxicity.

In contrast, compound 6.11 exhibits a significantly longer half-life (T _1/2λ ) compared to compound 6's much longer half-life than 6 hours after intraperitoneal (ip) administration.

Example 8: Biological Evaluation of Compound 8 Compound 8 was evaluated in several biological assays described above. The results are presented in Figures 7A-E. Compound 8 shows a potent dose-dependent inhibition of CHD1L-mediated TCF transcription compared to compound 6 (FIG. 7A). Similarly, compound 8 reverses EMT. This is evidenced by downregulation of vimentin and upregulation of E-cadherin promoter activity (FIGS. 7B and 7C, respectively). Compound 8 significantly inhibits clonogenic colony formation over 10 days (Fig. 7D). Compound 8 significantly inhibits HCT116 invasion ability over 48 hours (Fig. 7E).

Example 9: Method applied to examples herein
Additional Materials and Methods
antibody. Monoclonal mouse anti-TCF4 antibody was purchased from EMD Millipore (Billerica, Mass., USA) (catalog number 05-511) and a 1:1000 dilution was used for Western blots and 2 μg antibody per 300 μg protein was used for IP. Monoclonal rabbit anti-CHD1L antibody was purchased from Abcam (Cambridge, Mass., USA) (catalog number ab197019) and a 1:5000 dilution was used for Western blots and 1.5 μg antibody per 300 μg protein was used for IP. Monoclonal Rabbit Anti-Vimentin (Cat.#5741), Anti-Slug (Cat.#9585), Anti-E-Cadherin (Cat.#3195), Anti-ZO-1 (Cat.#8193), Anti-Histone H3 (Cat.#4620) from Cell Signaling (Danvers, Mass., USA) and mouse anti-α-tubulin (catalog number 3873) was purchased from Cell Signaling and a 1:1000 dilution was used for Western blotting. Monoclonal rabbit anti-β-catenin (catalog number 9582) was purchased from Cell Signaling and a 1:1000 dilution was used for Western blots. Monoclonal rabbit anti-phospho-β-catenin was purchased from Cell Signaling (catalog number 5651). Monoclonal rabbit anti-TCF4 (catalog #2569) and anti-histone H3 (catalog #4620) were purchased from Cell Signaling and 2 μg of antibody per mg of protein was used for ChIP. An anti-rabbit IgG HRP-conjugated secondary antibody (catalog number 7074) was purchased from Cell Signaling and a 1:3000 dilution was used for Western blotting. Anti-goat and anti-mouse IgG HRP-conjugated secondary antibodies (catalog numbers 805-035-180 and 115-035-003) were from Jackson ImmunoResearch (West Grove, PA) at a 1:10,000 dilution. was used for Western blots.

Clinicopathologic Characterization of CHD1L Transcriptome expression data of 585 CRC patients from the CIT cohort (GEO: GSE39582) were used for in silico validation (GSE39582). [Marisa et al., 2013]. Gene expression analysis was performed by Affymetrix GeneChip™ Human Genome U133 Plus 2.0 Array (Thermo Fisher Scientific, Waltham, Mass.). Robust multi-array analysis (RMA) was used for data preprocessing and ComBat (empirical Bayesian regression) was used for batch correction. Signal intensities were log2 normalized. CHD1L cutoffs for CRC risk stratification based on disease-specific survival were determined by receiver operating characteristic (ROC) curves. The cutoff for CHD1L expression was set at 6.45. Differences in OS were estimated by the Kaplan-Meier method and compared using the log-rank test. Fisher's exact test was used for comparison of categorical variables. The Mann-Whitney U test was used for two groups of continuous variables. When more than two groups, data were analyzed by the Kruskal-Wallis test. For all two-sided P-values, an unadjusted significance level of 0.05 was applied.

CHD1L cutoff and clinicopathologic characteristics were assessed by multiple cox regression analysis. Only significant variables were integrated into the cox regression model in univariate analysis using the Wald forward algorithm for significance determination. All variables containing more than 2 groups were classified, with graded entry criteria for covariates of P<0.05 and exclusion criteria of P>0.1. IBM® SPSS Statistics (IBM Corporation, Armonk, NY), Prism8 (GraphPad Software, Inc., San Diego, Calif.), JMP® (SAS Institute, Inc., Cary, NC), and RStudio™ IDE ( Statistical analysis was performed using RStudio Inc., Boston, Mass.).

UCCC Patient Sample RNA-seq Analysis RNA-seq data from CRC patient tumor xenograft explants were obtained from the UCCC (University of Colorado Cancer Center) GI Tumor Tissue Bank and analyzed as previously described. [Scott et al., 2017]. Briefly, gene expression was Log2-normalized and measured by FPKM (number of fragments/kilobase of transcript/million reads mapped). The Wnt signaling pathway defined by the Kyoto Encyclopedia of Genes and Genomes (KEGG) was used as the gene set in this study. Samples with CHD1L expression <1 FPKM were considered low expression and removed from this study. Genes with significant Spearman correlation (P<0.05) were shown as a heatmap using matrix2png (gene-based Z normalization) [see Abbott et al., 2020 and its Supplementary Information. matter].

CHD1L Overexpression and shRNA Knockdown Full-length CHD1L was synthesized in the pGEX-6P-1 plasmid (GenScript, Piscataway, NJ). The CHD1L sequence flanked by EcoRI and NotI was digested and ligated into the lentiviral backbone to create the pCDH1-CMV-CHD1L-EF1-puro plasmid for overexpression of CHD1L in human CRC cells. Mission® shRNA (Sigma-Aldrich Co. LLC, St. Louis, Mo.) (scrambled) and CHD1L-specific TRCN0000013469 and TRCN0000013470 (sh69 and sh70) were obtained from Sigma-Aldrich (St. Louis, Mo.). Virus was produced in HEK293T cells using TransIT®-293 reagent (Mirus, Madison, Wis.) and plasmids pHRdelta8.9 and pVSV-G. CRC cells were transduced with overexpressing or shRNA knockdown viruses and selected with 2 μg/ml puromycin for 7 days.

CRC cell lines and homogenized tumor tissue samples from Western blot mice were lysed in RIPA lysis buffer (20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM Na ₂ EDTA, 1 mM EGTA, 1% NP-40, 1% Sodium deoxycholate, 2.5 mM sodium pyrophosphate, 1 mM b-glycerophosphate, 1 mM Na ₃ VO ₄ , 0.1 mM PMSF). Protein concentrations were determined using the Pierce™ BCA Protein Assay Kit (ThermoFisher, Waltham, Mass.). A 40 microgram sample was run on a 10% Bis-Tris gel. After electrophoresis, proteins were transferred to a nitrocellulose membrane. Membranes were 5% delipidated in TBS/Tween® 20 (TBST contains 20 mM Tris, 150 mM NaCl, and 0.1% Tween® 20) (Croda International PLC, Snaith, UK) at room temperature. Block with milk for 1 hour. The membrane was washed 3 times with TBST. Blots were incubated overnight at 4° C. in 5% non-fat milk in TBST with appropriate primary antibodies. Membranes were washed three times with TBST and then incubated with appropriate secondary antibodies for 1 hour. The membrane was washed again with TBST three times. Blots were exposed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (ThermoFisher, Waltham, Mass.) and imaged using a ChemiDoc imaging system (Bio-Rad, Hercules, Calif.). [For western blots, see Abbott et al., 2020 and its Supplementary Information].

TOPflash TCF Transcriptional Reporter Assay The TOPflash assay (Millipore, Billerica, Mass.) was used to assess TCF transcriptional activity in CRC cells. A total of 20,000 cells per well were seeded in 96-well white plates and transfected with TransIT®-LT1 transfection reagent (Mirus, Madison, Wis.). Cells were incubated with transfection mix for 24 hours. Cells were then washed with phosphate buffer solution (PBS) and a 1:1 ratio of PBS:ONE-Glo™ luciferase reagent Promega (Madison, Wis.) was added and luminescence was detected within 10 minutes. Detected. Duplicate experiments were performed to measure cell viability using the CellTiter-Glo® Luminescent Cell Viability Assay (Promega, Madison, Wis.), which was used to measure TOPflash luminescence. Normalized to give fold change in TCF activity. Experiments were repeated twice (n=3 for each experiment).

Co-immunoprecipitation (Co-IP)
Nucleated cell lysates were produced from untreated SW620 cells. For input controls, 100 μL of 1 mg/mL nuclear extract was saved and used as input. Immunoprecipitation (IP) was performed using the Dynabeads™ Protein A IP Kit (ThermoScientific, Waltham, Mass.). Briefly, 300 μg of lysate incubated with 2 μg of anti-TCF4 and anti-CHD1L IP antibody, anti-rabbit IgG and anti-mouse IgG were used as non-specific binding controls and rotated at 4° C. for 2 hours. After pre-incubation, 50 μL of beads were transferred to the pre-incubated antibody/lysate mixture and then incubated overnight at 4°C. The flow-through was collected and the beads were washed 3 times with PBST. Proteins were eluted with 20 μL of 50 mM glycine (pH=2.8) at 70° C. for 10 minutes.

Chromatin immunoprecipitation (ChIP)
Cells were crosslinked with 1.42% formaldehyde for 15 minutes and quenched with 125 mM glycine for 5 minutes using the detailed method previously described [Zhou et al., 2016]. Cells were lysed with Szak's RIPA (radioimmunoprecipitation assay buffer) buffer and sonicated. The IP step was performed at 4° C. as follows: 50 μL of Protein A/G agarose beads were prewashed with cold Szak's RIPA buffer and incubated with 1 mg of lysate for 2 hours. 0.3 mg/mL salmon sperm DNA was added and incubated for 2 hours. Lysate (100 μL) was set aside as an input control. Anti-CHD1L (2 μg) was added to the remainder and incubated overnight. The beads were washed and the supernatant was aspirated to 100 μL followed by 200 μL of 1.5x Talianidis elution buffer (70 mM Tris-Cl pH 8.0, 1 mM EDTA pH 8.0, 1.5 w/v% SDS). added. To elute immunocomplexes and reverse cross-linking, 12 μL of 5 M NaCl was added and the mixture was incubated at 65° C. for 16 hours. Supernatants were mixed with 20 μg proteinase K and incubated at 37° C. for 30 minutes. DNA was extracted with phenol/chloroform and precipitated with ethanol. IP products were amplified with PowerUp™ SYBR™ Green Master Mix (Applied Biosystems, Austin, Tex.) using known published primers [Zhou et al., 2016].

Colony formation was assessed after CHD1L knockdown in SW620 cells or overexpression in DLD1 cells as previously described [Zhou et al., 2016; Abraham et al., 2019]. Cells were seeded at 1,000 cells/well in 6-well plates and medium was changed twice per week over a 10 day time course. Colony formation assays were also performed as previously described [Zhou et al., 2016; Abraham et al., 2019].

To assess CHD1L inhibitors for their ability to suppress CSC stemness, DLD1 cell lines overexpressing HCT-116 or CHD1L were treated in monolayer cultures with vehicle control (0.5% DMSO) or 2C were pretreated for 24 hours with the indicated concentrations of CHD1L inhibitors. Pretreated live cells were seeded at 1,000 cells/well in 6-well plates or 200 cells/well in 24-well plates. (The following parameters were used, modified from default settings: (1) For HCT116 cells, segmentation adjustment = 0.6; minimum area (μm ² ) = 3 x 10 ⁴ ; maximum area (μm ² ) = 1. 6×10 ⁶ ; maximum eccentricity=0.9; (2) for DLD1CHD1L OE cells, segmentation adjustment=1; minimum area (μm ² )=1×10 ⁴ ; maximum area was not defined; = 0.95) Colonies were analyzed using the IncuCyte® S3 2018A (Sartorius, France) software. Experiments were repeated twice (n=2 for each experiment).

Tumor Organoid Culture Cell lines were cultured as tumor organoids using phenol red-free RPMI-1640 with 5% FBS [Zhou et al., 2016; Abraham et al., 2019] and 5,000 cells/well were coated. Cell aggregation was facilitated by seeding into clean 96-well U-bottom Ultra Low Attachment Microplates (Perkin-Elmer, Hopkinton, Mass.) followed by centrifugation at 1,000 rpm for 15 minutes. Matrigel® matrix (Corning Incorporated, Corning, New York) was added to a final concentration of 2% and the tumor organoids were incubated (5% CO ₂ , 37° C., humidity) for 72 hours prior to treatment. were allowed to self-assemble and maintained under standard cell culture conditions for the duration of the treatment.

VimPro-GFP and EcadPro-RFP Reporter 3D High Content Imaging Assay Using the previously reported pCDH imPro-GFP-EF1-puro virus or pCDH-EcadPro-mCherry-EF1-puro virus, stable VimPro-GFP or EcadPro-RFP SW620 reporter cells were generated [Zhou et al., 2016; Abraham et al., 2019]. Stable fluorescently labeled reporter cells were used to produce the tumor organoids described herein. Tumor organoids were treated with CHD1L inhibitor at 10 μM for an additional 72 hours. After treatment, tumor organoids were stained with 16 μM Hoechst 33342 for 1 hour (nuclear staining). Images were taken using a 5x air objective. Z-stacks were set 26.5 μm apart for a total of 15 optical sections. Image processing and high content analysis were performed using Opera Phenix(TM) and Harmony(R) software (PerkinElmer, Hopkinton, Mass.). Nuclei were identified within each layer and cells were found in the GFP or mCherry channels. Fluorescence intensity for each channel was calculated and a threshold was set based on background intensity. Percentages of GFP or mCherry RFP positive cells were calculated and normalized to DMSO treated groups.

Tumor Organoid Cytotoxicity SW620 tumor organoids were cultured as described herein. CellTox™ Green Cytotoxicity Assay Solution was prepared according to the manufacturer's protocol (Promega, Madison, Wis.). Briefly, tumor organoids were treated with CellTox™ Green reagent (0.5×) and various doses of CHD1L inhibitor ranging from 0-100 μM for 72 hours. The organoids were imaged using an Operaphoenix™ 207 screening system (PerkinElmer Cellular Technologies, Hamburg, Germany) with excitation at 488 nm and emission at 500-550 nm. Average intensity across wells was used to calculate cytotoxicity with lysis buffer (Promega, Madison, Wis.) as 100% cytotoxicity control and 0.5% DMSO as 0% cytotoxicity control. Intensity values were normalized to these controls using Prism8 (GraphPad, San Diego, Calif.).

Invasion Assay HCT116 cells were seeded at 60,000 cells/well in IncuCyte® ImageLock 96-well plates (Sartorius, France) and allowed to adhere overnight. Wounds were made in all wells using the IncuCyte® WoundMaker and then washed twice with PBS. Plates were brought to 4° C. using a Corning XT Cool Core to avoid polymerization of the Matrigel® matrix (Corning Life Sciences, Inc., Corning, NY) during infiltration preparation. Wells were coated with 50 μL of 50% Matrigel® matrix in RPMI-1640 medium. Plates were centrifuged at 150 rpm for 3 minutes at 4° C. using a swinging bucket rotor to ensure even bubble-free matrix coating. Plates were then placed in a pre-warmed (5% _CO2 , 37°C, humidity) Corning XT CoolSink module for 10 minutes inside a cell culture incubator to uniformly polymerize the matrix, followed by 5% FBS. CHD1L inhibitor dissolved in 50 μL of RPMI-1640 medium was added. Finally, the plates were placed in an IncuCyte® S3 live cell imager (Sartorius, France) for 48 hours. Wounds were imaged hourly using the phase contrast channel and a 10x objective in wide mode.

Cloning and Purification of Recombinant Human CHD1L Cat-CHD1L (residues 16-61) and fl-CHD1L (residues 16-879) constructs were obtained from Helena Berglund of the Karolinska Institute, Department of Medical Biochemistry and Biophysics. was a generous gift of Proteins were expressed in Rosetta™ 2(DE3) pLysS cells (Novagen available from Sigma-Aldrich, St. Louis, Mo.) in Terrific Broth (ThermoFischer, Waltham, Mass.). Cultures were induced at OD600=2.0 with 0.2 mM IPTG for 16 hours at 18°C. Cells were harvested and placed in a buffer containing 20 mM HEPES (pH 7.5), 500 mM NaCl, 50 mM KCl, 20 mM imidazole, 10 mM MgCl ₂ , 1 mM TCEP (tris(2-carboxyethyl)phosphine), 10% glycerol and 500 μM PMSF. Resuspended in A. Cells were lysed by sonication and cell debris was removed by centrifugation. The supernatant was loaded onto a Ni-NTA resin column (Qiagen, Hilden, Germany). Proteins bound to the column were washed once with Buffer-A, once with Buffer-A containing 10 mM ATP, and an additional number of times with Buffer-A. Proteins were eluted with a gradient of 20→500 mM imidazole using buffer-B (buffer-A containing 500 mM imidazole). After affinity purification, Cat-CHD1L was dialyzed overnight against 50 mM Tris (pH 7.5), 200 mM NaCl, and 1 mM DTT. Similarly, fl-CHD1L was dialyzed overnight against 20 mM MES (pH 6.0), 300 mM NaCl, 10% glycerol, and 1 mM DTT. Proteins were then purified by ion exchange chromatography. cat-CHD1L was bound to a Q-Sepharose column (GE Healthcare, Chicago, Ill.), fl-CHD1L was bound to an S-Sepharose column (GE Healthcare, Chicago, Ill.), cat-CHD1L was bound to 0.2 Proteins were eluted using a 0.3-1 M NaCl gradient for ~1 M and fl-CHD1L. Pure fractions were pooled, concentrated and passed through a Superdex™ 200 column (GE Healthcare, Chicago, Ill.) with 20 mM HEPES (pH 7.5), 100 mM NaCl, 1 mM TCEP, and 10% glycerol. It was further purified by size exclusion chromatography using . Protein purification was performed using an ACTA Start FPLC (GE Healthcare, Chicago, Ill.).

All CHD1L ATPase assay reactions were performed using low volume non-binding surface 384-well plates (Corning Inc., Corning NY). cat-CHD1L or fl-CHD1L (100 nM) and 200 nM c-Myc DNA or mononucleosomes (Active Motif, Carlsbad, Calif.) in 50 mM Tris (pH 7.5), 50 mM NaCl, 1 mM DTT, 5% glycerol added to the buffer. Reactions were initiated by adding 10 μM ATP (New England Biolabs, Ipswich, Mass.) to a total volume of 10 μL and incubated at 37° C. for 1 hour. 500 nM Phosphate Sensor (Life Technologies, Carlsbad, Calif.) containing a labeled phosphate binding protein, specifically a binding protein labeled with the fluorophore MDCC, was added and excitation (430 nm) and emission (450 nm) were monitored with an EnVision ATPase activity was assayed by direct measurement on a TM™ plate reader (PerkinElmer, Hopkinton, Mass.). Fluorescence was converted to [Pi] using an inorganic phosphate calibration curve and enzymatic kinetics were determined using Prism8 (GraphPad Software, Inc., San Diego, Calif.).

The CHD1L inhibitor HTS drug discovery assay composition was the same as above using cat-CHD1L, except that the reaction mixture volume was modified to accommodate the addition of drug or DMSO. Selected amounts of compounds dissolved in 100% DMSO were added to 50 mM Tris (pH 7.5), 50 mM NaCl, 1 mM DTT, using a Janus® liquid handler (PerkinElmer, Hopkinton, Mass.). Mixed with 5% glycerol buffer to 200 μM in 10% DMSO. 1 μL of each compound was then added to the enzyme mixture to give a final concentration of 20 μM. Negative control used was 1% DMSO and 10 mM EDTA was used as positive control. Reactions were initiated by the addition of 10 μM ATP and incubated at 37° C. for 1 hour. ATPase activity was measured by fluorescence by adding 500 nM Phosphate Sensor. Cat-CHD1L was screened against a 20,000 compound diversity set from Life Chemicals (Woodbridge, Conn.) and a kinase inhibitor library from Selleck Chemicals (Houston, Tex.). Both libraries were prescreened prior to purchase to remove pan assay interfering compounds (PAINS), which tend to react non-specifically with more biological targets than selectively with the desired target. [Baell and Nissink, 2018; Baell and Holloway, 2010].

Patient-Derived Tumor Organoids (PDTO)) Culture and Viability Assay CRC patient tumor tissue was obtained from the UCCC GI tissue bank and extended following established protocols [Morin et al., 1997]. Briefly, cells were seeded at 5,000 cells per well in 96-well plates and cultured by established methods [Franken et al., 2006] allowing PDTO formation for 72 hours. PDTO was treated with DMSO (0.5%) or various concentrations of Compound 6 for an additional 72 hours to obtain a dose response. PDTO cell viability was measured using CellTiter-Blue® reagent (Promega, Madison, Wis.). Media (80 μL) was aspirated from the wells, 80 μL of reagent was added, incubated for 4 hours, and cell viability was measured by fluorescence intensity using excitation (560 nm) and emission (590 nm).

Apoptosis Assessment SW620 cells were seeded in 96-well plates at 30,000 cells/well. Cells were treated with DMSO (negative control), SN-38 (apoptosis positive control), and Compound 6 at the indicated concentrations for 12 hours. Cells were then rinsed twice with cold PBS and once with cold annexin-V staining buffer (10 mM HEPES (pH 7.4), 140 mM NaCl, 2.5 mM CaCl ₂ ), followed by annexin-V FITC 1:100. for 30 minutes in the dark. Cells were then rinsed twice with Annexin-V staining buffer and FITC intensity was measured using an EnVision® plate reader (PerkinElmer, Hopkinton, Mass.).

γ-H2AX Assessment of DNA Damage DLD1 ^CHD1L-OE cells were seeded in 96-well PerkinElmer Cell Carrier plates and allowed to adhere overnight. Cells were then treated with the appropriate compound at 10 μM (0.5% DMSO) or with a combination of CHD1L inhibitor and SN-38 (1 μM), oxaliplatin (10 μM), and etoposide (10 μM). bottom. Cells were treated for 6 hours. Medium was aspirated and cells were washed with cold PBS. Cells were then fixed with 3% paraformaldehyde for 15 minutes at room temperature and fixed cells were washed three times with PBS. Cells were blocked for 1 hour in 5% BSA, 0.3% Triton X-100 in PBS at room temperature. Cells were then immunostained overnight at 4° C. with phospho-(S139)-g-H2AX rabbit mAb using a 1:800 dilution in 1% BSA, 0.3% Triton X-100 in PBS. Primary antibody was aspirated and cells were washed with PBS. Cells were incubated with goat anti-rabbit Alexa Fluor Plus™ 647 fluorescent secondary antibody at room temperature for 2 hours at a concentration of 5 μg/mL in 1% BSA, 0.3% Triton X-100 in PBS. Cells were then washed with PBS and Hoechst 33342 stain was diluted to a concentration of 1:1000 in PBS and added to cells for 15 minutes at room temperature. Cells were then imaged with an Operaphoenix™ HCS imaging system (PerkinElmer) using a 20× water objective. Synergies were determined using the drug interaction index (CDI) equation CDI=(A+B)/(AB). Synergy was defined at CDI<0.8 in these experiments. Additivity was 0.8-1.2 and antagonism was defined by CDI>1.2.

Aqueous solubility and CLogP
Aqueous solubility was determined for compound 6 using a recently reported detailed method [Abraham et al., 2019]. PBS UV absorption spectra were compared to fully saturated solutions in 1-propanol and linear regression analysis was used to determine the solubility of compound 6 in 10% DMSO in PBS (pH 7.4). Solubility measurements in PBS were performed in duplicate experiments. Consensus LogP (CLogP) values were obtained using the SwissADME web tool [Daina et al., 2017].

Microsomal Stability Studies Microsomal stability of Compound 6 was determined using female CD-1 mouse microsomes (M1500) purchased from Sekisui XenoTech (Kansas City, KS) according to a recently reported method [Abraham et al., 2019. ]. Samples were centrifuged at 20,000 g for 10 minutes and supernatants were transferred to autosampler vials for LCMS analysis. The following mass transitions (m/z, amu) were monitored for compound 6 (MW=393.5).

All in vivo pharmacology animal studies were performed on animals approved by the Institutional Animal Care and Use Committee (IACUC) of the University of Colorado Anschutz Medical Campus (Aurora, CO) and Colorado State University (Fort Collins, CO). Performed according to protocol procedures.

Pharmacokinetics Nine-week-old female CD-1 mice purchased from Charles River (Wilmington, Mass.) were used for PK studies [Abraham et al., 2019] using a recently reported method [Abraham et al., 2019]. . Briefly, PK studies were designed to cover a range of 0.25-24 hours for a total of 21 mice/compound 6 with 3 mice/time point. Each mouse received a single intraperitoneal injection of compound 6 at 50 mg/kg prepared in 100% DMSO. Whole blood was collected at specified time points and separated plasma was stored frozen at -80°C or used for LC-MS/MS analysis.

Potency and Hepatotoxicity Two million VimPro-GFP SW620 cells suspended in 100 μL of a 1:1 mixture of Matrigel® matrix (Corning Life Sciences, Corning, NY) and RPMI 1640 were injected into the 9-week-old female thymus. Deficient nude mice (Nude-Foxn1nu (069)) (Envigo, Huntingdon, Cambridgeshire, UK) were injected into the flanks. Growth was monitored by caliper measurements three times per week. At 4 weeks, mice were randomized into two groups and treated with 50 mg/kg Compound 6 in 200 μL vehicle (10% DMSO, 40% PEG 400, 50% PBS pH=7.4) or vehicle control. Treatments were administered intraperitoneally once daily for 5 days. Mice were sacrificed 2 hours after the final dose on day 5 of treatment. Tumors and livers were harvested for analysis of Compound 6 accumulation as measured by LCMS and Western blot analysis to determine effects on EMT and apoptosis as well as hepatotoxicity.

Statistical analysis Data were subjected to an unpaired two-tailed Student's t-test with Welch's corrected statistical analysis or as described elsewhere using Prism 8 (GraphPad, LaJolla, Calif.). All experiments were repeated three times (n=3) or as described in Methods.

Example 10: Additional Experimental Methods for Assessment of Compound Activity
Microsomal stability . CD-1 mouse microsomes were purchased commercially and reactions were performed as previously described. Briefly, a master mix was prepared as follows: Microsomes (0.5 mg/mL), 10 μM CHD1Li (0.1%) solubilized in DMSO, 5 mM in 100 mM phosphate buffer (pH 7.4). UDPGA, 25 μg Alamethicin, and 1 mM MgCl ₂ . The master mix was pre-incubated for 5 minutes at 37°C, then 1 mM NADPH was added to initiate the microsomal activity reaction and maintained at 37°C throughout the time course. Reactions were stopped at 0, 5, 15, 30, 45, and 60 minutes by adding 200 μL of acetonitrile and analyzed by mass spectrometry. Appropriate microsome controls were also run under the same reaction conditions.

γ-H2AX DNA damage combinatorial studies with irinotecan (SN38) . CHD1L inhibitor 6 alone and in combination with SN38 were assessed for DNA damage as previously reported [Abbott et al., 2020; Abraham et al., 2019]. Immunofluorescence of γ-H2AX, an established biomarker of DNA damage, was measured using DLD1 colorectal cancer cells with low CHD1L endogenous expression and DLD1 cells engineered to overexpress CHD1L. , performed DNA damage studies [Ji et al., 2017; Ivashkevich et al., 2012]. Cells were seeded as monolayers in 96-well plates and treated with 10 μM (0.5% DMSO) of compound 6 or SN-38 (1 μM), or a combination of 6 and SN38 for 6 hours. Medium was aspirated and cells were washed with cold PBS. Cells were then fixed with 3% paraformaldehyde for 15 minutes at room temperature and washed three times with PBS. Cells were blocked for 1 hour in 5% BSA, 0.3% Triton X-100 in PBS at room temperature. Cells were then immunostained overnight at 4° C. with phospho-(S139)-γ-H2AX rabbit mAb using a 1:800 dilution in 1% BSA, 0.3% Triton X-100 in PBS. Primary antibody was aspirated and cells were washed with PBS. Cells were incubated with goat anti-rabbit Alexa Fluor Plus™ 647 fluorescent secondary antibody at room temperature for 2 hours at a concentration of 5 μg/mL in 1% BSA, 0.3% Triton X-100 in PBS. Cells were then washed with PBS and Hoechst 33342 stain was diluted to a concentration of 1:1000 in PBS and added to cells for 15 minutes at room temperature. Cells were then imaged on a PerkinElmer Phenix HCS imaging system using a 20x water objective. We observed synergy between compound 6 and SN38 in inducing injury in DLD1 cells overexpressing CHD1L, determined using the drug interaction index (CDI) equation. CDI=(A+B)/(AB), synergy was determined by CDI<0.8 in these experiments, additivity was between 0.8 and 1.2, antagonism was defined by CDI>1.2 bottom. Welch's t-test statistical analysis was used to determine significance (**=P≦0.01).

Cell-based cytotoxicity dose-response and combinatorial studies . The CHD1L inhibitor and SN38, the active pharmacophore of irinotecan, were assessed for antitumor activity against colorectal cancer cell lines alone or in combination. Cell lines were cultured as monolayers or 3D tumor organoids using RPMI-1640 with 5% fetal bovine serum as previously reported [Abbott et al., 2020]. For 3D SW620 tumor organoid cytotoxicity studies, 2,000 cells in 100 μL were seeded into each well of 96-well U-bottom Ultra Low Attachment Microplates (Corning Inc., Corning, NY, USA). Plates were centrifuged at 1,000 rpm for 15 minutes to promote cell aggregation. A final 2% Matrigel concentration was reached by coating the centrifuged cells with 25 μL of 10% Matrigel per well. Plates were then incubated for 3 days prior to treatment. 3D organoids were treated with 25 μL of various concentrations of drug. After 3 days of treatment, organoids were manually transferred to 96-well white non-porous bottom plates with 40 μL of medium. An equal volume of Celltiter-glo 3D (Promega) was added and the plate was kept on a plate shaker at 400 rpm for 45 minutes before reading luminescence on an Envision plate reader (PerkinElmer). In combination studies, Combinefit analysis was used to determine synergy scores [De Veroli et al., 2016].

In vivo study . The CHD1L inhibitors, compounds 6 and 6.11, were pharmacokinetically assessed to determine plasma half-lives in 9-week-old female CD-1 mice as previously reported [Abbott et al. 2020]. Compound 6 alone and in combination with irinotecan was further assessed for antitumor activity against SW620 tumor xenografts in athymic nude mice. Xenografts were generated using a previously reported method [Zhou et al., 2016]. Briefly, Compound 6 was administered by intraperitoneal injection (ip) at 5 mg/kg twice/day, 7 days/week for a total of 5 weeks. After the first week of compound 6 treatment, irinotecan was started at 60 mg/kg, once/week, ip for 3 weeks. Body weight and tumor volume were monitored twice/week. Mice were sacrificed and tissues were collected when a single tumor reached 2000 mm ³ or total tumor volume reached 3000 mm [Ji et al., 2017]. Compound 6.11 alone and in combination with irinotecan was similarly assessed for antitumor activity against SW620 tumor xenografts. Recently, it was reported that the CRC M-phenotype is significantly more tumorigenic than other CRC EMT-phenotypes, and that the M-phenotype also has significantly higher TCF transcription [Esquer et al., 2021 and supplementary information]. The xenografts used in this study were generated using dual reporter mesenchymal cells (M-phenotype) isolated as described in Esquer et al., 2021. The half-life of compound 6.11 is 8 hours in CD-1 mice and is 2.7-fold more stable than compound 6 (half-life = 3 hours). Therefore, the number of treatments was reduced from 2 times/day to 1 time/day. In addition, irinotecan was administered intraperitoneally at 50 mg/kg.

Figures 7A and 7B depict representative single-agent cytotoxicity dose-response studies in SW620 colorectal cancer (CRC) tumor organoids and provide _IC50s for the indicated exemplary compounds. Tables 4A and 4B below provide summary cytotoxicity data for exemplary compounds. Table 4A provides representative single compound cytotoxicity data in several different CRC tumor organoids.

Table 4B provides the results of combination therapy with the indicated representative CHD1L inhibitor (CHD1Li) and SN38 or Olaparib. Treatments were performed on four different CRC tumor organoid types. The concentration of CHD1L inhibitor is varied as indicated. The _IC50 for combination treatment is generally reduced compared to SN38 and olaparib alone.

FIG. 8B presents a graph of γ-H2AX intensity (vs. DMSO) of compound 6 alone, irinotecan (SN38) alone, and the combination of the two in DLD1 empty vector (EV) and DLD1(OE) overexpressing cells. . FIG. 8A is a Western blot showing relative expression of CHD1L in DLD1(EV) cells compared to DLD1(OE) cells compared to control expression of α-tubulin in these cells. CHD1L is known to be essential for PARP-1-mediated DNA repair and causes resistance to DNA-damaging chemotherapy [Ahel et al., 2009; Tsuda et al., 2017]. The data in Figure 8B show the 'on-target' effect of the CHD1L inhibitor, showing synergy with SN38 in inducing DNA damage.

Figures 9A-9C depict the results of synergy studies using exemplary CHD1L inhibitors 6, 6.3, 6.9 and 6.11 in SW620 colorectal cancer (CRC) tumor organoids. The combination of SN38 with 6 and 6.3 is 50-fold and 150-fold more potent than SN38 alone in killing colonic SW620 tumor organoids, respectively. The combination of SN38 with 6.9 and 6.11 are both over 100 times more potent than SN38 alone. Each of compounds 6, 6.3, 6.9 and 6.11 show synergy with irinotecan (and SN38) in killing SW620 tumor organoids.

Synergy scores for exemplary CHD1L inhibitors, where scores are determined as described in De Veroli et al., 2016, are listed in Table 5. To interpret the synergy score values, SynergyFinder normalized the input data as percentage inhibition so that they can be interpreted directly as the proportion of cellular responses that can be attributed to drug interactions (e.g., a synergy score of 20 corresponds to 20% response above expected). In our experience, synergy scores close to 0 give limited confidence in synergy or antagonism. When the synergy score is less than -10, the interaction between the two drugs is likely antagonism;
-10 to 10, the interaction between the two drugs is likely to be additive;
When greater than 10, the interaction between the two drugs is likely synergistic.

FIG. 10 contains graphs depicting tumor volume (fold) of SW620 tumor xenografts as a function of days (up to 28 days) of treatment with compound 6 alone, irinotecan alone or in combination. The combination of irinotecan and Compound 6 significantly inhibits colonic SW620 tumor xenografts to near zero tumor volume within 28 days of treatment compared to single-agent treatment groups or controls.

FIG. 11 contains graphs representing tumor volume (fold) of SW620 tumor xenografts as a function of days (up to 28 days) of treatment with irinotecan alone (1) or a combination of Compound 6 and irinotecan (2). The combination of irinotecan and compound 6 significantly inhibits colonic SW620 tumors to near zero tumor volume past the last treatment compared to irinotecan alone. Within 2 weeks of the last irinotecan-only treatment, tumor volume increased beyond that of the last treatment, signifying tumor recurrence. In contrast, the combination maintained smaller tumor volumes.

Figure 12 shows that compound 6 alone and in combination with irinotecan (4) significantly increases survival of CRC tumor-bearing mice compared to vehicle (1), compound 6 alone (2) and irinotecan alone (3). indicate.

FIG. 13 contains graphs representing tumor volume (fold) of SW620 tumor xenografts as a function of days (up to 20 days) of treatment with compound 6.11 alone, irinotecan alone or in combination. The combination of irinotecan and compound 6.11 significantly inhibits colorectal cancer SW620 tumor xenografts compared to irinotecan alone or control.

FIG. 14 contains graphs representing tumor volume (fold) of SW620 tumor xenografts as a function of days of treatment (up to 33 days) with treatment with irinotecan alone or in combination with compound 6.11 and irinotecan. The combination of irinotecan and compound 6.11 significantly inhibits colorectal SW620 tumors past the last treatment (day 33) compared to irinotecan alone. After 8 days of treatment (Tx release), tumor volume with irinotecan treatment alone increased approximately 3-fold, signifying tumor recurrence. Conversely, tumor volume treated with a combination of 6.11 and irinotecan decreased (approximately 1.5-fold) after treatment. After 8 days of treatment, there is a 3.4-fold difference in tumor volume between treatment with irinotecan alone and treatment with the combination of 6.11 and irinotecan.

Figure 15 shows that the combination of compound 6.11 and irinotecan significantly increases survival of CRC tumor-bearing mice compared to irinotecan alone and controls.

Ｂ環では、環は、６員芳香族環又は６，６員縮合芳香族環であり、Ｘは両方ともＮであることが現在好ましい。第２の縮合環が存在する場合それは、１個又は２個の追加のＮを含むことができる。好ましいＲ_Ｂ（Ｂ環置換）が存在する場合それは、電気陰性基以外である。好ましいＲ_Ｂは、水素又はＣ１～Ｃ３アルキルである。好ましいＡ環は、任意選択で置換されているフェニルであり、非置換フェニル（Ｒ_Ａが水素である）がより好ましい。Ｒ_Ｐ基は、水溶性に関連していると考えられ、－Ｎ（Ｒ_２）（Ｒ_３）基が、一般に好ましく、さらに特に好ましいのは任意選択で置換されているＮ含有ヘテロ環であり、Ｒ_２とＲ_３はそれらが結合しているＮと一緒になって、５員～８員の環を形成し、その環は、１個若しくは複数の追加のヘテロ原子を含むことができ、飽和（二重結合を含まない）であり、又は１個若しくは複数の二重結合を含むことができる。Ｒ_Ｈは、活性及び効力並びに代謝安定性に関連していると考えられる。Ｒ_Ｈは、好ましくは芳香族基、さらに詳細にはヘテロ芳香族基であり、芳香族環又はヘテロ芳香族環を安定化する環置換を含む。好ましいＹはＮＲであり、Ｒが水素であることがより好ましい。好ましくは、ｘは０である。好ましいＺは－ＣＯ－ＮＨ－である。好ましいＬ_２は、－ＣＨ_２－又は－ＣＨ_２－ＣＨ_２－である。 Example 11 Summary of Presently Preferred Structure-Activity Relationships for Inhibitors The presently preferred structure-activity relationships based on Formula I for the CDH1L inhibitors of the invention are as follows:

In ring B, it is presently preferred that the ring is a 6-membered aromatic ring or a 6,6-membered fused aromatic ring and X is both N. If a second fused ring is present, it may contain 1 or 2 additional N's. A preferred R _B (B-ring substitution), if present, is other than an electronegative group. Preferred R _B is hydrogen or C1-C3 alkyl. A preferred A ring is optionally substituted phenyl, more preferably unsubstituted phenyl (where _RA is hydrogen). The R _P group is believed to be associated with water solubility, and the —N(R ₂ )(R ₃ ) group is generally preferred, and even more particularly preferred is an optionally substituted N-containing heterocycle. , R ₂ and R ₃ together with the N to which they are attached form a 5- to 8-membered ring, which ring may contain one or more additional heteroatoms; It can be saturated (containing no double bonds) or can contain one or more double bonds. _RH is believed to be associated with activity and potency as well as metabolic stability. _RH is preferably an aromatic group, more particularly a heteroaromatic group, containing ring substitutions that stabilize the aromatic or heteroaromatic ring. Preferred Y is NR, more preferably R is hydrogen. Preferably x is 0. Preferred Z is -CO-NH-. Preferred L ₂ is -CH ₂ - or -CH ₂ -CH ₂ -.

［式中、Ｒ_１～Ｒ_９は、水素又は任意選択の置換基を表し、Ｒ_１０は、効力に関連していると考えられる部分であり、Ｒ_Ｎは、溶解度などの物理化学的特性に関連していると考えられる部分である］。実施形態において、Ｒ_５は、代謝安定性に関連していると考えられる、水素以外の置換基である。具体的な実施形態において、Ｒ_５は、ハロゲン、特にＦ又はＣｌ、Ｃ１～Ｃ３アルキル基、特にメチル基である。実施形態において、Ｒ_４は、水素以外の置換基、特にはＣ１～Ｃ３アルキル基、さらに詳細にはメチル基である。具体的な実施形態において、Ｒ_５はＦであり、Ｒ_４はメチルである。実施形態において、Ｒ_６～Ｒ_９は、水素、Ｃ１～Ｃ３－アルキル、ハロゲン、ヒドロキシル、Ｃ１～Ｃ３アルコキシ、ホルミル、又はＣ１～Ｃ_３アシルから選択される。実施形態において、Ｒ_６～Ｒ_９のうちの１個又は２個は、水素以外の部分である。実施形態において、Ｒ_６～Ｒ_９のうちの１個は、ハロゲン、特にフッ素である。具体的な実施形態において、Ｒ_６～Ｒ_９のすべては水素である。実施形態において、Ｒ_Ｎはアミノ部分－Ｎ（Ｒ_２）（Ｒ_３）である。具体的な実施形態において、Ｒ_Ｎは、第２のヘテロ原子（Ｎ、Ｓ又はＯ）を任意選択で含む５員～７員環を有する、任意選択で置換されているヘテロ環式基である。実施形態において、Ｒ_Ｎは、任意選択で置換されているピロリジン－１－イル、ピペリジン－１－イル、アゼパン－１－イル、ピペラジン－１－イル、又はモルホリノである。実施形態において、Ｒ_Ｎは、Ｃ１～Ｃ３アルキル、ホルミル、Ｃ１～Ｃ３アシル（特にアセチル）、ヒドロキシル、ハロゲン（特にＦ又はＣｌ）、ヒドロキシＣ１～Ｃ３アルキル（特に－ＣＨ_２－ＣＨ_２－ＯＨ）から選択される１個の置換基で置換されている。実施形態において、Ｒ_Ｎは、非置換のピロリジン－１－イル、ピペリジン－１－イル、アゼパン－１－イル、ピペラジン－１－イル、又はモルホリノである。 In embodiments, HTS screening of CHD1L identified phenylaminopyrimidine pharmacophores and salts thereof depicted in Formula XX:

[wherein R ₁ -R ₉ represent hydrogen or optional substituents, R ₁₀ is a moiety believed to be related to potency, and R _N This is the part that is considered relevant]. In embodiments, R ₅ is a substituent other than hydrogen that is believed to be relevant for metabolic stability. In specific embodiments, R ₅ is halogen, especially F or Cl, a C1-C3 alkyl group, especially a methyl group. In embodiments, R ₄ is a non-hydrogen substituent, particularly a C1-C3 alkyl group, more particularly a methyl group. In a specific embodiment, _R5 is F and _R4 is methyl. In embodiments, R ₆ -R ₉ are selected from hydrogen, C1-C3-alkyl, halogen, hydroxyl, C1-C3 alkoxy, formyl, or C1- _C3 acyl. In embodiments, one or two of R ₆ -R ₉ are moieties other than hydrogen. In embodiments, one of R ₆ -R ₉ is halogen, especially fluorine. In specific embodiments, all of R ₆ -R ₉ are hydrogen. In embodiments, R _N is an amino moiety -N(R ₂ )(R ₃ ). In specific embodiments, R _N is an optionally substituted heterocyclic group having a 5- to 7-membered ring optionally containing a second heteroatom (N, S or O) . In embodiments, R _N is optionally substituted pyrrolidin-1-yl, piperidin-1-yl, azepan-1-yl, piperazin-1-yl, or morpholino. In embodiments, R _N is C1-C3 alkyl, formyl, C1-C3 acyl (especially acetyl), hydroxyl, halogen (especially F or Cl), hydroxy C1-C3 alkyl (especially -CH ₂ -CH ₂ -OH) substituted with one substituent selected from In embodiments, R _N is unsubstituted pyrrolidin-1-yl, piperidin-1-yl, azepan-1-yl, piperazin-1-yl, or morpholino.

In embodiments, R ₁₀ is -NRy-CO-(L ₂ )y-R ₁₂ or -CO-NRy-(L ₂ )y-R ₁₂ where y is 0 or 1 , the presence or absence of _L2 , an optional linker group of 1-6 carbon atoms, wherein the linker is optionally substituted, wherein 1 or 2 carbons of the linker are O, NH, NRy or optionally substituted with S, Ry is hydrogen or alkyl of 1 to 3 carbons, R ₁₂ is an aryl group, cycloalkyl group, heterocyclic group, or heteroaryl group; Each is optionally substituted. In embodiments, y is one. L ₂ is -(CH ₂ )p- and p is 0-3. In embodiments, R ₁₂ is thiophen-2-yl, thiophen-3-yl, furan-2-yl, furan-3-yl, pyrrol-2-yl, pyrrol-3-yl, oxazol-4-yl, oxazol-5-yl, oxazol-2-yl, indol-2-yl, indol-3-yl, benzofuran-2-yl, benzofuran-3-yl, benzo[b]thiophen-2-yl, benzo[b] thiophen-3-yl, isobenzofuran-1-yl, isoindol-1-yl, or benzo[c]thiophen-1-yl. In embodiments, R ₁ is hydrogen or methyl. In embodiments, R ₁₂ is thiophen-2-yl, furan-2-yl, pyrrol-2-yl, oxazol-4-yl, indol-2-yl, benzofuran-2-yl, or benzo[b]thiophene -2-yl. In embodiments, R ₁₂ is thiophen-2-yl or indol-2-yl. In embodiments, R ₁ is hydrogen or methyl.
Exemplary compounds of the invention are illustrated in Scheme 1.

Exemplary R _P and —N(R ₂ )(R ₃ ) groups are illustrated in Scheme 2.

Exemplary R ₁₂ and R _H groups are illustrated in Scheme 3.

An exemplary B ring of Formula I is illustrated in Scheme 4.

wherein X ¹ and X ² are selected from CH and N, at least one of X ¹ and X ² is N, and X ³ -X ⁶ are CH ₂ , CH, O, S, N, and NH, the B ring is saturated, partially unsaturated or aromatic, depending on the selection of X ³ to X ⁶ , and R _B at the ring carbon and/or ring nitrogen is represented by formula I Represents an optional substitution defined for

Example 12 Exemplary Synthetic Methods Compounds of Formula XX and many other compounds of the invention are prepared, for example, by the methods illustrated in Scheme 5 (variables are as defined above). This three-step synthesis involves 2,4-dichloro-pyrimidine A (e.g., 2,4-dichloro-6-methylpyrimidine, where R ₄ is methyl or 2,4-dichloro-5-fluoropyrimidine, and R ₅ is fluorine) with p-phenylenediamine B at the 4-position to form intermediate C. Exemplary reaction conditions are shown in Scheme 5 where the reaction is added ice-cold ethanol with trimethylamine and stirred at room temperature for 15 hours [Kumar et al., 2014; Odingo et al., 2014]. Chlorinated intermediate C is then reacted with either amine HNR ₂ R ₃ D by amination to give intermediate E. Exemplary amination conditions are shown in Scheme 5 where the reactants are reacted in _DMF at elevated temperature in the presence of _K2CO3 . In step 3, an acid F employing an _R10 group is coupled to intermediate E. Various known synthetic methods such as cross-coupling, click chemistry or substitution reactions (e.g. SN2, aromatic, electrophilic) can be employed to introduce selected _R10 groups [Li et al., 2014a Li et al., 2014b; LaBarbera et al., 2007]. Scheme 5 illustrates the coupling of the amine group of E with selected carboxylic acids F to form R ₁₀ which is —NH—CO—R ₁₂ in compound G. Exemplary R ₁₂ are aryl, aryl-substituted alkyl, heteroaryl and heteroaryl-substituted alkyl. Exemplary coupling conditions are shown in Scheme 5 where the coupling proceeds at room temperature in the presence of propylphosphonic anhydride (T3P) and triethylamine to form the desired compound G. The illustrated method was used to prepare compounds 6 and 8, for example (see Scheme 6).

Various substituted starting materials A, B, D and F are commercially available or can be prepared using known methods. In embodiments, an aniline derivative (B') already substituted with R ₁₀ can be used in place of the p-phenylenediamine derivative B to form the corresponding R ₁₀ -substituted intermediate C'. Step 2 of the illustrated reaction is carried out by reacting intermediate C' with D to give the corresponding desired compound G' (where R ₁₀ is replaced by R ₁₂ --CO--NH--). As will be appreciated by those skilled in the art, it may be helpful or necessary to protect certain groups in starting materials or intermediates during any given reaction to prevent undesired side reactions. For example, ring N in reactant F can be protected with a suitable amine protecting group. The use of suitable protecting groups is generally routine in the art. Various primary or secondary amines (D) are commercially available or can be prepared by well-known methods. Alternatively, the chlorinated intermediate C can be reacted with a suitable nucleophile to add a selected -NR ₂ R ₃ group at the 4-chloro position. For example, D can be a cyclic amine such as pyrrolidine. As another possible alternative, Suzuki coupling can be used to introduce amine-containing groups via CC bond formation [Li et al., 2014a]. As another possible alternative, the Buchwald-Hartwig cross-coupling can be used to form a carbon-amine bond to such an intermediate.

Detailed Synthesis of Compounds 6 and 8 (Scheme 6)
N-(4-Aminophenyl)-2-chloro-6-methyl-pyrimidin-4-amine (102). To 0.5 g (3.06 mmol) of 2,4-dichloro-6-methylpyrimidine (100) dissolved in 10 mL of ethanol at 0° C., 513.7 μL (1.2 eq, 372.5 mg, 3.68 mmol) of triethylamine (TEA) and 330.5 mg (3.06 mmol) of p-phenylenediamine (101) were added. The reaction mixture was warmed to room temperature and stirred at that temperature overnight. The solvent was removed in vacuo and the resulting residue was chromatographed on silica gel using 40% hexane/ethyl acetate as eluent to give 500 mg (70% yield) of pure product (102). ¹ H NMR: (400 MHz, CDCl ₃ ): δ 7.19 (broad s, 1H), 7.02 (d, J = 8.8 Hz, 2H), 6.70 (d, J = 8.8 Hz, 2H), 6.18 (s, 1H ), 3.77 (wide s, 2H), 2.26 (s, 3H).

N-(4-aminophenyl)-6-methyl-2-(pyrrolidin-1-yl)pyrimidin-4-amine (104). To 1.2 g of N-(4-aminophenyl)-2-chloro-6-methyl-pyrimidin-4-amine (102) dissolved in 120 mL of DMF was added 777.3 mg (5.62 mmol) of potassium carbonate at room temperature. and 3.63 mg (4.12 mL, 51.1 mmol) of pyrrolidine (103) were added. The reaction mixture was heated to 80° C. for 8 hours. The reaction was cooled to room temperature and diluted with water. The product was extracted with ethyl acetate (3x100 mL). The combined organic layers were washed with brine, then dried _{over Na2SO4} , filtered and concentrated to give an oily crude product. It was chromatographed on silica gel using 10% methanol/DCM (with a few drops of TEA) to give 1.31 g (96% yield) of pure product (104). ¹ H NMR: (400 MHz, CDCl3): δ 7.11 (d, J = 8.4 Hz, 2H), 6.67 (d, J = 8.4 Hz, 2H), 6.25 (broad s, 1H), 5.68 (s, 1H) , 3.63 (wide s, 2H), 3.56(t, J = 6.8 Hz, 4H), 2.19 (s, 3H), 1.93 (t, J = 6.8 Hz, 4H).

N-(4-((6-methyl-2-(pyrrolidin-1-yl)pyrimidin-4-yl)amino)phenyl)-2-(thiophen-2-yl)acetamide (6). To 700 mg (2.60 mmol) of N-(4-aminophenyl)-6-methyl-2-(pyrrolidin-1-yl)pyrimidin-4-amine (104) in 15 mL was added 406 mg (2.86 mmol) at 0°C. of 2-thiopheneacetic acid (105), 906.8 μL (657.5 mg, 6.50 mmol) of TEA, and 3.06 mL (1.65 mg, 5.20 mmol) of T3P (50% w/w solution in ethyl acetate) were added. bottom. The mixture was warmed to room temperature and stirred for 15 hours. The reaction was quenched by slow addition of water and the product was extracted with DCM (3x150 mL) and then washed with brine. The organic layers were combined, dried over _Na2SO4 , filtered _, and concentrated under reduced pressure to give crude product. It was purified using silica gel and 10% methanol in DCM to give 864.5 mg (84% yield) of pure product (6). ¹ H NMR: (400 MHz, DMSO-d6): δ 10.07 (s, 1H), 9.06 (broad s, 1H), 7.64 (d, J = 8.8 Hz, 2H), 7.49 (d, J = 9.2 Hz, 2H), 7.39 - 7.37 (m, 1H), 6.98 - 6.96 (m, 2H), 3.84 (s, 2H), 3.47 (s, 4H), 2.13 (s, 3H), 1.89 (t, J = 6.6Hz , 4H). HPLC: 98% pure.

The Boc-protected indole-3-carboxylic acid 106-boc was used in a peptide coupling method with compound 104 in the presence of T3P and TEA to achieve the synthesis of the boc-protected indole derivative 8-boc and the addition of the boc-protecting group. It was converted to the indole derivative 8 via TFA deprotection in good yield. The boc protecting group is -COO-t-butyl, K2Co3, KI, EtOH.

N-(4-{[6-methyl-2-(1-pyrrolidinyl)-4-pyrimidinyl]amino}phenyl)-1-{[(2-methyl-2-propanyl)oxy]carbonyl}-1H-indole- 3-carboxamide (8-boc). To 80 mg (0.297 mmol) of compound 104 in 8 mL of DCM was added 85.36 mg (0.3267 mmol of boc-protected indole-3-carboxylic acid (106-boc), 103.6 μL (75.13 mg, 0 .742 mmol) of TEA, 350 μL (189 mg, 0.594 mmol) of T3P were added.The mixture was warmed to room temperature and stirred for 20 h.The reaction was quenched by the slow addition of water and the product was quenched with DCM ( 3×50 mL), then washed with brine.The organic layers were combined, dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to give the crude product, which was Purification using silica gel and 10% methanol in DCM gave 76 mg (50% yield) of pure product (8-boc) ¹ H NMR: (400 MHz, CDCl ₃ ): δ 8.28. (wide s, 1H), 8.24 - 8.13 (m, 3H), 7.57(q, J = 4.8, 8.8 Hz, 4H), 7.40 -7.31 m, 3H), 5.92 (s, 1H), 3.54 (s, 4H) ), 2.23(s, 3H), 1.86 (s, 4H), 1.66 (s, 9H).

N-(4-{[6-methyl-2-(1-pyrrolidinyl)-4-pyrimidinyl]amino}phenyl)-1H-indole-3-carboxamide (8). 70 mg (0.136 mmol) of N-(4-{[6-methyl-2-(1-pyrrolidinyl)-4-pyrimidinyl]amino}phenyl)-1H-indole-3-carboxamide (8-boc) in DCM Dissolved in 25% TFA (5 mL). The solution was stirred at room temperature for 3 hours. The solvent was removed under reduced pressure and the crude product was purified using silica gel and 10% methanol in DCM to give 43.78 mg (78% yield) of pure product. ¹ H NMR: (400 MHz, DMSO-d6): δ 11.7 (s, 1H), 9.65 (s, 1H), 9.09 (s, 1H), 8.28 (broad s, 1H), 8.20 (d, J = 7.6 Hz, 1H), 7.67 (s, 4H), 7.47 (d, J = 8.0 Hz, 1H), 7.20 -7.12 (m, 2H), 5.88 (s, 1H), 3.50 (s, 4H), 2.14 (s , 3H), 1.91 (s,4H).

Scheme 7 illustrates an alternative synthetic method optimized for compound 6 yield. In this method, a t-butyl protected carbamate, such as compound 35, is reacted with a selected aromatic carboxylic acid, such as compound 36, to form a protected carbamate intermediate, such as compound 37. The intermediate is deprotected as known in the art, for example with trifluoroacetic acid (TFA), and the deprotected carbamate is removed from the primary or secondary amine group (e.g. pyrrolidinyl group). with a chlorinated heterocyclic group, such as compound 38, to form the desired compound of formula XX, such as compound 6. This method can also be employed to prepare various compounds of formula XX by selection of the starting aromatic carboxylic acid and chlorinated heterocyclic compound with a primary or secondary amine group.

In scheme 7 the reagents employed in the synthesis of compound 6 are shown. In the first reaction, DCC is N,N'-dicyclohexylcarbodiimide, DMAP is dimethylaminopyridine and solvent DCM is dichloromethane. The second reaction employs potassium carbonate and potassium iodide in ethanol after TFA deprotection. A person skilled in the art can readily adapt the reagents and reaction conditions employed to prepare the desired compound of formula XX.

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Claims (25)

A method of treating a TCF transcription-induced cancer comprising administering to a patient in need thereof an amount of a CHD1L inhibitor effective to inhibit CHD1L or to inhibit TCF transcriptional aberrations. the CHD1L inhibitor is a compound of Formula I, or a salt or solvate;

or a salt or solvate thereof,
[In the formula,
Ring B is a heteroaryl ring or ring system having 1, 2 or 3 5- or 6-membered rings, any 2 or 3 of which can be fused rings; the ring is a carbocyclic, heterocyclic, aryl or heteroaryl ring, at least one of said rings being heteroaryl;
in said B ring, each X is independently selected from N or CH, at least one X is N;
R _P is a primary or secondary amine group [—N(R ₂ )(R ₃ )] or a —(M) _x —P group, where P is —N(R ₂ )( R ₃ ) or an aryl or heteroaryl group, x is 0 or 1, indicating the presence or absence of M, and M is an optionally substituted linker —(CH ₂ ) _n — or —N( R)(CH ₂ ) _n —, where each n is independently an integer from 1 to 6 (inclusive);
Y is -O-, -S-, -N(R ₁ )-, -CON(R ₁ )-, -N(R ₁ )CO-, -SO ₂ N(R ₁ )-, or -N( R ₁ ) a divalent atom or group selected from the group consisting of SO ₂ —;
L ₁ is an optional 1-4 carbon linker (which can be cis or trans) that is optionally substituted, saturated, or contains a double bond, and x is 0 or 1, indicating the presence or absence of L ₁ ,
Ring A is a carbocyclic or heterocyclic ring or ring system having 1, 2 or 3 rings, 2 or 3 of which may be fused, each ring having 3 -10 carbon atoms, optionally having 1-6 heteroatoms, each ring optionally being saturated, unsaturated or aromatic;
Z is a divalent group containing at least one nitrogen substituted with an R'group;
In embodiments, Z is -N(R')-, -CON(R')-, -N(R')CO-, -CSN(R')-, -N(R')CS-, - N(R')CON(R')-, _-SO2N (R')-, -N(R') _SO2- , -CH( _CF3 )N(R')-, -N(R' )CH(CF ₃ )—, —N(R′)CH ₂ CON(R′)CH ₂ —, —N(R′)COCH ₂ N(R′)CH ₂ —,

or said divalent Z group is a 5- or 6-membered heterocyclic ring having at least one nitrogen ring member, for example

including
L ₂ is an optional 1-4 carbon linker (which can be cis or trans) that is optionally substituted, saturated, or contains a double bond, and z is 0 or 1, indicating the presence or absence of L ₁ ,
R is selected from the group consisting of hydrogen, aliphatic groups, carbocyclyl groups, aryl groups, heterocyclyl groups and heteroaryl groups, each of which groups are optionally substituted;
each R' is independently selected from the group consisting of hydrogen, aliphatic groups, carbocyclyl groups, aryl groups, heterocyclyl groups and heteroaryl groups, each of which groups is optionally substituted;
R ₁ is selected from the group consisting of hydrogen, aliphatic groups, carbocyclyl groups, aryl groups, heterocyclyl groups and heteroaryl groups, each of which groups are optionally substituted;
R ₂ and R ₃ are independently selected from the group consisting of hydrogen, aliphatic groups, carbocyclyl groups, aryl groups, heterocyclyl groups and heteroaryl groups, each of these groups being optionally substituted; or R ₂ and R ₃ together with the N to which they are attached form an optionally substituted 5- to 10-membered saturated, partially unsaturated or aromatic heterocyclic ring; ,
R _A and R _B represent hydrogen or from 1 to 10 non-hydrogen substituents on the indicated A and B rings or ring systems, respectively, where R _A and R _B substituents are hydrogen, halogen, hydroxyl, cyano , nitro, amino, mono- or disubstituted amino (-NR _C R _D ), alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, alkoxy, acyl, haloalkyl, -COOR _C , -OCOR _C , -CONR _C R _D , —OCONR _C R _D , —NR _C COR _D , —SR _C , —SOR _C , —SO ₂ R _C and —SO ₂ NR _C R _D independently selected from alkyl, alkenyl, cycloalkyl , cycloalkenyl, aryl, heterocyclyl, alkoxy, and acyl are optionally substituted;
R _C and R _D are each selected from hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl, each of these groups being one or more halogen, alkyl, alkenyl, haloalkyl optionally substituted with , alkoxy, aryl, heteroaryl, heterocyclyl, aryl-substituted alkyl, or heterocyclyl-substituted alkyl;
_RH is an optionally substituted aryl or heteroaryl group;
Optional substitutions are one or more of halogen, nitro, cyano, amino, mono- or di-C1-C3 alkyl substituted amino, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6- Cycloalkenyl, C1-C3 haloalkyl, C6-C12 aryl, C5-C12 heteroaryl, C3-C12 heterocyclyl, C1-C3 alkoxy, C1-C6 acyl, -COOR _E , -OCOR _E , -CONR _E R _F , -OCONR _{Alkyl , alkenyl , cycloalkyl , cycloalkenyl , aryl , _} heterocyclyl, alkoxy, and acyl are optionally substituted;
R _E and R _F are each hydrogen, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6-cycloalkenyl, C1-C3 haloalkyl, C6-C12 aryl, C5-C12 heteroaryl, C3 ~C12 heterocyclyl. selected from C1-C3 alkoxy, C1-C6 acyl, each of these groups being one or more of halogen, nitro, cyano, amino, mono- or di-C1-C3 alkyl substituted amino, C1-C3 alkyl, C2 ~C4 alkenyl, C3-C6 cycloalkyl, C3-C6-cycloalkenyl, C1-C3 haloalkyl, C6-C12 aryl, C5-C12 heteroaryl, C3-C12 heterocyclyl, C1-C3 alkoxy, and C1-C6 acyl optional replaced by selection]
The method of claim 1. 2. The method of claim 1, wherein the CHD1L inhibitor is not a PARP inhibitor, an inhibitor of topoisomerase, or an inhibitor of β-catenin. the CHD1L inhibitor is a compound, salt or solvate of Formula XX;

and a salt or solvate thereof,
[In the formula,
R ₁₀ is -NRy-CO-(L ₂ )y-R ₁₂ or -CO-NRy-(L ₂ )y-R ₁₂ , y is 0 or 1, optionally 1-6 indicates the presence or absence of _L2 , a linker group of 1 carbon atom, wherein the linker is optionally substituted, wherein 1 or 2 carbons of said linker are optionally O, NH, NRy or S substituted, Ry is hydrogen or alkyl of 1-3 carbons, R ₁₂ is an aryl group, cycloalkyl group, heterocyclic group, or heteroaryl group, each of which is substituted may have been
R _N is an amino moiety -N(R ₂ )(R ₃ ), R2 and R3 are hydrogen or optionally substituted alkyl groups, or R2 and R3 are the nitrogen to which they are attached. together form a saturated or partially unsaturated 5- to 10-membered heterocyclic ring,
R ₁ , R ₆ -R ₉ represent hydrogen or optional substituents]
The method of claim 1. effective in inhibiting CHD1L inhibition or TCF transcriptional abnormalities in patients in need of their prevention against tumor growth, invasion and/or metastasis in CHD1L-induced cancers, EMT-induced cancers or TCF transcription-induced cancers A method of prevention by administering an amount of a CHD1L inhibitor. 6. The method of claim 5, wherein the CHD1L inhibitor is a compound, salt or solvate of any one of Formula I or XX. 6. The method of claim 5, wherein the CHD1L inhibitor is not a PARP inhibitor, an inhibitor of topoisomerase, or an inhibitor of β-catenin. A method of treating CRC, including mCRC, comprising administering to a patient in need thereof an amount of a CHD1L inhibitor effective to inhibit TCF transcriptional abnormalities. 9. The method of claim 8, wherein the CHD1L inhibitor is a compound, salt or solvate of Formula I or XX. 9. The method of claim 8, wherein the CHD1L inhibitor is not a PARP inhibitor, an inhibitor of topoisomerase, or an inhibitor of β-catenin. A method of treating a drug-resistant cancer comprising administering to a patient in need thereof an amount of CHD1L effective to inhibit CHD1L or inhibit TCF transcriptional aberrations in combination with a known therapy to which said cancer has become resistant. A method comprising administering an inhibitor. 12. The method of claim 11, wherein the CHD1L inhibitor is a compound, salt or solvate of Formula I or XX. 12. The method of claim 11, wherein the CHD1L inhibitor is not a PARP inhibitor, an inhibitor of topoisomerase, or an inhibitor of β-catenin. 12. The method of claim 11, wherein the treatment the cancer has become resistant to is conventional chemotherapy. A combination method for the treatment of cancer comprising administering a CHD1L inhibitor in combination with a PARP inhibitor, a topoisomerase inhibitor, a platinum-based antineoplastic agent or a thymidylate synthase inhibitor. A method of identifying CHD1L inhibitors that inhibit CHD1L-dependent TCF transcription, comprising determining whether a selected compound inhibits CHD1L ATPase. A method of identifying a compound useful for treating cancer comprising determining whether the compound inhibits CHD1L ATPase. A pharmaceutical composition for the treatment of cancer comprising one or more compounds, salts or solvates of formula I or XX. Compounds, salts or solvates of Formula I

[In the formula,
Ring B is a heteroaryl ring or ring system having 1, 2 or 3 5- or 6-membered rings, any 2 or 3 of which can be fused rings; the ring is a carbocyclic, heterocyclic, aryl or heteroaryl ring, at least one of said rings being heteroaryl;
in said B ring, each X is independently selected from N or CH, at least one X is N;
R _P is a primary or secondary amine group [—N(R ₂ )(R ₃ )] or a —(M) _x —P group, where P is —N(R ₂ )( R ₃ ) or an aryl or heteroaryl group, x is 0 or 1, indicating the presence or absence of M, and M is an optionally substituted linker —(CH ₂ ) _n — or —N( R)(CH ₂ ) _n —, where each n is independently an integer from 1 to 6 (inclusive);
Y is -O-, -S-, -N(R ₁ )-, -CON(R ₁ )-, -N(R ₁ )CO-, -SO ₂ N(R ₁ )-, or -N( R ₁ ) a divalent atom or group selected from the group consisting of SO ₂ —;
L ₁ is an optional 1-4 carbon linker (which can be cis or trans) that is optionally substituted, saturated, or contains a double bond, and x is 0 or 1, indicating the presence or absence of L ₁ ,
Ring A is a carbocyclic or heterocyclic ring or ring system having 1, 2 or 3 rings, 2 or 3 of which may be fused, each ring having 3 -10 carbon atoms, optionally having 1-6 heteroatoms, each ring optionally being saturated, unsaturated or aromatic;
Z is a divalent group containing at least one nitrogen substituted with an R'group;
In embodiments, Z is -N(R')-, -CON(R')-, -N(R')CO-, -CSN(R')-, -N(R')CS-, - N(R')CON(R')-, _-SO2N (R')-, -N(R') _SO2- , -CH( _CF3 )N(R')-, -N(R' )CH(CF ₃ )—, —N(R′)CH ₂ CON(R′)CH ₂ —, —N(R′)COCH ₂ N(R′)CH ₂ —,

or said divalent Z group is a 5- or 6-membered heterocyclic ring having at least one nitrogen ring member, for example

including
L ₂ is an optional 1-4 carbon linker (which can be cis or trans) that is optionally substituted, saturated, or contains a double bond, and z is 0 or 1, indicating the presence or absence of L ₁ ,
R is selected from the group consisting of hydrogen, aliphatic groups, carbocyclyl groups, aryl groups, heterocyclyl groups and heteroaryl groups, each of which groups are optionally substituted;
each R' is independently selected from the group consisting of hydrogen, aliphatic groups, carbocyclyl groups, aryl groups, heterocyclyl groups and heteroaryl groups, each of which groups is optionally substituted;
R ₁ is selected from the group consisting of hydrogen, aliphatic groups, carbocyclyl groups, aryl groups, heterocyclyl groups and heteroaryl groups, each of which groups are optionally substituted;
R ₂ and R ₃ are independently selected from the group consisting of hydrogen, aliphatic groups, carbocyclyl groups, aryl groups, heterocyclyl groups and heteroaryl groups, each of these groups being optionally substituted; or R ₂ and R ₃ together with the N to which they are attached form an optionally substituted 5- to 10-membered saturated, partially unsaturated or aromatic heterocyclic ring; ,
R _A and R _B represent hydrogen or 1 to 10 non-hydrogen substituents of the indicated A and B rings or ring systems, respectively, where R _A and R _B substituents are hydrogen, halogen, hydroxyl, cyano, nitro, amino, mono- or disubstituted amino (-NR _C R _D ), alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, alkoxy, acyl, haloalkyl, -COOR _C , -OCOR _C , -CONR _C R _D , —OCONR _C R _D , —NR _C COR _D , —SR _C , —SOR _C , —SO ₂ R _C , and —SO ₂ NR _C R _D independently selected from alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, alkoxy, and acyl are optionally substituted;
R _C and R _D are each selected from hydrogen, alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocyclyl, aryl, or heteroaryl, each of these groups being one or more halogen, alkyl, alkenyl, haloalkyl optionally substituted with , alkoxy, aryl, heteroaryl, heterocyclyl, aryl-substituted alkyl, or heterocyclyl-substituted alkyl;
_RH is an optionally substituted aryl or heteroaryl group;
Optional substitutions are one or more of halogen, nitro, cyano, amino, mono- or di-C1-C3 alkyl substituted amino, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6- Cycloalkenyl, C1-C3 haloalkyl, C6-C12 aryl, C5-C12 heteroaryl, C3-C12 heterocyclyl, C1-C3 alkoxy, C1-C6 acyl, -COOR _E , -OCOR _E , -CONR _E R _F , -OCONR _{Alkyl , alkenyl , cycloalkyl , cycloalkenyl , aryl , _} heterocyclyl, alkoxy, and acyl are optionally substituted;
R _E and R _F are each hydrogen, C1-C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6-cycloalkenyl, C1-C3 haloalkyl, C6-C12 aryl, C5-C12 heteroaryl, C3 -C12 heterocyclyl, C1-C3 alkoxy, C1-C6 acyl, each of these groups being selected from one or more halogen, nitro, cyano, amino, mono- or di-C1-C3 alkyl-substituted amino, C1- C3 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C3-C6-cycloalkenyl, C1-C3 haloalkyl, C6-C12 aryl, C5-C12 heteroaryl, C3-C12 heterocyclyl, C1-C3 alkoxy, and C1- optionally substituted with C6 acyl], except said compound is not one of compounds 1-9. a compound of formula XX or a salt or solvate thereof;

and a salt or solvate thereof,
[In the formula,
R ₁₀ is -NRy-CO-(L ₂ )y-R ₁₂ or -CO-NRy-(L ₂ )y-R ₁₂ , y is 0 or 1, optionally 1-6 indicates the presence or absence of _L2 , a linker group of 1 carbon atom, wherein the linker is optionally substituted, wherein 1 or 2 carbons of said linker are optionally O, NH, NRy or S substituted, Ry is hydrogen or alkyl of 1-3 carbons, R ₁₂ is an aryl group, cycloalkyl group, heterocyclic group, or heteroaryl group, each of which is substituted may be
R _N is an amino moiety -N(R ₂ )(R ₃ ), R ₂ and R ₃ are hydrogen or optionally substituted alkyl groups, or R ₂ and R ₃ are together with each nitrogen atom form a saturated or partially unsaturated 5- to 10-membered heterocyclic ring,
R ₁ , R ₆ -R ₉ represent hydrogen or optional substituents], except that said compound is not one of compounds 1-9. A compound selected from compounds 6.5, 6.11 to 6.29 whose structures are shown in Scheme 1, or a salt or solvate thereof. A compound selected from compounds 6.5, 6.11, 6.16, 6.18, 6.20 and 6.21 whose structures are depicted in Scheme 1, or a salt or solvate thereof. A pharmaceutical composition comprising a compound selected from compounds 6.5, 6.11 to 6.29 or a salt or solvate thereof and a pharmaceutically acceptable carrier. Use of CHD1L inhibitors in the manufacture of cancer therapeutics, particularly CRC and mCRC therapeutics. A CHD1L inhibitor for use in the treatment of cancer, in particular for the treatment of CRC and mCRC.

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